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 BLOOM
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FAWCET1 A TEXTBOOK OF

msPcNUcXGY
DON W. FAWCETT, M.D.
Hersey Professor of Anatomy, Emeritus
Harvard Medical School
Boston, Massachusetts
Senior Scientist
International Laboratory for Research
on Animal Diseases
Nairobi, Kenya, Africa

ELEVENTH EDITION

1986
W. B. SAUNDERS COMPANY
Philadelphia D London D Toronto D Mexico City D Rio de Janeiro D Sydney D Tokyo D HongKong
W. B. Saunders Company: West Washington Square
Philadelphia, PA 19105

Library of Congress Cataloging in Publication Data

Fawcett, Don Wayne, 1917—

A textbook of histology.

At head of title: Bloom and Fawcett.

Rev. ed. of: Textbook of histology/William Bloom,

Don W. Fawcett. 10th ed. 1975.

Includes bibliographies and index.

1. Histology. I. Bloom, William, 1899-1972. Textbook of

histology. 10th ed. II. Title. III. Title: Histology.

QM551.F34 1986 61F.018 85-8218

ISBN 0-7216-1729-8

Listed here is the latest translated edition of this book together

with the language of the translation and the publisher.

Portuguese (10th Edition)—DISCOS CBS Industria e Comercio Ltda., Rio de Janeiro, Brazil

Japanese (10th Edition)—Hirokawa Publishing Co., Tokyo, Japan

Spanish (10th Edition)—Editorial Labor, S. A., Barcelona, Spain

Italian (10th Edition)—Piccin Editore, Padova, Italy

Editor: Dana Dreibelbis

Designer: Terri Siegel
Production Manager: Bob Butler
Manuscript Editor: David Harvey
Illustration Coordinator: Walt Verbitski
Indexer: Susan Thomas

A Textbook of Histology ISBN 0-7216-1729-8

© 1986 by W. B. Saunders Company. Copyright 1930, 1934, 1938, 1942, 1948, 1952, 1957, 1962,
1968 and 1975 by W. B. Saunders Company. Copyright under the Uniform Copyright Conven¬
tion. Simultaneously published in Canada. All rights reserved. This book is protected by
copyright. No part of it may be reproduced, stored in a retrieval system, or transmitted in any
form or by any means, electronic, mechanical, photocopying, recording, or otherwise, without
written permission from the publisher. Made in the United States of America. Press of W. B.
Saunders Company. Library of Congress catalog card number 85-8218.

Last digit is the print number: 9876543 2 1

 Preface

Advances in histology and cytology in the past decade have continued to

be impressive, requiring extensive revision of the Textbook of Histology. In this
period a number of smaller books have appeared that may seem to be more
commensurate with the diminishing time allotted to this subject in the curricula
of medicine, dentistry, and veterinary science. Nonetheless, we believe that
there is still a place for a book that treats this important body of knowledge
more thoroughly. The benefits of such a book may extend beyond the imme¬
diate needs of students and teachers to the future needs of those engaged in
biomedical research.
Research in many areas of biological science is now dominated by “reduc-
tionism”—the laudable attempt to understand complex living organisms by
studying the properties of their smallest components. The advancing frontier
of knowledge has rapidly progressed down to the cell and its subcellular
organelles and macromolecules. Much of current research is carried out on a
small number of cell types that can be grown in vitro. The contributions of
such studies to our understanding of fundamental biological processes have
been impressive. But in the longer term, the objective must be to understand
not only what cells can do in the controlled environment of the culture vessel,
but how cells function in the complex interactive systems that make up the
tissues and organs of the body. In the foreseeable future the pendulum will
surely swing back from reductionism. When it does, it is hoped that this book
will serve as a repository of useful information that would otherwise have to be
rediscovered by cellular and molecular biologists when they undertake to apply
their knowledge of isolated components to the functioning of the heterogeneous
associations of interacting cells characteristic of higher levels of organization in
whole animals.
In the productive period embraced by this revision, there have been
remarkable advances in our understanding of the structural basis of cell and
tissue function. New interpretations of the molecular organization of the cell
membrane have been put forward. New filamentous components of the cyto-
skeleton have been identified and their relationships to each other and to the
plasmalemma described. Receptor-mediated endocytosis has been found to be
a widespread cellular activity of great physiological importance. Gaps in our
understanding of the biosynthesis of proteins have been filled by further
elaboration of the signal hypothesis and elucidation of a number of molecular
events in the assembly and translocation of the nascent polypeptides. The
mechanism of action of hormones and neurotransmitters has been clarified by
identification and isolation of many of their receptors. The discovery of a host
of new biologically active peptides and their immunocytochemical localization
in cells widely dispersed in the gastrointestinal tract and brain has led to
recognition of a diffuse endocrine system hitherto overlooked. Atrial cardiac
muscle cells have been shown to have an endocrine as well as a contractile
mechanical function. The endothelium of arteries has been found to synthesize
iv • PREFACE

and release a clotting factor into the blood and the endothelium of the
pulmonary vessels has been shown to have surface enzymes that transform or
catabolize certain hormones. Functions have been assigned to a number of
other cell types previously considered to be relatively inert. Several genetically
distinct types of collagen have been identified in the extracellular matrix of
connective tissue. Fibronectin and laminin and other new structural proteins
have been characterized and their distribution and functions described. Much
has been learned about the properties of proteoglycans and their interactions
with the fibrous components of the extracellular matrix. Major changes have
taken place in interpretation of stem cell kinetics and cell lineages in hemopoesis
and the influence of humoral and microenvironmental factors in control of
blood cell differentiation.
We have endeavored to incorporate these and many other advances into
the book without substantial increase in its length. Outmoded interpretations
have been deleted and the sections on histogenesis have been omitted except
where essential to an understanding of the structure of the organ. In past
editions the opening chapter was devoted to Methods, for it was thought desirable
to give the student some insight into how new information in this field is
acquired. It seems unlikely that curricular time still permits presentation of
such material, and the techniques for analysis of structure and function have
become so diverse that their exposition, however brief, would occupy a dispro¬
portionate fraction of the book. Therefore, the chapter on Methods has been
omitted from this edition, and those pages have been used to expand and
update the material on The Cell. This chapter does not presume to cover the
burgeoning field of cell biology, but describes briefly the structure and function
of the cell organelles, and introduces terminology and concepts needed for an
understanding of later chapters.
A special effort has been made to update and improve the sections on the
histophysiology of the organs. Histology is a highly visual subject in which
illustrations often contribute as much to understanding as the text. As in
previous revisions, old photomicrographs and electron micrographs have been
replaced by new ones of better quality or more informational content wherever
these were available. Approximately 100 figures have been eliminated and 160
added, bringing the total to over 900. For the first time the references for each
chapter have been grouped under headings corresponding to those in the text,
to facilitate the task for those readers who may wish to consult the literature
for more detailed information on specific topics.
All books can be improved, and this book is no exception. I will welcome
constructive criticism from knowledgeable colleagues and students, and will
depend upon you, my readers, to point out items that need to be changed,
added, or deleted. With your help I look forward to a greatly improved next
edition.
I am indebted to Helen Deacon in the United States and Doris Churi in
Kenya for typing and retyping manuscript, and to Drs. Giuseppina and Elio
Raviola for sending countless xeroxes of literature not otherwise available to
me in East Africa. The patience of Mr. Dana Dreibelbis, Medical Editor, and
others at the W. B. Saunders Company was greatly appreciated.

Don W. Fawcett, M.D.

 Contents

1
THE CELL . 1
The Cell . 1
Cell Membrane . 3
Endoplasmic Reticulum. 6
Golgi Complex . 7
Mitochondria . 13
Lysosomes . 16
Annulate Lamellae . 20
Centrioles ..... 20
Cytoplasmic Inclusions. 22
Cytoskeleton . 24
Nucleus . 31
Cell Activities. 43

2
EPITHELIUM . 57
Classification of Epithelia. 58
Specializations of Epithelia . 64
Specializations of the Free Surface 72

3
GLANDS AND SECRETION . 83
Exocrine Glands . 63
Synthesis and Release of Protein Secretory Products . 83
Mechanisms For Release of Secretory Products. 91
Classification of Exocrine Glands. 92
Histological Organization of Exocrine Glands . 97
Control of Exocrine Secretion .
Endocrine Glands . 99
Cytology of Polypeptide-Secreting Endocrine Glands. 100
Cytology of Steroid-Secreting Endocrine Cells. 100
Storage and Secretion of Hormones. 103
Relation of Endocrine Cells to Blood and Lymph Vascular Systems . 104
Control Mechanisms and Interrelations within the Endocrine System
Receptors and Mechanism of Hormone Action on Target Organs ...
Neuroendocrine Cells, Paraneurones.

4
BLOOD . Ill
Erythrocytes ... 111
Platelets . 115
Leukocytes ... 117
Neutrophilic Leukocytes (Neutrophils) . 120
Eosinophilic Leukocytes (Eosinophils) . 125
Basophilic Leukocytes (Basophils) . 126
Lymphocytes . 128
Monocytes . 131
Other Components of Blood . 132
v
Vi • CONTENTS

5
CONNECTIVE TISSUE PROPER . 136
Loose Connective Tissue . 137
Extracellular Matrix . 137
Cells of Connective Tissue . 151
Free Cells of Connective Tissue ... 156
Serous Membranes . 164
Dense Connective Tissue . 164
Connective Tissue with Special Properties ... 167
Histophysiology of Connective Tissue . 169

6
ADIPOSE TISSUE ..... 174
Histological Characteristics of the Adipose Tissues ... 174
Histogenesis of Adipose Tissue. 181
Histophysiology of Adipose Tissue. 182

7
CARTILAGE . 188
Hyaline Cartilage . 188
Elastic Cartilage . 194
Fibrocartilage. 194
Other Varieties of Cartilage and Chondroid Tissue . 195
Regeneration of Cartilage . 196
Regressive Changes in Cartilage . 196
Histophysiology of Cartilage. 197

8
BONE . 199
Macroscopic Structure of Bones . 199
Microscopic Structure of Bones . 200
Bone Matrix .. 207
The Cells of Bone . 208
Histogenesis of Bone . 216
Histophysiology of Bone . 231
Joints and Synovial Membranes . 235

9
BONE MARROW AND BLOOD CELL FORMATION . 239
Prenatal Hemopoiesis . 239
Structural Organization of Bone Marrow. 241
Hemopoietic Stem Cells . 243
Erythropoiesis . 245
Granulopoiesis. 248
Monopoiesis . 254
Thrombopoiesis . 255
Lymphopoiesis . 259
Regulation of Hemopoiesis . 260

10
MUSCULAR TISSUE . 265
Smooth Muscle .. 265
Skeletal Muscle . 272
Cardiac Muscle . 296
 CONTENTS

11
THE NERVOUS TISSUE . 311
Jay B. Angevine

The Neuron . 312

Distribution and Diversity of Neurons . 325
The Nerve Fiber . 329
Peripheral Nerves. 335
Nerve Endings . 337
Autonomic Nervous System . 342
Neuroglia. 345
The Synapse and the Relationships of Neurons . 347
Development of the Neurons and of the Nervous Tissue . 355
Meninges, Ventricles, and Choroid Plexus . 357
Response of the Neuron to Injury. 362

12
BLOOD AND LYMPH VASCULAR SYSTEMS . 367
Arteries. 367
Physiological Implications of the Structure of Arteries . 378
Changes in Arteries with Age . 380
Capillaries . 381
Structural Basis of Trans endothelial Exchange . 384
Sinusoids. 390
Veins . 391
The Heart . 396
Impulse Conducting System . 398
Histogenesis of the Blood Vessels . 399
Lymphatic Vessels . 400

13
THE IMMUNE SYSTEM . 406
Elio Raviola

Cells of the Immune System. 409

Cytology of the Cells of the Immune System . 409
Histophysiology . 414
Lymphoid Tissue .. 425
Histophysiological Overview of the Immune System . 430
Lymphocyte Circulation . 430

14
THYMUS . 436
Elio Raviola

Histological Organization... 436

Normal, Accidental, and Experimental Involution . 443
Histophysiology . 445

15
LYMPH NODES . 449
Elio Raviola

Histological
O
Organization.
O
449
i M Q

Histophysiology of Lymph Nodes . 4oo

16
SPLEEN . 464
Elio Raviola

Histological Organization. 464

Histophysiology . ^'3
Vlli CONTENTS

17
HYPOPHYSIS ... 479
Pars Distalis . 482
Acidophils (Alpha Cells) . 483
Basophils (Beta Cells) . 483
Chromophobes (Reserve Cells) . 487
Follicular Cells (Stellate Cells) . 488
Nerves .,. 489
Blood Supply . 489
Histophysiology of the Pars Distalis . 490
Pars Intermedia . 492
Histophysiology of the Pars Intermedia . 493
Pars Infundibularis or Tuberalis ..... 494
Neurohypophysis . 494
Histophysiology of the Neural Lobe . 496

18
THE THYROID GLAND . 500
Histological Organization.%. 500
Histophysiology . 505
The Principal Cells . 505
The Parafollicular Cells . 508

19
PARATHYROID GLANDS . 511
Histophysiology . 513

20
ADRENAL GLANDS AND PARAGANGLIA . 516
The Adrenal Cortex .. 516
The Adrenal Medulla . 524
Blood Supply and Lymphatic Drainage of the Adrenal . 527
Nerves of the Adrenal. 528
Histophysiology of the Adrenal Cortex. 528
Histophysiology of the Adrenal Medulla . 530
Histogenesis of the Adrenal Glands . 531
The Paraganglia . 533

21
PINEAL GLAND . 535
Histological Organization. 535
Histophysiology . 538

22
SKIN . 543
The Epidermis . 543
Cell Types of the Epidermis . 552
Mucocutaneous Junctions. 558
The Dermis . 558
Hairs . 561
Nails. 567
Glands. 568
Blood and Lymphatic Vessels . 572
Nerves. 574
Histogenesis of the Skin and Its Accessories. 575
 CONTENTS • ix

23
ORAL CAVITY AND ASSOCIATED GLANDS . 579
The Oral Cavity . 579
The Tongue. 582
Glands of the Oral Cavity . 587
Histophysiology of the Salivary Glands . 597
Tonsils . 597
The Pharynx . 599

24
THE TEETH . 602
Histogenesis of the Teeth. 609

25
THE ESOPHAGUS AND STOMACH . 619
Esophagus . 619
Glands of the Esophagus . 619
Histophysiology of the Esophagus . 623
Stomach . 624
The Gastric Mucosa. 624
Submucosa . 636
Muscularis Externa . 636
Serosa . 636
Blood Supply . 636
Cell Renewal and Repair. 637
Histophysiology of the Stomach . 638

26
INTESTINES . 641
The Small Intestine . 641
The Intestinal Mucosa. 641
The Large Intestine . 660
The Appendix . 660
The Cecum and Colon. 660
The Rectum and Anus . 663
Histophysiology of the Intestine . 664
Blood Vessels of the Gastrointestinal Tract. 668
Lymph Vessels of the Gastrointestinal Tract . 669
Nerves of the Intestinal Tract . 671

27
THE LIVER AND GALLBLADDER . 679
Liver . 679
Histological Organization of the Liver . 679
Regeneration . 704
Histophysiology of the Liver. 705
Bile Ducts . 708
The Gallbladder . 708

28
PANCREAS . 716
The Exocrine Pancreas . 716
Histophysiology of the Exocrine Pancreas. 720
The Endocrine Pancreas . 721
Histophysiology of the Endocrine Pancreas . 725
The Duct System . 727
Blood Vessels, Lymphatics, and Nerves . 728
X • CONTENTS

29
RESPIRATORY SYSTEM .,. 731
The Nose. 731
Paranasal Sinuses . 734
The Larynx . 734
Conducting Portion of the Respiratory Tract. 735
Respiratory Portion of the Lungs. 739
The Pleura . 750
Innervation of the Lungs .. 750
Blood Supply . 751
Lymphatics . 751
Histophysiology of the Respiratory Tract. 752

30
THE URINARY SYSTEM . 755
Kidneys. 755
Uriniferous Tubules. 756
The Renal Interstitium . 775
Juxtaglomerular Complex ........ 776
Blood Supply . 779
Lymphatics . 780
Nerves . 782
Histophysiology of the Kidneys 782
Passages for the Excretion of Urine 787
Urethra ... 790

31
MALE REPRODUCTIVE SYSTEM . 796
Testis . 796
Seminiferous Tubules. 798
The Spermatozoon . 802
Spermatogenesis. 810
Interstitial Tissue .. 826
Blood Vessels and Lymphatics of the Testis . 828
Histophysiology of the Testis . 832
Excretory Ducts of the Testis. 834
Accessory Glands of the Male Reproductive Tract . 842
The Penis . 845
Semen . 848

32
FEMALE REPRODUCTIVE SYSTEM . 851
Ovary. 851
The Oviduct or Fallopian Tube . 873
Uterus ... 877
Endocrine Regulation of the Female Reproductive System 885
Implantation . 886
Placenta ... 888
Vagina. 896
External Genitalia .. 899

33
MAMMARY GLAND 901
 CONTENTS • XI

34
THE EYE . 913
Structure of the Eye in General . 913
Dimensions, Axes, Planes of Reference. 916
Fibrous Tunic .. 916
The Vascular Tunic: The Uvea . 924
Refractive Media of the Eye . 931
The Retina .. 936
Histophysiology of the Retina . 953
Blood Vessels of the Eye . 955
Lymph Spaces of the Eye. 955
Nerves of the Eye . 955
Accessory Organs of the Eye .... 955
Histogenesis of the Eye. 959

35
THE EAR . 961
External Ear. 961
Middle Ear . 961
Tympanic Cavity. 962
Tympanic Membrane . 963
Auditory Tube . 963
Internal Ear . 964
The Bony Labyrinth . 966
The Membranous (or Endolymphatic) Labyrinth . 966
The Perilymphatic Labyrinth . 980
Endolymph and Perilymph . 981
Nerves of the Labyrinth. 981
Blood Vessels of the Labyrinth. 983
Functional Considerations . 984

INDEX 987
\

'■*

*
 THE CELL

The living substance of plants and animals interests of contemporary histologists also
has traditionally been described by the gen¬ extend to histo- and cytochemistry.
eral term protoplasm. The smallest unit of
protoplasm capable of independent existence
is the cell. The simplest animals consist of a
single cell. The higher animals can be THE CELL
thought of as a colony or complex society of
interdependent cells of many kinds, special¬ Hundreds of microscopically distinguisha¬
ized in various ways to carry out the functions ble kinds of cells are found in the body, but
essential to survival and reproduction of the they all have certain structural features in
animal as a whole. Cells serving the same common. This chapter does not refer to a
general function are grouped together and particular cell type but undertakes to describe
united by varying amounts of intercellular the structural components of cells in general.
substance to form the basic tissues of the body, The cell is partitioned into two major com¬
such as blood, bone, muscle, nervous tissue, partments: the nucleus, composed of nucleo¬
connective tissue and so forth. Two or more plasm (karyoplasm), and the cytoplasm, which
tissues are usually combined to form the surrounds the nucleus and makes up the rest
larger functional units called organs: e.g, skin, of the cell body. Both of these compartments
kidney, blood vessels, glands, and so on. contain structural components of character¬
Several organs whose functions are interre¬ istic form and staining properties that can be
lated constitute an organ system; examples are recognized with the light microscope (Fig. 1—
the respiratory system (comprising the nose, 1). On the basis of the generality of their
larynx, trachea, and lungs) and the urinary occurrence and certain assumptions as to
system (composed of the kidneys, ureters, their functional significance, these have been
urinary bladder, and urethra). classified in two categories: organelles and
The etymological derivation of the term inclusions.
histology may suggest that this is a subdivision Organelles are structures common to
of morphological science that deals only with nearly all cell types and are regarded as
the tissues, but it is in fact equally concerned metabolically active internal organs carrying
with the cellular and extracellular compo¬ out essential specific functions. Inclusions, on
nents and the patterns of their association to the other hand, are considered to be meta¬
form organs. In this broader sense histology bolically inert accumulations of metabolites
is synonymous with microscopic anatomy. The or cell products such as stored carbohydrate,
boundaries of the held were formerly limited protein or lipid and various crystals, pigment
by the resolving power of the light micro¬ deposits, secretory granules, and the like. In
scope. In the past 30 years the increasing use contrast to organelles, inclusions are re¬
of the electron microscope by histologists has garded as dispensable and often temporary
greatly extended the scope of the subject so constituents of cells. Although the distinction
that it now includes tissue ultrastructure and between organelles and inclusions is still use¬
much of cell biology, and thus embraces ful, the assignment of cell components to
biological structure at all levels of organiza¬ these two categories was originally made by
tion from the lower limit of direct visual histologists at a time when too little was
inspection down to the structure of large known about their ultrastructure and chem¬
molecules. For the localization of specific ical nature to make a valid judgment as to
macromolecules and enzymatic activities, the whether they were metabolically active or

1
2 • THE CELL

Figure 1-1. In the center of this figure is a drawing of the cell and its organelles and inclusions as they appear by light
microscopy. Around the periphery are representations of these same components as seen in electron micrographs.
What was called ergastoplasm by light microscopists is now called the rough endoplasmic reticulum. The illustration of
the plasma membrane encircled by an interrupted line is not directly visualized but represents the bimolecular layer of
phospholipids inferred from indirect methods of analysis.

inert, essential or dispensable. As knowledge that escaped detection with the light micro¬
of cell biology has advanced, the list of cell scope are not defined as organelles or inclu¬
organelles has lengthened, and the tradi¬ sions but are grouped in a third category:
tional classification of some components as cytoskeletal components.
inclusions has become debatable. For exam¬ Control of the differentiation and synthetic
ple, some pigment granules formerly consid¬ functions of the cell resides in the nucleus
ered to be inert inclusions are now identified while most of the responding synthetic and
as melanosomes, organelles with a highly metabolic activities are functions of the cyto¬
organized internal structure and specific en¬ plasm. The cell organelles and inclusions are
zymatic activity. Similarly, secretory granules suspended in a cytoplasmic matrix that appears
formerly regarded as accumulations of stored structureless with the light microscope, but
cell product are now found to be limited by which consists of a fluid phase occupying the
an enzymatically active membrane and hence interstices of a complex three-dimensional
can qualify as organelles. A number of the meshwork of fibrillar components constitut¬
fibrillar elements of the cytoplasmic matrix ing the cytoskeleton.
 THE CELL • 3

Although the organelles visible with the graphs usually appears as a thin dense line
light microscope (mitochondria, Golgi appa¬ 8.5 to 10 nm thick. Although microscopically
ratus, ergastoplasm) were originally inter¬ unimpressive, it is remarkably complex in its
preted as granular or lamellar solid struc¬ molecular organization and carries out many
tures, the electron microscope has revealed essential functions. It regulates the traffic in
that these and other newly described organ¬ ions and macromolecules in and out of the
elles are hollow structures bounded by thin cell. It possesses devices for cell attachment
membranes of lipid and protein and often and cell-to-cell communication; antigenic
having a complex internal structure. One of molecules that are the basis of cell recognition
the most important contributions of electron and tissue specificity; ion pumps for regulat¬
microscopy has been to make us aware of the ing the internal environment of the cell;
importance of compartmentation of the cell receptors for hormones; and mechanisms for
by membranes. generating messenger molecules that activate
Most of the important physiological proc¬ the cell’s physiological responses to stimula¬
esses take place at surfaces and interfaces. tion.
Hundreds of enzymes that catalyze the chem¬ In electron micrographs of high magnifi¬
ical transformations occurring in cells are cation, three parallel lines can be discerned
strategically located in the membranes at the in thin sections of the cell membrane—two
interface between the cell and its environ¬ electron-dense layers (2.5—3.0 nm) separated
ment or between the organelles and the cy¬ by an electron-lucent intermediate layer (3.5—
toplasmic matrix. The internal partitioning 4 nm) (Fig. 1—2). With minor dimensional
of the cytoplasm achieved by membrane- variations, all membranous components of
bounded organelles promotes the efficiency cells have the same appearance. This does
of countless complex chemical reactions by not reflect a trilaminar structure at the mo¬
amplifying the area of the physiologically lecular level, but is a characteristic pattern of
active interfaces within the cell. The parti¬ binding of the osmium used as a stain for
tioning of the cytoplasm is also essential to electron microscopy. Membranes are com¬
the control mechanisms that govern cell me¬ posed of lipids and proteins, some of which
tabolism. The membranes enable the cell to possess terminal polysaccharides and hence
maintain a separation of enzymes and sub¬ are glycolipids and glycoproteins. The lipid
strates at some times, and at other times to is responsible for the form and many of the
permit their controlled interaction by varying permeability properties of membranes, while
permeability of a particular membrane or the their enzymatic activities reside in their pro¬
rate of active transport across it. If there tein constituents.
were unlimited diffusion and interaction All biological membranes consist of a bi-
within the cell it would be impossible to molecular layer of mixed phospholipids ori¬
maintain the high degree of chemical heter¬ ented with their hydrophilic ends at the outer
ogeneity characteristic of the cytoplasm. En¬ surfaces and their hydrophobic chains pro¬
zymes would attack their substrates, and all jecting toward the middle of the bilayer.
the potential interactions of the countless Deposition of osmium in the hydrophilic
chemical constituents of the cell would race ends of the phospholipid molecules is be¬
out of control. This does not occur. The cell lieved to result in the two dense lines seen in
is able to regulate its activities and to hold in electron micrographs, while the intervening
reserve a large repertoire of unexpressed pale line represents the unstained hydrocar¬
biochemical reactions. It can call each of these bon chains of the lipid molecules. The integral
into play at the proper time and at the rate proteins of the membrane occur as globular
appropriate to the needs of the organism as particles of varying size distributed in the
a whole. The fact that this is possible is due lipid bilayer (Fig. 1—3). They occupy different
in large measure to the prevalence of mem¬ positions within the bilayer depending on the
brane-bounded organelles in the cytoplasm. distribution of the hydrophilic and hydro-
phobic regions in the protein molecules.
Some having their polar amino acids at one
CELL MEMBRANE end will be exposed on an outer surface and
have their nonpolar region in the hydropho¬
The outer boundary of all cells is the plas- bic interior of the membrane. Others with
malemma or cell membrane. It is invisible with polar regions at either end and a nonpolar
the light microscope, and in electron micro¬ segment in the middle extend through the
4 • THE CELL

Figure 1-2. Electron micrograph of microvillous processes of an intestinal epithelial cell in cross section. Each microvillus
is enclosed by an extension of the cel! membrane. The membrane appears trilaminar—two dense layers separated by
a light intermediate layer. The dense lines are due to deposition of the osmium stain in the hydrophilic ends of the
phospholipid molecules of a bimolecular layer of lipid. The hydrocarbon chains remain unstained and form the light
middle line. On the outer aspect of the membrane is a moderately dense surface coat or glycocalyx consisting of
terminal polysaccharides of the integral glycoproteins embedded in the lipid bilayer of the membrane. (Micrograph
courtesy of Atsushi Ichikawa.)

Figure 1-3. Schematic representation of the fluid mosaic model of the cell membrane. Globular protein molecules are
positioned in the lipid bilayer at different depths depending on the distribution of their hydrophilic and hydrophobic
groups. Some extend entirely through the bilayer (transmembrane proteins). The lipid bilayer is fluid and the integral
proteins are free to diffuse laterally within the plane of the membrane if not restrained by binding to peripheral proteins
in the underlying cytoplasm. Terminal oligosaccharides of the glycoproteins extend outward contributing to the surface
coat or glycocalyx.
 THE CELL • 5

Figure 1-4. In the freeze-fracture method of specimen preparation, the fracture plane follows the hydrophobic region
of the membrane exposing two fracture faces—the E-face, the inwardly facing outer half, and the P-face, the outwardly
facing inner half, which usually contains the majority of integral protein particles.

entire thickness of the lipid bilayer and are The concentration of particles seen in
described as transmembrane proteins. freeze-fracture replicas of various cellular
The proteins are not identifiable in thin membranes correlates well with their bio¬
sections of membranes, but they can be vis¬ chemically determined protein content. Since
ualized in tissues prepared by the freeze- their distinctive enzymatic properties reside
fracture method. In this procedure, tissues in their proteins, the membranes of very
are rapidly frozen in liquid Freon or liquid active cell organelles have a higher concen¬
nitrogen and then cracked under vacuum by tration of membrane particles than those that
impact of a blade cooled to - 196° C. The are metabolically relatively inert.
plane of fracture through the frozen tissue At body temperature, the lipid bilayer is
preferentially follows the path of least resis¬ fluid and many of the integral protein parti¬
tance through the hydrophobic region of the cles are free to move laterally within the plane
lipid bilayer and thus cleaves cell membranes of the membrane. In freeze-fracture prepa¬
in half, exposing extensive areas of their rations, the seemingly random pattern of
interior (Fig. 1—4). A replica of the exposed particles in unspecialized regions of the
surface is then made by evaporation of a plasma membrane gives a misleading impres¬
heavy metal, such as platinum, from a source sion of uniformity of membrane structure
at an acute angle to the fracture surface. and function over the entire cell. It is known,
Carbon is then deposited uniformly over the however, that the luminal, lateral, and basal
surface by evaporation from a separate elec¬ membranes of many epithelial cells differ in
trode directly over the specimen, forming a their enzymatic and other properties. There¬
coherent and stable replica of all irregulari¬ fore, despite the lateral mobility of some
ties on the fracture face. The tissue is then membrane proteins, the cell is able to main¬
digested away and the replica recovered for tain regional differences in the population of
examination with the electron microscope. certain specific protein particles. How this is
In such preparations, the cleaved cell mem¬ accomplished is not entirely clear, but it is
branes present two distinct appearances. The speculated that mobility of some transmem¬
outwardly facing inner half-membrane, brane proteins is restricted by their binding
called the P-face, shows numerous randomly to peripheral membrane proteins—polypeptide
distributed 6- to 9-nm particles that represent polymers in the underlying cytoplasm asso¬
the integral membrane proteins (Fig. 1—5). ciated with the inner aspect of the plasma
The inwardly facing outer half-membrane, membrane.
called the E-face, is relatively smooth but may In addition to these regional differences in
contain occasional particles and numerous biochemical properties of the cell membrane,
shallow pits corresponding in distribution to which are not reflected in obvious differences
the particles on the opposing P-face. Why in distribution of intramembrane particles,
the majority of protein particles remain with there are local specializations of the mem¬
the inner half-membrane is not clear. brane for cell-to-cell communication and for
6 • THE CELL

Figure 1-5. Freeze-fracture preparation of the plasma membranes of two adjacent cells. The fracture plane has broken
across from one to the other, exposing the E-face of one and the P-face of the other.

cell-to-cell attachment. At these sites, integral erties probably confer some degree of selec¬
protein particles are aggregated into closely tivity with respect to naturally occurring sub¬
packed plaques or into linear arrays that stances that can bind to the cell surface.
form circumferential bands around the cell
apex. These junctional specializations of the
membrane will be discussed in more detail in ENDOPLASMIC RETICULUM
Chapter 2.
The polysaccharide components of the in¬ The cytoplasm of many cell types is per¬
tegral glycoproteins and glycolipids project meated by an extensive system of membrane-
from the outer aspect of the lipid bilayer and bounded canaliculi making up the endoplasmic
contribute to the formation of a carbohy¬ reticulum. This organelle consists of a loose
drate-rich surface coat or glycocalyx. This is network of branching and anastomosing tu¬
present on the free surface of nearly all cells, bules throughout the cytoplasm, but the tu¬
but is most highly developed on the luminal bules may be expanded locally into broad flat
surface of certain epithelia. It is especially saccules called cisternae. Where cisternae are
conspicuous on the brush border of the ab¬ abundant they tend to become arranged in
sorptive cells of the intestinal epithelium parallel array. The endoplasmic reticulum is
where it appears as a mat of delicate, branch¬ a closed intracellular canalicular system that
ing, 2- to 5-nm polysaccharide filaments. Ion¬ does not normally open onto the cell surface.
ized carboxyl and sulfate groups of the acid Its limiting membrane is, however, continu¬
polysaccharides give the glycocalyx a strong ous with the outer membrane of the nuclear
negative charge, and it avidly binds cationic envelope. Thus, the space between the two
dyes such as Alcian blue, ruthenium red, and nuclear membranes may be considered a per¬
cationic ferritin. This layer acts as a protective inuclear cistern of the endoplasmic reticu¬
mechanical barrier, and its polyanionic prop¬ lum.
 THE CELL • 7

This organelle occurs in two forms: the two molecules of RNA and about 40 associ¬
rough endoplasmic reticulum and the smooth en¬ ated proteins. Ribosomes may occur singly
doplasmic reticulum (Figs. 1—6, 1—7). In the free in the cytoplasmic matrix but in this
former the outer surface of the limiting form they are nonfunctional. To participate
membrane is studded with dense granules of in protein synthesis, they must become asso¬
ribonucleoprotein, the ribosomes, which are ciated with a molecule of mRNA, which car¬
involved in the synthesis of proteins. The ries the codons determining the sequence of
newly synthesized protein accumulates in the assembly of amino acids into polypeptides.
lumen of the reticulum and is transported In synthetically active cells, the great majority
through it to the supranuclear region for of ribosomes occur in clusters of 10 to 20
further processing in the Golgi complex, an linked together by their common attachment
organelle concerned with concentrating and to a long molecule of mRNA. These assem¬
packaging cell products for secretion. blies are called polyribosomes or polysomes.
The smooth reticulum predominates in Polyribosomes that occur free in the cyto¬
cells involved in synthesis of triglycerides, plasm are engaged in synthesis of the integral
cholestrol, or steroid hormones. The mem¬ proteins of the cell. Those that are bound to
branes contain a versatile complement of the membrane of the rough endoplasmic
enzymes involved in conjugation, oxidation, reticulum are involved in synthesis of protein
and methylation. The smooth reticulum in destined for export from the cell as a secre¬
liver cells plays an important role in synthesis tory product. When seen in surface view on
of plasma lipoproteins, and in the metabolism the membranes, they appear as beaded
and detoxification of exogenous lipid-soluble chains or rosettes. The connecting strand of
drugs. A special form of smooth reticulum, mRNA usually cannot be resolved in micro¬
the sarcoplasmic reticulum, surrounds the graphs of thin sections. The ribosomes bind
myofibrils in striated muscle. Here its prin¬ to receptors in the membrane by their large
cipal function is the sequestration and release subunit. The nascent polypeptide chains
of the calcium ions that control muscular come off vectorially, extending through the
contraction and relaxation (see Chapter 10). membrane into the lumen of the reticulum.
The ribonucleoprotein of the ribosomes is The newly synthesized protein is channeled
largely responsible for the affinity of the through the reticulum to the Golgi region
cytoplasm for basic dyes. Aggregations of where it is transferred in small transport
rough endoplasmic reticulum appear under vesicles to the Golgi for further processing
the light microscope as coarse basophilic bod¬ and packaging for export. The participation
ies of irregular shape and varying size. When of the rough endoplasmic reticulum in pro¬
ceils are homogenized for biochemical anal¬ tein synthesis will be considered in greater
ysis, the endoplasmic reticulum is broken up detail in a subsequent chapter on glands and
and the membranes of the fragments close secretion.
to form small ribosome studded vesicles that The smooth endoplasmic reticulum has no
make up the microsome fraction recovered by associated ribosomes and usually takes the
differential centrifugation (Fig. 1-8). Thus, form of a closer-meshed three-dimensional
microsomes, as such, do not occur in the network of tubules (Fig. 1—7). Smooth-sur¬
living cell but are fragments of the endo¬ faced cisternae are rarely seen. Where rough
plasmic reticulum resulting from homogeni¬ and smooth reticulum occur in the same cell
zation of cells. If the microsome fraction is they are continuous with one another but
treated with lipid solvents to remove the represent regional differentiation of the or¬
membranes, recentrifugation yields a ribosome ganelle for different functions. The relative
fraction. Much of our present understanding proportions of the two vary with the cell type.
of the biosynthesis of proteins is based on The rough endoplasmic reticulum is most
the study of such fractions. Upon addition of highly developed in glandular cells synthesiz¬
messenger RNA (mRNA) and cofactors, micro¬ ing a large volume of a protein-rich secretory
somes will synthesize specific proteins in vi¬ product.
tro.
The ribosomes measure 15 by 25 nm and
are composed of a smaller and larger subunit. GOLGI COMPLEX
The small subunit contains a single large
molecule of ribonucleic acid (RNA) and some The Golgi apparatus or Golgi complex is an
30 associated small proteins. The larger sub¬ organelle found in nearly all eukaryotic cells.
unit adjacent to the membrane consists of It is best developed and has been most thor-
8 • THE CELL

Figure 1-6. Electron micrograph of a portion of a human plasma cell containing an extensive rough endoplasmic
reticulum consisting of cisternae studded with ribosomes. The moderately dense material in the lumen of the cisternae
represents newly synthesized protein.

Figure 1-7. Electron micrograph of an area of cytoplasm rich in smooth endoplasmic reticulum. The elements of the
reticulum are branching and anastomosing tubules. This form of the organelle is common in cells engaged in steroid
synthesis and lipid metabolism. The round dense body is a droplet of lipid.
 THE CELL • 9

Endoplasmic Reticulum freed from the membranes

o * •v
• V#» • », ••

Solubilization • ........
of membranes
with deoxvchoiate
*■ •
.s. •
• •
•
• •.
•• ••••:
:'•••• r.::‘ ••
: •v
and further :• v >:“
centrifugation

Figure 1-8. Diagrammatic representation of the three-dimensional configuration of the granular or rough endoplasmic
reticulum. When cells are homogenated for biochemical analysis, the reticulum is fragmented into small closed vesicles
called microsomes. The membranes of the microsome fraction can be solubilized and the ribosomes of the reticulum
can be recovered by further centrifugation.

oughly studied in glandular cells, where it is 1-9, 1-10). In section, the lumen of the
the site of concentration, chemical modifica¬ cisternae is narrow in its central portion but
tion, and packaging of the secretory products somewhat expanded toward the ends where
synthesized on the rough endoplasmic retic¬ the profile of the cistern may be interrupted
ulum. It has limited synthetic activity of its by fenestrations. There is no membrane con¬
own for complex carbohydrates, and plays tinuity between the successive cisternae. The
an important role in renewal of the plasma cisternae are often curved so that the orga¬
membrane of the cell and its carbohydrate- nelle as a whole has a convex and a concave
rich surface coat. surface. These curved assemblages of parallel
The organelle is not seen in routine histo¬ cisternae correspond to the arciform struc¬
logical preparations, but can be impregnated tures seen in stained preparations and called
with silver or osmium by several classical dictyosomes by classical cytologists.
staining procedures developed 75 to 100 A structural and functional polarity is evi¬
years ago. It is also stained by histochemical dent in the organization of the Golgi com¬
reactions for localizing certain enzymes. plex. The lumen of the cisternae is narrow
In electron micrographs, it consists of sev¬ at the convex face of the stack and becomes
eral membrane-limited flattened saccules or wider in the successive saccules toward the
cisternae, stacked in parallel array (Figs. concave face, and in actively secreting glan-
10 • THE CELL

Transport
vesicles

Figure 1-9. Three-dimensional drawing of the Golgi complex sectioned vertically. Small transport vesicles originating
from the endoplasmic reticulum are abundant at its convex or forming face. In glandular cells, secretory granules are
associated with its concave or maturing face.

Figure 1-10. Electron micrograph of the Golgi complex of a cell in the epithelium of the vas deferens. (Microoraph
courtesy of Daniel Friend.)
 THE CELL • 11

dular cells the density of the cisternal con¬ that membrane proteins characteristic of the
tents increases progressively from the outer reticulum are excluded and those destined
convex to the inner concave surface. There for incorporation in the Golgi are clustered
is also histochemical evidence of regional in the transitional region from which the
functional diversity. When subject to pro¬ transport vesicles arise by budding. More¬
longed osmication, the metal is deposited over, if the cisternae move through the stack
preferentially in one or two cisternae at the in the continual renewal of the organelle, the
convex face of the stack. The intermediate consistent regional differences in enzymatic
cisternae are selectively stained for nicotina¬ properties demonstrated histochemically
mide adenine dinucleotide phosphatase would be difficult to explain in an organelle
(NADPase), and the reaction product of the lacking the capacity for protein synthesis.
histochemical method for thiamine pyro¬ An alternative interpretation favored by
phosphatase is confined to one or two cister¬ some cell biologists assumes that the trans¬
nae on the concave inner side of the orga¬ port vesicles go directly from the transitional
nelle. region of the reticulum to the condensing
Proteins synthesized on the rough endo¬ vacuoles and in certain conditions to the
plasmic reticulum are transported in small expanded rims of cisternae near the secretory
transitional vesicles that bud off from ribo¬ face of the Golgi (Fig. 1-115). Thus, the bulk
some-free areas of the reticulum near the of the Golgi complex would be bypassed and
convex surface of the Golgi complex. The would not participate in condensation or
exact destination of these transport vesicles modification of the product. Such a scheme
in this organelle is not settled. According to would avoid the problem of mixing and di¬
the most widely accepted view, the vesicles lution of Golgi-specific membrane compo¬
fuse with one another to form the outermost nents by contributions of membrane from
cistern of the Golgi (Fig. 1-11A). This saccule the reticulum and would make the mainte¬
with its content of secretory product is nance of regional distribution of specific en¬
thought to progress through the organelle to zymes more understandable. On the other
the opposite face as successive cisternae are hand, it leaves unexplained the gradient in
formed behind it. The convex side of the product density in cisternae from the convex
stack is therefore called the forming face and to the concave side of the organelle, and
the concave side the maturing or secretory face. offers no mechanism for replacement of the
In this interpretation, all the cisternae in the continual loss of Golgi membrane from the
stack are involved in processing the secretory secretory face through formation of the con¬
product. In its transit through the organelle, densing vacuoles. The contradictions in both
the protein is concentrated and glycosylated interpretations remain unresolved, and
by glycosyl transferases present in the Golgi clearly much remains to be learned about the
membranes. Upon reaching the secretory function of this important organelle in the
face, the cistern rounds up into a number of secretory process and its role in renewal and
small vacuoles containing the concentrated recycling of components of the plasma-
protein. These subsequently coalesce to form lemma.
a single large condensing vacuole. After further A group of structures in close proximity to
concentration of its contents, this vacuole the Golgi complex, but considered by some
becomes a dense, spherical secretory granule. to be a distinct organelle, is the Golgi-associated
This interpretation accounts for the gra¬ Endoplasmic Reticulum from which Lysosomes
dient in density of the content of the cisternae form—commonly referred to by the acronym
from one side of the stack to the other, but GERL. When glandular cells are staining with
it involves a number of assumptions that are the cytochemical method for the enzyme acid
yet to be validated. If the transport vesicles phosphatase, the condensing vacuoles, a cis¬
budded off from the reticulum are continu¬ tern near the secretory face of the Golgi
ously incorporated in the Golgi membranes, complex, and contiguous tubules of the en¬
then with time the lipid and enzymatic com¬ doplasmic reticulum all give a positive reac¬
position of the two organelles should become tion. It has been proposed that these struc¬
the same. Biochemical studies provide no tures constitute a specialized complex for the
evidence of mixing of either the lipid or transfer of hydrolases directly from their site
protein components of the two compart¬ of synthesis in the endoplasmic reticulum to
ments. To explain the maintenance of their lysosomes forming near the inner face of the
specific properties, it is necessary to assume Golgi. Because condensing vacuoles and im-
12 • THE CELL

Endoplasmic
reticulum Figure 1-11. Alternative interpretations of
the path of secretory products through the
Golgi complex. A, Transport vesicles fuse
O T Intermediate at convex surface into a cistern, which
\ // vesicles
progresses through the stack as new ones
O
form behind it. B, transport vesicles fuse
with the periphery of relatively static cis¬
ternae or directly with the condensing vac¬
uoles at the secretory face of the orga¬
nelle. C, Transport vesicles fuse directly
with condensing vacuoles arising not from
Golgi
cisternae
the Golgi but from Golgi-associated en¬
doplasmic reticulum (GERL) from which
Secretory lysosomes also arise.
granule
Condensing
B vacuoles
 THE CELL • 13

mature secretory granules also exhibit phos¬ tor activities of cells. They are present in all
phatase activity, it is argued that these arise eukaryotic cells in numbers ranging from a
from GERL and not from the Golgi complex few to several thousand, depending on the
(Fig. 1—11C). If this is true, the secretory energy requirements for the specific func¬
pathway would bypass the Golgi, and the tions of the cell.
condensing vacuoles would receive secretory In living cells, the mitochondria appear as
proteins from the reticulum via intermediate slender rods 0.2 to 0.3 pm in diameter and
vesicles, and hydrolases directly from acid 2 to 6 pm long. They may be randomly
phosphatase—positive elements of the adja¬ distributed in the cytoplasm or concentrated
cent reticulum. Although this interpretation at sites of high energy utilization. In electron
has recently gained some measure of accept¬ micrographs of thin sections, mitochondria
ance, it remains controversial. The Golgi are seen to be limited by a smooth contoured
complex has long been assigned a major role outer membrane about 7 nm thick and a slightly
in the packaging of secretory product in a thinner inner membrane that is generally par¬
membrane capable of fusing with the plas- allel to the limiting membrane but exhibits a
malemma during exocytosis. The suggestion number of slender folds called cristae, which
that condensing vacuoles do not arise from project some distance into the interior of the
the Golgi but are derived from the GERL, a organelle (Fig. 1-12). The cristae are a device
specialized region of the endoplasmic retic¬ for amplifying the surface area of this en¬
ulum, is difficult to bring into accord with zyme-rich membrane to increase the effi¬
the widely accepted view of Golgi function. ciency of the organelle in generating energy.
In cells with high energy requirements, the
number of cristae in the mitochondria is
MITOCHONDRIA greater than in cells with less metabolic activ¬
ity.
Mitochondria are the organelles that pro¬ The mitochondrial membranes delimit two
vide the energy for the biosynthetic and mo¬ compartments: a large intercristal space com-

Fjgure 1—12. Electron micrograph of a typical mitochondrion from the pancreas of a bat, showing the cristae, matrix,
and matrix granules. Endoplasmic reticulum is seen at upper left and lysosomes at lower right. (Micrograph courtesy of
K. R. Porter.)
14 • THE CELL

prising all of the area within the inner mem¬ reside in the inner membrane subunits pro¬
brane, and a smaller membrane space consisting jecting into the mitochondrial matrix.
of the narrow cleft between outer and inner Mitochondria are self-duplicating organ¬
membranes and its inward extensions be¬ elles that increase their number by undergo¬
tween the leaves of the cristae. These latter ing division that resembles the binary fission
clefts are sometimes referred to as intracristal of bacteria. Indeed, mitochondria are be¬
spaces, but it is clear that they are not func¬ lieved to have evolved from symbiotic bacte¬
tionally distinct from the rest of the mem¬ ria very early in the development of living
brane space. This space usually is only 10 to forms when the earth’s atmosphere was still
20 nm across, and because it contains little relatively poor in oxygen and most of the
protein precipitable by chemical fixatives, it primitive unicellular organisms depended on
generally appears empty in electron micro¬ anaerobic fermentation of organic molecules.
graphs. The large intercristal space, on the It is speculated that a small bacterium that
other hand, is occupied by a moderately had evolved the ability to utilize oxygen in¬
electron dense mitochondrial matrix. Although vaded a larger anaerobic cell type, and a
the outer and inner membranes are similar symbiotic relationship developed with the
in appearance in electron micrographs of aerobic symbiont generating energy in return
thin sections, they differ in their physiological for protection and nutrients from the larger
properties, the outer being freely permeable host cell. With the passage of time, the
to small molecules and the inner having the interdependence of the two increased and
osmotic properties of a semipermeable mem¬ the parasite became an indispensable cell
brane. The inner membrane also has a distin¬ organelle passed on by the host cell to its
guishing morphological feature that can be progeny. The mitochondria retain a mode
demonstrated only by negative staining of division similar to that of their ancestral sym¬
disrupted isolated mitochondria. In such biotic respiring bacteria.
preparations, its inner surface is studded with Mitochondria not only have their own
numerous minute particles. These inner mem¬ DNA, but also have ribosomal RNA, transfer
brane subunits consist of a globular head 9 to RNA, and messenger RNAs, the informa¬
10 nm in diameter connected to the under¬ tional macromolecules involved in translation
lying membrane by a slender stem 3 to 4 nm of the information encoded in DNA. Their
thick and about 5 nm long. DNA is not enclosed in a nuclear membrane
The energy-generating function of the mi¬ but is visible as loose aggregations of slender
tochondria depends on oxygen and products filaments in electron-lucent areas of the mi¬
of the digestion of the protein, carbohydrate, tochondrial matrix (Fig. 1—13). When isolated
and lipid in the food ingested by the orga¬ from centrifugal fractions of mitochondria
nism. These are delivered to the cell in the and examined in negatively stained prepa¬
form of amino acids (from proteins), glucose rations, it closely resembles the DNA of bac¬
(from carbohydrates), and fatty acids (from teria—a double helix of naked DNA in the
lipids). A multienzyme system in the mito¬ form of a circle with a circumference of about
chondria oxidizes these substrates in a series 5.5 (Jim (Fig. 1—14). It differs from the DNA
of chemical reactions called the tricarboxylic of the cell nucleus in its circular form and in
acid cycle or Krebs cycle. The hydrogen atoms its much lower molecular weight. Although
removed in the successive steps of oxidation mitochondria have their own genome, they
are passed along a chain of respiratory en¬ are not genetically self-sufficient. Their DNA
zyme complexes in the inner mitochondrial consists of only about 15,000 nucleotides and
membrane and utilized in the conversion of has a very limited coding capacity. It deter¬
adenosine diphosphate (ADP) to adenosine mines the RNA of mitochondrial ribosomes,
triphosphate (ATP), the ubiquitous energy¬ which are visible as 12-nm granules distrib¬
carrying molecule required for transport uted throughout the matrix, and transfer
across membranes, for all synthetic processes RNAs, but it makes messenger RNAs that
throughout the cells, and for the mechanical code for only a few elements of the respira¬
work involved in the motor activities of cells. tory enzyme complexes in the inner mem¬
The succinoxidase, cytochrome oxidase, and brane. The nuclear DNA encodes the rest of
other enzymes concerned with electron trans¬ the inner membrane proteins, those of the
port are in the inner mitochondrial mem¬ outer membrane, and those of the matrix.
brane, while the enzymes of oxidative phos¬ These proteins are all synthesized on ribo¬
phorylation of ADP and hydrolysis of ATP somes in the cytoplasm and imported into
 THE CELL • 15

Figure 1-13. Electron micrograph of a mitochondrion from a plant cell showing filaments of DNA and ribosome-like
granules of ribonucleoprotein in the matrix. Similar components are demonstrable in mitochondria of animal cells.
(Micrograph courtesy of H. Swift.)

the mitochondria. All the enzymes needed to

replicate DNA and transcribe it into RNA
also depend on nuclear genes. Thus, in evolv¬
ing from free-living bacteria over a billion
years, mitochondria have given over to the
nucleus and cytoplasm of the host cell the
coding and synthesis of most of their struc¬
tural and functional proteins. Interestingly
enough, however, mitochondrial protein syn¬
thesis is blocked by antibacterial antibiotics
that do not affect synthesis elsewhere in the
cell. Although an evolutionary origin of mi¬
tochondria from symbiotic bacteria is an at¬
tractive speculation, the fact that mitochon¬
drial DNA encodes information in a manner
differing somewhat from that of the univer¬
sal genetic code casts some doubt upon this
hypothesis.
The most conspicuous of the matrix com¬
ponents are the matrix-granules—dense spher¬
ical or irregular bodies 30 to 50 nm in di¬
ameter occurring anywhere in the intercristal
Figure 1-14. When mitochondria of Xenopus oocytes are space but often closely associated with the
spread upon the surface film of fluid, the organelles are membranes of the cristae. Their density
disrupted and the DNA strands can be collected on a tends to obscure their internal structure, but
specimen grid and examined with the electron micro¬ in very thin sections they appear to be sub¬
scope. Mitochondrial DNA is in the form of circles 5.6 to
divided by septa into minute loculi or com¬
6 [xm in contour length. (Micrograph courtesy of I. B.
Dawid and D. R. Wolstenholme.) partments. The composition and function of
16 • THE CELL

the matrix granules continues to be a subject They may be ovoid or irregular in outline
of controversy. When calcium or other diva¬ (Fig,. 1—15). Their content may be homoge¬
lent cations are present in high concentration neous or may include aggregations of dense
in the fluid bathing isolated mitochondria, granules in a less dense matrix. They may
the size and density of the matrix granules is contain crystalline inclusions or concentric
increased. It was initially suggested that these systems of lamellae interpreted as myelin
granules function in the internal ionic regu¬ forms of phospholipid. The lysosome cannot
lation of the mitochondrion. This interpre¬ be confidently identified from morphological
tation has been weakened by x-ray microan¬ criteria alone. Histochemical demonstration
alyses of glutaraldehyde-hxed mitochondria of acid phosphatase or some other hydrolase
from noncalcifying tissues that failed to re¬ is required for verification.
veal calcium in matrix granules. However, Some 50 enzymes have now been identified
when fixed in osmium in the presence of in lysosomes. These include a number of
calcium, they bound calcium in detectable proteases, glycosidases, nucleases, phospha¬
amounts. Their affinity for osmium and their tases, phospholipases, and sulfatases. The pH
membrane-like ultrastructure suggest that optima of nearly all of these are in the acid
their major component is lipid. Biochemical range. In the normal cell, these hydrolytic
analysis of fractions enriched in matrix gran¬ enzymes are safety contained within the lim¬
ules indicates that they are composed of iting membrane of. the lysosome. In various
phospholipoprotein. There is no convincing pathological conditions, however, the perme¬
evidence that the matrix granules in tissues ability of the membrane may be increased,
other than bone and cartilage are involved allowing the enzymes to escape and digest or
in calcium sequestration in vivo. Their close lyse the cell. The term lysosome is descriptive
association with cristae and their membrane¬ of this property.
like composition suggest instead that they Cell digestion by lysosomes is not confined
may play some role in assembly of the mito¬ to pathological states. Programmed cell death
chondrial inner membrane, but the evidence is a normal part of the embryonic develop¬
for this is largely circumstantial. ment of some organs. Also, in postnatal life,
certain organs may undergo massive reduc¬
tion in volume, as in the regression of the
LYSOSOMES mammary gland after weaning of the off¬
spring. Under these physiological circum¬
The lysosome was added to the roster of stances, lysosomal membranes are somehow
common cell organelles in 1955 when De- altered, permitting escape of acid hydrolases
Duve and associates isolated from cell ho¬ and destruction of the cells.
mogenates a fraction of particles, distinct Lysosomes probably play their most impor¬
from mitochondria, that were rich in hydro¬ tant role in defense of the organism against
lytic enzymes active at acid pH. Since sub¬ microbial invasion. Lysosomes are abundant
strates were accessible to the enzymes only in the cytoplasm of white blood cells and
after disruption of the particles or their treat¬ tissue macrophages that are specialized for
ment with surface active agents, it was in¬ phagocytosis—ingestion of bacteria and other
ferred that they were enclosed by a mem¬ extracellular particles. The lysosomes consti¬
brane, and this was subsequently confirmed tute an intracellular digestive system that
by electron microscopy. Historically, cell or¬ enables these cells to break down the ingested
ganelles have first been described by micros- material and dispose of the degradation
copists, and their isolation and biochemical products. When bacteria are engulfed, they
characterization followed many years later. are taken into the cell in membrane-bounded
But lysosomes went unrecognized by mor¬ phagocytosis vacuoles or phagosomes. Lysosomes
phologists because, unlike other membrane- then gather around the phagosome, and
bounded organelles, they do not have a con¬ their membrane fuses with its membrane,
sistent and characteristic form. They are a releasing hydrolytic enzymes into the phag¬
highly heterogeneous group of bodies so di¬ osome to kill and digest the ingested bacteria
verse in size, shape, and internal organization (Fig. 1-16).
that no single description encompasses all of Intracellular digestion of extracellular ma¬
their variations. terial taken into the cell by phagocytosis is
In general, they are dense bodies 0.25 to referred to as heterophagy, and the phago¬
0.5 pm in diameter limited by a membrane. somes to which lysosomes have contributed
 THE CELL • 17

Figure 1-15. Micrograph of a cluster of lysosomes in the Golgi region of a cell from the adrenal cortex.

Figure 1-16. Schematic depiction of the role of lysosomes in heterophagy (lower right) and in autophagy (upper left).
Bacteria may be taken up by phagocytosis; primary lysosomes fuse with these heterophagic vacuoles and their enzymes
digest the contents. On the other hand, membranes may be formed around excess organelles or inclusions, and
lysosomes then fuse with these autophagic vacuoles. The end products of either process may be recognized in cells
as residual bodies or lipofuscin pigment.
18 • THE CELL

their enzymes are described as heterophagic

vacuoles. Some kinds of ingested material are
completely broken down. Others may leave
indigestible residues that persist in the form
of membrane-bounded structures called re¬
sidual bodies. These vary greatly in the ap¬
pearance of their content. It has been sug¬
gested that cells are capable of divesting
themselves of this indigestible matter by exo-
cytosis—fusion of the limiting membrane of
residual body, with the plasmalemma and
discharge of its contents into the extracellular
space. This is well documented for amebae,
but examples of such behavior in mammalian
cells have only rarely been recorded and
some cell biologists remain unconvinced of
its occurrence.
The lysosomal digestive system of the cell
is also involved in the elimination of cell
organelles and inclusions in the course of the
normal turnover of cell components and in
the reorganization of the cytoplasm that is
associated with rapid changes in physiological
activity. Mitochondria, elements of the en¬
doplasmic reticulum, or secretory granules
that are damaged or present in excess of the
functional requirements are enveloped in a Figure 1-17. Examples of autophagic vacuoles. A, A
membrane to form an autophagic vacuole. Ly- peroxisome and a mitochondrion enclosed in the same
vacuole. B, Elements of rough endoplasmic reticulum in
sosomes then fuse with this vacuole, releasing
two adjacent autophagic vacuoles.
into it enzymes that digest its content (Fig.
1-17). This process of controlled degrada¬
tion of organelles in an otherwise healthy cell and segregated and packaged in the Golgi
is called autophagy, in contradistinction to region. A considerable body of evidence now
heterophagy, which is the digestion of exoge¬ indicates that they may be formed in a special
nous material imported into the cell by differentiated region of the endoplasmic re¬
phagocytosis. The residual bodies resulting ticulum that is closely associated with the
from the two processes are readily distin¬ secretory face of the Golgi complex—an or¬
guishable. The indigestible residues of auto¬ ganelle designated GERL (Fig. 1—11C).
phagic activity that accumulate in cells of Clinical interest in lysosomes has been stim¬
aging animals have traditionally been called ulated by the discovery that 20 or more rare
wear-and-tear pigment or lipofuscin. “storage diseases” of children are due to an
The development of the concept of an inherited defect in the synthesis of lysosomal
intracellular digestive system has led to a enzymes (Table 1—1). When a lysosomal en¬
proliferation of terms to describe its compo¬ zyme is lacking, its normal substrate accu¬
nents. The term primary lysosomes is often used mulates in large membrane-bounded inclu¬
for those that have not yet become engaged sions in the cytoplasm that are, in effect,
in digestive activity, and the term secondary autophagic vacuoles that are unable to digest
lysosomes is applied to vacuolar structures that their contents. Massive accumulation of such
are the sites of current or past digestive material in the cells of liver and other organs
activity. The latter term thus encompasses seriously impairs their other functions and
autophagic and heterophagic vacuoles, resid¬ leads to early death.
ual bodies, and lipofuscin pigment deposits. Although lysosomes normally carry out
Some uncertainty prevails as to the mode their functions intracellularly, there are situ¬
of formation of primary lysosomes. They are ations in which lysosomal enzymes are re¬
believed to arise in much the same manner leased from cells. At sites of acute inflam¬
as secretory granules, their enzymes being mation where leukocytes are actively
synthesized on the endoplasmic reticulum phagocytizing bacteria, their lysosomal gran-
 THE CELL • 19

Table 1-1. INBORN LYSOSOMAL DISEASES

Substance Accumulating
Disease in Cells Enzyme Defect

Glycogen storage disease II Glycogen a-Glucosidase

(POMPE)
Gaucher’s disease Glucocerebroside (3-Glucosidase
Niemann-Pick disease Sphingomyelin Sphingomyelinase
Krabbe’s disease Galactocerebroside (3-Galactosidase
Metachromatic leukodystrophy Sulfatide Sulfatidase
Fabry’s disease Ceramide trihexoside a-Galactosidase
Tay-Sachs disease Ganglioside GM2 A’-Acetyl-fB-glucosaminidase A
Generalized gangliosidoses Ganglioside GMt p-Galactosidase
Fucosidosis H-isoantigen a-Fucosidase

ules may fuse with a phagosome before its 1—18). The nucleoids have been isolated and
closure and enzymes may escape into the identified as crystals of urate oxidase. The
surrounding connective tissue, destroying peroxisomes of human liver are devoid of
collagen, elastin, or cartilage matrix. It is now urate oxidase and therefore lack a nucleoid.
realized that much of the lasting damage to The matrix of peroxisomes is believed to
joints, kidney, and lungs following protracted consist mainly of catalase. Peroxisomes stain
inflammation is a consequence of this leakage selectively and intensely with the diamino
of acid hydrolases from phagocytic cells mo¬ benzidine reaction for peroxidase, and this
bilized in the defenses against invading bac¬ serves to distinguish them from primary ly-
teria. sosomes.
The origin of peroxisomes remains unset¬
tled. Some investigators report that their lim¬
Multivesicular Body
iting membrane is continuous at some point
Membrane-bounded vacuoles containing on its circumference with that of the smooth
numerous small vesicles are observed in endoplasmic reticulum and insist that they
many cell types. These multivesicular bodies
are stained with histochemical reactions for
acid phosphatase and are usually interpreted
as a form of lysosome. Their origin and
relationship to the more common dense ly-
sosomes remain to be established.

Peroxisome
With centrifugation procedures of im¬
proved resolution, it is possible to separate
from a crude lysosome fraction a class of
particles that lack acid hydrolases but are rich
in urate oxidase, D-amino acid oxidase, and
catalase. These are called peroxisomes because
of the ability of two of their enzymes to
generate hydrogen peroxide. It has become
apparent that these particles correspond to
spherical, membrane-limited bodies 0.2 to 0.4
pm in diameter observed by microscopists a
decade earlier and called microbodies. This old
term has now been abandoned in favor of
peroxisome. This organelle is found in the
cytoplasm of a number of cell types, espe¬
cially those of the liver and the kidney. In
some species, those in the liver contain a Figure 1-18. Electron micrograph of three peroxisomes
dense inclusion, the nucleoid, suspended in a from rat liver. Two contain a conspicuous dense nucleoid.
homogeneous matrix of lower density (Fig. (Micrograph courtesy of Daniel Friend.)
20 • THE CELL

arise as evaginations from that organelle.

Others believe that their enzymes are synthe¬
sized on ribosomes and are released into the
cytoplasmic matrix, where they form aggre¬
gates that subsequently acquire a membrane.
The function of peroxisomes in the metab¬
olism of the cell is not well understood.
Administration of drugs that induce hypo-
cholesterolemia causes a striking increase in
the number of hepatic peroxisomes, but this
observation has not led to any valid inference
as to their function.

ANNULATE LAMELLAE

Some cell types have in their cytoplasm

membrane structures that resemble segments
of nuclear envelope. These annulate lamellae
consist of a pair of parallel membranes en¬
closing a cisternal cavity 30 to 50 nm wide.
At regular intervals of 100 to 200 nm, the
membranes are continuous with one another
around circular fenestrations or pores in the
cistern about 50 nm in diameter. These are
lined by dense material forming a cylindrical
annulus within the pore that projects a few
nanometers on either side of the pair of Figure 1-19. Micrograph of annulate lamellae. Notice
membranes. Thus, annulate lamellae are their close resemblance to the nuclear envelope at left of
membrane-bounded cisternae traversed by figure (Nuc.). The lamellae are continuous above with
uniformly spaced pore complexes identical cisternae of rough endoplasmic reticulum. (Micrograph
courtesy of S. Ito.)
in appearance to those of the nuclear enve¬
lope (Fig, 1—19). They are frequently stacked
in parallel array with the pores of neighbor¬
ing lamellae in register. At their ends lamel¬ with this interpretation is the finding that in
lae are often continuous with cisternae of such cells they are often associated with ac¬
rough endoplasmic reticulum. Annulate la¬ cumulations of ribonucleoprotein-containing
mellae were previously believed to form by material that may represent deposits of ri-
delamination from the nuclear envelope but bosomal subunits or long-lived messenger
are now known to arise independently in the RNA. Until more evidence becomes available,
cytoplasm, often at some distance from the the function of annulate lamellae will remain
nucleus. conjectural.
The functional significance of this orga¬
nelle remains unclear. It has been suggested
that the pore complexes of the nuclear en¬ CENTRIOLES
velope may be involved in formation of pol¬
yribosomes or in processing subunits of ri¬ In cells that have been suitably stained, a
bosomes and mRNA as these leave the nu¬ pair of short rods, the centrioles, is visible with
cleus. It is possible that annulate lamellae the light microscope. These are centrally
may have a comparable function in assembly placed in a specially differentiated region of
or activation of informational macromole¬ the juxtanuclear cytoplasm called the centro-
cules stored in the cytoplasm. The hypothesis some or cell center. In secretory epithelial cells,
derives some indirect support from the fact the centrosome with its pair of centrioles
that annulate lamellae are most consistently (diplosome) is usually located in the supra¬
found in the early male and female germ nuclear cytoplasm and partially surrounded
cells where there is a long delay between by the Golgi complex. In other epithelial
transcription and translation. Also consistent cells, the centrioles may be immediately be-
 THE CELL • 21

neath the plasma membrane at the free sur¬ its respective tangent. This oblique orienta¬
face of the cell. tion of the triplets results in a cross-sectional
The centrioles are self-duplicating organ¬ pattern reminiscent of the vanes of a turbine
elles, ensuring their continuity from one cell or the charges of a pyrotechnic “pinwheel”
generation to the next. Immediately before (Fig. 1-20, inset).
cell division, they replicate and the two re¬ When centrioles replicate, they do not di¬
sulting pairs migrate to opposite poles of the vide. The new centriole develops in an end-
nucleus. When the nuclear envelope breaks to-side relationship to a specific region on
down and the mitotic spindle forms, a pair the preexisting centriole but separated from
of centrioles is found at each pole of the it by a narrow electron-lucent space. The
spindle. This association of the centriole with anlage, called the procentriole, is an annular
spindle poles suggested that they served as condensation of dense material about the
organizing centers or sites of nucleation of same diameter as a centriole but initially
the spindle microtubules. It is now known devoid of microtubules. The procentriole
that small dense components of the centro- elongates by accretion of material to its free
some called centriolar satellites are the micro- end and polymerization of tubulin to form
tubule-initiating centers. the triplets incorporated in its wall. The
In electron micrographs, the centrioles are newly formed centriole separates from the
short cylindrical structures about 0.2 jim in parent centriole but maintains its perpendic¬
diameter and 0.5 jam long with a dense wall ular orientation to it (Fig. 1—21). After du¬
surrounding an electron-lucent central cavity plication the two members of the original
(Fig. 1-20). Embedded in the wall are nine diplosome separate and each, with its newly
evenly spaced triplet microtubules. Each trip¬ formed daughter centriole, moves to one
let is set at a constant angle of about 40° to pole of the division figure. Centrioles are not

Fiqure 1—20. Micrograph of a diplosome near the free surface of an epithelial cell. The long axes of the two centrioles
are usually perpendicular. (Micrograph courtesy of S. Sorokin.) Inset shows a centriole in croSfe section at higher
magnification. The wall is composed of nine triplet microtubules (with subunits a, b, c). (Micrograph courtesy of J.
Andre.)
22 • THE CELL

immediately beneath and perpendicular to

the apical cell membrane. Two members of
each triplet microtubule in the wall of the
centriole then become sites of nucleation for
rapid polymerization of tubulin to form the
doublet microtubules that make up the axo-
neme of the developing cilium. Elongation
of the cilium to its definitive length requires
less than 50 minutes. The centrioles thus
serve a template function in ciliogenesis, im¬
posing upon the axonemes the ninefold ra¬
dial symmetry expressed in the arrangement
of triplets in their wall. Rootlet-like appen¬
dages may develop at the inner ends of each
centriole to serve as anchoring devices to
stabilize the base of the cilium.

CYTOPLASMIC INCLUSIONS

Glycogen
Carbohydrate is stored in animal cells in
the form of the polysaccharide glycogen. It is
broken down as needed to yield glucose. The
enzymatic degradation of glucose in turn
provides energy and short-chain carbon skel¬
etons that are reused in the synthesis of
Figure 1-21. Before the onset of cell division, each
member of the diplosome becomes a site of nucleation of
various components of protoplasm. The ma¬
a new centriole. Its anlage, the procentriole, is a ringlike jor sites of carbohydrate storage are the liver
structure devoid of microtubule elements. It elongates by and skeletal muscle, but glycogen is found in
accretion to its distal end, and microtubules assemble smaller amounts in a great many other cell
within its wall. Each member of the original diplosome,
types.
together with its newly formed daughter centriole, migrates
to one pole of the division figure. Glycogen occurs in the form of dense iso-
diametric 20- to 30-nm particles, often
slightly irregular in outline. These so-called
essential for mitosis. In plants, acentriolar beta particles of glycogen occur singly in some
divisions are common. The association of cell types, while in others they form larger
centrioles with the poles of the spindle ap¬ aggregates of varying size called alpha particles
pears to have evolved as a device for ensuring (Fig. 1—22). Glycogen is usually confined to
that both daughter cells receive a pair of the cytoplasm but in diabetes and the herit¬
centrioles. able glycogen storage diseases, it may also
Centrioles are essential for the formation accumulate in the nucleus.
of cilia and flagella. In the cells of ciliated
epithelia, the large numbers of centrioles that Lipid
serve as basal bodies for the cilia do not arise
simply by successive replications from the Cells frequently contain lipid stored as
original pair. Another mechanism has spherical droplets of triglycerides that are
evolved for their more rapid generation. liquid at body temperature. In routine his¬
Dense spherical bodies of unknown prove¬ tological preparations, the lipid is extracted
nance appear in the cytoplasm, and multiple by the solvents used in dehydration, leaving
procentrioles arise radially around each of behind round clear vacuoles. In tissues fixed
these procentriole organizers of deuterosomes. with glutaraldehyde and osmium tetroxide
When the assembly of multiple centrioles for electron microscopy, the lipid is pre¬
around each deuterosome is completed, they served as spherical globules varying in den¬
dissociate from it and move to a position sity from gray to black, depending in part on
 THE CELL • 23

Figure 1-22. Micrographs of liver glycogen of two different species at the same magnification. In the salamander liver
(A), the rosettes or alpha particles are considerably smaller than in the hamster (B). In both the glycogen particles are
aggregates of smaller subunits.

the degree of unsaturation of the constituent occurs complexed with the structural protein
fatty acids (Fig. 1—23). of dense ellipsoidal granules about 0.3 p by
The lipid droplets serve as an energy store 0.7 (jl, called melanosomes (Fig. 1—24). These
and source of short carbon chains that can form in the Golgi and become distributed in
be utilized in the synthesis of membranes and the cytoplasm of melanocytes. When trans¬
other lipid-rich structural components. In fat ferred to other cells, especially those of the
cells specialized for its storage, lipid in a basal layer of the epidermis, clusters of mel¬
single very large droplet makes up the bulk anosomes tend to occur enclosed in a limiting
of the cell volume. It is also abundant in cells membrane derived from the plasmalemma
that synthesize and secrete steroid hormones. of the donor melanocyte.
More widely distributed in cells throughout
the body are deposits of a yellowish-brown
Pigment substance called lipofuscin or lipochrome pig¬
The principal pigment of mammalian cells ment (Fig. 1-25). It occurs in coarse, irregu¬
is melanin, the brown pigment of skin and larly shaped granules that exhibit a golden-
hair; the choroid, ciliary body, and iris of the brown fluorescence in ultraviolet light. They
eye; and the leptomeninges and substantia stain lightly with lipid-soluble dyes, but are
nigra of the brain. Melanin is synthesized in insoluble in acid and alkali and in most lipid
specialized cells called melanocytes, which arise solvents. Lipofuscin deposits are rare in cells
in the neural crest of the embryo and subse¬ from young animals but increase in number
quently migrate to the skin and other sites with advancing age—an observation that led
mentioned above. Melanin is not confined to to their designation as “wear-and-tear pig¬
the melanocytes but may be transferred to ment.” Since the discovery of lysosomes and
the cytoplasm of other cell types that lack the their digestive properties, lipofuscin deposits
capacity to synthesize melanin. The pigment have come to be regarded as the end stages
24 • THE CELL

somes are probably more appropriately in¬

cluded among the cell organelles.

Crystalline Inclusions
In a few cell types, conspicuous crystals
normally occur free in the cytoplasmic matrix
or within the lumen of the endoplasmic re¬
ticulum. Most of these have not been isolated
and chemically characterized, but their stain¬
ing and solubility properties suggest that they
may be a storage form of protein. Crystals of
guanine occur in cells of the epidermis of
amphibia, and precisely oriented, rodlike in¬
tracellular crystals form a mirror-like layer,
the tapetum lucidum, behind the retina of
cats. In general, crystalline inclusions as nor¬
mal cell components are rare, but viruses
often form crystalline nuclear or cytoplasmic
inclusions in infected cells.

CYTOSKELETON

The term cytoskeleton is now commonly

used to describe a structural framework of

Figure 1-23. Lipid droplets in an electron micrograph of

an osmium-fixed cell. After glutaraldehyde fixation, they
are less osmiophilic and often appear pale gray.

of autophagic activity—accumulations of un-

digestible residues not completely degraded
by lysosomal hydrolases.
Cells in the liver, spleen, and bone marrow
may contain membrane-bounded deposits of
a golden-brown, iron-containing pigment, he¬
mosiderin. It usually occurs in the cytoplasm
of phagocytic cells that participate in the
degradation of the hemoglobin of senescent
erythrocytes. The amount of this pigment is
greatly augmented in diseases that involve an
increase in the rate of destruction of the red
blood cells. Hemosiderin can be distin¬
guished from lipofuscin and melanin by
staining reactions for iron. In electron micro¬
graphs, hemosiderin appears as dense masses
of 9-nm particles of ferritin, an iron storing
protein.
Deposits of lipofuscin and hemosiderin are
metabolically inert inclusions. Melanosomes,
on the other hand, are rich in the enzyme
tyrosinase, which catalyzes the oxidation of
tyrosine to dihydroxyphenylalanine, which is
Figure 1-24. Micrograph of portion of an invertebrate
an intermediate in the synthesis of melanin. melanocyte containing very dense melanosomes. In ver¬
Because of this metabolic activity, melano- tebrates, they are less dense and more fusiform.
 THE CELL • 25

the cell composed of several filamentous com¬

ponents—microtubules (25 nm), microfilaments
(6 nm), intermediate filaments (8—10 nm), and
a microtrabecular lattice. These components of
the cytoplasm are visible with the light micro¬
scope only when aggregated into coarse bun¬
dles. In transmission electron micrographs
segments of filaments and microtubules of
varying length are seen, but their three-di¬
mensional organization cannot be satisfactor¬
ily studied in thin sections. The architecture
of the cytoskeleton is best demonstrated in
thinly spread cells in culture that have been
extracted with non-ionic detergents to re¬
move membranes and soluble matrix com¬
ponents. The general distribution of the
several filamentous components of the cyto¬
skeleton can be studied at lower resolution
with the light microscope if tissue culture
cells are stained with fluorescent antibodies
against specific polypeptide subunits of the
filaments (Fig. 1-26).
The realization that many of the dynamic
processes of cells depend on interactions be¬
tween the plasma membrane and the cyto¬
skeleton has stimulated intense interest in the
filamentous components of the cytoplasm.
Maintenance of cell shape; stabilization of
Figure 1-25. Micrograph of two lipofuscin pigment gran¬ cell attachments; endocytosis; movements of
ules in a cell of the human adrenal cortex. local specializations of the cell surface; and

Figure 1-26. Immunofluorescence photomicrographs of cells treated with fluorescein-labeled antibody to cytoskeletal
components. A, Hamster cells (NILS) stained for 58K polypeptide of 10 nm filaments. (Micrograph from Hynes, R. O.
Cell 73:151, 1978.) B, Epithelial cell (PtK2) reacted with antibody against prekeratin. (Micrograph from Franke, W. W.
Proc. Natl. Acad. Sci. 75:5034, 1978.)
26 • THE CELL

cell motility in general all involve integrated bodies associated with the centrioles. These
interaction of the cytoskeleton and the cell centriolar satellites serve as nucleation sites for
membrane. polymerization of tubulin to form microtu¬
bules. Developing microtubules exhibit a po¬
larity, tubulin being added preferentially at
Microtubules
one end—usually the end away from the
The cytoplasm of most eukaryotic cells initial site of nucleation. The polarity mani¬
contains slender straight microtubules that fested in their directional polymerization is
can be traced in electron micrographs for thought to be related to the ability of micro¬
several micrometers before they pass out of tubules to function in promoting directed
the plane of section (Fig. 1-27). They are movements of cytoplasmic organelles.
circular in transverse section with a diameter When tubulin is extracted from brain, the
of about 25 nm and a wall 5 to 7 nm thick. purified preparation contains about 15 per
The cylindrical wall is composed of 13 par¬ cent of two other proteins of 270,000 and
allel protofilaments having a center-to-center 340,000 M.W. These microtubule associated pro¬
spacing of 5.5 nm. The protofilaments are teins (MAPs) form slender lateral projections
linear polymers of tubulin, a 55,000 M.W. that are observed in electron micrographs at
protein that occurs in two forms, a-tubulin regular 32-nm intervals along the length of
and (Ttubulin. These form 110,000 M.W. het¬ microtubules. The function of these filamen¬
erodimers that are subunits of the protofila¬ tous projections is not entirely clear, but it is
ments in the wall of the microtubule (Fig. 1— speculated that they may form stabilizing
28). cross links where microtubules are laterally
Microtubules may be found anywhere in associated in organized arrays as they are in
the cytoplasm and in any orientation, but the the mitotic spindle.
majority tend to converge upon the centro- The number of microtubules present in
some and often terminate in small dense the cytoplasm varies. They appear to be tran-

Frigure 1-27. Micrograph of the cytoplasm of a Sertoli cell showing numerous microtubules (at arrows). (Microaraoh
courtesy of W. Vogl.)
 THE CELL • 27

Figure 1-28. Schematic depiction of the Tubulin dimers of protofilaments

iinear polymers of tubulin that form the
protofilaments in the wall of the micro¬
tubule. Below are three configurations in
which the protofilaments may be assem¬
bled: cytoplasmic singlet microtubule,
doublet in ciliary and flagellar axonemes,
and triplet in the wall of centrioles.

Cytoplasmic
microtubule Ciliary Centriolar triplet
doublet

sient structures being disassembled, assem¬ circular cross section. However, in the axo¬
bled, and redeployed in new patterns as the nemes of cilia and flagella, tubulin polymer¬
cytoskeletal requirements of the cell change. izes to form nine doublets—pairs of conjoined
Thus, they are believed to be in dynamic microtubules that share a segment of the wall
equilibrium with a large reserve of their un¬ of one and therefore have a figure 8 cross
polymerized subunits in the cytoplasmic ma¬ section. And in centrioles, the wall is formed
trix. They are most conspicuous in dividing of nine triplets (Fig. 1-28). Although similar
cells where very large numbers of microtu¬ in subunit composition, these doublets and
bules are assembled to form the mitotic spin¬ triplets are much more stable than the singlet
dle. Colchicine, a drug that arrests cell divi¬ microtubules of the cytoplasm. In addition to
sion, binds to tubulin, preventing its this difference, the doublets possess paired
polymerization to form the microtubules of lateral appendages, the dynein arms regularly
the spindle. spaced at 24-nm intervals along one member
The relatively ephemeral microtubules of of the doublet. Dynein is a protein with
the cell cytoplasm are single tubules with a ATPase activity essential for ciliary and fla¬
gellar motility.
As major components of the cytoskeleton,
Table 1-2. COMPONENTS AND DISTRIBUTION
microtubules function in maintenance of cell
OF MICROTUBULES
shape, and their directional polymerization
Peptide and orientation plays a significant role in
Subunit
shape changes. They also participate in
Protein Mol. Wt. Distribution
movement of particles within the cell by de¬
Tubulin 55,000 Mitotic spindle; termining the direction along which cyto¬
53,000 axonemes of cilia
and flagella; cy¬
plasmic streaming occurs. Whether they con¬
toplasm of nearly tribute to the generation of force for
all cell types translocation of particles remains unsettled.
Microtubule associ¬ However, in the case of the specialized doub¬
ated proteins let microtubules of axonemes, it is generally
(MAPs)
MAPj 340,000 Associated with cy¬ accepted that the dynein arms generate slid¬
toplasmic micro¬ ing movements responsible for the oscillatory
tubules of neu¬ motion of cilia and the propagated waves of
rons and most
bending in flagella.
other cell types
map2 270,000 Associated with cy¬
toplasmic micro¬ Microfilaments
tubules of neu¬
rons and most The contractile proteins, actin and myosin,
other cell types
were formerly thought to occur principally
28 • THE CELL

in the myofibrils of striated muscle where Myosin filaments are a conspicuous feature
they form highly ordered interdigitating sets of striated and smooth muscle. In nonmuscle
of filaments that interact to cause muscle cells the forces required for cellular motility
shortening. It is now known that both of are low and in contrast to actin, myosin is a
these proteins are present in nearly all cell minor protein of the cytoplasm. Moreover,
types and are responsible for the viscoelastic myosin cannot be identified in filamentous
properties and contractility of the cytoplasm. form in these cells by conventional light or
Indeed, actin is one of the most abundant electron microscopy. It can be detected by
cellular proteins accounting for 10 to 15 per fluorescein-labeled antibodies and can be ex¬
cent of the total protein. tracted for biochemical analysis.
Demonstration of the distribution of actin Whereas actins from different cell types
within cells depends on its staining with fluo¬ and different species are very similar, myo¬
rescein-labeled antibody or upon use of sins vary in molecular size and configuration.
heavy meromyosin fragments of the myosin They all have in common ATPase activity
molecule that bind specifically to actin fila¬ and the ability to bind reversibly to actin
ments, decorating them with regularly spaced filaments, and they can be induced to form
barbs that result in a characteristic arrowhead bipolar filaments in vitro. The molecules con¬
configuration. Both these methods identify sist of a rod-shaped “tail” and two globular
as actin, the 6-nm microfilaments seen in “heads” that contain the actin-binding sites
electron micrographs of cytoplasm. These and ATPase activity. The thick myosin fila¬
filaments are usually widely dispersed and ments of muscle are 15 |xm long and 15 nm
have a sinuous course through the cytoplasm, wide and are composed of 300 to 400 mole¬
but in cells grown in vitro they often occur cules. The filaments formed in vitro from
in bundles that are visible with the light the myosins of nonmuscle cells are all very
microscope and have been called “stress fi¬ much smaller.
bers.” The interaction of actin and myosin fila¬
Actin extracted from tissues is in the form ments to generate tension in striated muscle
of globular molecules (G-actin), but under has been thoroughly studied and the mech¬
appropriate conditions it will polymerize in anism is quite clear (see Chapter 10). How¬
vitro to form 6-nm filaments (F-actin). These ever, in nonmuscle cells the arrangement of
consist of the globular molecules arranged in actin, myosin, a actinin, and tropomyosin in
a double helix with a pitch length of 37 nm. the cytoplasm has not been worked out and
Electron micrographs give a misleading the mechanism of their interaction in cell
impression that microfilaments are present motility is not understood.
in rather low concentration. This can be
attrributed to the fact that actin filaments are
Intermediate Filaments
dissolved by prolonged exposure to conven¬
tional fixatives, or are transformed to net¬ The early studies of the filamentous com¬
works of very thin microfilaments that usually ponents of the cytoplasm concentrated upon
go undetected in thin sections. When cells the microtubules (25 nm) and actin filaments
are rapidly frozen with liquid helium, deeply (6 nm) as the principal components of the
etched, and rotary shadowed, their cytoplasm cytoskeleton. But it soon became apparent
is seen to be crowded with actin filaments. that there was another major category of
They are present in highest concentration at filaments (8—11 nm) present in the cytoplasm
the peripfiery of the cell where they form a of most eukaryotic cells. Because their cross-
dense meshwork immediately beneath the sectional dimensions fell between those of
plasma membrane. Cytoplasmic organelles microtubules and actin, they came to be
are usually excluded from this zone, which called intermediate filaments. When these were
was called the ectoplasm or cell cortex by clas¬ isolated from various cell types and their
sical cytologists. polypeptides characterized, the intermediate
Actin has a dual function in the cell. A filaments proved to be not a single protein
cross-linked lattice of actin filaments is the but a family of ultrastructurally similar but
component of the cytoskeleton largely re¬ chemically distinct filaments composed of
sponsible for the gelatinous consistency of subunits of differing molecular weight. Em¬
the cytoplasmic matrix, and its local interac¬ ploying fluorescent antibodies raised against
tion with myosin generates the forces neces¬ their purifed subunits, five major classes of
sary for cell motility. intermediate filaments can now be identified
 THE CELL • 29

Table 1-3. COMPONENTS AND DISTRIBUTION OF INTERMEDIATE FILAMENTS

Peptide Subunit
Protein Mol. Wt. Distribution

Prekeratin 40,000-80,000 Keratinizing and

nonkeratinizing
epithelia
Vimentin —52,000 Mesenchymal cells and
their derivatives
Desmin -55,000 Skeletal, cardiac, and
smooth muscle
Neurofilament protein 68,000 Neurons
160,000
210,000
Glial fibrillary acidic protein 51,000 Astrocytes

immunocytochemically; keratins, desmin, vi- are not found in cells of mesenchymal origin.
mentin, neurofilaments, and glial filaments (Ta¬ Keratins are composed of six or more ma¬
ble 1—3). Most of the cell types in adult jor polypeptides with molecular weights
animals contain only a single intermediate ranging from 47,000 to 58,000. The solubi¬
filament type, but a few contain two types. lized subunits can be repolymerized in vitro
Keratin Filaments. Intermediate filaments to form filaments (8 nm) of the same appear¬
of this class are characteristic of epithelial ance as the native filaments. Although all
cells (Fig. 1—29). They are most abundant in keratin filaments are morphologically similar,
stratified squamous epithelia such as the ep¬ the keratins in different epidermal tissues
idermis where they occupy the bulk of the and those formed at successive stages of dif¬
cytoplasm of the fully differentiated superfi¬ ferentiation of the same cell type may differ
cial cells. Present in other epithelial cells in slightly in the proportions of their several
smaller numbers, they form bundles of tono- polypeptide subunits.
hlaments that terminate in desmosomes at Desmin Filaments. The cytoskeleton of
sites of cell-to-cell adhesion. Keratin filaments smooth, skeletal, and cardiac muscle is com-

Figure 1-29. Micrograph or an area of cytoplasm from cultured Sertoli cell showing numerous intermediate filaments.
The individual filaments are not resolved by the light microscope, but bundles of filaments such as those shown here
are visible when treated with fluorescent antibody to filament peptides. (Micrograph courtesy of W. Vogl.)
30 • THE CELL

prised of intermediate filaments of desmin, a brillary acidic protein (GFA), a polypeptide of

protein of 50,000 to 55,000 M.W. In striated 51,000 M.W.
muscle, a delicate framework of these fila¬ The intermediate filaments are morpho¬
ments links the myofibrils together laterally logically indistinguishable, and to divide
and serves to keep the sarcomeres in register. them into five types on the basis of immu¬
At each Z band, desmin filaments form a nological differences in their polypeptide
planar network that surrounds the myofibrils subunits may seem to the student an aca¬
and extends laterally to the plasma mem¬ demic exercise of little value, but it has had
brane. In smooth muscle, desmin filaments unexpected applications. Most cell types in
form a three-dimensional network linking the body contain only one of the several kinds
together cytoplasmic dense bodies that are of intermediate filaments. Specific antibodies
sites of atachment of the actin filaments of against the major subunits of each are now
the cell’s contractile apparatus. The lattice of available. Immunofluorescence microscopy
desmin intermediate filaments is also an¬ has therefore proved a valuable tool for dis¬
chored to dense plaques distributed over the tinguishing cell types in pathological condi¬
inner aspect of the cell membrane. Thus, in tions where morphological criteria alone are
all three types of muscle cells, desmin fila¬ inadequate for identification. For example,
ments provide a cytoskeletal framework for in malignant tumors the cytological charac¬
attachment and mechanical integration of the teristics of the cells may be so altered that it
contractile proteins. is difficult to determine the tissue of origin.
Vimentin Filaments. Connective tissue fi¬ In such cases, immunocytochemical identifi¬
broblasts and many other cell types derived cation of the type of intermediate filaments
from the embryonic mesenchyme contain in¬ often makes it possible to determine whether
termediate filaments whose major subunit is the tumor is of epithelial, mesenchymal,
a 52,000 M.W. polypeptide called vimentin. neural, or glial origin.
Although filaments of this type may occur
throughout the cytoplasm, they are most con¬
Microtrabecular Lattice
sistently in intimate association with the nu¬
clear envelope and may provide the nucleus When thinly spread cells in tissue culture
with mechanical support or stabilize its posi¬ are fixed, dried by the critical point method,
tion in the cell. In cell types that contain and examined with the high-voltage electron
more than one kind of intermediate filament, microscope, a three-dimensional lattice of
vimentin is always one of them. Thus, it may slender strands (10—12 nm) is seen through¬
be associated with desmin in vascular smooth out the ground substance of the cytoplasm.
muscle cells; with glial filament acidic protein This provocative observation has led to the
in astrocytes of the brain; and so forth. suggestion that the cytoplasmic matrix is not
Neurofilaments. Neurofilaments (10 nm) a homogeneous protein solution, but a gel
are a conspicuous feature of the perikaryon, with a highly structured microtrabecular lattice
dendrites, and axon of nerve cells. They tend forming its solid phase, and linking together
to be uniformly spaced and oriented parallel the other filamentous components and or¬
to the long axis of the cell processes. They ganelles into a single structural and func¬
have thin lateral appendages not found on tional unit called the cytoplast. The molecular
other intermediate filaments. Neurofilaments structure and composition of the trabecular
consist of three major polypeptides with mo¬ lattice have not been worked out in any detail
lecular weights of 210,000, 160,000, and and its existence in the living cell has not yet
68,000. They appear to provide internal sup¬ been accorded general acceptance. Some cell
port for the long nerve cell processes and are biologists contend that the preparative pro¬
probably essential for maintaining the gel- cedures necessary for its demonstration pre¬
ated state of the cytoplasm. Freshly extruded cipitate soluble proteins of the cytoplasm on
axoplasm is a firm gel but rapidly becomes a a three-dimensional network of actin fila¬
sol in the presence of Ca2+, which permits ments. On the other hand, those who accept
proteolytic degradation of the neurofila¬ its reality attribute the control of cell shape
ments. and cell movement to the integrated func¬
Glial Filaments. The intermediate fila¬ tioning of the cytoskeletal elements and the
ments (8 nm) of the nonneuronal cells of the microtrabecular lattice, and speculate that
central nervous system—astrocytes, oligoden¬ enzymes incorporated in it may be spatially
drocytes, and microglia—consist of glial fi¬ coordinated in such a way as to favor their
 THE CELL • 31

Figure 1-30. Schematic presentation of the microtrabecular lattice in the cytoplasm of a thinly spread cell in culture as
interpreted from high-voltage electron micrographs. The trabeculae are joined at thicker nodal points. The lattice is
traversed by microtubules, and the trabeculae appear to attach to these and to the cell membrane. (Drawing courtesy
of K. R. Porter.)

sequential interaction with their substrates bonucleic acid (DNA), the chromatin is col¬
rather than relying on diffusion and random ored but the nucleolus, which is rich in ribo¬
collision. The viscoelastic properties of the nucleic acid (RNA), is not stained.
ground substance and the directional move¬ The nucleus is the archive of the cell, the
ments of visible particles within the cytoplasm repository of its genetic material. Encoded in
are consistent with a highly structured ma¬ its DNA is the information necessary for
trix, but the concept of a microtrabecular synthesis of all the integral proteins of the
lattice distinct from other filamentous com¬ cell and all its potential secretory products.
ponents of the cytoplasm remains controver¬ It is the source of the informational macro¬
sial. molecules ribosomal ribonucleic acid (rRNA),
messenger ribonucleic acid (mRNA), and transfer
ribonucleic acid (tRNA), which direct the pro¬
NUCLEUS tein synthetic activities of the cell cytoplasm.

The nucleus stained with hematoxylin or Nuclear Envelope

other basic dyes is the most conspicuous or¬
gan in the cell. It is centrally situated and The nucleus is enclosed by two parallel
usually round or elliptical but, in some cell membranes that bound a narrow space, the
types, may be deeply infolded or lobulated. perinuclear cistern. The inner and outer mem¬
Chromatin, the nucleoprotein that binds the branes are continuous with one another
basic dyes, occurs in aggregations of varying around the nuclear pore channels (Fig. 1—31)
size called karyosomes. These are scattered that traverse the nuclear envelope and pro¬
throughout the nucleoplasm but are usually vide avenues of communication between the
more concentrated at its periphery. The nu¬ nucleoplasm and cytoplasm. The outer nu¬
cleolus, a prominent intensely staining nuclear clear membrane has associated ribosomes
organelle, is centrally or eccentrically placed and is occasionally continuous with tubules
in the nucleoplasm. In preparations stained or cisternae of the endoplasmic reticulum
by the Feulgen reaction specific for deoxyri¬ (Fig. 1—31). Thus, the lumen of the perinu-
32 • THE CELL
v
Nuclear pores

( heterochromatin )

Condensed
chromatin

Nucleolus Figure 1-31. Schematic interpreta¬

tion of the state of the chromatin in
the interphase nucleus. The con¬
densed portions of the chromosomes
(heterochromatin) are thought to be
relatively inactive. The extended or
uncoiled segments (euchromatin) are
sites of active transcription. Also
shown is the nuclear envelope delim¬
iting a perinuclear cistern, which is
continuous with the endoplasmic re¬
ticulum.

Dispersed
extended
chromatin
(euchromatin)

Nuclear Endoplasmic
envelope reticulum

clear cistern is continuous with that of an plex. Associated with the perimeter of the
extensive system of membrane-bounded can- ring facing the cytoplasm are eight particles
aliculi in the cytoplasm. The nuclear enve¬ identified as ribosomes (Fig. 1—33).
lope is therefore regarded as a specialized The structural framework of the complex,
portion of the cell’s endoplasmic reticulum. consisting of the rings, the spokes, and their
The term nuclear pore originally referred connecting links, thus possesses octagonal
only to the membrane-bounded channel symmetry around an axis perpendicular to
from nucleus to cytoplasm; it was subse¬ the planes of the membranes. In freeze-
quently found that there are several discrete, fracture replicas, and in negatively stained
nonmembranous structures associated with preparations of the intact nuclear envelope,
the pore. The term nuclear pore complex is now the pores occasionally have an octagonal out¬
used to embrace all constituents of this com¬ line, presumably imposed by the octagonal
plex organelle. A hollow cylinder of dense symmetry of the associated nonmembranous
material extending through the pore has long components. How this highly organized com¬
been called the annulus, and a thin transverse plex functions to permit and regulate the
diaphragm with a central thickening was de¬ passage of molecules between the nucleus
scribed as extending across the annulus at its and cytoplasm is not known.
midpoint (Fig. 1-32). By image processing of The number and distribution of nuclear
micrographs of negatively stained pore com¬ pores varies with cell type and in different
plexes, the annulus has now been resolved states of differentiation of the same cell type
into two distinct coaxial rings attached to the (Fig. 1—34). Most detailed studies have been
nuclear membranes at the inner and outer carried out on oocytes in which nuclear pores
rims of the pore, so that one faces the nu¬ are exceptionally abundant.
cleoplasm and the other the cytoplasm. Ex¬ On the inner aspect of the nuclear enve¬
tending inward from these rings, in the mid¬ lope, a thin meshwork of interwoven fine
dle of the pore, are eight radially arranged filaments is interposed between the mem¬
structures called spokes. These approach a brane and the peripheral chromatin (Fig. 1—
central spherical granule or plug. Together 32). This fibrous lamina has a supporting func¬
these components correspond to the pore tion stabilizing the inner nuclear membrane
diaphragm and its central thickening de¬ and providing sites of attachment for other
scribed from thin sections of the pore com¬ structural components of the nucleoplasm. It
 THE CELL • 33

Figure 1-32. Micrograph of a typical nucleus showing a prominent nucleolus and large aggregations of heterochromatin
against the nuclear envelope, which is traversed by numerous pores (at arrows). Inset (upper left) shows two nuclear
pores traversed by pore diaphragms. Inset (lower right) shows the fibrous lamina present on the inner aspect of the
nuclear envelope.
 34 • THE CELL

a . iNcycuiveiy siainea preparation of nore mm

Plexes isolated from an amphibian oocyte The central
Tnn£hanlei9ht surrounclin9 "Spokes” are clearly visible
T*!!r heSe comPrise subunits of what was formerly
called the pore diaphragm in micrographs of thin sectionsy

The Journal CofUceell N' Z1982

ReProd“ced from
ine journal of Cell Biology 93:63, by coovriaht
permission of The Rockefeller University Press.) 9

pores. In most cell types'"''they^rdTs't'ributed3morf untolmVove°La sperma!ocyte showing the nuclear

pores are separated by wide pore-free areas and nTe ? lUrface' but ln ,his ceil dense aggregations o
spermatids. (From FawL, D. as the diffeSteinto
 THE CELL • 35

varies greatly in its thickness in different cell types in the body vary greatly in their range
types and in the same cell type in different of synthetic activities and in the proportion
species, but it is now believed to be a ubiq¬ of chromatin that is in the active dispersed
uitous component of the eukaryotic nuclear state. The depth and pattern of chromatin
envelope. It can be isolated as a thin coherent staining differs, therefore, from cell type to
lamina also containing portions of the nu¬ cell type and may provide useful criteria for
clear pore complexes. It is composed of three their identification.
major polypeptides with molecular weights Analysis of the ultrastructural components
between 60,000 and 70,000. When fluores¬ of the nucleoplasm progressed less rapidly
cent antibody against these is applied to ceils than that of the cytoplasm. Electron micro¬
in culture, the staining is confined to the graphs of the nucleus in thin sections gave
periphery of the nuclei, suggesting that the no hint of the high degree of order that
fibrous lamina is biochemically distinct from genetic considerations had led us to expect.
a general structural framework of the nuclear Heterochromatin appeared as poorly defined
matrix that has been postulated by some cell dense aggregations of 20- to 30-nm subunits.
biologists. When the nuclear envelope is dis¬ Whether these represented small granules of
persed during cell division, the antigenic nucleoprotein or cross sections of highly con¬
polypeptides of the fibrous lamina become voluted fibrils could not be determined by
diffusely distributed throughout the cyto¬ study of thin sections. However, if isolated
plasm, and in telophase they become local¬ nuclei are disrupted and their content spread
ized at the site of reconstruction of the nu¬ on the surface of water, the dissociated chro¬
clear envelope around the aggregated matin can be picked up on specimen grids,
chromosomes of the daughter cells. The spe¬ dried, and examined directly. In micro¬
cific association of these polypeptides with graphs of such preparations, the chromatin
the inner aspect of the nuclear envelope appears as a tangle of 20- to 30-nm fibrils
suggests a significant biochemical difference (Fig. 1—35A ). When nuclei are ruptured and
between the inner and outer membranes. spread on the surface of low-salt buffers, the
chromatin is dissociated into 10-nm fila¬
ments, and if the shearing forces generated
Chromatin in outflow of the chromatin are strong
enough, these are further extended and ap¬
The genetic material of the nucleus is con¬ pear as delicate beaded strands composed of
fined to the chromosomes, which are seen as regularly repeating discoid subunits about 11
discrete rodlike organelles only during one nm in diameter connected by an extremely
stage of nuclear division. When the nucleus thin (4-nm) filament interpreted as double-
is reformed after division, major segments of stranded DNA (Fig. 1—355). The discoid
the chromosomes become uncoiled and dis¬ subunits, now called nucleosomes, have a core
persed in the nucleoplasm. In this extended composed of two molecules each of the his¬
state the genetic material has little or no tones H4, H3, H2A, and H2B. A segment of
affinity for histological stains, and in electron DNA, 140 base pairs in length, is coiled
micrographs its subunits are indistinguisha¬ around this histone core, and spacer seg¬
ble from other finely granular and flocculent ments of variable length (10—70 base pairs)
constituents of the nucleoplasm. Other seg¬ extend between successive nucleosomes. His¬
ments of the chromosomes remain con¬ tone H, is somehow associated with the con¬
densed and do bind basic dyes. These stain- necting segments of DNA (Fig. 1—36). When
able portions of the interphase chromosomes not artificially extended, the nucleosomes
constitute the visible chromatin of the nucleus. and their associated DNA are believed to
This condensed chromatin is termed hetero¬ form the 10-nm filaments seen in micro¬
chromatin and the invisible dispersed form is graphs of dissociated chromatin. In their
referred to as euchromatin (Fig 1-31). Differ¬ native state, these are helically coiled around
ing degrees of synthetic activity are associated a central channel to form the 20- to 30-nm
with these two states of aggregation of the nucleoprotein fibers making up the hetero¬
genetic material. The euchromatin is the por¬ chromatin of the nucleus. The dense subunits
tion being transcribed to direct protein syn¬ seen in thin sections of chromatin, formerly
thesis in the cytoplasm, and the heterochro¬ called “granules,” are now interpreted as
matin contains that portion of the genome transverse and oblique sections of 20- to 30-
not being expressed. The numerous cell nm nucleoprotein fibers.
 36 • THE CELL

W’ Cbromfn from salamander erythrocyte spread upon water, fixed in formalin critical ooint driPd an
shadowed with carbon-platinum appears as a tangled mass of 20- to 30-nm fibrils. (Microqraoh courtesv n? RkS
B, If chromatin fibrils are extended sufficiently by the shear forces generated in the outflow of chromatin the fihri

“SS.rrand8, COnSiStin9 °f nUCleOSOmeS COnneCted by SegmentS 0f doub,e-strande

Chromosomes
contributed by the sperm and the other by
the ovum. One pair, the sex chromosomes, dif¬
Chromosomes are the rodlike or threadlike fers from the other 22 in males. One mem¬
organelles that become visible in the nucleus
ber, the Y chromosome, is shorter than its
of cells when all of the chromatin reverts to
partner, the X chromosome. Since the homol¬
its condensed form during cell division. The
ogous pairs separate during formation of the
number of chromosomes is constant in all
haploid germ cells, the male produces, in
somatic cells and is characteristic for each
equal numbers, spermatozoa containing a Y
species. For the human somatic cells, the
chromosome and spermatozoa bearing an X
number is 46—the diploid number. The germ
chromosome. In humans, the male is the
cells have half as many, 23—the haploid num¬
heterogametic sex and its chromosome comple¬
ber. At fertilization, the fusion of the haploid
ment is designated 44XY. The female is the
sets of chromosomes of the sperm and egg
homogametic sex (44XX) having two X chro¬
restores the diploid number in the cells of
mosomes and producing ova all containing
the zygote. Abnormal cells that contain more
an X chromosome. Their fertilization by a
than two sets of chromosomes are said to be
22X spermatozoon results in a female off¬
polyploid. Cells exhibiting departures from the
spring (44XX); fertilization by a 22Y sper¬
diploid number that do not involve whole
matozoon results in a male (44XY).
sets of chromosomes are described as aneu-
ploid. The study of chromosome morphology
seemed to be purely of academic interest
Each of the chromosomes in a haploid set
until it was discovered 25 years ago that the
has its own characteristic size and shape, and
cells of children with mongolism (Down’s
a diploid cell has two chromosomes of each
syndrome) had an extra chromosome. The
kind forming homologous pairs, one member
discovery of a specific chromosomal abnor-
 THE CELL • 37

Figure 1-36. Diagrammatic representation of a 30-nm chromatin fiber showing its postulated helical structure around a
central channel. In lower part of figure, the fiber is drawn out to maximal extension to illustrate the nucleosome core of
histones with the DNA double helix wrapped around it. Successive nucieosomes are joined by spacer segments of DNA
and their associated histone (Figure redrawn after Bradbury, LaRecherche 9.644, 1978, Worcel, A., and Benyajati,
C., The Cell 72:88, 1977; Olins, A. L., and Olins, D. E. Am. Scientist 66:704, 1978.)
38 • THE CELL

mality in a common congenital disorder This permits accumulation of cells in meta¬

aroused great interest in the examination of phase of mitotic division. The cultures are
human chromosomes. This has led to iden¬ then fixed and stained for chromosomal anal¬
tification of a growing list of examples of ysis.
disease states that are associated with visible The metaphase chromosomes in such
chromosomal abnormalities. Cytogenetic preparations each consist of two identical
analysis has now become a routine laboratory parallel strands, the chromatids, joined to¬
procedure on pediatric and obstetrical serv¬ gether at a narrow region, the primary con¬
ices of hospitals. It is possible in some in¬ striction. This contains a pale-staining centro¬
stances to diagnose congenital disorders be¬ mere or kinetochore, which is the site of
fore birth from chromosomal analysis of cells attachment of the spindle fiber (fusal micro¬
obtained by aspiration of amniotic fluid (am¬ tubules) to that chromosome. Some chromo¬
niocentesis). It is necessary, therefore, for somes have another narrow segment, a sec¬
students of medicine to acquire some knowl¬ ondary constriction, on one of the arms. The
edge of the basic terminology of cytogenetics. small terminal portion separated from the
Since chromosomes are accessible for anal¬ rest of the chromosome by the secondary
ysis only at metaphase of mitotic cell division, constriction is called a satellite.
a source of dividing cells is needed (Fig. 1— Successful cytogenetic analysis requires
37). Leukocytes separated by centrifugation that the 23 chromosomes be individually
of a blood sample are cultured for a few days identifiable. Since chromosomes differ in
after exposure to phytohemagglutinin, a sub¬ length and in the position of their centro¬
stance of plant origin that has the property mere or primary constriction, these features
of stimulating proliferation of lymphoblasts. are useful in their recognition. If the cen¬
Cultures are then treated with colchicine, a tromere is in the middle and the arms of the
plant alkaloid that blocks the formation of chromatids are of about equal length, the
the mitotic spindle and arrests cell division. chromosome is described as metacentric. If the

Figure 1-37. Polar view of chromosomes on the equatorial plate of a dividing lymphoblast as seen in an electron
micrograph. Micrographs of thin sections are not useful for cytogenetic analysis because whole chromosomes are not
included in the section and their relative lengths cannot be determined.
 THE CELL • 39

centromere is between the midpoint and one Primary constriction

end, the chromosome is said to be submetacen- (Kinetoehore)
Chromatid
tric. If it is quite near one end, the chromo¬
some is termed acrocentric, and if at the very
end, it is telocentric (Fig. 1—38).
To facilitate systematic analysis, the chro¬
Secondary constriction
mosomes are cut out of a photomicrograph;
the homologous pairs are identified, juxta¬
posed, and arranged in groups on the basis
of differences in length, and position of the
centromere (Fig. 1—39). Such an ordered Metacentric
arrangement of the chromosomes of any an¬
imal species is referred to as its karyotype.
The human karyotype is composed of
Submetacentric
seven groups within each of which the mem¬
bers are arranged in order of decreasing size.
Group A consists of large metacentric chro¬
mosomes, numbers 1 to 3; Group B includes Acrocentric
large submetacentrics 6 to 12 and the X
chromosome; group D includes medium¬
sized acrocentric chromosomes 13 to 15;
Group E consists of short metacentrics 19 Telocentric
and 20; and Group G are very short acrocen- Figure 1-38. Diagram presenting the descriptive terms
trics 21 and 22, and the Y chromosome. Some for chromosomes based on the location of the primary
cytogeneticists include the X and Y chromo¬ constriction or kinetoehore.
somes in groups with autosomes according to
their size; others prefer to set them apart as mosome (44X0) leads to a phenotypic female
a separate group as in Figure 1—39. whose ovaries are vestigial and usually devoid
Within the seven groups, it is difficult to of germ cells. An extra X chromosome in a
identify individual chromosomes by number, phenotypic male (44XXY) results in Klinefel¬
but this has been facilitated by staining with ter’s syndrome, characterized by underdevel¬
a modified Giemsa technique or with a flu¬ oped testicles, sterility, and breast develop¬
orescent quinacrine compound. These meth¬ ment (gynecomastia).
ods bring out characteristic patterns of cross¬ In the cells of females, one of the X chro¬
banding that provide additional criteria for mosomes remains condensed (heterchro-
identification of individual chromosomes. matic) during interphase and appears as a
The hypothetical units of inheritance, the small clump of intensely staining chromatin
genes, are known to be arranged in linear usually located adjacent to the inner surface
sequence along the length of the chromo¬ of the nuclear envelope. This sex chromatin
somes. Although these cannot be seen with (Barr body) is not found in the nuclei of
the light microscope, the approximate posi¬ normal males. This sexual dimorphism of
tion of a particular gene (gene locus) can the interphase nuclei makes it possible to
sometimes be described in relation to one of determine the genetic sex of an individual by
the bands characteristic of that chromosome. light microscopic examination of squamous
The cytogenetic anomalies of clinical sig¬ cells scraped from the inside of the cheek.
nificance involve aneuploidy of one or more This simple test is clinically useful in the
chromosomes, loss of a portion of a chro¬ differential diagnosis of abnormalities of sex
mosome (deletion), or transfer of a segment development.
to a nonhomologous chromosome (transloca¬ Ultrastructure of Chromosomes. Analysis
tion). In Down’s syndrome, for example, of the ultrastructural organization of chro¬
there is an extra chromosome 21 (trisomy 21), mosomes continues to be one of the more
owing to failure of one of the members of challenging problems in cell biology. Any
that homologous pair to go to the appropri¬ acceptable model must be compatible with
ate spindle pole in meiotic division during what is known about the replication of the
formation of one of the gametes. Aneuploidy genome, gene transcription, chromatid ex¬
involving the sex chromosomes may result in changes, chiasma formation in meiosis, and
Turner’s syndrome, in which loss of an X chro- other well-documented aspects of chromo-
40 • THE CELL
\ \

Figure 1-39. Photomicrograph of hu¬

man metaphase chromosomes of a di¬
viding lymphoblast (above). In lower
half of figure is the human karyotype
constructed by cutting out the chro¬
mosomes from a photograph like that
above and arranging them in groups
according to their size and the location
of the primary constriction. The 22 pairs
of autosomes are identified by number
KARYOTYPE and the six chromosomes by X and Y.
(Photomicrograph courtesy of L. Lisco

n
and H. Lisco.)

lift
4 5
1 2 3

14 K a IIS A if
6 7 8 9
H
io
ft 8
11
88
12

Tnft ft a o a'
13 14 15
X ft
16
OS lit'
17 18
, T7—
'* v * *'
21
* *
22
A ft •
X Y
19 20

some behavior. No model proposed for the High-voltage electron microscopic (Fig. 1—
mammalian chromosome has yet been ac¬ 40) and biochemical studies on isolated mam¬
corded general acceptance, and current malian chromosomes suggest that they too
interpretations depend on analogy with the are made up of closely packed 0.5 to 2 pim
chromosomes of amphibian oocytes, which long loops of nucleoprotein fibers arranged
lend themselves to analysis because of their radially around a central axis. The mitotic
large size and relative simplicity. They consist chromosomes are transcriptionally inactive
of two chromatids each made up of two very and the loops consist of the same 20- to 30-
long DNA molecules that are folded to form nm fibers described above in the heterochro¬
hundreds of symmetrical lateral loops that matin of interphase nuclei. When the his¬
radiate from the long axis of the chromo¬ tones of isolated chromosomes are chemically
some. These lateral loops are responsible for extracted and the residue spread and exam¬
the name “lamp-brush” or “bottle-brush” ined with the electron microscope, a remark¬
chromosomes. The loops have been shown able transformation is found to have taken
to be sites of active RNA synthesis and rep¬ place. The DNA of the nucleoprotein loops,
resent functional units of the DNA molecule now depleted of histones, has uncoiled and
called replicons. The central axis of the chro¬ extended, forming a broad halo around an
mosome seems to consist of contracted inac¬ axial protein framework that retains the orig¬
tive segments of the DNA molecule. inal linear dimension of the chromosome
 THE CELL • 41

by a rim of reactive nucleolus-associated chro¬

matin.
In electron micrographs the nucleolus is
quite variable in its appearance from one cell
type to another, and within the same cell type
its size and organizational pattern may
change in different functional states. Com¬
mon to all, however, is a three-dimensional
network of anastomosing dense strands 60 to
80 nm thick, called the nucleolonema (Fig. 1—
42). It is composed of 12-nm dense granules
in a matrix of fine filaments. The nucleolo¬
nema exhibits local variations in the state of
aggregation of its filamentous component.
Regions rich in granules tend to be less
dense with the filaments more loosely orga¬
nized, whereas in segments free of granules
the filaments are closely compacted, result¬
ing in greater electron density. The filamen¬
tous and granular components of the nu¬
Figure 1-40. Electron micrograph of unsectioned meta¬ cleolonema are ribonucleoprotein. The pale
phase chromosome from a cultured cell, showing the
chromatids, primary constriction, and what appear to be interstices between the meshes of the nucleo¬
closely packed loops of filaments radially arranged around lonema are occupied by material indistin¬
the long axis of the chromatids. (Micrograph courtesy of guishable from the surrounding nucleo¬
H. Ris.) plasm. The presence of DNA in some of the
interstices can be demonstrated autoradio-
graphically.
The nucleolus disperses during cell divi¬
(Fig. 1-41). The DNA still seems to be in the sion and is reformed in the daughter cells
form of loops but now many times longer during reconstitution of their nuclei. The
(20—24 pm) than when associated with his¬ nucleolus is reformed at particular sites on
tone and coiled into 20- to 30-nm nucleopro- one or more of the chromosomes. The nu¬
tein fibers. cleolus organizer region of the chromosomes is
A typical mammalian cell nucleus 4 to 5 often recognizable with the light microscope
(Jim in diameter contains a quantity of DNA as a narrower and less intensely stained seg¬
that if fully extended would be nearly 1 meter ment. These are also called secondary con¬
in length. Its coiling and condensation into strictions to distinguish them from the pri¬
the small volume occupied by the mitotic mary constriction (kinetochore or centromere),
chromosomes depends on a highly specific which is the site of attachment of spindle
association with histones and involves an microtubules to the chromosome. The genes
8000-fold shortening and compaction. How coding for ribosomal RNA are located in the
this orderly process is controlled at each cell nucleolus organizer region, and the nucleolus
division so as to maintain the same chromo¬ remains in close topographical relation to this
somal form and arrangement of genes along segment of the chromosome.
their length continues to defy explanation. The number of nucleolus organizer re¬
gions in the chromosome set determines the
number of nucleoli formed. This number
Nucleolus should theoretically be constant for a given
The most conspicuous organelle of the species. However, the number actually found
nucleus is the nucleolus. It is visible in the may depart somewhat from the expected
living cell as a rounded refractile body ec¬ number. The number in most somatic cells
centrically placed in the nucleus. In histolog¬ ranges from one to four, but polyploidy may
ical sections it is intensely stained with basic result in multiples of the basic number, and
dyes. Its affinity for basic dyes is due to its coalescence of nucleoli may result in fewer
content of ribonucleoprotein. It is usually than the expected number.
unstained by the Feulgen reaction for deox- The nucleolus is an essential nuclear or¬
yribonucleoprotein but is often surrounded ganelle that functions as a site of processing
42 • THE CELL

Figure 1-41. If isolated chromosomes are treated with dextran sulfate and heparin, the histones are extracted, permitting
the DNA to uncoil forming a broad halo around the core structural proteins. These can be spread upon water and picked
up on electron microscope grids (see inset). If an area such as that in the rectangle is examined at higher magnification,
the halo is revealed as a labyrinthine pattern of 4-nm filaments of DNA. (Micrograph from Paulsen, J. R., and U. K.
Laemmli. Cell 72:817, 1977.)
 THE CELL • 43

Figure 1-42. Electron micrographs of nucleoli from two different cell types illustrating variations in the reticular pattern
of the nucleolonema.

and partial assembly of subunits of the ribo¬ somai RNA. Thus, it is apparent that the
somes. Biochemical analyses of isolated nu¬ protein-synthetic activities of the cytoplasm
cleoli have identified three major classes of are dependent on the functional integrity of
RNA with sedimentation constants of 45S, the nucleolus.
35S, and 28S. Labeled precursors are incor¬
porated most rapidly into the 45S fraction
and appear later in the others. It is believed CELL ACTIVITIES
that ribosomal RNAs are first synthesized as
long 45S molecules that subsequently Having defined the principal structural
undergo cleavage to yield smaller fragments. components of the cell, it may facilitate dis¬
Among these an 18S RNA combines with cussion of tissue and organ functions in later
specific proteins and passes rapidly into the chapters if certain activities common to most
cytoplasm where it is incorporated into the cell types are briefly described here.
smaller subunits of the ribosomes. A 35S
fragment conjugated with protein forms Cell Division
small particles that may correspond to the
granular component seen in electron micro¬ Growth, renewal, and repair in all multi¬
graphs of the nucleolonema. The 35S RNA cellular organisms depend on formation of
is ultimately cleaved to yield a 28S RNA, new cells by repeated division of preexisting
which is incorporated with its associated pro¬ cells. Two processes of cell division are distin¬
teins into the large subunit of the ribosomes. guished, mitosis, which occurs in somatic cell
Destruction of the nucleolus of living cells types, and meiosis, which takes place only in
by bombardment with a laser beam results in the development of germ cells in the ovary
cessation of incorporation of RNA precursors and testis. The two have many features in
into ribosomes. In a mutant of the African common but differ in the behavior of the
frog Xenopus, development of embryos that chromosomes during the early stages of di¬
lack nucleoli is arrested at an early stage vision. Two events occur in succession in the
because of their inability to synthesize ribo- course of cell division—division of the nu-
44 • THE CELL

cleus (karyokinesis), and division of the cyto¬ Anaphase is initiated by division or redu¬
plasm (cytokinesis). plication of the kinetochore of the chromo¬
Mitosis. Mitotic division results in distri¬ somes so that each chromatid has its own.
bution of identical copies of the original cell’s The sister chromatids, no longer attached,
genome to the two daughter cells. Before the are then free to migrate to opposite poles of
onset of division the DNA is replicated so the spindle as separate chromosomes (Fig. 1—
that the cell enters mitosis with double the 431, J). During these anaphase movements
normal diploid complement. Cell division ex¬ of the chromosomes, the kinetochore regions
tends over a period of 30 to 60 minutes in to which the microtubules attach usually lead
mammals but may take considerably longer and the arms trail behind. The mechanism
in cold-blooded vertebrates. The period be¬ of anaphase movements is still a subject of
tween successive episodes of cell division is debate. The chromosomal fibers appear to
called interphase. The continuous train of shorten, possibly by depolymerization of the
events in mitosis is arbitrarily divided, for microtubules at their polar ends, while the
descriptive purposes, into four stages—pro- continuous fibers appear to lengthen, moving
phase, metaphase, anaphase, and telophase. the spindle poles apart. It is not yet clear
During interphase the only structure visi¬ whether movement of the chromosomes de¬
ble in the living nucleus is the nucleolus, but pends solely upon changes in length of the
in stained preparations a few clumps of chro¬ two sets of microtubules by polar addition of
matin can be seen in the nucleoplasm, usually tubulin to one and depolymerization of the
concentrated around the periphery adjacent other, or whether there is a sliding mecha¬
to the nuclear envelope. Except for these nism involving interaction of chromosomal
condensed segments, the chromosomes are and continuous microtubules and their dis¬
in the extended euchromatic state and are placement relative to one another. In either
not visually identifiable. After replication of case, at the end of anaphase, the two groups
the DNA the euchromatic regions condense, of chromosomes are clustered at the spindle
so that the chromosomes become visible in poles.
both living and fixed material as slender At telophase, segments of nuclear envelope
threadlike structures pursuing a meandering begin to reform around the chromosomes,
course throughout the nucleoplasm (Figs. 1 — and these uncoil and extend, losing their
43, 1-44). stainability except in those regions destined
The first appearance of the chromosomes to remain condensed as the heterochromatin
marks the beginning of prophase (Fig. 1— of the interphase nucleus. Nucleoli are re¬
43D, E, F). Throughout this stage they con¬ formed and the initially discontinuous seg¬
tinue to condense, becoming shorter and ments of nuclear envelope coalesce to form
thicker. Each consists of two parallel strands a complete perinuclear cistern (Fig. 1-43L-
called chromatids joined to one another at the O). With these events, karyokinesis is con¬
centromere or kinetochore, a constricted seg¬ cluded and the two identical daughter nuclei
ment common to both. Other events occur¬ have attained their interphase appearance.
ring during prophase include disappearance While the reconstitution of the nuclei is in
of the nucleoli; migration of the centrioles progress, a constriction of the cytoplasm oc¬
(which replicate just before onset of division) curs midway between them. This cleavage
to opposite poles; and the breakdown of the furrow deepens until it encounters a bundle
nuclear envelope. Disappearance of the nu¬ of the continuous fibers of the spindle. The
clear envelope marks tfie end of prophase. cell bodies of the two daughter cells remain
Metaphase begins with the alignment of connected for a short time in telophase by a
the chromosomes in the same plane in the slender cytoplasmic bridge filled with spindle
middle of the cell to form the equatorial plate microtubules that are held together at their
(Fig. 1-43H). This ordered assembly of the midpoint by dense amorphous material,
chromosomes is associated with the develop¬ which is visible with the light microscope as
ment of the mitotic spindle—a fusiform array a dark dot called the midbody. After a brief
of microtubules extending between the cen¬ delay the microtubules depolymerize, the
trioles located at the poles of the division spindle bridge gives way at one side of the
figure. Some of these microtubules go from midbody, and the two halves retract into their
pole-to-pole as the continuous fibers of the respective daughter cells to complete cytoki¬
spindle while others, called the chromosomal nesis.
fibers, extend only from the poles to the The precise replication of the DNA prior
kinetochore of each chromosome. to cell division, and the longitudinal splitting
 THE CELL • 45

Late stages of nuclear

reconstruction

Anaphase

J
Late anaphase

C D
Interphase Early prophase

Advancing reconstruction

Wk
m
^;
s:
V

Metaphase

Prometaphase E. Bohlman

Figure 1-43. Drawings of changes in mitosis of mesothelial cells of Amblystoma in culture as seen in fixed and stained
material. Compare with phase-contrast photomicrographs of living cells in Figure 1-44.
46 • THE CELL

10:53J
■P-

Figure 1-44. Phase-contrast photomicrographs of the same Amblystoma mesothelial cell in successive stages of
mitosis. Large white arrow points to the nucleolus: black arrows indicate centrioles. A to E, Prophase; F, G, H,
metaphase; i, J, K, anaphase; L, telophase. The duration of three hours is much longer than required for mitosis in
warm-blooded vertebrates.
 THE CELL • 47

of each chromosome into two identical chro¬ phase plate. The kinetochore of the bivalents
matids, which separate and are distributed to does not divide as it does in mitosis. Conse¬
opposite poles, ensures that the daughter quently, in anaphase of the first meiotic divi¬
cells are of the same genetic make-up as the sion, whole chromosomes, not sister chro¬
cell from which they were derived. matids, separate and move to the opposite
Meiosis. Meiosis occurs early in the devel¬ poles.
opment of ova and spermatozoa, and consists The nuclei in mammals are reconstituted
of two successive divisions with only one in telophase for a brief interval, but in some
replication of the chromosomes. It results in other taxa the chromosomes may proceed
separation of the two members of each pair directly into the events of the second division
of homologous chromosomes in the first di¬ without full reconstitution of the nucleus. In
vision, thus reducing the chromosomes in the second meiotic division, the haploid nu¬
each daughter cell to half the normal num¬ clei divide by a mechanism essentially iden¬
ber. The second meiotic division involves tical to that seen in mitosis. The chromo¬
separation of the two chromatids of each somes split longitudinally, the kinetochores
chromosome and results in four nuclei with divide, and sister chromatids move to oppo¬
half the normal number of chromosomes. site poles, resulting in formation of four cells
The gametes that differentiate following with haploid nuclei. The reduction in chro¬
meiosis are thus described as haploid, and mosome number occurs in the first meiotic
when a male and female gamete unite at division. Therefore, this is also called the
fertilization, the normal diploid number of reductional division, while the second is called
chromosomes is restored in the fertilized egg, the equational division.
now called the zygote. The biological significance of meiosis is
Meiosis is characterized by a very long twofold. It ensures constancy of chromosome
prophase, which is divided into five stages. number from generation to generation by
In the earliest of these, called leptotene, the producing haploid gametes whose fusion re¬
chromosomes become visible as long, thin, stores the original diploid number. It also
single strands. In zygotene, the homologous provides for genetic variability upon which
chromosomes begin to come together in close evolution depends. The members of each
lateral apposition with corresponding sites pair of homologous chromosomes in the dip¬
along their length in register. This pairing is loid spermatogonia and oogonia come from
also referred to as synapsis. The chromosomes the two parents of the individual. As a result
then begin to coil, becoming shorter and of the exchange of segments between ho¬
thicker in pachytene. In this stage, because of mologous chromosomes in prophase of the
the close apposition of homologues, the chro¬ first meiotic division, the chromatids emerg¬
mosomes may appear to be present in only ing are different genetically from the chro¬
the haploid number. The pairs are referred mosomes that entered meiosis. The random¬
to as bivalents. ness of distribution of the homologues to the
In diplotene, the paired chromosomes sep¬ daughter nuclei during reduction division
arate along their length. Their coiled nature contributes further to the genetic diversity of
is evident, and it becomes apparent that each the gametes.
is split longitudinally. Each bivalent therefore
consists of four chromatids. At certain points Cell Cycle
along their length, the homologous half¬
chromosomes cross one another and ex¬ The frequency of cell division varies
change segments. The sites of crossing are greatly among the many cell types in the
called chiasmata and are the morphological body. In some tissues (nervous tissue, cardiac
expression of the genetic phenomenon of muscle), the cells are very long-lived and
crossing over. The shortening and thickening there is no cell division in postnatal life to
of the chromosomes continues and they tend replace those lost through age or injury. In
to clump in the center. The nucleolus, which other organs such as the liver, there is nor¬
remained during earlier stages of prophase, mally a very slow renewal with cell divisions
begins to fragment in diplotene and later seen only rarely. However, after removal of
disappears altogether. part of the organ, the remaining cells are
In metaphase, the nuclear envelope is dis¬ stimulated to proliferate rapidly until the
persed and the spindle forms as it does in original volume of liver has been restored.
mitosis. The bivalents gather on the meta¬ In a number of epithelia such as those of the
48 • THE CELL
\

gastrointestinal tract and the skin where 30 tq 40 per cent of the cycle length and is
there is a continual loss of fully differentiated referred to as the S-phase. It is followed by a
superficial cells, a population of relatively short G2-phase, occupying 10 to 15 per cent
undifferentiated cells retains the capacity for of the cycle during which other preparations
rapid proliferation and provides for contin¬ for division take place. The succeeding M-
ual replacement commensurate with the rate phase includes the visible morphological
of cell loss. events of mitosis. This is followed by the Gr
For cell types that proliferate rapidly in phase, a period of active RNA and protein
vivo or in vitro with a constant doubling time, synthesis during which the daughter cells
it is possible to define a repeating sequence increase in size. It extends for 30 to 40 per
of biochemical and morphological events cent of the cycle length (Fig. 1-45). The
known as the cell cycle. The key biochemical duration of the cell cycle varies with the cell
event of the cycle is replication of the strands type and animal species. In rapidly prolifer¬
of DNA that form the chromosomes. This ating cells of laboratory rodents, it may be
occurs during interphase while the chromo¬ only one third of the cycle length of the same
somes are in the extended condition, and it cell type in humans.
is not attended by any microscopically visible The study of these phases in synchronized
change in the cell nucleus. DNA replication cultures has made important contributions to
can be detected, however, by autoradiogra¬ our understanding of the molecular biology
phy following administration of radiolabeled of cell proliferation and growth, but it is
thymidine, a precursor that is incorporated important to note that these definitions re¬
into the newly synthesized DNA molecules. quire that the cells of the population are
The period of DNA synthesis extends over rapidly growing, with a constant doubling

Interphase M itosi s

Figure 1-45. Schematic representation of the phases of the cycle and their approximate duration in the case of
proliferating bone cells with a cycle length of 25 hours. Cycle length varies greatly among different cell types, Gt being
the most variable phase. (Redrawn after Junqueira, L. C., and Carneiro, J. Basic Histology. Lange Medical Publishers,
1983.)
 THE CELL • 49

time. This type of growth is seldom found in oughly studied example, the ameboid stage
normal adult tissues. Cells may remain for of the fungus Dictyostelium, the chemotactic
weeks or months in interphase and then substance is cyclic AMP. A number of the
undergo one or more division cycles. Some white blood cells of mammals respond to
authors refer to these long phases of rest as chemotactic agents liberated at sites of tissue
a G0-phase, but this is not properly a part of injury or bacterial invasion. The chemical
the cell division cycle. characterization of the attractant for these
cells is still incomplete.
Cell Motility Connective tissue cells and epithelial cells
in culture have a different behavior. They
All cells are capable of some degree of spread and flatten against the substrate.
motility. Free cells that are not organized into Their thin, advancing end forms thin folds
compact tissues wander through interstices or ruffles that exhibit an active undulating
in the connective tissues of the body with an motion while the rest of the cell remains
active ameboid locomotion. Where cells are quiescent. This ruffling edge advances over
in close contact as in epithelia, their motility the surface and the cell body follows. In
may be limited to changes of surface config¬ neurons in culture, the cell body remains at
uration by extension and retraction of micro¬ the site of initial adherence and the active
villi, and to streaming movements within the ruffling portion moves away, drawing out
cytoplasm that continually alter the distribu¬ very long slender processes simulating the
tion of their organelles. Even such relatively axons and dendrites of such cells in the brain.
sessile cell types undergo changes of shape, Thus, the shape and pattern of movement of
rounding up at the onset of division and cells is not determined entirely by environ¬
forming a cleavage furrow during cytokine¬ mental stimuli but also depends on endoge¬
sis, and then returning to their original form. nous determinants for particular cell lin¬
During repair after injury, they migrate and eages. Where the endogenous determinants
flatten to cover denuded areas of the basal are located in the cell; how they are trans¬
lamina. mitted to daughter cells; or how they exercise
Exploration of the mechanisms of loco¬ their control over cell form are unknown.
motion and of the streaming movements that Information is now rapidly accumulating
occur within cells has proved to be one of on the structural components determining
the most challenging problems in cell biology, cell form, and on the changes induced by
and some understanding of its molecular extrinsic stimuli. The architecture of the cy-
basis is only beginning to emerge. Ameboid toskeleton can be revealed in three dimen¬
movements of migratory cells have been most sions by detergent extraction of the mem¬
thoroughly studied. In such cells there is an branes and soluble components of the
actin-rich cortical zone of cytoplasm that has cytoplasm followed by examination of the
a gel-like consistency. The initial event in unextracted filamentous framework in stereo
movement appears to involve local conver¬ micrographs. The several categories of cyto¬
sion of the gel to a sol, making this a weak plasmic filaments can be identified in situ by
point. Contraction of the cortex at the other their decoration with specific monoclonal an¬
end of the cell forces the solated cytoplasm tibodies. The organization of the cytoskeleton
forward, extending a blunt process or pseu¬ can be altered experimentally by drugs that
dopod that adheres to the substrate in a more prevent or promote the polymerization of
advanced position while the contracting tail tubulin to form microtubules. Drugs are
region detaches. The process is repeated with available that similarly affect the polymeri¬
extension of a new pseudopod, and the cell, zation and depolymerization of actin that
thus slowing, crawls over the substrate. takes place in gel-sol transformation and in
The movement may be random, or its assembly of contractile associations of actin
direction may be determined by extracellular and myosin. Polypeptides that cross-link actin
signals such as the topography of the sub¬ and stabilize the cytoskeletai lattice have been
strate, or by diffusible chemical signals in the identified and others that mediate the attach¬
medium called chemotactic agents. In chemo- ment of actin to integral proteins of the cell
taxis, pseudopods are induced at the site of membrane—a linkage essential to any mode
contact with the attractant and the cell con¬ of cell locomotion.
tinues to move up the concentration gradient Although at present we are still in the stage
of the chemotactic agent. In the most thor¬ of analyzing motile behavior and compiling
50 • THE CELL

a catalogue of the visible components and of the capsule of pathogenic bacteria, making
the macromolecules involved, these descrip¬ them vulnerable to ingestion by ensuring
tive and analytic approaches may soon pro¬ their attachment to specific receptors in the
vide the basis for valid explanations of cell membranes of phagocytes.
motility. Phagocytosis is considered to occur in two
steps: (1) attachment, which involves binding
of ligands on the particle to receptors in the
Endocytosis membrane of the phagocyte; and (2) intenor-
Endocytosis is a general term applied to the ization of the particle. In the case of patho¬
bulk uptake of materials by cells. Fluid is genic bacteria referred to above, the immune
taken in from the extracellular environment globulin (IgG) and a component of comple¬
by local invagination of the plasma mem¬ ment (C3) bind selectively to the surface of
brane followed by fusion of the neck of the the bacterium and serve as ligands, which
invagination to form a closed, fluid-filled vac¬ interact with specihc receptors (Fc) on the
uole. This is then separated from the cell membrane of the phagocytic cell (Fig. 1—46).
membrane and moved into the cytoplasm. This attachment phase of the process can
This process is called pinocytosis (drinking by occur at 1° to 3° C in vitro and thus is not
cells). All eukaryotic cells probably engage in energy dependent. Jnteriorization, however,
this activity, but in varying degrees. requires expenditure of energy. At the point
Solid particles may be taken into cells by of initial contact, a zipper-like process of
formation of pseudopods, blunt processes of ligand-to-receptor binding is initiated and
the cell surface that progressively elongate spreads laterally, tightly apposing the cell
and ultimately encircle the object. Fusion of membrane to the surface of the particle. This
the tips of the engulfing pseudopods beyond interaction is believed to generate a signal
the particle then detaches a membrane- that results in assembly of actin filaments and
bounded vacuole from the plasmalemma, myosin in the cell cortex, leading to forma¬
carrying the ingested matter into the cyto¬ tion of pseudopods. Their extension in¬
plasm. This mechanism for ingestion of solid creases the area of contact with the bacter¬
objects, 1 pm or more in diameter, is called ium, entailing further ligand-receptor
phagocytosis (eating by cells). Although some binding, which in turn induces more exten¬
epithelial cells are capable of phagocytosis, it sive aggregation of contractile proteins. This
is most highly developed in white blood cells self-propagating process continues until the
and tissue macrophages, which are special¬ tightly adhering pseudopods meet and fuse
ized for defense of the organism against beyond the bacterium to complete its enclo¬
microbial invasion. Cells exhibiting this activ¬ sure in a phagocytic vacuole or phagosome. Dis¬
ity are referred to as phagocytes. They not charge of lysosomal hydrolytic enzymes into
only ingest and destroy invading microorga¬ the vacuole and activation of other bacteri¬
nisms, but also play an important role in cidal mechanisms of the cell then results in
tissue maintenance by disposing of dead cells destruction of the invader.
and cellular debris. For example, some 3 X In addition to receptors for IgG and C3,
1011 senescent red blood cells are removed some phagocytes evidently have nonspecific
from the blood each day and destroyed by receptors that enable them to ingest a variety
the phagocytic cells in the spleen. of nonbacterial foreign particles without the
Living cells and normal extracellular ele¬ cooperation of the immune system.
ments are generally immune to phagocytosis, In pinocytosis, imbibition of fluid occurs in
but degenerating cells with altered mem¬ vacuoles of two size categories whose forma¬
branes and tissue components whose proteins tion involves somewhat different activities of
are partially denatured are readily ingested. the cell surface. In macropinocytosis, sizeable
Most nonpathogenic bacteria are rapidly droplets of fluid (0.5-1.5 pm) that are visible
phagocytized. On the other hand, a number by phase-contrast microscopy are enveloped
of pathogenic bacteria have capsules that by thin, undulating folds (lamellipodia) on the
resist binding to the surface of phagocytes. cell surface and are drawn into the cytoplasm.
Their ingestion requires the presence of im¬ In micropinocytosis, fluid is taken up in minute
mune globulin (antibody) produced by cells of invaginations of the cell membrane that bud
the body’s immune system, and/or the pres¬ off to form vesicles visible only in electron
ence of a protein constituent of blood plasma micrographs.
called complement. These substances bind to Two functionally distinct forms of micro-
 THE CELL • 51

Figure 1-46. Schematic representation of the mechanism of phagocytosis. In the attachment phase, ligands on the
surface of the particle bind to receptors on the phagocyte membrane. Progressive circumferential binding spreads from
the site of initial contact, “zippering” the membranes together. This triggers polymerization of actin in the underlying
cytoplasm and extension of engulfing pseudopods that bring the phagocyte membrane into contact for binding around
the entire circumference of the particle. When the pseudopods meet and fuse, interiorization is complete.

pinocytosis are recognized: fluid-phase pinocy- enclosed in a basket-like lattice composed

tosis and adsorptive or receptor-mediated pinocy¬ mainly of an 180,000 M.W. protein called
tosis. In the former, fluid, solutes, and clathrin (Fig. 1-48). The presence of the
submicroscopic particulates in suspension are clathrin basket gives the coated vesicles of re¬
taken up in small vesicles whose limiting ceptor-mediated pinocytosis a characteristic
membrane is unspecialized (Fig. 1—47). This spiny appearance that serves to distinguish
process is nonselective. The latter is selective in them from the smooth vesicles of fluid-phase
that certain substances are bound to specific pinocytosis.
receptors clustered in areas of the membrane Nonselective, fluid-phase pinocytosis is ap¬
that then invaginate to form pits and later parently a device for imbibition of fluid and
detach as free vesicles in the cytoplasm. The solutes or for their transport across epithelia.
luminal aspect of these vesicles has a visible Receptor-mediated endocytosis is now a sub¬
surface coat and the cytoplasmic surface is ject of intense interest because of its potential

Figure 1-47 In fluid-phase micropinocytosis, minute invaginations of the cell membrane detach and move into the
cytoplasm as smooth vesicles. In some epithelia they may transport materials across the cell. In endothelium, chains of
communicating vesicles may form transient channels from lumen to cell base. This form of pinocytosis is nonselective.
52 • THE CELL

vrCfl V HTYtt

Figure 1-48. A, In receptor-mediated pinocytosis the vesicles are lined by a thin carbohydrate-rich cell coat and their
cytoplasmic surface is enclosed in a basket-like lattice of clathrin.
B, Receptors are randomly distributed in the cell membrane. Binding of specific macromolecules causes clustering of
the receptors, and this portion of the membrane forms a coated pit that subsequently separates from the membrane as
a coated vesicle. (Redrawn and modified from Goldstein et al. Nature 279:679, 1979.)

Figure 1-49. Electron micrograph of the cytoplasmic side of the plasmalemma of a liver cell prepared by rapid-freezing,
fracturing, and deep-etching. Filaments in the cortical cytoplasm are visible and the clathrin lattice of many forming
coated pits and vesicles. (Micrograph courtesy of N. Hirokawa and J. Heuser.)
 THE CELL • 53

physiological importance in selective uptake Fawcett, D. W.: The Cell. 2nd ed. Philadelphia, W. B.
of nutritional and regulatory proteins (e.g., Saunders Co., 1981.
Flickinger, C. J., J. C. Brown, H. C. Kutchai, and J. W.
hormones such as insulin and transport pro¬
Ogilvie: Medical Cell Biology. Philadelphia, W. B.
teins such as transferrin). There is also strong Saunders Co., 1979.
evidence for the involvement of clathrin- Karp, G.: Cell Biology. New York, McGraw-Hill Book
coated vesicles in retrieval and recycling of Co., 1979.
Watson, J. D., ed.: Organization of the Cytoplasm. Cold
membrane added to the plasmalemma dur¬
Spring Harbor Symp. Quant. Biol. Vol. 46, 1982.
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rotransmitters. Cell Membrane
Branton, D.: Freeze-etching studies of membrane struc¬
ture. Trans. R. Soc. Fond. (Biol.) 261:133, 1971.
Exocytosis Finian, j. B.: The development of ideas on membrane
structure. Subcell. Biochem. 7:363, 1977.
The term exocytosis describes the release of Hendler, R. W.: Biological membrane ultrastructure.
secretory products of the cell into the extra¬ Physiol. Rev. 57:66, 1971.
cellular compartment. The product of glan¬ Ito, G.: The surface coat of enteric microvilli. J. Cell
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dular cells is accumulated and concentrated
Marchesi, V. T., H. Furtlmayr, and M. Tomita: The red
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membrane that is capable of fusing with the of a cell coat rich in carbohydrates at the surface of
apical plasma membrane. The secretory cells in the rat. Anat. Rec. 754:41, 1966.
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product of such cells may be stored for some
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receives an appropriate stimulus, the gran¬ model of the structure of the cell membranes. Sci¬
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Steck, T. F.: The organization of proteins in human red
fuses with the plasmalemma, permitting their
blood cell membrane. J. Cell Biol. 62:1, 1974.
contents to flow out. The product thus leaves
the cell without creating any breach in the Endoplasmic Reticulum

cell membrane. The membrane of the secre¬ Blobel, G., and B. Dobberstein: Transfer of proteins
tory granules that is added to the plasma across membranes. I. Presence of proteolytically
processed and unprocessed nascent immunoglobu¬
membrane during exocytosis is subsequently
lin light chains on membrane-bound ribosomes of
recovered in small vesicles that invaginate, murine myeloma. J. Cell Biol. 67:835, 1975.
bud off, and are recycled through the Golgi Blobel, G., and B. Dobberstein: Transfer of proteins
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is best exemplified in glandular cells that
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granules, but are released continuously in
Porter, K. R.: Observations on a submicroscopic baso¬
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of peptides released by puromycin from attached
indistinguishable from pinocytosis vesicles.
ribosomes. Proc. Natl. Acad. Sci. USA 56:608, 1966.
The secretory activity of such cells went un¬ Sabatini, D. D., and G. Blobel: Controlled proteolysis of
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Smooth Endoplasmic Reticulum

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54 • THE CELL
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Dallner, G., and L. Ernster: Subfractionation and com¬ Nass, M. M. K., and S. Nass: Intramitochondrial fibers
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 —— *
EPITHELIUM

In the past decade, increasing emphasis epithelial, and most of the epithelial organs
has been placed on the biology of the cell, of the body are derived from these germ
often to the neglect of higher levels of orga¬ layers. For example, the epidermis of the
nization. Much of the research in this held is skin and the epithelium of the cornea, which
now done upon a small number of cell types together cover the entire external surface of
that can be grown in isolation in vitro. Valu¬ the body, develop from the ectoderm. By
able as those studies are, the student should invagination and proliferation, this outer cov¬
be aware that isolated cells are seldom en¬ ering epithelium gives rise to tubes or solid
countered in the body. The great majority cords that form the glandular appendages of
are assembled in coherent groupings bound the skin—the sudoriparous, sebaceous, and
together by cell junctions and extracellular mammary glands. Similarly, the alimentary
matrix to form tissues. Each of the tissues is tract is lined by epithelium of endodermal
specialized for a particular function and has origin, and liver, pancreas, gastric glands,
a distinctive pattern of organization. and intestinal glands all arise in the embryo
The first chapter was devoted to the cell— by invagination, proliferation, and speciali¬
the fundamental unit of living matter. This zation of epithelial outgrowths from the lin¬
chapter, and four following, will present the ing of the primitive gut. Each exocrine gland
identifying characteristics of the basic tis¬ of the adult communicates with an internal
sues—epithelium, connective tissue, blood, mus¬ cavity or an external surface by way of ducts
cular tissue, and nervous tissue. Later chapters that open onto the epithelium of the inner
will describe the patterns in which these tis¬ or outer surface layer from which it devel¬
sues are combined to form the larger func¬ oped during embryonic life. The endocrine
tional units called organs. glands, on the other hand, usually lose their
Epithelium is a tissue composed of cells in connection with the surface epithelium from
close apposition over a large portion of their which they originally develop.
surface and with little or no intercellular In addition to the epithelial structures that
substance. In its simplest form an epithelium develop from the ectoderm and endoderm,
consists of a single coherent layer of identical there are a few organs composed of epithelia
cells covering an external, or lining an inter¬ that arise from mesoderm. Examples are the
nal, surface. But the cells may differentiate kidney and the lining of the male and female
into two or more functionally distinct types reproductive tracts.
to meet specific local requirements, and may The linings of the peritoneal cavity and of
take on various shapes and become arranged other serous cavities, and the linings of blood
in multiple layers. Several categories of epi- and lymph vessels, are all derivatives of mes¬
thelia are defined and assigned different enchyme. These are in all respects typical
terms to lend precision and consistency to epithelia but it is convenient and customary
the descriptive literature of histology and to refer to the lining of blood and lymph
pathology (Fig. 2—1). It is of some importance vessels as endothelium and to the lining of
therefore for students to learn the classifica¬ serous cavities as mesothelium.
tion and terminology of the several kinds of Epithelia are specialized for many different
epithelia. functions—absorption, secretion, transport, ex¬
cretion, protection, and sensory reception. Those
that form the outer surface of the body are
Origin and Distribution of Epithelium
adapted for protection of the organism
Two of the primary germ layers of the against mechanical damage and loss of mois¬
early embryo, the ectoderm and endoderm, are ture. They also play a role in sensory recep-

57
58 • EPITHELIUM

Figure 2-1. Drawings illustrating

the shape and arrangement of cells
in the principal types of epithelium.

Pseudostratif ied columnar

Simple columnar

Stratified squamous Transitional

tion, containing nerve endings that provide shape of the cells. If there is one layer of
the warning of pain that make the organism cells the epithelium is described as simple; if
avoid injury. Others contain neural elements, there are two or more layers it is said to be
such as taste buds and olfactory cells, spe¬ stratified. The superficial cells can usually be
cialized to function as chemical receptors. All described as squamous, cuboidal, or columnar.
substances received or given off by the body Thus, a single layer of flat cells is a simple
must traverse an epithelium; thus, many of squamous epithelium. A single layer of tall pris¬
the epithelia lining internal surfaces are mod¬ matic cells is a simple columnar epithelium. The
ified for absorption or secretion. Those con¬ corresponding multilayered epithelia are
cerned with secretion may contain only scat¬ called stratified squamous epithelium and strati¬
tered individual secretory cells, or the fied columnar epithelium.
epithelium may give rise to a gland in which Within the same general category of epi¬
most of the cells are specialized for elabora¬ thelium the cells may or may not have motile
tion of a particular product. Other epithelia, cell processes called cilia on their free sur¬
concerned with excretion, become modified faces. In the interest of more precise descrip¬
to increase their efficiency in transport of tion, it is customary to make note of this
solutes and water or in elimination of sub¬ surface specialization in naming the epithe¬
stances from the body. lium. When such a border is present, the
tissue is described as a ciliated simple columnar
epithelium or a ciliated stratified columnar epithe¬
CLASSIFICATION
lium. Similarly in stratified squamous epithe¬
OF EPITHELIA lium, the superficial cells in some cases ac¬
cumulate in their cytoplasm the fibrous
Epithelia are named and classified accord¬ protein keratin and are reduced to scalelike
ing to the number of cell layers and the lifeless residues of cells. To distinguish such
 EPITHELIUM • 59

epithelia from others in which the superficial Simple Squamous Epithelium

cells do not undergo this change, it is custom¬
ary to describe them as keratinized stratified Thin platelike cells are arranged in a single
squamous epithelium. layer and adhere closely to one another by
Among the simple columnar epithelia one their edges (Fig. 2—1). When examined in
category is described as pseudostratified be¬ surface view, especially after the cell limits
cause the cells are so arranged that the nuclei are stained with silver nitrate, a characteristic
occur at two or more levels and the epithe¬ mosaic pattern is seen (Fig. 2-2). The indi¬
lium thus appears to be stratified. However, vidual cells have polygonal or irregular wavy
it can be shown by maceration that there is outlines. In sections perpendicular to the
actually only one layer, all the cells being epithelium the cells, in profile, appear as
fixed to the basal lamina (basement mem¬ plump spindles or thin rectangles (Fig. 2-3).
brane) but only some of them reaching the Because of the large area covered by each
free surface. In truly stratified epithelia, only flattened cell, a given plane of section will
the ceils of the lowermost laver are in contact
j
pass through the nucleus of only a fraction
with the substrate. of the cells transected.
Among the stratified epithelia there are Epithelia of this variety are found on the
two types that cannot adequately be described inner surface of the wall of the membranous
by reference to the shape of their surface labyrinth and on the inner surface of the
cells. These are the transitional epithelium of tympanic membrane of the inner ear; in the
the urinary tract and the germinal epithelium parietal layer of Bowman’s capsule and in
of the male gonad. Their distinguishing char¬ the thin segment of the loop of Henle in the
acteristics will be described later. kidney; in the rete testis; and in the smallest

Figure 2-2. A thin spread of guinea-pig mesentery treated with silver nitrate and subsequently stained with May-
Grunwald and Giemsa stains. The limits of the simple squamous mesothelial cells have been blackened by the silver,
revealing their polygonal outlines. The large oval nuclei are those of the mesothelial cells. The darker ellipsoidal nuclei
are those of fibroblasts beneath the mesothelium.
60 • EPITHELIUM
\ \ X \

Figure 2-3. Epithelium of a collecting duct of the dog kidney is flattened to form a thick squamous epithelium here, but
the cells in other species may be cuboidal.

excretory ducts of many glands. Also in¬ simple columnar epithelium (Figs. 2-4E and
cluded in this category are the mesothelium 2-5) is found in the uterus and oviducts, in
lining the pleural, peritoneal, and other ser¬ the bronchi of the lung, in some of the
ous cavities and the endothelium lining the paranasal sinuses, and in the central canal of
walls of the blood and lymph vessels. the spinal cord.

Simple Cuboidal Epithelium Stratified Squamous Epithelium

On surface view this epithelium also ap¬ The epithelial sheet is thick, and a vertical
pears as a mosaic of small, usually hexagonal section shows the cells to vary in shape from
polygons, and in vertical section the sheet of base to free surface (Figs. 2—4E, 2-6). The
cells appears as a row of square or rectan¬ layer next to the basal lamina consists of
gular profiles (Fig. 2-1). plump cuboidal or even columnar cells with
Epithelium of this kind is found in many rounded or beveled upper ends. Above this
glands, as in the thyroid; on the free surface basal layer are additional layers of irregularly
of the ovary; on the choroid plexus; on the polyhedral cells. The nearer to the free sur¬
capsule of the lens; in the ducts of glands, face they occur, the more the cells are flat¬
and as the pigmented epithelium of the ret¬ tened (Fig. 2-6.). The superficial layers are
ina. Secreting epithelia in the terminal por¬ composed of thin squamous cells.
tions of many glands can often be placed in Epithelium of this kind is found in the
this class, although the cells of glandular acini epidermis, the mouth, the esophagus, part of
or tubules are usually pyramidal rather than the epiglottis, the conjunctiva, the cornea,
cuboidal. and the vagina and in a portion of the female
urethra. When stratified squamous epithe¬
Simple Columnar Epithelium lium occurs on the exposed outer surface of
the body, the superficial cells have lost their
Sections parallel to the surface of this type nuclei and the cytoplasm has been largely
of epithelium present a mosaic pattern much replaced by the scleroprotein keratin. The
like that in other simple epithelia, but one in cells are dry, devitalized, scalelike structures.
which the polygonal outlines of the cells are Such epithelium is described as keratinized
considerably smaller (Fig. 2-1). In sections stratified squamous epithelium. On the inner
perpendicular to the surface the rectangular moist surfaces of the body, the superficial
outlines of the cells may be only a little taller cells of this type of epithelium are viable
than those of cuboidal epithelium (Fig. 2— nucleated elements, not unlike the deeper
4A), or they may be very tall and slender, lying cells except for their shape. In these
standing upright like columns or fence pal¬ sites the tissue is described as a nonkeratinized
ings (Fig. 2—4C). In many examples of col¬ stratified squamous epithelium (Fig. 2—4T).
umnar epithelium, all the nuclei are at ap¬
proximately the same level (Fig. 2—4E).
Stratified Columnar Epithelium
Simple columnar epithelium lines the surface
of the digestive tract from the cardia of the The deeper layer or layers consist of small,
stomach to the anus and is also common in irregularly polyhedral cells that do not reach
the excretory ducts of many glands. Ciliated the free surface. The superficial cells are
 EPITHELIUM • 61

Figure 2-4. Photomicrographs of various types of epithelium. A, Simple low columnar epithelium of the papillary duct
of the dog kidney. Mallory trichrome stain.
B, Stratified columnar epithelium from a large salivary gland duct. Mallory-azan stain.
C, Simple columnar epithelium from the intestinal mucosa of a cat. The epithelium has a striated border. Notice the
presence of a single goblet cell among the columnar cells. Heidenhain’s azan stain.
D, Simple columnar epithelium of mucous cells from the stomach. The mucus is stained a deep magenta. PAS-
hematoxylin stain.
E, Simple ciliate columnar epithelium from the typhlosole of a mollusc. Notice the long cone of rootlets extending from
the basal bodies of the cilia downward into the apical cytoplasm. Chrome alum-hematoxylin-phloxine stain.
F, Stratified squamous epithelium from the esophagus of a macaque. Hematoxylin and eosin stain.
G, Ciliated pseudostratified columnar epithelium from the human trachea. Mallory-azan stain.
H, Keratinized stratified squamous epithelium of the epidermis of the sole of the foot. Notice the very thick superficial
stratum of fully keratinized devitalized cells.
62 • EPITHELIUM
\

Figure 2-5. Simple columnar ciliated epithelium from the alimentary tract of a fresh water muscle. Notice the dark row
of ciliary basal bodies just beneath the free surface of the cells and the cones of fibrous rootlets that extend downward
from the basal bodies into the apical cytoplasm. Observe also the distinct “basement membrane” upon which the
epithelium rests.

columnar in form. Stratified columnar epi¬ the basally situated cells to attach to the basal
thelium is relatively uncommon. It is found lamina. Since the nucleus in both categories
in the fornix of the conjunctiva, in the cav¬ is in the widest portion of the cell, nuclei are
ernous urethra, in some small areas of the found aligned at two or more levels in this
anal mucous membrane, in the pharynx, on class of epithelium, giving a specious appear¬
the epiglottis, and in the large excretory ducts ance of stratification—hence the term pseu¬
of some glands (Figs. 2—1, 2—4B, 2—7). Cil¬ dostratified.
iated stratified columnar epithelium is found on Pseudostratified epithelium occurs in the
the nasal surface of the soft palate, in the large excretory ducts of the parotid and sev¬
larynx, and transiently in the fetal esophagus. eral other glands and in the male urethra.
Ciliated pseudostratified columnar epithelium lines
the greater part of the trachea and bronchi
Pseudostratified Columnar (Fig. 2-4G), the eustachian tube, a part of
Epithelium the tympanic cavity, and the lacrimal sac.
In pseudostratified columnar epithelium,
all the cells are in contact with the basal Transitional Epithelium
lamina but not all of them reach the surface
(Fig. 2-1). The cells are quite variable in This was originally interpreted as an inter¬
shape: some have fairly broad attachment at mediate or transitional form between strati¬
the base, narrow rapidly, and extend upward fied squamous and columnar epithelium.
through only a fraction of the thickness of The term transitional epithelium persists, even
the epithelium. Others extend throughout though its implication of change from one
the thickness of the epithelium, but are wid¬ type to another is no longer considered ap¬
est near the free surface and have long slen¬ propriate. This kind of epithelium varies
der processes that extend downward between greatly in appearance depending on the con-
 EPITHELIUM • 63

Figure 2-6. Photomicrograph of stratified squamous epithelium from the gingiva of a kitten. Observe the darker-staining
cuboidal cells of the basal layer and the progressive flattening of cells in the more superficial layers.

Figure 2-7. Photomicrograph of stratified columnar epithelium from the human male urethra. Notice the columnar form
of the superficial layer of cells and the multiple layers of nuclei not attributable to obliquity of section.
64 • EPITHELIUM

ditions under which it is fixed. It is found acteristic of the particular organ, and under
lining the urinary bladder, which is subject normal conditions it does not change. How¬
to great changes owing to emptying and ever, in chronic inflammation or in the de¬
distention. In the contracted condition it con¬ velopment of tumors one type of epithelium
sists of many cell layers (Fig. 2-1 and 2-8). may change into another, a process called
The deepest cells have a cuboidal or even a metaplasia. For example, under certain path¬
columnar shape; above these are several lay¬ ological conditions the ciliated pseudostrati-
ers of irregularly polyhedral cells, and the fied epithelium of the bronchi may change
superficial layer consists of large ceils with a to stratified squamous epithelium—a change
characteristic rounded free surface. In the described as squamous metaplasia. A similar
stretched condition the interrelations of the transformation can be induced experimen¬
cells change to accommodate to the distention tally. If one nostril of a dog is closed sur¬
of the organ, and usually only two layers can gically, the greater evaporative loss of mois¬
then be distinguished—a superficial layer of ture that accompanies the increased
large squamous elements over a layer of more ventilation through the remaining nasal pas¬
or less cuboidal cells. This type of epithelium sage causes its pseudostratified ciliated col¬
is characteristic of the mucosa of the excre¬ umnar epithelium to transform into stratified
tory passages of the urinary system from the squamous epithelium.
renal calyces to the urethra and is sometimes
called uroepithelium.
The classification of epithelia that has been SPECIALIZATIONS OF
presented here applies primarily to the
higher vertebrates. Other categories would
EPITHELIA
be necessary to describe adequately the pat¬
terns of cell association found in inverte¬ A fundamental property of epithelial cells
brates and lower vertebrates. In the adult is their inherent tendency to maintain exten¬
mammal a given type of epithelium is char¬ sive contacts with one another and thus to

Figure 2-8. Photomicrograph of stratified transitional epithelium from the renal pelvis of a monkey, showing the
characteristic superficial layer of large rounded cells, often binucleate.
 EPITHELIUM • 65

form coherent sheets covering surfaces and was referred to as the intercellular cement. It
lining cavities. As structural correlates of this was stainable by the periodic acid-Schiff re¬
property, there are specializations of the plas- action and hence appeared to have a carbo¬
malemma on the lateral surfaces of the cells hydrate component. It also was a site of
that serve to maintain close cell-to-cell con¬ selective deposition of metallic silver when
tact. The free surfaces of the superficial cells tissues immersed in silver nitrate solution
may also be modified in various ways to were subsequently exposed to sunlight. This
increase the efficiency of the epithelium in method was often used to demonstrate the
carrying out the functions of absorption or cell boundaries of epithelia (Fig. 2-2).
transport. Because all substances entering or Certain epithelial cells, particularly those
leaving the body must traverse an epithelium of epidermal stratified squamous epithelium,
in a direction perpendicular to its surface, appeared with the light microscope to adhere
the cells are structurally and functionally po¬ to one another by many small processes dis¬
larized—i.e., the distal end, toward the free tributed over the entire cell surface. These
surface, differs from the proximal end, to¬ seemed to extend from cell to cell and were
ward the underlying connective tissue. This called “intercellular bridges.” Between the
polarity is evident not only in the specializa¬ bridges a labyrinthine system of expanded
tion of the respective surfaces but also in the intercellular spaces called interfacial canals was
arrangement of organelles in the interior of described. At the midpoint of each bridge
the cell. The centrosome and Golgi apparatus was a densely staining dot called a desmosome.
are usually in an adluminal or supranuclear Some histologists insisted that there was pro¬
position. An imaginary line passing through toplasmic continuity from cell to cell through
the centrosome and the center of the nucleus the bridges, but others interpreted the des-
defines the cell axis, and this is usually vertical mosomes as special sites of attachment at end-
or perpendicular to the basement lamina. to-end junctions of short processes on the
The long filamentous mitochondria of col¬ neighboring cells. With special preparative
umnar epithelia tend to be oriented parallel procedures, desmosomes could also be dem¬
to the cell axis and are often mobilized in onstrated on the lateral surfaces of columnar
greater numbers in the apical cytoplasm. epithelial cells that had no obvious intercel¬
The evidence of cell polarity is less obvious lular bridges.
in stratified squamous epithelia, which are The nature of the contacts between epithe¬
more concerned with protection than with lial cells was greatly clarified with the advent
absorption or secretion. In these, the surface of electron microscopy. In micrographs of
specializations for cell attachment are espe¬ simple squamous, cuboidal, and columnar
cially well developed, and the cytoplasm con¬ epithelia, the lateral membranes of adjacent
tains a conspicuous internal reinforcement or cells are usually separated by a clear intercel¬
cytoskeleton composed of filaments, aggre¬ lular cleft only 15 to 20 nm wide. If this
gated in bundles, to form tonofibrils visible space is occupied in vivo by an intercellular
with the light microscope. Such an internal substance, it is extracted during specimen
supporting framework is not a prominent preparation. The existence of an adhesive
feature of columnar epithelia, but in many “intercellular cement” has not been substan¬
of these there is a terminal web, consisting of tiated, but some authors would assign such a
a feltwork of fine filaments immediately be¬ role to the recently described glycoprotein
neath the free surface, where it apparently cell product, hbronectin. In columnar ab¬
provides for stiffening and mechanical sup¬ sorptive epithelia the lateral cell membranes
port for the striated or ciliated border of the often diverge toward the base, creating wider
epithelium. intercellular spaces. The existence of an ex¬
tensive system of interfacial canals in strati¬
Specializations for Ceil Attachment fied squamous epithelia is confirmed in elec¬
and Communication tron micrographs.
On the boundary between adjacent col¬
Adjacent epithelial cells cohere so tightly umnar epithelial cells, immediately subjacent
that a relatively strong mechanical force must to their free surface, dark dots can be seen
be applied to separate them. According to with the light microscope in vertical sections,
the traditional interpretation, a thin layer of or dense bars in horizontal subtangential sec¬
an interstitial substance between cells acted tions. In the latter they appear to form a
as an adhesive. This hypothetical substance continuous band around the polygonal pe-
66 • EPITHELIUM

microscope in columnar epithelia, the elec

rimeter of each cell. These were formerly
tron microscope reveals a junctional complex
called terminal bars and were intei preted as
consisting of three distinct components—the
local accumulations of intercellular cement.
zonula occludens, the zonula adherens, and the
They were assumed to seal or close the inter¬
macula adherens or desmosome. Immediately
cellular spaces at the free surface of the
below the free surface of the epithelium is
epithelium.
the zonula occludens, a region of surface
Electron microscopy now permits a moie
specialization where the membranes of ad¬
precise definition of the contact surface spe
joining cells converge and appear to fuse
cializations of epithelia.
(Fig. 2-9). Within this region, which may
Zonula Occludens or Tight Junction.
occupy 0.1 to 0.3 jam of the lateral cell
Where terminal bars were seen with the light

Zonula occludens

Microvilli
Zonula adherens

Desmosome
\fmoca/a adherens)

Zymogen granule

Gap junction
(nexus)

Intercellular
canaliculus
Desmosome

Intercellular
canaliculus

Zonula occludens

Basement lamina

Figure 2-9. Drawing of a typical epithelial cell illustrating the basement lamina, microvilli on the free surface, desmosomes
on the lateral surfaces, and a juxtaluminal junctional complex. The latter consists of a zonula occludens, zonula
adherens, and macula adherens. In addition a gap junction is depicted near the junctional complex. These communicating
junctions may occur anywhere on the opposing lateral surfaces of epithelial cells. (Drawing from Hay, E., In Greep, R.
O., ed.: Histology. 2nd ed. New York, McGraw-Hill Book Co., 1965.)
 EPITHELIUM • 67

boundary, there may be multiple sites of ing osmotic gradient that serves to move
fusion separated by short regions in which water across the epithelium, thereby concen¬
the opposing membranes are separated by trating the luminal contents.
10 to 15 nm. This juxtaluminal region of The zonula occludens also has a mechani¬
membrane fusion extends in a belt around cal role in maintaining the structural integrity
the perimeter of the cell and serves to close of the epithelium. The cells are more firmly
the intercellular space. attached in this region of membrane fusion
When this region is examined in freeze- than anywhere else on their surface.
cleaved preparations, it presents a very dis¬ The Zonula Adherens. On the epithelial
tinctive reticular pattern (Fig. 2-10). Each cell boundaries just below the zonula occlu¬
membrane appears to contain straight, rod¬ dens, the membranes diverge to a distance
like structures that branch and join to form of 15 to 20 nm. The opposing unit mem¬
a rectilinear network of ridges on the P-face branes are reinforced on their cytoplasmic
and a complementary pattern of shallow surface by a dense mat of fine filamentous
grooves on the E-face of the cleaved mem¬ material that forms a continuous band
brane. These intramembranous rods are be¬ around the cell parallel to the zonula occlu¬
lieved to coincide with the lines of membrane dens (Fig. 2—9). This junctional element is
fusion seen in this region of the cell mem¬ not identifiable in freeze-cleaved prepara¬
brane in thin sections. tions because there seems to be no speciali¬
As a consequence of the obliteration of the zation within the plane of the membrane that
intercellular space along these circumferen¬ distinguishes this from other parts of the
tial bands at the apex of the cells, large plasmalemma. The associated condensation
molecules cannot traverse the epithelium of cytoplasmic filaments has ill-defined limits,
from the lumen via an intercellular route. and in some epithelia it blends with a trans¬
Occluding junctions of this kind are espe¬ verse zone of filament-rich cytoplasm called
cially important in a transporting epithelium the terminal web. No structural elements are
such as that of the gallbladder (see Chapter seen traversing the intercellular space at the
27). They make it possible for the cells to zonula adherens and the nature of the forces
pump solute actively through their lateral that hold the membranes more tightly to¬
membranes into the intercellular cleft below gether at this site is not known. Nevertheless,
the zonula occludens, creating there a stand¬ as the name implies, the structure is generally

Figure 2-10. Electron micrograph of a freeze-fracture replica of the zonula occludens of intestinal epithelium. A reticular
pattern of anastomosing intramembrane strands is seen on the P-face of the cleaved lateral cell membrane just below
the brush border. (Micrograph courtesy of J. P. Revel.)
68 • EPITHELIUM

believed to be a band of firm adhesion be¬ zonula adherens, freeze-cleaved preparations

tween neighboring cells. show no distinctive internal differentiation of
Owing to the affinity of its filamentous the cell membrane at desmosomes.
component for stains, it is probable that it In thin sections, a slender intermediate
was mainly this portion of the epithelial junc¬ dense line is often seen in the middle of the
tional complex that was identified by light intercellular space between the two halves of
microscopists as the terminal bar. the desmosome (Fig. 2—11). Occasionally,
The Macula Adherens or Desmosome. with intense heavy-metal staining, there is
The third component of the typical junc¬ also a suggestion of delicate striations trav¬
tional complex of columnar epithelium is the ersing the intercellular space. The integrity
macula adherens, which corresponds to the of desmosomes is dependent on the presence
desmosome of classical histology and is com¬ of calcium. Perfusion of tissue with calcium-
monly referred to by this name. These ap¬ free media or immersion in a chelating agent
pear with the light microscope merely as results in separation of the two halves.
dense dots or fusiform thickenings of the cell In simple cuboidal or columnar epithelia,
boundaries, but they are resolved in electron there is often a circumferential row of des¬
micrographs as bipartite structures consisting mosomes below the zonula adherens forming
of plaquelike local differentiations of the op¬ the third element .of the typical junctional
posing membranes. The cell surfaces are 15 complex, but additional desmosomes may be
to 20 nm apart, and the opposing membranes found scattered at random over the lateral
are of normal dimensions but appear thick¬ cell surfaces. In stratified squamous epithelia,
ened owing to the presence of a thin but very zonulae occludentes and zonulae adherentes
dense layer closely applied to their cyto¬ may be absent, but desmosomes are unu¬
plasmic surface (Fig. 2—11). Immediately sub¬ sually abundant, attaching the ends of the
jacent to this dense plaque is a thicker layer short processes that were erroneously inter¬
consisting of a feltwork of fine filaments. preted by light microscopists as intercellular
Tonofilaments in the cytoplasm converge bridges.
upon the desmosomes and appear to termi¬ The formation of one half of a desmosome
nate in their inner layer. High-resolution evidently induces simultaneous formation of
micrographs suggest that many of the tono¬ the complementary half by the neighboring
filaments form hairpin loops in the matt of cell. Hemidesmosomes are not seen on the
filaments associated with the dense plaque boundaries between cells. They are found,
and turn back into the cytoplasm. The des¬ however, on the basal surface of stratified
mosomes are thus sites of attachment of the squamous epithelia where the cells are ex¬
cytoskeleton to the cell surface as well as sites posed to the underlying basal lamina (Fig.
of cell-to-cell adhesion. As in the case of the 2-12).

Figure 2-11. A, Electron micrograph of a desmosome from amphibian epidermis showing the attachment plaque and
tonofilaments forming recurving loops in the adjacent cytoplasm. (Micrograph courtesy of D. Kelly.)
B, Schematic representation of the structure of the desmosome. (Redrawn from Staehelin, L. A., and B. E. Hill. Sci.
Am. 238:146, 1978. Copyright Scientific American, Inc. All rights reserved.)
 EPITHELIUM • 69

Figure 2-12. Micrograph of hemidesmosomes along the basal cell membrane of an amphibian epidermal cell. Notice
the tonofilaments converging on the hemidesmosomes. At lower left are cross sections of collagen fibers in the
connective tissue underlying the epithelium. (Micrograph courtesy of D. Kelly.)

The Nexus or Gap Junction. A junctional When cells some distance apart in an epi¬
specialization that went undetected by light thelial sheet are impaled with microelec¬
microscopy is the gap junction, also called the trodes, they can be shown to be electrically
nexus, or communicating junction. At these sites coupled. In replicas of high resolution a
the intercellular space is reduced to a narrow minute central depression can be detected in
slit about 2 nm wide. This intercellular gap each 8-nm intramembrane particle on the P-
is of constant width throughout and at no face of the junction. This corresponds to a
point do the opposing membranes appear to 1.5- to 2-nm central pore seen better in
be fused. When epithelial tissues are exposed negatively stained isolated gap junctions
to an extracellular electron-opaque tracer, where the contrast medium penetrates the
such as lanthanum, it penetrates the intercel¬ pore (Fig. 2—13 C). The globular particles
lular space and is visible as a thin dense line in such preparations appear to be composed
about 2 nm wide between the opposing mem¬ of six subunits arranged radially around
branes. Where the plane of section passes the pore. The junctional particles extend
tangentially to the cell membrane providing through the lipid bilayer and project into the
an en face view of the interspace, a highly intercellular gap where they attach to corre¬
ordered pattern is seen in the specialized sponding particles in the opposing mem¬
portions of the opposing membranes. The brane. The alignment and end-to-end bond¬
lanthanum outlines hexagonally packed glob¬ ing of the junctional particles forms units
ular subunits with a center-to-center spacing called connexons.
of 9 nm. With the freeze-fracturing tech¬ The flow of ions through the hydrophilic
nique, the replica of the outwardly directed channels of the connexons explains the elec¬
inner half-membrane (P-face) shows a high trical coupling of cells throughout an epithe¬
concentration of closely packed particles lium. Microinjected fluorescent dyes of low
within the membrane (Fig. 2-13 B). molecular weight can also be shown to pass-
70 • EPITHELIUM

mm

ll§l§§
tEaggs
lllll
L-rL** 9 vm. at iS.*.! * ,*

Figure 2-13. A, Micrograph of a gap junction as it appears in thin sections. (Micrograph courtesy of Kiyoshi Hama.) B,
Freeze-fracture’replica of the P-face of a gap junction. (Micrograph courtesy of D. Albertini.) C, Isolated gap junctions
as they appear in negatively stained preparations. The connexons have a central dot representing the central 1.5- to 2-
nm channel, filled with the contrast medium. (Micrograph courtesy of B. Gilula.)
 EPITHELIUM • 71

from cell to cell via gap junctions. Cyclic AMP The basal lamina consists of two zones—
and other small molecules can also pass one of low density adjacent to the cell mem¬
through these communicating junctions and brane called the lamina rara or lamina lucida,
may play an important role in coordinating and one of greater density adjacent to the
the responses of groups of epithelial cells to connective tissue matrix called the lamina
hormonal and neural stimuli. Proteins, nu¬ densa. The two are of about equal thickness,
cleic acids, and other macromolecules greater 40 to 60 nm, but in a few exceptional exam¬
than 1200 M.W. do not pass through the ples the lamina densa may be two to three
connexons. times thicker. Substructure is difficult to re¬
Junctional specializations of this type are solve in the lamina rara, which generally
not confined to epithelia but are also seen at appears as a clear zone. The lamina densa
the so-called electrical synapses in the inver¬ consists of a dense meshwork of randomly
tebrate central nervous system and between oriented filaments approximately 4 nm in
cellular units of smooth and cardiac muscle. diameter. The basal and external laminae are
There is compelling evidence that in these formed by the cells with which they are
latter the nexuses or gap junctions are the associated, and consist largely of Type IV
low-resistance pathways through which exci¬ collagen, laminin, and proteoglycan rich in
tation passes rapidly from cell to cell, per¬ heparan sulfate.
mitting the muscle to function as if it were a Type IV collagen to date has been found
syncytium (see Chapter 10). exclusively in basal laminae. It consists of
three alpha chains richer in carbohydrate
The Basal Surface of Epithelia side chains than those of other collagens, and
these alpha chains retain the propeptide ex¬
The basal surface of most epithelia is tensions at the ends of the molecule that are
smooth ^contoured and unspecialized. How¬ usually cleaved off in the extracellular proc¬
ever, in certain transporting epithelia (espe¬ essing of collagen. Their persistence is
cially those of the renal tubules, and the thought to prevent assembly of Type IV
ciliary epithelium of the eye) the cell mem¬ collagen into cross-banded fibers. Instead,
brane may be deeply infolded, partitioning the molecules become cross-linked to form a
the basal cytoplasm into multiple narrow al¬ resilient three-dimensional meshwork.
coves containing mitochondria. Some of Laminin is a glycoprotein of 900,000 M.W.
these are open to the cytoplasm above. Oth¬ with a distinctive crosslike molecular config¬
ers appear to be closed compartments and uration. It seems to be localized exclusively
are in fact undermining processes extending in basal laminae where it is bound to Type
laterally from the base of neighboring cells. IV collagen in the lamina densa and less
The effect of this basal specialization is to abundantly in the lamina rara.
create a labyrinthine system of extracellular The proteoglycans of basal laminae vary in
clefts that greatly increase the area of the cell their glycosaminoglycan composition in dif¬
surface engaged in pumping of solute to ferent tissues. Proteoglycans are commonly
generate an extracellular osmotic gradient rich in heparan sulfate, which gives the basal
that moves water and electrolytes across the lamina a strong anionic charge that probably
epithelium. plays a role in cell attachment and in its
At the boundary between epithelia and the function as a selective filter.
underlying connective tissue is a supporting In addition to these components that are
structure called the basal lamina or basement synthesized and secreted by the associated
membrane. The latter term commonly used in cells, there may be varying small amounts of
descriptions based on light microscopy often hbronectin on the connective tissue face of
includes other components of the extracel¬ the basal lamina and this may have a role in
lular matrix and is therefore less specific than binding it to the underlying stroma.
the term basal lamina, which describes the The primary function of the basal lamina
continuous layer seen underlying epithelia in is to provide a physical support for the epi¬
electron micrographs. A similar layer sur¬ thelium. Its structural framework of Type
rounds smooth, skeletal, and cardiac muscle IV collagen gives it considerable tensile
cells, Schwann cells, and certain other sessile strength, and at the same time it is flexible
cells of mesenchymal origin. The term exter¬ enough to permit stretching and recoil in
nal lamina is more appropriate for this cir¬ epithelia lining hollow organs subject to
cumferential investment than basal lamina or changes of caliber. The basal lamina also
basement membrane. provides for cell attachment possibly by spe-
72 • EPITHELIUM

cific binding sites in the cell membrane for

its constituent proteins. A third function, best
exemplified in the basal lamina of capillary
endothelium, is ultrafiltration. This has been
most thoroughly investigated in the kidney,
where the urine is formed as an ultrafiltrate
of the blood circulating through the capillar¬
ies of the glomerulus. There the basal lamina
acts as a sieve holding back molecules on the
basis of their size, shape, and electrostatic
charge. During embryonic development the
basal laminae act as a substrate for cell mi¬
gration and influence differentiation^ of the
overlying cells by inductive interaction with
the cell membranes. In repair after injury to
an epithelium in postnatal life, the persisting
basal lamina serves as a substrate guiding the Figure 2-14. Schematic representation of the connexons
migration of new cells from the margins of and their subunits in a-portion of a gap junction. The
the wound to reestablish epithelial continuity. hydrophilic pores through the connexons permit passage
of ions and small molecules such as cyclic AMP or the
dye fluorescein, but exclude larger molecules. (Redrawn
after Tagawa, B., and T. Lowenstein. In Weismann, G.,
and R. Claiborne, eds.: Cell Membranes: Biochemistry,
SPECIALIZATIONS OF THE Cell Biology and Pathology. New York, H. P. Publishing
FREE SURFACE Co., 1975.)

Striated Border ments are cross-linked into a bundle by a

Under the light microscope a number of polypeptide called villin, and at regular inter¬
absorptive columnar epithelia have a refrac- vals of 33 nm along their length fine cross
tile border at the free surface that exhibits a filaments project laterally, linking the core
fine vertical striation at high magnifications. bundle to the membrane of the microvillus.
This striated or brush border is found in elec¬ The actin filaments extending downward
tron micrographs to consist of microvilli, slen¬ from the bases of the microvilli are anchored
der cylindrical cell processes 80 to 90 nm in in a zone of transversely oriented interwoven
diameter and 1 to 2 pim long (Fig. 2—15). filaments making up the terminal web of the
The microvilli stand erect and parallel and cell. This structure has a number of compo¬
are closely packed, with approximately 60 nents and a complex organization that is still
per square micrometer of epithelial surface. being worked out. It contains myosin, tro¬
Each is enclosed in an extension of the pomyosin, and several smaller polypeptides
plasma membrane and collectively they result associated with a dense meshwork of 6-nm
in a 15- to 30-fold increase in surface area actin filaments and 10-nm intermediate fila¬
exposed to the lumen compared with that of ments. A peripheral band of circumferen¬
a flat apical surface. At the tips of the micro¬ tially oriented actin filaments is closely bound
villi, delicate branching filaments 3 to 5 nm to the zonula adherens of the junctional com¬
in diameter project from the membrane plex.
forming a conspicuous surface coat or glyco- It was formerly thought that interaction of
calyx at the luminal surface of the epithelium actin filaments of the rootlets with myosin in
(Fig. 2-15). The fine filaments making up the terminal web might actively shorten the
this layer are terminal oligosaccharides of microvilli. The core filaments are now be¬
integral membrane proteins. Staining histo¬ lieved to have a cytoskeletal function, stiff¬
logical sections with cytochemical methods ening the microvilli and helping to maintain
for carbohydrates renders the glycocalyx vis¬ their parallel array. Contraction of the ter¬
ible at the light microscope level. minal web can, however, decrease the diam¬
The cytoplasm in the interior of each mi¬ eter of the cell apex, making its surface bulge
crovillus contains a bundle of 25 to 35 actin into the lumen and thereby spreading apart
filaments, which are attached to the mem¬ the tips of the microvilli and expanding the
brane at the tip and extend downward into intervillous spaces.
the apical cytoplasm (Fig. 2-16). The fila¬ Microvilli occur on the free surface of most
 EPITHELIUM 73

Glycocalyx Microvilli

■ ■

Figure 2-15. Electron micrograph of the brush border of an intestinal epithelial cell showing the closely packed microvilli.
The layer on the tips of the microvilli is the glycocalyx, a mat of fine filaments that are terminal oligosaccharides of
membrane glycoproteins. (Micrograph courtesy of S. Ito.)

Figure 2-16. Micrograph of transverse sections of the microvilli in the brush border of intestinal epithelium showing the
thinner glycocalyx on the sides of the microvilli and the actin filaments in their core. (Micrograph courtesy of A. Ichikawa.)
74 • EPITHELIUM

epithelia, but they form a typical striated the individual processes within this tuft are
border only where they are very numerous resolved as slender, hairlike structures as
and closely packed as they are on the absorp¬ long as cilia but more slender. Because they
tive epithelium of the intestine. Biochemical were nonmotile they were named stereocilia
analysis of brush borders isolated from this to distinguish them from the motile kinocilia.
epithelium has shown that their membrane In electron micrographs they have the ap¬
is rich in enzymes that carry out the terminal pearance of very long, flexible microvilli but
steps in digestion of carbohydrates. Thus, somewhat thicker at the base and tapering to
the intestinal brush border is a structural a narrower tip. They have a core bundle of
adaptation for increasing the digestive and actin filaments that extends into and through
absorptive efficiency of the epithelium by a moderately well developed terminal web.
greatly amplifying the surface area of mem¬ The stereocilia are parallel at their base but
brane exposed to nutrients in the intestinal become increasingly sinuous and entwined
lumen. In epithelia that are not highly spe¬ toward their tips. Their function is not en¬
cialized for absorption or transport the mi¬ tirely clear but the epididymal epithelium is
crovilli are relatively sparse, shorter, and known to absorb 90 per cent of the original
more variable in their orientation. In such volume of fluid secreted by the testis, and
cells the terminal web and actin cores of the the increased surface provided by the ster¬
microvilli are less well developed. eocilia no doubt contributes to the efficiency
of the epithelium in concentrating the sem¬
inal plasma in its passage through the long
Stereocilia epididymal duct.
The pseudostratihed columnar epithelium
lining the epididymal duct in the male has a Cilia
unique surface specialization. Under the light
microscope a pyriform tuft of cell processes Cells specialized for transport of fluid or a
projects from each cell several microns into him of mucus along the surface of an epithe¬
the lumen (Fig. 2-17). At high magnifications lium have motile, hairlike cell processes called

Figure 2-17. Photomicrograph of pseudostratified columnar epithelium of the human epididymis. Notice the tufts of
long flexible stereocilia projecting from each cell.
 epithelium * 75

Figure 2-18. Scanning micrograph of the luminal surface of the epithelium of the human oviduct, which contains ciliated
and nonciliated cells. The latter bear many short microvilli. (Micrograph from Gaddum-Rosse, P. R. Blandau, and R.
Tiersch. Am. J. Anat. 138:269, 1973.)
76 • EPITHELIUM

cilia. These execute rapid to-and-fro oscilla¬ a whole. When viewed from above with the
tions in the direction of movement of the microscope, this activity is reminiscent of the
luminal contents. There are from 50 to 100 waves that run before the wind across a held
or more per cell, arranged in rows with of grain. The metachronal waves of ciliated
microvilli between the rows. They are 7 to epithelia, however, are regular in the peri¬
10 |xm long and 0.2 |xm in diameter and odicity of their occurrence and constant in
easily resolved with the light microscope (Fig. direction. The effect of this coordinated ac¬
2-5). Their form and arrangement are re¬ tivity of the cilia is to move a blanket of
vealed more dramatically in the three-dimen¬ mucus slowly over the epithelium or to pro¬
sional images provided by scanning electron pel fluid and particulate matter through the
microscopy (Fig. 2—18). lumen of a tubular organ.
On living cells cilia beat rapidly in a consis¬ With the light microscope, no internal
tent direction. If the rate of beat is slowed structure is discernible in cilia to account for
down in cinematographic analysis, each cil- their movement, but under the electron mi¬
ium is observed to stiffen on the more rapid croscope cilia are found to have a core struc¬
forward or effective stroke and to become flex¬ ture called the axoneme, consisting of longi¬
ible on the slower recovery stroke. Cilia may tudinal microtubules that have a constant
have an isochronal rhythm, in which all cilia number and precise arrangement. In trans¬
beat together, but more commonly those on verse sections, two single microtubules are
ciliated epithelia exhibit a metachronal rhythm, located in the center of the axoneme, with
in which the successive cilia in each row start nine doublet tubules uniformly spaced
their beat in sequence so that each is slightly around them (Figs. 2—19 and 2—20). At the
more advanced in its cycle than the preceding base of each cilium is a cylindrical basal body
one. This sequential activation of the cilia with a structure identical to that of a cen-
results in the formation of waves that sweep triole, with nine triplet microtubules in char¬
slowly over the surface of the epithelium as acteristic pinwheel arrangement making up

Figure 2-19. Micrograph parallel to the surface of tracheal epithelium, affording a comparison of the cross-sectional
appearance of the cilia and the intervening microvilli. (Micrograph from Simionescu, M., and N. Simionescu. Reproduced
from The Journal of Cell Biology 70:608, 1976 by copyright permission of The Rockefeller University Press.)
 EPITHELIUM • 77

Outer fiber Outer arm

(doublet)

Membrane
Inner arm

Figure 2-20. Diagram of the cross-

sectional appearance of the compo¬
Nexin
nents in the axoneme of a cilium.
bridge
(Redrawn after R. Linck.) Central
fiber

Central
Radial link sheath
head
Radial
link

the wall of the hollow organelle. The micro¬ The doublet microtubles of the axoneme
tubules of the axoneme extend throughout are the structures responsible for movement
the length of the ciliary shaft. The central of the cilium. It was originally thought that
pair terminate at the base of the cilium, but they were contractile. If they were, those on
the nine peripheral doublets are continuous the concave side during the effective stroke
with the two inner subunits of the nine trip¬ would be expected to shorten. Instead, it has
lets in the wall of the basal body. been shown that these doublets actually ex¬
The central pair of microtubules in the tend further into the ciliary tip than those on
axoneme of cilia have much in common with the convex side (Fig. 2-21). Bending there¬
the microtubules found in the cytoplasm of fore occurs without shortening of any of the
nearly all cells. The doublets are rather dif¬ microtubules. It is now generally accepted
ferent. They are not composed of two similar that ciliary motion involves a sliding-micro-
tubules adherent along one side. Instead, tuble mechanism comparable with the slid¬
there is one complete tubule with a circular ing-filament mechanism of muscle contrac¬
cross section, subunit A, and an incomplete tion. The ATPase activity of the dynein arms
tubule, subunit B, which has a C-shaped cross liberates the energy necessary to generate
section. The latter is fused to subunit A along sliding of the microtubules, and the radial
its edges so that the resulting doublet has a spokes attaching to the central microtubule
figure-eight cross section with a segment of complex provide the resistance to shear that
the wall of tubule A closing the defect in the is needed to convert microtubule sliding into
wall of tubule B (Fig. 2-20). The wall of the bending.
microtubules is composed of tubulin hetero¬ Convincing evidence for this interpretation
dimers arranged end-to-end to form protofi¬ has come from the observation that in mutant
laments. The wall of subunit A of the doublets protozoa that fail to form the radial spokes,
consists of 13 proto filaments while subunit B the cilia do not execute bending movements.
has 10 and shares 3 with subunit A. Each The essential role of dynein in energizing
doublet has two rows of short arms that pro¬ microtubule sliding has found confirmation
ject from subunit A toward the next doublet in a rare congenital anomaly of humans in
in the row. The arms diverge slightly and are which the arms on the doublets are lacking
directed clockwise from the point of view of and the cilia and sperm flagella are immobile.
an observer looking along the ciliary shaft Classical cytologists believed that the for¬
from base toward tip. Each arm has three mation of cilia was preceded by repeated
subunits and contains the protein dynein, division of an initial pair of centrioles until
which has adenosine triphosphatase activity. the required number of basal bodies was
Radial spokes extend inward from subunit A produced. These then became arranged in
to a sheath around the central pair of micro¬ rows beneath the cell surface, and each gave
tubules (Fig. 2-20). rise to a cilium. More recent studies of cili-
78 • EPITHELIUM
V \

perpetuating—the ends of the doublets that

polymerize on the ends of the triplets in the
wall of the basal body then become sites for
deposition of additional subunits until the
cilium has attained the normal length (Fig.
2-22).

Fiagella
Flagella differ from cilia in their greater
length, the character of their movements,
and the number per cell—their internal
structure is the same as that of cilia. In their
most typical form, flagella occur singly or in
pairs on free-swimming cells. They are the
motor organs of many protozoa reaching
Figure 2-21. A simplified diagram illustrating the sliding
microtubule hypothesis of ciliary motility. At center, cilium lengths of 15 to 30 jam. They are located at
C is straight and the doublets of the axoneme terminate the anterior pole and have an undulatory
at the same level. A transverse section near the tip would movement in which waves of bending are
show all nine peripheral doublets. At left, cilium L is bent propagated along the flagellum, pulling the
toward doublets 5 and 6, which project farthest toward
organism through its fluid medium. T he
the tip. Doublet 1 terminates first and is missing from
cross sections near the tip. Doublets become single near spermatozoa of nearly all multicellular ani¬
their termination; hence, 9 and 2 are shown as singlets. mals are propelled by a flagellum that ex¬
At right, cilium R ijS bent toward doublet 1, which now tends posteriorly and moves the cell body
projects farthest. Doublets on the concave side of the forward. In marine invertebrates the internal
cilium are present in transverse sections, while 5 and 6
on the convex side are missing. If the microtubules structure of the sperm flagellum is identical
shortened to produce bending, the relative lengths of to that of a cilium. In higher forms in which
those on the convex and concave side would be the the sperm flagellum may reach a length of
reverse of that shown here. The observations therefore several hundred micrometers, there is an
favor a sliding mechanism rather than contractile ele¬
additional row of nine dense longitudinal
ments. (Figure modified after Sleigh, M. A. Endeavour
30:11, 1971.) fibers peripheral to the nine doublets of the
axoneme. Multiple flagella occur on epithelial
cells in certain segments of the nephron in
ogenesis have shown that the basal bodies lower vertebrates, but when flagella occur on
arise in two ways. (1) They may originate in epithelia in mammals there is only one per
the same manner in which centrioles are cell; it may be no longer than a cilium and
duplicated in the mitotic cycle—a dense, ring¬ probably has a similar pattern of movement.
like procentriole arising in end-to-side rela¬ Such vestigial flagella are found on cells of
tionship to a preexisting centriole (Fig. 2- the renal tubules; in the ducts of many
22). The centriole seems to act merely as a glands; in the rete testis; and on the noncil-
site of induction or nucleation for a process iated cells of uterine epithelium (Fig. 2-23).
of self-assembly in which one or several ra¬ What their function may be is not apparent
dially arranged new centrioles form from unless simple agitation of the fluid in the
hbrogranular precursor material. (2) Multi¬ lumen may have some desirable physiological
ple basal bodies may arise de novo without consequences. Still more puzzling is the ob¬
participation of a preexisting centriole. In servation that single short flagella occur on
this case, they develop around dense spheri¬ the cells in some epithelial organs that lack a
cal bodies variously called procentriole organ¬ lumen, such as the anterior lobe of the hy¬
izers or deuterosomes. Clusters of small, round, pophysis and the islets of Langerhans. Here
fibrous granules (hlosomes) appear to be the flagella simply project into the intercel¬
their precursor material. Multiple procentrioles lular spaces or into the connective tissue
develop around the organizer center and stroma, where their agitation would seem to
grow by accretion at their ends until they serve no useful purpose. Similar abortive
attain the appropriate length. The newly flagella have been described occasionally on
formed centrioles move to the cell surface smooth muscle cells, on the stromal cells of
where they function as basal bodies, initiating the endometrium, and on mesenchymal de¬
polymerization of tubulin to form the axo- rivatives in many other organs. Some of these
nemes of the cilia. Once initiated, the assem¬ cilia or flagella have a normal appearing
bly of the ciliary shaft is rapid and self- axoneme; others lack the central pair of tu-
 EPITHELIUM • 79

Cilia Cilia

Basal
bodies

L *
FV
\L
cif>

* //x
New centrioles
New centrioles
u
Figure 2-22. Schematic depiction *>
of the alternative origins of basal
bodies during ciliogenesis. New
^ ^

basal bodies may form around v: #

one or both of the preexisting cen-
trioles, as indicated at left, or they i Procentrioles
may arise de novo from fibrogran-
ular precursors coalescing around ♦
centers of organization called pro¬ Procentrioles
centriole organizers or deutero- X X/W *
somes, as shown at right. The
latter mechanism accounts for Deuterosome
most of the basal bodies formed.
(From Fawcett, D. W. In Beatty,
R. A., and S. Glueckson-Waelsch, Centrioles
eds.: Genetics of Spermatozoa.
Fibrous granules
Edinburgh, 1972.)

f; ’-'X
Nucleus

bules. Because some sensory epithelia employ vessels. The nutritive substances from the
modified cilia or flagella as receptor organ¬ blood vessels of the underlying connective
elles, it has been speculated that the nonmo- tissue reach the epithelial elements after pass¬
tile flagella found sporadically on a wide ing through the basal lamina and the narrow
range of cell types may also have a sensory intercellular spaces between the epithelial
function, but there is no experimental evi¬ cells. If the epithelium is unusually thick, as
dence to support this. in the skin, the underlying connective tissue
It is an interesting example of the unity of usually forms vascular papillae, which project
nature that the same basic structural organi¬ into the deep surface of the epithelium.
zation is found in the axoneme of cilia and These facilitate nutrition by shortening the
flagella throughout the plant and animal diffusion distance to the cells in the superfi¬
kingdoms. Those that enable the protozoa to cial layers. In a few sites such as the stria
swim about in a drop of pond water have vascularis of the cochlea and the maternal
exactly the same cross-sectional appearance layer of some epitheliochorial placentae,
as those that help to remove dust and bacteria loops of blood capillaries may actually pene¬
from the sinuses and respiratory passages of trate among the cells of the epithelium. Such
humans. intraepithelial capillaries are rare in other
organs.
In the epidermis, olfactory mucosa, and
BLOOD VESSELS AND many other epithelia, numerous terminal
NERVES branches of sensory nerve fibers pierce the
basement lamina and run in the interstices
As a rule, epithelia covering surfaces or among the epithelial cells. The epithelia of
lining cavities are not penetrated by blood the stomach and the cervix of the uterus, on
80 • EPITHELIUM
\

Figure 2-23 Scanning micrograph of a spermatozoan lying on the surface of the uterine epithelium. The illustration
permits comparison of the length of the sperm flagellum with that of the cilia on three ciliated cells in this field. Each of
the nonciliated cells has a single centrally placed vestigial flagellum (at arrows). (From Motta, P., P. Andrews, and K.
R. Porter: Microanatomy of Cell and Tissue Surfaces. Philadelphia, Lea & Febiger, 1977, p. 173.)

the other hand, seem to lack sensory nerve great number of leukocytes of various kinds
endings, and the mucous membranes of these migrate through the vaginal epithelium. It is
organs can be rubbed or cauterized in the not surprising that these actively motile cells
unanesthetized patient without discomfort. can insinuate themselves between the sessile
epithelial cells, but how they breach the base¬
ment lamina and separate the desmosomes
EXTRANEOUS CELLS and even the occluding junctions of epithe¬
lium, and how these latter are restored to
Lymphocytes normally invade the epithe¬ their previous relations after the migratory
lium from the connective tissue in some or¬ cell has passed, are problems that remain
gans. For example, individual lymphocytes unsolved.
are very often found in the epithelium of the
intestinal tract. Peyer’s patches in the intes¬
tinal submucosa are large accumulations of RENEWAL AND
lymphoid cells, and the overlying epithelium REGENERATION OF
is often infiltrated by a multitude of lympho¬ EPITHELIUM
cytes that may push aside and distort the
epithelial cells. Similarly, the epithelium over- Epithelia, especially those that cover the
lying the tonsils is extensively infiltrated by outer surface of the body and line the intes¬
lymphocytes. In all these examples the lym¬ tinal tract, are subject to constant mechanical
phocytes represent a part of the body’s im¬ and other trauma. Under physiological con¬
munological defenses against invasion by mi¬ ditions, their cells continuously perish and
croorganisms from the environment. At are shed. This is especially manifest in the
certain phases of the reproductive cycle of stratified squamous epithelium of the epider¬
rodents, and to a lesser extent in humans, a mis, where the superficial cells undergo ker-
 EPITHELIUM • 81

atinization, a peculiar kind of differentiadon Lowenstein, W. R.: Intercellular communication. Sci.

that leads to death and desquamation of the Am. 222:79, 1970.
superficial cells. In the gastrointestinal tract, Lowenstein, W. R.: Cellular communication through
membrane junctions. Arch. Intern. Med. 729:299,
cells are continually exfoliated at the tips of
1972.
the villi. On the other hand, in the respiratory Pitts, J. D. and J. W. Simms: Permeability of junctions
passages and especially in most of the glands, between animal cells. Intercellular transfer of nu¬
degeneration of the epithelium is rare, and cleotides but not macromolecules. Exp. Cell. Res.
794:153, 1977.
the cells are correspondingly long-lived.
Staehelin, A.: Structure and functions of intercellular
The physiological loss of cells in the epi¬ junctions. Int. Rev. Cytol. 39:191, 1974.
thelium is balanced by a corresponding re¬
BASAL LAMINA
generation. The keratinized cells lost from
stratified squamous epithelium are replaced Dodson, J. D., and E. D. Hay: Secretion of collagenous
stroma by epithelium grown in vitro. Exp. Cell. Res.
by mitotic proliferation of relatively undiffer¬ 65:215, 1971.
entiated cells in the deeper layers near the Hay, E. D., and J. P. Revel: Autoradiographic studies of
base of the epithelium. These cells differen¬ the origin of the basement lamella in Ambystroma.
tiate and become keratinized during their Dev. Biol. 7:152, 1963.
Kanwar, Y. S., and M. G. Farquhar: Anionic sites in the
ascent to the epithelial surface. The simple
glomerular basement membrane: in vivo and in vitro
columnar epithelia of the stomach and the localization in the laminae rarae by cationic probes.
intestine are regenerated from proliferating J. Cell Biol 57:137, 1979.
undifferentiated cells in the neck of the gas¬ Kanwar, Y. S., and M. G. Farquhar: Presence of heparan
tric glands or in the intestinal crypts of Lie- sulfate in the glomerular basement membrane. Proc.
Natl. Acad. Sci. USA 76:1303, 1979.
berkuhn. The rate of normal physiological Kefalides, N. A., A. Alper, and C. C. Clark: Biochemistry
loss and replacement is so great that the and metabolism of basement membranes. Int. Rev.
epithelial covering of the intestinal villi is Cytol. 67:167, 1979.
entirely replaced every few days (see Chapter Martinez-Hernadez, A., and P. S. Amenta: The base¬
26). ment membrane in pathology. Lab. Invest. 48:656,
1983.
The epithelial cells are nonmotile as a rule. Pierce, G. B., T. F. Beals, J. Sri Ram, and A. R. Midgley:
In healing wounds, however, epithelial cells Basement membranes. IV. Epithelial origin and
flatten out into a thin sheet that rapidly immunological cross reactions. Am. J. Pathol.,
45:929, 1964.
spreads to cover large denuded areas of con¬
Pierce, G. B., A. R. Midgley, and J. Sri Ram: Histogenesis
nective tissue. In the initial stages of this of basement membrane. J. Exp. Med. 77 7:339,
repair there is no mitotic activity, but prolif¬ 1963.
eration begins later at the margins of the
SPECIALIZATIONS OF THE FREE SURFACE
wound, providing the cells necessary to re¬
Afzelius, B. Q.: A human syndrome caused by immotile
store the covering epithelium to its normal
cilia. Science 193:317, 1976.
thickness. Bennett, H. S. : Morphological aspects of extracellular
polysaccharides. J. Histochem. Chyochem. 77:2,
1963.
REFERENCES
Fawcett, D. W.: Cilia and flagella. In Brachet, J., and A.
JUNCTIONAL SPECIALIZATIONS E. Mirsky, eds.: The Cell: Biochemistry, Physiology,
Morphology. Vol. II. New York, Academic Press,
Barr, L., W. Berger, and M. M. Barr: Electrical trans¬ 1961.
mission at the nexus between smooth muscle cells. Fawcett, D. W., and K. R. Porter: A study of the fine
J. Gen. Physiol. 51:347, 1968. structure of ciliated epithelia. J. Morphol. 94:221,
Brightman, M. W., and T. S. Reese: Junctions between 1954.
intimately apposed cell membranes in vertebrate Gibbons, I. R., and A. J. Rowe: Dynein: a protein with
brain. J. Cell Biol. 40:648, 1969. adenosine triphosphatase activity from cilia. Science
Farquhar, M. G., and G. E. Palade: Junctional complexes 749:424, 1965.
in various epithelia. J. Cell Biol. 77:375, 1963. Ito, S.: The surface coat of enteric microvilli. J. Cell
Gilula, N. B., M. L. Epstein, and W. H. Beers: Cell-to- Biol. 27:475, 1965.
cell communication and ovulation. A study of the Pedersen, H., and H. Rebbe: Absence of arms on the
cumulus-oocyte complex. J. Cell Biol. 75:58, 1978. axoneme of immobile human spermatozoa. Biol.
Gilula, N. B., O. R. Reeves, and A. Steinbach: Metabolic Reprod. 72:541, 1975.
coupling, ionic coupling and cell contracts. Nature Sadr, P.: Studies on cilia. III. Further studies on the
235:262, 1972. cilium tip and a “sliding filament” model of ciliary
Kelly, D.: Fine structure of desmosomes, hemidesmo- motility. J. Cell Biol. 39:77, 1968.
somes and an adepidermal globular layer in devel¬ Sleigh, M. A.: Cilia. Endeavour 30:11, 1971.
oping newt epidermis. J. Cell Biol. 28:51, 1966. Warner, F. D.., and P. Sadr: The structural basis of
Leblond, C. P., H. Puchtler, and Y. Clermont: Structures ciliary bend formation. Radial spoke positional
corresponding to terminal bars and terminal web in changes accompanying microtubule sliding. J. Cell
many types of cells. Nature 786:764, 1960. Biol. 63:35, 1974. '
82 • EPITHELIUM

the renewal of stratified squamous epithelia. In

Witman, G. B., J. Plummer, and G. Sander: Chlamydo-
monas flagellar mutants lacking radial spokes and Montagna, W., and R. E. Billingham, eds.: Advances
central tubules. Structure, composition and function in Biology of the Skin. Vol. 5. New York, Pergamon
of specific axonemal components. J. Cell Biol. Press, 1964.
Leblond, C. P., and B. E. Walker: Renewal of cell
76:729, 1978.
populations. Physiol. Rev. 36:255, 1956.
REGENERATION AND RENEWAL Lipkin, M., P. Serlock, and B. Bell: Cell proliferation
Arey, L. B.: Wound healing. Physiol. Rev. 76:327, 1936. kinetics in the gastrointestinal tract of man. Gastro¬
Bertalanffy, F. D., and K. P. Nagy: Mitotic activity and enterology 45:721, 1963.
renewal rate of the epithelial cells of human duo¬ Messier, B., and C. P. Leblond: Cell proliferation and
migration as revealed by radioautography after in¬
denum. Acta Anat. 45:362, 1961.
Leblond, C. P., R. G. Greulich, and J. P. M. Pereira: jection of thymidine-H3 into male rats and mice.
Relationship of cell formation and cell migration in Am. J. Anat. 106:247, 1960.
 3
GLANDS AND
SECRETION

EXOCRINE GLANDS Much has been learned in recent years

about the intracellular sites of synthesis and
Secretion is the process by which some cells the mechanisms of secretion by correlating
take up small molecules from the blood and electron microscopic and biochemical obser¬
transform them by intracellular biosynthetic vations. These primary events at the subcel-
mechanisms into a more complex product lular level will be reviewed before we proceed
that is then released from the cell. The chem¬ to a discussion of the histological organization
ical transformations involved in secretion are and classification of glands.
active processes consuming energy in con¬
trast to excretion as exemplified by the passive
diffusion of carbon dioxide across the epithe¬ SYNTHESIS AND RELEASE OF
lium lining the lung, or the filtration of blood PROTEIN SECRETORY PRODUCTS
in the kidney to form urine.
Cells and associations of cells specialized Glandular cells often contain granules or
for secretion are called glands. We define droplets that represent intracellular accu¬
three major categories of glands on the basis mulation and storage of their secretory prod¬
of the path of release of their products. ucts. Just as all cells are continually perform¬
Those that deliver their product into a system ing chemical work in maintaining their
of ducts opening onto an external or internal structural integrity and internal organization,
surface are called exocrine glands. Those that most glandular cells are synthesizing and
release their product into the blood or lymph secreting their products continually at mini¬
for transport to target cells in another part mal levels. They can often be stimulated to
of the body are called endocrine glands. Those increase the rate of delivery of their product,
that release their product into the extracel¬ and much of the information on secretion
lular space for simple diffusion to target cells gained by traditional histologists depended
in the immediate vicinity are called paracrine on observation of the cytological changes
glands or paraneurones. associated with stimulated or exaggerated ac¬
Secretion has traditionally been regarded tivity. It was observed, for example, that
as a function of epithelial cells because it was when glandular cells were stimulated, the
in these that accumulation and liberation of number of secretory granules in their cyto¬
the product could be observed directly with plasm decreased concomitantly with the in¬
the light microscope. However, since indirect crease in outflow of secretion from the ducts.
autoradiographic methods have become After depletion of the granules, the baso¬
available for tracing uptake of precursors and philic substance of the cytoplasm seemed to
discharge of product, it has become obvious become more prominent, the Golgi appara¬
that the traditional definition must be ex¬ tus hypertrophied, the nucleus increased
tended to include many nonepithelial mes¬ slightly in volume, and the nucleolus en¬
enchymal derivatives such as fibroblasts, larged. Each of these organelles was there¬
chondroblasts, and osteoblasts that release fore thought to be involved in some way in
substances into the extracellular space to the synthetic activities of the cell. As the
form the fibrous and amorphous components secretory granules began to reaccumulate,
of the connective tissues. they first appeared in very close association
83
84 . GLANDS AND SECRLTION

with the Golgi apparatus and this organelle which sequences in the -DNA molecules
was therefore thought to be the site of con¬ (genes) are exposed and available for tran¬
centration of material synthesized elsewhere scription of information. The cell compo¬
in the cytoplasm. nents that are capable of utilizing this infor¬
Ultrastructural and biochemical studies mation for synthesis of protein reside in the
have now substantiated these classical obser¬ cytoplasm. There must therefore be inter¬
vations of light microscopists and have made mediaries or messenger molecules to carry
it possible to define more precisely the re¬ the information from the nucleus to the cy¬
spective roles of the various cell organelles in toplasm, and there must be avenues of com¬
the secretory process. The cell type that has munication between these two compartments
been most thoroughly studied is the pan¬ of the cell. Three classes of ribonucleic acid
creatic acinar cell (Fig. 3—1), which secretes (RNA) molecules are essential for the trans¬
several protein enzymes into the digestive fer of genetic information and its translation
juice. The description of the cellular mecha¬ during protein synthesis. These are messenger
nisms of protein synthesis and of the intra¬ RNA (mRNA), transfer RNA (tRNA), and ri-
cellular secretory pathways that follows is bosomal RNA (rRNA). All three are formed
based largely on studies of this cell but it by transcription of specific segments of the
applies equally to a great variety of other chromosomal DNA, molecule. A region of
glandular cells producing secretions rich in one of the chromosomes that is concerned
protein. with elaboration of ribosomal RNA is closely
The chemical nature of a cell product is associated with the nucleolus. This organelle
determined in the nucleus. The information is the site of union of ribosomal RNA with
or “blueprint” for construction of the protein protein to form nucleoproteins that then pass
products of cells is encoded in the sequence into the cytoplasm to form ribosomes. The
of nucleotides in the DNA of the chromo¬ pathway that all three types of RNA take to
somes. The protein synthesized by a cell the cytoplasm is through the pores in the
depends on which regions of its chromo¬ nuclear envelope.
somes are active at the time—in other words, In the cytoplasm, the ribosomes become

Secretory
granules

Zonula occludens

Desmosomes

Golgi complex

Figure 3-1. Drawing of the fine

Mitochondria structure of a pancreatic acinar
cell, which can be considered typ¬
ical of exocrine glandular cells se¬
creting a protein product.

Basa Endoplasmic
reticulum
 GLANDS AND SECRETION • 85

associated with the site of initiation of protein tron micrographs either free in the cyto¬
synthesis at one end of the long molecules of plasmic matrix or attached to the outer sur¬
mRNA and subsequently move along the face of the limiting membrane of the
length of the molecule, “reading” in each endoplasmic reticulum (Figs. 3-2, 3-3, 3-4).
successive set of three nucleotides (codons) The free polyribosomes are concerned with
the instructions that determine the sequence synthesis of the integral proteins of the cell,
of assembly of amino acids in the protein while those attached to the endoplasmic re¬
being synthesized. The appropriate amino ticulum are involved in synthesis of secretory
acid is brought to the site of assembly on the proteins for export from the cell. The vast
ribosome by a molecule of tRNA. There are majority of ribosomes in glandular cells are
specific tRNAs for each of the 20 amino acids. associated with the reticulum.
A molecule of tRNA carrying its specific The mRNAs for secretory proteins have a
amino acid recognizes and attaches itself to special sequence of nucleotides, the signal
the appropriate complementary site on the codons, that is lacking on messengers for in¬
mRNA. Its amino acid is then inserted into tegral proteins. When these codons have
the protein molecule being developed on the been read, the resulting signal peptide binds
ribosome. The ribosome moves along the in the cytoplasmic matrix to a signal recogni¬
mRNA to the next codon, and the first tRNA tion particle consisting of polypeptides and a
molecule is released. Other ribosomes in turn small 7s RNA molecule. This particle serves
become attached to the vacated initiation site as an adapter binding the ribosome and the
and follow along after the first, reading the emerging signal peptide to receptor proteins
same message and synthesizing the same kind within the membrane of the reticulum. In¬
of protein molecule. In this way numerous teraction of the signal receptor particle and
ribosomes become associated with the same signal sequence with the membrane is be¬
mRNA molecule to form a chain of intercon¬ lieved to cause aggregation of ribosome receptor
nected ribosomes. These assemblages, called proteins in such a way as to form a transmem¬
polyribosomes or polysomes, are found in elec¬ brane channel through which the lengthen-

Figure 3-2. An area of cytoplasm from a glandular cell at higher magnification, showing the ribosomes attached to the
outer surfaces of several closely spaced cisternae of the endoplasmic reticulum.
86 • GLANDS AND SECRETION

Figure 3-3. Electron micrograph of an oblique section passing tangentially to several parallel cisternae of the
endoplasmic reticulum, showing numerous spiral and rosette configurations of polyribosomes. (Courtesy of E. Yamada.)
Insets show positively stained, isolated ribosomes connected by a thin strand (drrows) representing the messenger
RNA. (Courtesy of C. Hall and A. Rich.)
 GLANDS AND SECRETION • 87

Messenger RNA

Code reading
bases(anticodons) Peptide about to be joined to amino
acid brought in by t-RNA on left

Lumen of endoplasmic reticulum

Figure 3-4. A, Drawing of the three-dimensional configuration of the cisternae and tubules making up the rough
endoplasmic reticulum. Ribosomes occur in spirals and rosettes on the outer surface of the membrane.
B, Schematic depiction of a strand of messenger RNA and its associated ribosomes on the membrane of the
endoplasmic reticulum. Polypeptide chains of increasing length from the 5' end (right) to the 3' end (left) pass through
channels formed by assembly of integral receptor proteins in the underlying membrane. The nascent polypeptides thus
are translocated into the lumen. (Redrawn after E. C. Jordan, Oxford/Carolina Biological Reader, 45-9616, Carolina
Biological Supply Co., Burlington, North Carolina.)
88 • GLANDS AND SECRETION

5 end

qrhPmatic reoresentation of the signal hypothesis and the elements involved in attachment of the
ooNribosom'es to the membrane of the reticulum and translocation of the nascent polypeptide into its lumen. Codons at
tPhPV pnd 0f tde mRNA determine an initial amino acid sequence called the “signal.” When the nascent polypeptide
chain enierges^om ttie riboSmeJts signal sequence binds an Us signal recognition protein (SRP). This m turn.binds
to an SRPmcepZ in the membrane. Interaction of ribosome receptor and SRP receptor forms a transmembranei pore
throuah which the lenqthening polypeptide chain is translocated into the lumen of the reticulum. The signal segment o
the°po!ypeptide^s of. by a signal peptidese located on the inner aspect o he=g membrane
nf thp rpticulum (Redrawn after Blobel, G. In International Cell Biology 1976-1977, p. 320, and waiter, r., anci u.
Blobel. J. Cell Biol. 91:557, 1981. Reproduced from The Journal of Cell Biology by copyright permission o
Rockefeller University Press.)

Figure 3-6. Diagram of the Golgi regioji of

a glandular cell, showing smooth-surfaced
intermediate vesicles budding from the en¬
doplasmic reticulum. The product carried in
these vesicles to the forming face of the
Golgi complex emerges from its secretory
face in condensing vacuoles. These concen¬
trate the product to form zymogen granules.
 GLANDS AND SECRETION • 89

ing polypeptide chain extends into the lumen evaginations of its membrane pinch off as
of the reticulum. When elongation of the free vesicles each containing a small quantity
nascent polypeptide chain has moved the of the newly synthesized protein. These inter¬
signal sequence into the cistern, a signal pep¬ mediate or transport vesicles carry the product
tidase on the inner aspect of the membrane
from the endoplasmic reticulum to the Golgi
cleaves off the signal sequence. After the complex. The site of fusion of the transport
entire message has been translated and syn¬ vesicles with the membranes of the Golgi
thesis of the polypeptide chain terminated, complex is debated but it is the prevailing
the ribosome separates from the reticulum, view that the vesicles fuse with one another
and it is assumed that the underlying channel to form a flattened saccule or cistern at the
in the membrane is obliterated by dissociation convex surface of the organelle, commonly
of the signal receptor particle and lateral referred to as its forming face. This cistern
diffusion of the ribosome receptor proteins. with its content of cell product is then
The signal sequence coding on mRNA for thought to progress through the organelle to
secretory protein and the occurrence of sig¬ its concave secretory face as successive cisternae
nal recognition particles in the cytoplasm are formed behind it. According to this inter¬
capable of binding to receptor in the mem¬ pretation, all cisternae in the stack constitut¬
brane ensures that the translation machinery ing the Golgi complex are involved in proc¬
of the cell is coupled to the translocation essing the product. In the outermost
machinery in the endoplasmic reticulum. cisternae the product is dilute and, owing to
The newly synthesized molecules of secretory extraction during specimen preparation,
protein are thus segregated within the retic¬ these cisternae usually appear empty in elec¬
ulum and transported in its fluid contents to tron micrographs. However, the protein is
the supranuclear Golgi region of the cell. progressively concentrated as the cistern
On ribosome-free regions of the reticulum moves through the organelle and it is usually
in the vicinity of the Golgi complex, small preserved in the cisternae at the secretory

Figure 3-7. Golgi region of an actively secreting glandular cell showing transport vesicles and concentration of the
product in the inner cisternae of the Golgi complex and in a condensing vacuole.
90 • GLANDS AND SECRETION

face as a moderately dense content (Fig. 3- The excess membrane added to the plas¬
malemma during exocytosis is retrieved from
7). The innermost cistern at the secretory
face of the Golgi rounds up into small con¬ the cell surface and recycled through the
densing vacuoles, which subsequently coalesce Golgi complex. Small patches of surface
with each other and with contributions from membrane invaginate to form clathrin-coated
the succeeding cistern to form a single large vesicles that move into the cytoplasm and
condensing vacuole. With further concentra¬ fuse with the distended peripheral portions
tion of its contents, this vacuole becomes a of the Golgi cisternae or with condensing
typical dense secretory granule (Fig. 3-8). In vacuoles. Other vesicles fuse with lysosomes
its transit through the Golgi complex, the and are degraded.
secretory protein not only undergoes a 20 to In recapitulation, secretory protein is syn¬
25 per cent concentration but is also chemi¬ thesized on polyribosomes associated with the
cally modified by formation of disulfide endoplasmic reticulum. It is sequestered in
bonds and addition of polysaccharide chains. the lumen of this organelle and transported
The secretory granules accumulate in the through it to the Golgi region of the cell.
apical cytoplasm for transient storage. When There, transport vesicles transfer it to the
the glandular cell is stimulated to release its Golgi complex, which concentrates and pack¬
product, the granules are moved into contact ages the product in granules limited by a
with plasmalemma. Their limiting membrane membrane capable of fusing with the cell
fuses with the cell membrane at the point of membrane to release the secretion by exocy¬
contact and the contents flow into the lumen tosis. The mitochondria participate in the
(Fig. 3-9). As the granule is evacuated, its process by providing ATP for the energy-
membrane is incorporated in the cell surface. requiring steps of synthesis and transport.
Thus, the secretory product leaves the cell Incorporated amino acids traverse this intra¬
without any discontinuity being produced in cellular biosynthetic pathway much more
the plasmalemma. This process of release of rapidly than was previously imagined. The
material from cells is called exocytosis. mean transit time of exportable protein in

Figure 3-8. Juxtanuclear region of a glandular cell showing fully condensed secretory granules.
 GLANDS AND SECRETION • 91

Figure 3 9. Electron micrograph of the lumen of an acinus and the apical portions of four acinar cells. Large dense
zymogen droplets or granules are found in the cell apex. The limiting membrane of one of these has fused with the cell
membrane and its zymogen is being discharged into the lumen.

the pancreatic acinar cells is now estimated MECHANISMS FOR RELEASE OF

to be about 50 minutes. SECRETORY PRODUCTS
In glandular cells whose secretion is rich
in carbohydrate as well as protein, the Golgi
complex plays an active synthetic role in Histologists have traditionally distin¬
addition to concentrating and packaging the guished three mechanisms by which cells dis¬
product. The assembly of the carbohydrate charge their secretory products.
component of such secretions has been 1. Merocrine secretion was defined as release
shown to depend on glycosyltransferases lo¬ through the cell membrane with the cell
cated in the membranes of the Golgi cister- remaining intact. The limited resolution of
nae. In cells secreting mucus and other gly¬ the light microscope did not reveal how this
coproteins, the Golgi complex appears to be was accomplished but it was assumed either
the principal site of synthesis of the polysac¬ that the secretory material diffused through
charides and their conjugation with protein an intact membrane or, more likely, that
previously synthesized on the ribosomes of whole granules passed out through transient
the endoplasmic reticulum. discontinuities in the membrane. Electron
Much more is known about the transcrip¬ microscopic observations on exocytosis have
tion and translation of genetic information now shown how a product can be released in
and the regulation of protein synthesis than bulk without the creation of discontinuities
has been indicated here. Some of this will in the membrane. Merocrine secretion is now
already be familiar to students well prepared understood to consist of release by the proc¬
in cell and molecular biology. Others may ess of exocytosis, involving fusion of the lim¬
wish to consult a modern textbook of bio¬ iting membrane of the secretory granule with
chemistry for additional details. the cell membrane.
92 • GLANDS AND SECRETION
\ V

2. Apocrine secretion was believed to involve the form of the cell, which has an expanded
cup-shaped rim of cytoplasm called the theca
loss of part of the apical cytoplasm along with
filled with secretory droplets, and a thin base,
the material secreted. Despite this loss, the
like the stem of a goblet, extending to the
cell was believed to be able to lestore conti¬
nuity of its surface and reaccumulate prod¬ basal lamina of the epithelium (Fig. 3-10).
The mucigen droplets tend to swell and
uct. This form of secretion is less common
and has been less thoroughly studied. Elec¬ coalesce during specimen preparation and
tron microscopic observations on the Mam¬ are seldom resolved as separate entities. They
mary gland, which is generally accepted as are better preserved by freeze-drying and
an apocrine gland, substantiate the belief that they stain well with the periodic acid—Schiff
reaction because of their polysaccharide con¬
some of the cell is lost, but this loss involves
only a segment of the membrane and a thin tent.
rim of cytoplasm around the lipid component The finer structure of goblet cells is diffi¬
of the secretion—certainly a less drastic loss cult to study because of the degree to which
of cell substance than was envisioned by light their organelles are compressed into the basal
cytoplasm. The basophilia of the cytoplasm
microscopists.
3. Holocrine secretion consists of release of results from the abundance of free and at¬
whole cells into the excretory ducts or total tached ribosomes. The latter are deployed
discharge of the contents of cells, leading to on the surface of the cisternae arranged
their complete destruction. In sebaceous roughly parallel to the cell surface in the
glands, the cells break down with an out¬ paranuclear and basal cytoplasm. The Golgi
pouring of their cytoplasm and accumulated complex is well developed and located be¬
lipid. Release of spermatozoa from the sem¬ tween the compressed nucleus and the mu¬
iniferous epithelium of the testis is regarded cigen droplets in the theca. The individual
as a form of holocrine secretion in which droplets of mucigen are enveloped by ex¬
tremely delicate membranes that are often
living cells are the product.
Although the traditional terms merocrine, broken in preparation of the specimen.
apocrine, and holocrine have required some The synthesis of mucigen involves the syn¬
redefinition, they will continue to be used thesis of protein in the manner previously
until their mechanisms are better understood
and a more precise terminology evolves.

Mucigen
CLASSIFICATION OF EXOCRINE droplets
GLANDS Theca

Exocrine glands may be unicellular or mul¬

ticellular. The latter are further classified,
according to the organization and geometry
of their epithelial component, as tubular, al¬
veolar, tubuloalveolar, saccular, and so forth.

Unicellular Glands
In mammals, the most common and indeed
virtually the only example of a unicellular
gland is the mucous or goblet cell found scat¬
tered among the columnar cells of the epi¬
thelium on many mucous membranes. It se¬
cretes mucin, a glycoprotein, which upon
hydration forms a lubricating solution called Nucleus
mucus. A fully developed cell of this type has
Endoplasmic
an expanded apical end filled with pale drop¬ reticulum
lets of mucigen, and a slender basal end con¬
taining a compressed nucleus and a small Figure 3-10. Drawing of the fine structure of an intestinal
amount of deeply staining basophilic cyto¬ goblet cell. (From Lentz, T. L. Cell Fine Structure. Phila¬
plasm. The term globlet cell is descriptive of delphia, W. B. Saunders Co., 1971.)
 GLANDS AND SECRETION • 93

described for protein-secreting cells in gen¬

eral. However, autoradiography after admin¬
istration of 3oS or 3H-glucose shows that the
label goes directly to the supranuclear region,
thus establishing that the synthesis of the
carbohydrate moiety and sulfation of the
glycoprotein of the mucigen take place in the
Golgi complex. In discharging the secretion,
the membrane of individual secretory drop¬
lets or of groups of coalesced droplets fuses
with and becomes part of the plasma mem¬
brane, permitting the mucus to pour out onto
the surface.
The secretion of mucus proceeds more or
less continuously and the cell retains its goblet
form for most of its life span—which is only Figure 3—12. Intraepithelial gland from the pseudostrati-
two to four days in the intestinal mucosa. fied ciliated epithelium of the laryngeal surface. (After V.
Patzelt, from Schaffer.)
Although goblet cells normally pass through
only one long secretory cycle, they may be
made to expel nearly all of their secretion at
mogeneous population of secretory cells (Fig.
once. Under these conditions, they soon re¬
3-11). The surface epithelium of the gastric
sume mucosynthesis and refill their theca.
mucosa and of the uterine lining at certain
stages belong to this category, sometimes de¬
Multicellular Glands scribed as a secretory sheet.
Intraepithelial glands are intermediate be¬
The simplest form of multicellular gland
tween a secretory sheet and a simple tubular
is a sheet of epithelium consisting of a ho-
gland. They are small accumulations of glan¬
dular cells (usually mucous) that lie wholly
within the thickness of the epithelium but
are arranged around a small lumen of their
own (Fig. 3-12). In man, examples are found
in the pseudostratihed columnar epithelium
of the nasal mucosa and in the ductuli effer-
entes and urethra of the male reproductive
tract.
All other multicellular glands arise as tu¬
bular invaginations of an epithelial sheet and
extend into the underlying connective tissue
(Fig. 3-11). The glandular cells are usually
confined to the terminal or secretory portions
of the tubular invagination. Secretion elabo¬
rated by the gland cells reaches the surface
directly, or through an excretory duct consist¬
ing of less specialized, nonsecretory cells.
In many glands, the surface available for
release of secretion is further increased by
development of many extremely hne canals,
the secretory canaliculi, which extend from the
lumen between glandular cells. These slender
Figure 3-11. Diagram of unicellular and multicellular
glands, a, Granular; glandular cells are scattered singly extracellular passages are often branched,
among clear, common epithelial cells, b, Glandular cells and they end blindly before reaching the
arranged in a continuous sheet—secretory epithelial sur¬ basal lamina. They have no walls of their own
face. c, Simplest type of multicellular gland; the area lined but are formed by apposition of groovelike
with glandular cells forms a saclike invagination into the
subjacent tissue, d, Multicellular gland of greater com¬
excavations in the surface of adjoining cells,
plexity; the glandular spaces are lined partly with glandular d hey are usually lined by numerous micro¬
cells (terminal portions), partly with common epithelium villi. The parietal cells of the gastric glands
(excretory ducts). are exceptional in appearing to have an in-
94 • GLANDS AND SECRETION

tracellular system of canaliculi. Electron more branches, which may be somewhat

microscopy has shown, however, that these coiled near their ends (Fig. 3-13d). An excre¬
so-called intracellular canaliculi are not ac¬ tory duct may be absent, as in the glands of
tually within the cytoplasm but are deep the stomach and uterus, or there may be a
invaginations of the cell surface. They are short excretory duct, as in some of the small
limited by the plasma membrane and their glands of the oral cavity, the tongue, and the
lumen is therefore actually extracellular, like esophagus, and in the glands of Brunner in
that of the more common intercellular canalic¬ the duodenum.
In the simple acinar (or simple alveolar)
uli.
Simple Exocrine Glands. The elaborate glands, the terminal portion is expanded to
scheme of classification of glands may seem form a spherical or elongated sac. If only
to the student unnecessarily complex but it one acinus is associated with one excretory
has the advantage of permitting more precise duct, the gland is a simple acinar gland (Fig.
description of the great variety of patterns of 3_13£). This type is thought not to occur in
association of secretory cells found in the mammals. If the acinus is subdivided by par¬
body. Multicellular glands are designated as titions into several smaller compartments, or
simple or compound depending on whether or if several acini are arranged along a duct, it
not their avenue of communication with the is a simple branched acinar gland (Fig. 3-13/,
surface is branched. A simple exocrine gland is g). Examples are the sebaceous glands of the
one in which the functional unit is connected skin and the meibomian glands of the eyelids.
directly to the surface epithelium via an un¬ Compound Exocrine Glands. The duct of
branched duct (Fig. 3—13). Glands fulfilling a compound exocrine gland branches re¬
this criterion are further categorized on the peatedly. Such a gland can be thought of as
basis of the configuration of their terminal consisting of a variable number of simple
or secretory portion. Thus, they may be de¬ glands at the ends of an arborescent sy stem
scribed as simple tubular, simple coiled tubular, of ducts of progressively diminishing caliber
simple branched tubular, and simple acinar (or (Fig. 3-14). Thus, there are compound tubular,
alveolar).
and compound acinar glands.
In simple tubular glands, there is no excre¬ In compound tubular glands, the terminal
tory duct and the terminal portion is a portions of the smallest lobules are more or
straight tubule that opens directly onto the less coiled tubules, usually branching. To this
epithelial surface. The intestinal glands of category belong the pure mucous glands of
Lieberkuhn are examples (Fig. 3—13a). the oral cavity, glands of the gastric cardia,
In simple coiled tubular glands, the terminal some of the glands of Brunner (Fig. 3-15),
portion is a long coiled tubule connected to the bulbourethral glands, and the renal tu¬
the surface by an unbranched excretory duct bules. In special cases, the terminal coils an¬
(Fig. 3—136). The common sweat glands be¬ astomose.
long to this category. In the larger apocrine In compound acinar glands (also called com¬
sweat glands, the terminal portions do pound alveolar glands), the terminal portions
have been described as occurring in the form
branch.
In simple branched tubular glands, the tubules of spherical or pear-shaped units with a small
of the terminal portion bifurcate into two or lumen (Fig. 3-14a). As a rule, however, the

Fiaure 3-13. Diagrams of simple exocrine glands, a, Simple tubular; b, simple coiled tubular, c and d, simple branched
tubular; e, simple alveolar; f and g, simple branched acinar. Secretory portions black; ducts double-contoured.
 GLANDS AND SECRETION • 95

Figure 3-14. Diagram of com¬

pound exocrine glands, a, Mixed
compound-tubular and tubuloaci¬
nar; b, compound acinar. Secre¬
tory portions black; ducts double-
contoured.

form is that of irregularly branched tubules much larger size, and particularly in the
with numerous acinar lateral outgrowths larger lumen of their secretory end pieces.
from the wall and on the blind ends. These The examples commonly cited are the mam¬
glands would be more corrrectly designated mary gland and the prostate gland. Other
compound tubuloacinar (tubuloalveolar). To this authors, however, include these organs
group belong most of the larger exocrine among the compound tubuloacinar glands.
glands—the salivary glands, glands of the In some cases the excretory ducts do not
respiratory passages, and the pancreas (Fig. all join into a single main duct but open
3—16). independently on a restricted area of a free
Some authors add another category called epithelial surface. Such is the case with the
compound saccular glands, which differ from lacrimal, mammary, and prostate glands.
the compound alveolar type only in their In addition to classification according to

Figure 3-15. Photomicrograph of an intestinal submucosal gland, an example of a simple branched tubular gland. The
duct, seen penetrating the muscularis mucosae at the top of the micrograph, is unbranched, but the secretory portion
branches repeatedly and presents many cross-sectional profiles when sectioned.
96 • GLANDS AND SECRETION

Figure 3-16. Photomicrograph of the pancreas, a compound tubuloacinar gland. A small duct is sectioned longitudinally
at right of figure. The duct branches to the several acini that are clustered around it.

Figure 3-17. Photomicrographs illustrating the contrasting appearance of a mucous gland (A) and a serous gland (B)
both from the tongue.
 GLANDS AND SECRETION • 97

histological organization, compound glands alveoli. The cells of the demilunes are in
are often classified according to the nature communication with the lumen via intercel¬
of the secretion they produce. Thus, they lular secretory canaliculi (Figs. 3-18, 3-19).
may be designated as mucous, serous, or mixed.
Mucous glands secrete a viscous material with
a lubricating or protective function (Fig. 3- HISTOLOGICAL ORGANIZATION OF
17A); serous glands have a watery secretion EXOCRINE GLANDS
often rich in enzymes (Fig. 4-175).
In mucous glands the major portion of the There are certain common features in the
cell is occupied by mucigen droplets, and organization of the larger glands. They are
appears pale and highly vacuolated in histo¬ usually enclosed in a condensation of con¬
logical sections. The nucleus is displaced far nective tissue that forms the capsule of the
to the base and is often greatly flattened by organ. Septa of connective tissue extending
the accumulated mucigen droplets. In serous inward from the capsule divide the gland
glands the cells are generally smaller and into grossly visible subdivisions called lobes.
their nucleus is spherical and situated in the These in turn are partitioned by thinner
basal half of the cell surrounded by deeply septa into smaller units called lobules, still
basophilic cytoplasm. The apex of the cell visible with the naked eye (Fig. 3-20). These
may be clear, owing to extraction of the are separated to some extent into microscopic
secretory material (Fig. 3—175), or, where lobules of glandular units, but as a rule colla¬
the secretory product is preserved, it may genous connective tissue penetrates for only
stain as discrete granules. The ultrastructure a short distance into the lobule before giving
of serous cells is similar to that of the pan¬ way to a delicate network of reticular fibers
creatic acinar cell but with somewhat less surrounding the terminal ducts and secretory
extensive development of the endoplasmic acini or tubules. The connective tissue com¬
reticulum. ponents of a gland are collectively called the
Mixed glands contain both mucous and stroma, and its epithelial portion, the paren¬
serous cells. Mucous cells often make up the chyma.
greater part of the gland, with somewhat Blood vessels, lymphatics, and nerves of
flattened serous cells forming crescentic caps, glands usually show a pattern of distribution
called serous demilunes, over the ends of the similar to that of the connective tissue. They

Serous cells Mucous cells

Figure 3-18. Drawing of the end piece of a tubuloalveolar salivary gland showing the mucous cells and the serous
demilune. (Modified from Krstic, R. V. Gewebe des Menschen und der Saugetiere. Heidelberg, Springer-Verlag, 1978.)
98 • GLANDS AND SECRETION

while the ducts of the microscopic lobules are

called intralobular ducts. The latter are contin¬
uous with the intercalary ducts, whose branches
communicate with the secretory acini eithei
directly, via intercellular canaliculi, or by a
combination of these arrangements. The epi¬
thelium of the largest ducts may be simple
or stratified columnar. As the duct becomes
smaller, the epithelium is first simple colum¬
nar, then cuboidal, and finally squamous.

CONTROL OF EXOCRINE SECRETION

%

Many glands secrete continuously at a low

level but are stimulated under certain con¬
ditions to secrete more abundantly. The
mechanisms for physiological control of se¬
cretion vary greatly from gland to gland. In
many the stimulation is mediated solely via
the autonomic nervous system. In other
glands the stimulus is hormonal, and in some
there is a dual mechanism. It is well known,
for example, that the sight or smell of food
will increase the secretion of acid, mucus,
and digestive enzymes in the stomach. These
psychic stimuli are mediated via the \ agus

Figure 3-19. Photomicrograph of the terminal portions of

the submandibular gland. This is an example of a mixed
gland; some of the terminal elements are purely mucous, Acini
others have crescentic caps of serous cells called demi¬
lunes. The relationship is seen to better advantage in
longitudinal section (A) than in transverse section (B).

Collagenous
connective
penetrate the capsule and follow the colla¬ tissue
genous septa and the thinner partitions be¬
tween the lobules, and from there send
branches to the parenchyma. Within the lob¬
ule they are ultimately enclosed by reticular
connective tissue. The blood and lymph cap¬ Mierolobule
illaries form networks around small groups
of acini and terminal ducts. The major vas¬
cular supply is supplemented in most glands Intralobular' duct

by a collateral circulation mediated through

capsular vessels of small caliber. The terminal -A—Interlobular duct5

nerve fibers branch, and their final divisions Intercalary duct

end on the surfaces of the acini.
The duct system of a complex exocrine Figure 3-20. Diagram showing branches of duct system
gland conducts the product of the gland cells and their relationship to secretory portion in a lobule of a
to a free external or internal body surface. compound tubuloacinar gland. Collagenous stroma sep¬
arates (often incompletely) the microscopic lobules. The
The ducts may also modify the secretion
main duct shown is a branch of the interlobular duct. The
during its passage. The main duct of the gland interlobular duct branches into intralobular ducts of several
divides in the connective tissue to form lobar orders. These are continuous with fine terminal intercalary
ducts. Their further branchings in the septa ducts that end in the secretory portion. (Modified from
between lobules are called interlobular ducts, Heidenhain.)
 GLANDS AND SECRETION • 99

nerves and are abolished if these are cut. On two integrative systems have been supple¬
the other hand, food placed in the stomach mented by development of ductless endocrine
also initiates gastric and pancreatic secretion glands whose cells synthesize chemical agents
even if it has not previously been seen or called hormones that are carried in the circu¬
tasted. Secretion in this case depends on the lating blood to distant parts of the body,
intrinsic nerves in the organ and upon locally where they act upon specific target organs.
produced hormones. Activation of nerves in These chemical messengers tend to have a
the gastric mucosa releases the transmitter longer latent period because they are distrib¬
acetylcholine, which stimulates release of the uted by the blood, but they produce more
hormone gastrin from endocrine cells in the sustained effects than the signals carried by
mucosa, and this in turn activates secretion nerves.
by the gastric glands. Gastrin-secreting cells Endocrine glands arise in the embryo as
may also respond directly to the presence of tubular evaginations or solid outgrowths
food as a result of its detection by modified from lining epithelia, but later in their de¬
microvilli that are believed to have a chemo- velopment their connection with the surface
receptor function. is lost. They are penetrated by blood vessels,
There are no visible morphological criteria which form a very rich capillary plexus in
that enable the histologist to determine intimate relationship to the cords, follicles,
whether a given gland is under control of or acini of the endocrine glands. The close
hormones, but nervous control can be in¬ proximity of the cells to a dense vascular bed
ferred from the observation of nerve endings favors release of secretory product into the
in close contact with the secretory cells. In blood.
the pancreas, nerve endings can be found The fully developed endocrine gland is
inside the basal lamina of the acini, in close usually completely dissociated from exocrine
contact with the base of the exocrine cells, glandular tissue, but in a few examples there
i hese cells are also known to be responsive is relatively little morphological separation of
to the hormone gastrin, produced in the stom¬ the endocrine tissue from an exocrine gland.
ach, and secretin and cholecystokinin, secreted Thus, the small islets of Langerhans, the
in the duodenum. Thus, multiple factors are endocrine component of the pancreas, are
involved in the control of pancreatic secre¬ scattered throughout the much greater bulk
tion. of the exocrine portion of the gland (see
Chapter 28). Similarly, in the testis, the Ley-
dig cells secreting male sex hormone are
ENDOCRINE GLANDS located in interstitial tissue between the tu¬
bules comprising the exocrine portion of the
Phylogeneticaily, three main mechanisms organ (see Chapter 31). Thus, in these mixed
have developed in animals to integrate the glands, one group of cells secretes into a duct
functions of their different tissues and or¬ system while another group delivers its secre¬
gans. To some degree these are recapitulated tion into the blood. In the unique case of the
in vertebrate ontogeny. The earliest mecha¬ liver, the cells secrete bile into the intercel¬
nism to appear involves substances that sim¬ lular terminations of a duct system, but the
ply diffuse through the intercellular spaces same cells also release internal secretions into
to influence the behavior or function of other the blood flowing through sinusoids between
cells at a limited distance from the source the sheets of hepatic cells.
(paracrine secretion). Integration by simple The principal endocrine glands are the
diffusion of chemical messengers is slow, hypophysis, thyroid, parathyroid, pancreas, adre¬
poorly controlled, and of limited usefulness nals, pineal, testes, and placenta. These are so
in larger metazoa. This primitive humoral diverse in their architecture that they do not
mechanism was later supplemented by devel¬ lend themselves to classification on the basis
opment of a nervous system consisting of of their histological organization. The chem¬
cells that had acquired the capacity to re¬ ical nature of their hormones is also varied,
spond to external stimuli and to rapidly con¬ including modified amino acids, peptides,
duct a signal over the surface of their long proteins, glycoproteins, and steroids. It is not
cell processes (axons) to affect other cells. surprising, therefore, that one cannot de¬
The nervous system increased in complexity scribe cytological features common to all en¬
and became highly efficient in dealing with docrine cells. It is possible, however, to assign
elaborate integrated patterns of behavior in¬ them to categories related to the chemical
volving precise and rapid motor events. The nature of their product.
100* GLANDS AND SECRETION

CYTOLOGY OF POLYPEPTIDE-
SECRETING ENDOCRINE GLANDS

As might be expected, endocrine cells se¬

creting peptide and glycoprotein hormones
have many ultrastructural features in com¬
mon with the protein-secreting exocrine cells
described previously, but there is a significant
difference in the degree of development of
the organelles concerned with protein syn¬
thesis. The granular endoplasmic reticulum
is much less extensive. This is consistent with
the great difference in the volume of product
produced. The acinar cells of the exocrine
pancreas produce over 1 liter of enzyme-rich
digestive juice per day, whereas the output
of a polypeptide- or glycoprotein- secreting
endocrine gland would be measured in mil¬
ligram or microgram quantities.
The beta cell of the pancreatic islets, which
secretes the hormone insulin, can be consid¬
ered representative of this category of en¬
docrine cells. Electron micrographs of these
cells show a few meandering profiles of
rough endoplasmic reticulum, and clustei s
of free ribosomes in a cytoplasmic matrix of
Figure 3-21. Schematic representation of the mechanism
low density. There is a small Golgi apparatus of release in endocrine cells producing protein or peptide
and numerous membrane-limited granules hormones. Membranes bounding the granules coalesce
200 to 300 nm. in diameter. As in exocrine with the cell membrane. The exteriorized granule disin¬
glandular cells, the granules are formed m tegrates and the hormone diffuses into the blood through
the fenestrated endothelium of an adjacent capillary or
the Golgi complex. They tend to be some¬
sinusoid.
what more numerous at the vascular pole of
the cell but occur in considerable numbers
cells are arranged in a simple cuboidal epi¬
throughout the cytoplasm (Fig. 3—21). In
thelium bounding spherical follicles with a
humans insulin occurs in the form of pleo¬
morphic crystals within membrane-limited central cavity. The cells have an extensive
endoplasmic reticulum in the form of cistei -
secretory vesicles, but in common laboiatory
nae distended with the proteinaceous precur¬
animals the secretory granules are uniformly
dense and homogeneous. With minor differ¬ sor of the secretory product (Fig. 3-22). In
the Golgi complex, this material is packaged
ences in cytology and granule size, this same
in membrane-limited secretory vesicles that
description would apply to the alpha cells of
do not accumulate in the cytoplasm but pass
the pancreas, which secrete glucagon; the so¬
matotrophs, thyrotrophs, gonadotrophs, and directly to the apical surface and dischaige
corticotrophs of the hypophysis (secreting their content into the lumen of the follicle by
growth hormone, thyroid stimulating hormone, exocytosis.
gonadotrophic hormone, and adrenocorticotrophic
hormone)] and the ultimobranchial oi C cells
of the thyroid (secreting calcitonin). In all of
CYTOLOGY OF STEROID-SECRETING
these, it is clear that the intracellular secre¬ ENDOCRINE CELLS
tory pathway involves synthesis on ribosomes,
The steroid-secreting endocrine cells of the
segregation in the reticulum, concentration
in the Golgi complex, and storage in mem¬ ovary, testis, and adrenal gland are all quite
similar to their ultrastructure, and are very
brane limited granules.
The thyroid gland belongs to the category different from protein and peptide-secreting
of protein-secreting endocrine glands, but cells. They have little granular endoplasmic
differs from the others in that its product, reticulum and relatively few free ribosomes.
thyroglobulin, is stored extracellularly. The Their most characteristic feature is a remark-
 GLANDS AND SECRETION • 101

Figure 3-22. Diagram depicting

(at left) the ultrastructure of the
thyroid epithelial cell with secre¬
tory droplets of thyroglobulin
being formed in the Golgi zone
and discharged at the cell apex
for extracellular storage. In the
TSH-stimulated cell (at right),
droplets of colloid are taken up by
pinocytosis. Lysosomes coalesce
with these and their hydrolytic en¬
zymes degrade thyroglobulin to
release thyroxin, which diffuses
into perifollicular capillaries. (From
Fawcett, D. W., et al. Advances in
Hormone Research. Voi. 25. New
York, Academic Press, 1969.)

Lipid

Golgi complex

Agranular
endoplasmic
reticulum
Granular
reticulum

Lysosome

Granular reticulum

Lipochrome
pigment
Figure 3-23. Schematic drawing of the characteristic cytologic features of a steroid-secreting cell. Most notable are the
large Golgi complex and extensive smooth-surfaced endoplasmic reticulum. (From Fawcett, D. W., et al. Advances in
Hormone Research. Vol. 25. New York, Academic Press, 1969.)
102 • GLANDS AND SECRETION

ably extensive smooth-surfaced endoplasmic step in conversion of cholesterol to steroid

reticulum in the form of a close-meshed hormones is cleavage of its side chain by an
network of branching and anastomosing tu¬ enzyme in the mitochondria. Several subse¬
bules (Figs. 3-23, 3-24). The juxtanuclear quent steps in steroidogenesis involve en¬
Golgi complex is very large but has no asso¬ zymes in the reticulum. In the case of the
ciated secretory granules. Lipid droplets are adrenal steroids, there are additional steps in
present in the cytoplasm in greater or lesser the synthesis that take place in the mitochon¬
numbers depending on the organ and the dria. Little is known about how cholesterol
species. Mitochondria are numerous and of and the intermediate products in steroido¬
variable size and often have an unusual in¬ genesis are moved back and forth between
ternal structure, with tubular or vesicular mitochondria and the reticulum to accom¬
amplifications of the internal membrane in¬ plish the successive biosynthetic steps. Since
stead of the usual lamellar or foliate cristae. there is no appreciable storage of hormone,
These cells also contain lysosomes and per¬ these cells must maintain the organelles
oxisomes and have a tendency to accumulate needed to synthesize steroids on demand.
deposits of lipochrome pigment. The extensive development of the smooth
Steroid-secreting cells store very little hor¬ endoplasmic reticulum thus represents a spe¬
mone but may store a precursor, cholesterol. cialization to ensure the presence of the en¬
The lipid droplets, when present, contain zymes necessary for rapid synthesis of steroid
cholesterol esters as well as triglycerides. The hormone. The observation that the lipid con¬
steroid-secreting cells in some species depend tent of steroid-secreting cells diminishes on
mainly on cholesterol from the blood, while stimulation is interpreted as mobilization and
those of other species synthesize much of the depletion of stored precursors during en¬
cholesterol they utilize for hormone synthe¬ hanced hormone production. It is not yet
sis. The enzymes for synthesis of cholesterol known what role is played by the prominent
reside mainly in the membranes of the Golgi complex, but the fact that it increases
smooth endoplasmic reticulum. The initial in size in response to trophic hormone stim-

Figure 3-24. Electron micrograph of a small area of cytoplasm from a cell of the human fetal adrenal cortex. A few
cisternae of granular endoplasmic reticulum are present, but most of the cytoplasm is occupied by branching tubules of
the smooth endoplasmic reticulum. (Micrograph courtesy of N. S. McNutt.)
 GLANDS AND SECRETION • 103

ulation indicates that it is involved in some this mode of secretion that the soluble pro¬
way in the secretory process. No consistent tein and other components of the granules
morphological changes associated with the are discharged as well as the hormones. Their
release of steroid have been reported, and fate is poorly understood. The biologically
nothing can be said at present about the active hormones evidently become disso¬
mechanism of release or its regulation. ciated from the other constituents extracel-
lularly and are free to diffuse into the blood
vessels.
STORAGE AND SECRETION OF Active hormone secretion requires a mech¬
HORMONES anism not only for release of the membrane-
limited granules (exocytosis) but also for
Endocrine glands differ greatly in the transport of the granules to the cell surface.
amount of hormone stored and in the site of In glands, whether the cells are polarized
storage. As previously noted, the steroid- toward the lumen of an acinus or toward a
secreting endocrine glands have no visible blood vessel, this transport must be selective
secretory granules and they store little or no and directional. Recent studies indicate that
hormone. They are evidently able to vary microtubules and possibly the microfilaments
their rate of synthesis and release to keep of the cytoplasm may be involved in this
pace with current needs. The thyroid gland, phase of the secretory process. The evidence
on the other hand, is unique among endo¬ for this is more pharmacological than mor¬
crine glands in that it stores hormone extra- phological, and rests upon the demonstration
cellularly in the lumen of the follicles and that colchicine and other alkaloids that are
ordinarily contains enough to meet normal known to prevent polymerization of tubulin
needs for several weeks. The several endo¬ to form microtubules also block the release
crine glands that secrete protein, glycopro¬ of secretory products. A relation between
tein, or polypeptide hormones, as well as microtubules and the secretory process was
those that secrete catecholamines, store their suggested first by the observation that colchi¬
product intracellularly in membrane-limited cine prevents release of insulin from the
granules in sufficient numbers to represent pancreatic beta cells. Similar observations
one to three weeks’ supply. This latter cate¬ have been made for release of catecholamines
gory of endocrine glands with intracellular from the adrenal medulla and for secretion
storage has been the most thoroughly studied of thyroxin by the thyroid gland.
with respect to the composition of the gran¬ In the case of the thyroid, there is also
ules and their mechanism of release. evidence for involvement of the microhla-
The neurohypophyseal hormones (vaso¬ ments of the cytoplasm. The fungal metabo¬
pressin and oxytocin) are stored in the neu¬ lite cytochaiasin B is known to cause the dis¬
rosecretory granules together with specific appearance of microfilaments and to block a
soluble proteins called neurophysins. These number of cellular processes involving pro¬
proteins seem to serve as “carriers” for the toplasmic movement. When this substance is
hormones in the hypothalamo-neurohypo- added to the medium in which mouse thyroid
physeal tract. The binding of the hormones glands are being maintained in vitro, the
to them may prevent diffusion of the biolog¬ normal response to hormonal stimulation is
ically active molecules out of the vesicles in blocked. Both colloid droplet uptake from
which they are stored. Zymogen granules, the follicle lumen and release of hormone
alpha-cell granules of the pancreas, calci¬ into the medium are suppressed. The action
tonin-containing granules, granules of var¬ of cytochaiasin B on the secretory process
ious pituitary cell types, and the granules of suggests that microfilaments, as well as mi¬
the adrenal medulla all appear dense in elec¬ crotubules, are involved. Since uptake of col¬
tron micrographs and have similar histo- loid by macropinocytosis is an initial step in
chemical staining reactions. The presump¬ thyroxin release, it may be that the motility
tion is that in all of these the hormones are of the ectoplasm necessary for formation of
bound to or at least associated with a class of the engulfing pseudopods is the process that
proteins comparable to the neurophysins. All requires microfilaments. A word of caution
these glands release their secretion by exo- may be in order. The conclusions drawn
cytosis entirely comparable with that de¬ from these observations assume that the ac¬
scribed above for the exocrine pancreas but tion of colchicine and of cytochaiasin is spe¬
less easily observed because the quantities of cific for microtubules and for microfilaments,
material released are smaller. It follows from respectively. The possibility that either or
104 • GLANDS AND SECRETION

trated. The only exception is the interstitial

both may have toxic effects on other com¬
tissue of the testis, which has unfenestrated
ponents or activities of the cell has not been
capillaries. In all cases the diffusion distance
ruled out.
In the case of the thyroid, which stores its between the secretory cells and the blood is
product, thyroglobulin, extracellularly, the short, and the intervening structural com¬
mechanism for release of hormone into the ponents to be traversed are (1) a thin basal
blood is more complex. Droplets of colloid lamina around the endocrine cells, (2) a nar¬
are taken up by pinocytosis from the lumen row perivascular space, (3) the basal lamina
of the follicle. Within the cell the droplets of the capillary endothelium, and (4) the thin
fuse with lysosomes, and the thyroglobulin is diaphragms of the capillary pores. None of
degraded by hydrolytic enzymes liberating these appears to constitute a significant bar¬
thyroxin, which diffuses through the base of rier to access of the hormones to the blood.
the cell into the perifollicular capillaries. In addition to the microscopic structure of
Thus, participation of lysosomes is an inte¬ the vascular bed, a consideration of the pat¬
gral part of the normal secretory process in tern of drainage of blood from an endocrine
this particular endocrine gland but occurs in gland, at a somewhat grosser level, may be
relevant to an understanding of its function.
few others (Fig. 3-22).
In a number of endocrine cells, lysosomes For example, the hypothalamic releasing hor¬
do play a role in disposal of unneeded stores mones are liberated into capillaries of the
of secretion. For example, when the young median eminence of the hypothalamus,
of a lactating rat are weaned, there is a large which are drained via special hypophyseo-
excess of secretory granules in the mammo- portal vessels to responsive target cells im¬
trophic cells of the anterior pituitary. These mediately downstream in the anterior lobe of
coalesce with lysosomes and are degraded by the pituitary. Similarly, the mammalian ad¬
autophagy. Endoplasmic reticulum and ri¬ renal gland consists of two portions arranged
bosomes no longer required for minimal concentrically—an outer cortex that secietes
rates of hormone secretion suffer a similar steroid hormones and an inner medulla com¬
posed of cells that synthesize the catechol¬
fate.
amines epinephrine and norepinephrine. No cap¬
sule or sharp boundary separates the two
RELATION OF ENDOCRINE CELLS TO zones, and much of the blood supply flows
BLOOD AND LYMPH VASCULAR centripetally from the cortex, carrying ste¬
roid hormones downstream to the cells of
SYSTEMS
the medulla. It now appears that a relatively
Despite their histological and cytological high concentration of cortical steroids may
diversity, a common feature of endocrine be necessary for induction and maintenance
glands is their great vascularity. Nearly every of an enzyme in the medulla that is essential
cell is in close relation to one or more thin- for epinephrine synthesis. Thus, the local
walled vessels of a rich vascular bed. In some “downstream” effects of hormones may be as
glands, the vessels are typical capillaries; in important as their effects exerted at a dis¬
others, they are more appropriately de¬ tance via the general circulation.
scribed as sinusoids. The latter are generally In most endocrine organs the hormones
larger than true capillaries and are more are released exclusively into the blood, but
variable in shape—often conforming to the recent physiological studies indicate that in a
contours of the interstices they occupy among few instances the lymphatics may also be
the plates or cords of epithelial cells that significant pathways for egress of hormones.
constitute the parenchyma of the endocrine This is particularly true of the perifollicular
gland. In the pituitary and adrenal glands, lymphatics of the thyroid and the intertubu¬
the endothelium of the sinusoids was for¬ lar lymphatics of the rodent testis.
merly believed to be phagocytic to colloidal
dyes, and it was traditionally included in the
reticuloendothelial system. This interpreta¬ CONTROL MECHANISMS AND
tion has not been substantiated in ultrastruc- INTERRELATIONS WITHIN THE
tural studies, which show that the phagocytic ENDOCRINE SYSTEM
potential resides in perivascular cells rather
than in the endothelium. Endocrine glands modify the function of
Whether the vessels of endocrine glands specific target organs. In the course of evo¬
are true capillaries or sinusoids, the lining lution, some have also developed the capacity
endothelium is extremely thin and fenes¬ to sense changes in the concentration of me-
 GLANDS AND SECRETION • 105

tabolites or cell products in the body fluid; Brain

others have become specifically responsive to
the hormones of other endocrine glands, and
still others are stimulated by secretory prod¬
ucts of the central nervous system. The activ¬
ity of the brain and of the endocrine glands
has become so closely integrated that changes
in one are reflected in alterations in function
of the other. On the basis of these interac¬
tions, a variety of control mechanisms has
evolved that ensure the coordinated func¬
tioning of organs situated at a distance from
one another. These mechanisms serve to
maintain the constancy of the internal envi¬
ronment of the organism.
One of the simplest forms of control is the
case in which a hormone acts upon a target
organ, causing its cells to discharge a sub¬
stance into the extracellular compartment.
The resulting change then acts back upon
the endocrine gland to decrease its output of
hormone. This is commonly referred to as a
negative feedback mechanism. The release of the Figure 3-26. Illustration of the neuroendocrine interrela¬
hormone insulin from the beta cells of the tionships involved in the suckling reflex. Stimulation of the
pancreas drives glucose into cells and lowers nipples generates sensory impulses that pass to the
central nervous system via the dorsal root ganglia. In the
blood sugar (Fig. 3—25). The lowered concen¬
brain these impulses are relayed to the hypothalamus,
tration of blood sugar, in turn, acts back where they activate neurosecretory cells whose processes
upon the beta cells to diminish insulin re¬ extend into the neural lobe of the hypophysis. Stimulation
lease. Similarly, parathyroid hormone acts of these cells results in release of the hormone oxytocin,
upon bone cells to mobilize calcium, and the which is carried in the blood to the breast, where it causes
contraction of myoepithelial cells around acini of the
elevated blood calcium, by negative feedback, mammary gland, expelling milk. No feedback mechanism
depresses release of parathyroid hormone. is involved.
A somewhat greater degree of complexity
is encountered when the endocrine gland is

under control of the nervous system. The

milk ejection reflex is a good example of a
PANCREAS simple neuroendocrine mechanism in which
the train of integrated events begins with a
peripheral sensory stimulus. In this case, the
tactile stimulus to the nipple, involved in
suckling, is conducted over afferent neural
pathways via the dorsal nerve roots and
Low Blood Sugar spinal cord to the brain and thence to neu¬
rosecretory cell bodies in the hypothalamus
(Fig. 3-26). Stimulation of these cells results
in release of the hormone oxytocin from their
terminations in the posterior lobe of the
glucose:
pituitary. This hormone carried back to the

ifsi
'Siii&ti&iiiiiV-*'
breast via the bloodstream causes contraction
of myoepithelial cells around the glandular
acini, resulting in ejection of milk. Feedback
CELLS control is not a feature of this neuroendo¬
Figure 3-25. Drawing of a simple endocrine feedback crine reflex.
mechanism. Insulin, the hormone from the islets of Lan- The neuroendocrine response to stress is
gerhans in the pancreas, promotes entry of glucose into
more complex, involving more steps in the
other cells of the body. The resulting low blood sugar acts
back upon the alpha cells in the pancreas to reduce the sequence of events; it depends on a special
release of insulin. vascular pathway from hypothalamus to pi-
106 • GLANDS AND SECRETION

back” on the cells of the hypothalamus,

tuitary and is under negative feedback con¬
diminishing their output of ACTH-RH.
trol. A painful stimulus reaching the hypo¬
The complex system of neuroendocrine
thalamus over afferent neural pathways
interrelations that control the female repro¬
stimulates cell bodies whose axons end in
ductive cycle will be discussed in Chapter 32.
close relation to small blood vessels in the
The few examples cited here may provide
median eminence of the hypothalamus (Fig.
sufficient introduction to the mode of oper¬
3-27). Adrenocorticotrophic hormone releasing
ation of the endocrine system to enable the
hormone (ACTH-RH) is liberated at the nerve
student to appreciate the correlations of
ending and carried in the blood via the spe¬
structure and function that appear in later
cial hypophyseoportal system of blood vessels
to the anterior lobe of the hypophysis, where chapters.
The transmitter substances released at the
it causes specifically responsive cells to release
nerve terminals have a very transient exist¬
adrenocorticotrophic hormone (ACTH) into the
ence, being inactivated in a matter of seconds
general circulation. When this hormone
by specific enzymes at the endings. In the
reaches the adrenal cortex, steroid hor¬
slower-acting endocrine integrative system,
mones, called glucocorticoids, are released.
the hormones are quite variable in their half
These reach all the cells in the body and
lives in the circulation, which range from a
modify their function in various ways that
few minutes to several days. Some hormones
increase the ability of the organism to tolerate
are transported in the blood in combination
prolonged muscular activity, trauma, infec¬
with specific carrier proteins. Inactivation or
tion, or intoxications. Adrenal steroids car¬
degradation of the hormone may take place
ried back to the brain exert a “negative feed-
at the target organ or in the liver or kidney.
Some hormones enter their target cells to
exert their effects; others are evidently able
Brain to act simply by binding to specific receptor
sites on the cell membrane.

RECEPTORS AND MECHANISM OF

HORMONE ACTION ON TARGET
ORGANS

Hormones circulating in the blood reach

all of the tissues. The selectivity of their
action on particular organs depends on the
presence in the membranes of the target cells
of specific receptors having a high binding
affinity for that hormone. The receptors are
integral protein molecules of the cell mem¬
brane. Their number varies in different tar¬
get organs but may be of the order of 10,000
molecules per cell. They are not static com¬
ponents of the plasma membrane but are in
a dynamic state of turnover. There is nor¬
mally a preprogrammed rate of receptor syn¬
thesis and insertion into the membrane, but
Stressful
stimulus their number may change with the state of
Figure 3-27. More complex neuroendocrine relationships cell differentiation and can be influenced by
involved in response to stress. A painful peripheral stim¬ exposure to unphysiological levels of the spe¬
ulus reaching the brain is relayed to neurosecretory cells cific hormone (homospecific regulation). The
in the hypothalamus. These liberate ACTH releasing action of one hormone on a cell may induce
hormone (ACTH-RH) into the vessels of the hypophyso-
portal system. Carried downstream to the anterior lobe, the appearance of receptors for a second
this hormone stimulates corticotrophs to release ACTH. hormone (heterospecific regulation). The re¬
The ACTH borne by the blood to the adrenal cortex sponsiveness of target cells is a function of
causes release of corticosteroids (ACH) that are carried the number of available receptors. Tissues
to cells throughout the body, inducing protective metabolic
that do not respond to a given hormone
responses. Adrenocortical hormones act back upon the
hypothalamus to suppress liberation of ACTH-RH. invariably lack receptors for that hormone.
 GLANDS AND SECRETION • 107
The biochemical mechanisms by which in which this mechanism is operative, the
hormones elicit a response in their target action of the hormone can be duplicated by
cells have been a subject of intensive investi¬ application of exogenous cyclic AMP to the
gation. Two principal mechanisms have target cells.
emerged. In the case of steroid hormones, This mode of coupling a stimulus to its
the hormone—receptor complex is translo¬ response is involved in the action of the
cated to the cell nucleus where it becomes
hypophyseal hormones adrenocorticotropin,
associated with acceptor sites on the chro¬ thyrotropin, and gonadotropins, and in the
matin. There it activates gene transcription, release of calcitonin from the parafollicular
which in turn directs the appropriate syn¬ cells of the thyroid. It also mediates those
thetic activities of the responding cell. examples of exocrine secretion that are ini¬
The action of other hormones is mediated tiated by the neurotransmitter norepineph-
by a mechanism discovered in studies on the line. An explanation of the remarkable di¬
effects of norepinephrine and glucagon on versity in the responses of various target cells
the liver (Fig. 3-28). In binding to receptors, to a single second messenger is still being
these molecules become associated with an sought. All the effects of cyclic AMP appear
enzyme, adenyl cyclase, in the cell membrane. to be mediated by controlling the activity of
The activated enzyme catalyzes the formation protein kinases that catalyze the transfer of
of cyclic AMP from adenosine triphosphate phosphate from ATP to protein. The diver¬
(ATP). Intracellular accumulation of cyclic sity of the responses of target cells, therefore,
AMP activates protein kinases, enzymes that seems to depend, not on the control system,
set in motion a train of events leading to the but on the distinctive proteins available and
characteristic response of the target cell to on their varied biological activities when
the hormone. These findings led to the “sec¬ phosphorylated.
ond messenger” concept. Briefly stated, a The second messenger, cyclic AMP, is com¬
hormone, the “first messenger,” is carried in monly involved in the category of nonexcitable
the blood and activates adenyl cyclase in the tissues such as liver, other glands, and adipose
target cell membrane, resulting in an increase tissue. Another mechanism by which stimuli
in the “second messenger,” cyclic AMP, which evoke cellular responses utilizes an ionic mes¬
induces a specific response. In all instances senger, the divalent cation Ca2+. Calcium is
principally involved in coupling stimulus to
response in excitable tissues such as nerve and
muscle, where ion currents lead to action
potentials propagated over the cell surface
from the site of the initial stimulus. Stimulus-
induced fluxes in free intracellular Ca2+ are
controlled by an ubiquitous regulatory pro¬
tein, calmodulin, which can activate a number
of enzymes involved in the physiological re¬
sponses of cells. Among its other functions,
calmodulin plays a role in activation of adenyl
cyclase by modulating the local concentration
of Ca2+. The validity of the sharp distinction
often made between the ionic and cyclic AMP
mechanisms of stimulus-response coupling is
now in doubt since it has been shown that
the two often interact in achieving a cellular
response.
Although our understanding of the mech¬
anisms by which binding of hormones to
Figure 3-28. Diagram of the mechanism of action of
hormones. The hormone epinephrine (first messenger) receptors results in propagation of signals to
carried to the liver cell membrane in the blood activates the appropriate synthetic pathways within the
the enzyme adenylate cyclase in the cell membrane, cell is still incomplete, enough is known to
causing it to convert some of the ATP of the cytoplasm explain several diseases whose pathogenesis
into cyclic AMP (second messenger). This then activates
was previously obscure. The congenital dis¬
a protein kinase, which activates a second kinase. The
second kinase initiates a four-step sequence that converts order testicular feminization is now known to
glycogen into glucose, which then passes out into the be a genetic abnormality of receptor func¬
bloodstream in the hepatic sinusoids. tion. Individuals with a male chromosome
108 • GLANDS AND SECRETION % ^

linkage between the nervous and endocrine

complement (46 XY) at conception fail to
system and initiated a new held of research
develop testicular function and male external
called neuroendocrinology.
genitalia because of receptor anomalies that
The traditional distinction between neural
result in complete insensitivity of the target
and endocrine systems was further clouded
tissues to the hormone dihydrotestosterone.
by the discovery that the neurotransmitter
In some patients the receptors are completely
norepinephrine is also a hormone i eleased into
absent. In others they are present but the
the circulation by endocrine cells in the ad¬
hormone—receptor complex lacks the ability
renal medulla. Conversely, the peptide hor¬
to activate gene transcription in the nucleus.
mone vasopressin, released in the neurohy¬
Comparable receptor abnormalities are re¬
pophysis, has been found to serve as a
sponsible for familial male pseudohermaphrodi¬
neurotransmitter for certain cells in the hy¬
tism. . pothalamus. A dozen or more peptides have
Endocrine disturbances may arise m adults
now been found to relay signals between
as a consequence of development of auto¬
endocrine cells and their targets. Cells releas¬
antibodies that bind to receptors on specific
ing these peptides occur singly or in small
target organs. Patients with Graves disease
groups in various regions of the brain, and
(thyrotoxicosis) have an abnormal immuno¬
are widely distributed in the gastrointestinal
globulin in their serum that binds to the
tract. In contrast to the compact endocrine
receptor for thyrotropic hormone and acti¬
glands, these dispersed cells collectively make
vates adenyl cyclase, resulting in continual
up a diffuse neuroendocrine system, which can
stimulation of the cells to produce excess
be regarded as a third system for coordinat¬
thyroid hormone. By competing with thyro¬
ing cellular activities using chemical messen¬
tropic hormone for binding sites, the immu¬
gers commonly referred to as neuropeptides.
noglobulin overrides the endocrine control
These isolated cells, possessing properties in
mechanisms that normally govern thyroid
common with both neurones and endocrine
function. Similarly, rare cases of diabetes are
cells, are now the focus of much investigative
attributable to an abnormal immunoglobulin
interest. Twenty or more morphologically
that competes with insulin for receptor sites.
similar but functionally distinct cell types
In this condition, the immunoglobulin is not
have been identified in the epithelium oi the
stimulatory and therefore acts as an insulin
gut and its associated glands. These have
antagonist.
been called the entero-endocrine cells or gastro-
enteropancreatic (GEP) endocrine cells (Chapter
26). But these terms are not sufficiently in¬
NEUROENDOCRINE CELLS,
clusive, for cells with the same peptide secre¬
PARANEURONES
tory products have also been found widely
dispersed in the central nervous system.
As stated earlier in this chapter, two sys¬
Some, but not all, of these cells have the
tems for coordinating the activities of cells in
capacity for amine precursor uptake and de¬
the body have long been recognized: (1) the
carboxylation and have been grouped to¬
nervous system, providing rapid point-to-
gether by some investigators under the ac¬
point communication and using chemical neu¬
rotransmitters released at special synaptic sites ronym APUD cells.
Thus, the traditional view of histologists
on long cell processes; and (2) the endocrine
that neurones were so differentiated in struc¬
system, acting more slowly and using hormones
ture and function that they could easily be
transported to distant target cells by the
identified on the basis of shape and posses¬
blood. The neurotransmitters are commonly
sion of certain distinctive organelles has been
monoamines or amino acids; the hormones
progressively eroded. The electron micro¬
are small glycoproteins, polypeptides, or ste¬
scope showed that some of the structural
roids.
components believed to be unique to neu¬
The separation of neurones from endo¬
rones were common to many cell types. “Neu¬
crine cells was narrowed in the 1930s by the
rofilaments” and “neurotubules” do not dif¬
description of cells in the nervous system that
fer significantly from the cytoskeletal
have the form of neurones but contain secre¬
filaments and microtubules of cells in gen¬
tory granules that are transported along their
axonal processes, releasing their contents into eral. The “Nissl bodies” are simply parallel
perivascular spaces to be carried in the blood arrays of cisternae of the endoplasmic retic¬
ulum and associated polyribosomes. And the
to their target cells. The discovery of these
neurosecretory cells established an important defining physiological property of membrane
 GLANDS AND SECRETION • 109

depolarization is now known to be shared lial cells or adjacent nerve cells. This local
with several endocrine cells. A consensus has action is described as paracrine secretion to
gradually developed that there are overlap¬ distinguish it from blood-borne distribution
ping properties and a broad continuity be¬ of hormones from the endocrine glands. Cy-
tween neurones and endocrine cells. tological criteria alone do not permit unam¬
It has been suggested that the numerous biguous identification of the cells releasing a
cell types of the diffuse endocrine system particular peptide. Such identification must
should be regarded as closely related to neu¬ be accomplished immunocytochemically by
rones and might appropriately be called par- using fluorescein-labeled antibodies against
aneurones. They all have in common the pres¬ each of the biologically active peptides.
ence of small secretory granules or vesicles Included in the paraneurone category are
concentrated at the base or vascular pole of the several endocrine cell types of the gut
the cell. The cytoplasm is usually electron exemplified by the gastrin cell, which has
lucent, the endoplasmic reticulum sparse, microvilli serving a chemoreceptor function,
and the Golgi apparatus relatively small. and basal granules that release hormone (Fig.
They secrete substances identical with or 3—29A). Similar in structure and sensory
closely related to known neurosecretions or function are the gustatory cells of the lingual
neurotransmitters (peptides and sometimes taste buds (Fig. 3—29B); the basal granular
monoamines). They have the morphological cells of the bronchial epithelium that sense
characteristics of cells specialized for recep¬ hypoxia of the respired air (Fig. 3-29C); the
tion of a stimulus and release of a secretion principal cells of the carotid body (Fig.
in response. The products of some are re¬ 3—29D); and the olfactory cells of the nasal
leased into the bloodstream for transport to epithelium (Fig. 3—29E). In addition to these
distant target cells. The products of many cells with a chemoreceptor function, there
others simply diffuse to neighboring epithe¬ are mechanoreceptor cells such as the hair

Figure 3-29. Principal types of paraneurons. A, Endocrine cell of gut; B, gustatory cell of taste bud; C, Basal granulated
cell of bronchus; D, chief cell oflhe carotid body; E, olfactory cell; F, hair cell of the inner ear; G, Merkel cell of the skin;
H, avian pinealocyte; I, visual cell of retina; J, adrenal chromaffin cell; K, endocrine cell of adenohypophysis, parafollicular
ceil of thyroid, or pancreatic islet cell. (Modified after Fujita, T., et al. In Farner, D., and K. Lederis, eds.: Neurosecretion:
Molecules, Cells, Systems. New York, Plenum Press, 1982.)
110 • GLANDS AND SECRETION
Farquahar, M. D.: Recovery of surface membrane in
cells of the inner ear (Fig. 3-29F) and the anterior pituitary cells. Variations in traffic detected
Merkel cells of the epidermis (Fig. 3-29G). with ionic and cationic ferritin. J. Cell Biol.
Possibly qualifying as paraneurones are the 77-.R35-R42, 1978.
avian pinealocytes that retain some photore¬ Herzog, V., and H. Reggio: Pathways of endocytosis
from luminal plasma membrane in rat exocrine
ceptor function (Fig. 3-29H) and the phylo-
pancreas. Eur. J. Cell Biol. 21.141, 1980.
genetically related visual cells of the mam Jamieson, J. D„ and G. E. Palade: Production of secre¬
malian retina (Fig. 3-29/). Finally, there are tory proteins in animal cells. In Brinkley, B. B., ana
noninnervated endocrine cells such as those K. R. Porter, eds.: International Cell Biology. New
of the adenohypophysis, the parafollicular York, Rockefeller University Press, 1977.
Leblond, C. P., and G. Bennett: Role °f the Golg1
cells of the thyroid, and the pancreatic islet apparatus in terminal glycosylation. In Brinkley, B.
cells (Fig. 3-29)J). B and K. R. Porter, eds.: International Cell Biol¬
The paraneurone concept is still evolving, ogy. New York, Rockefeller University Press, 1977.
but is a convenient generalization that en¬ Neutra, M., and C. P. Leblond: Synthesis of the carbo¬
hydrate of mucus in the Golgi complex as shown by
compasses several more narrowly defined cat¬ electron microscope radioautography of goblet cells
egories and brings some oi der out of the from rats injected with glucose-H3. J. Cell Biol.
terminological chaos that has resulted from 30:119, 1966. i ,
the rapid discovery of numerous types of Palade, G. E.: Intracellular aspects of the process ol
protein secretion. Science 759:347, 1975.
granulated cells secreting a bewildering va¬
Walter, P., I. Ibrahimi, and G. Blobel: Translocation ol
riety of hormones or transmitters. The find¬ proteins across the endoplasmic reticulum. J. Cell
ing of isolated cells with a sensory apex and Biol. 97:545, 1981.
a secretory base in hydroids and other inver¬
ENDOCRINE SECRETION
tebrates suggests that paraneurones and their
Fawcett, D. W„ J. A. Long, and A. L Jones. The
secretory products have been highly con¬ ultrastructure of the endocrine glands. Recent Prog.
served in evolution and may well have ante¬ Hormone Res. 25:315, 1969.
dated development of the neural and endo¬ Grossman, M. I.: Integration of neural and hormonal
crine coordinating systems. control of gastric secretion. Physiologist 6:349, 19b5
O’Malley, B. W., and W. T. Schrader. Thejeceptors ol
steroid hormones. Sci. Am, 234.32, 1976.
Schally, A. U., A. J. Kastin, and A. Anmura: Hypotha¬
REFERENCES lamic hormones: the link between brain and body.
Am. Scientist 65:712, 1977.
EXOCRINE GLANDS, GENERAL Smith, A. D.: Storage and secretion of hormones. Sci.
Basis Med. 74:102, 1972.
Bowen, R. H.: The cytology of glandular secretion. Q.
Turner, C. D., and J. T. Bagnara: General Endocrinol¬
Rev. Biol. 4:299, 484, 1929. ogy. 5th ed. Philadelphia, W. B. Saunders Co., 1971.
Nassonov, D.: Das Golgische Binnennetz und seine Be-
ziehungen zu Sekretion. Morphologische und ex- CONTROL OF SECRETION
perimentelle Untersuchungen an einigei Saugetiei
Rasmussen, H.: Cell communication, calcium ion and
drusen. Arch. Mikr. Anat. 7(90:433, 1924. cyclic adenosine monophosphate. Science 7 79.404,
Scharrer, E„ and B. Scharrer: Neurosekretion. In von
Mollendorff, W., and W. Bargman, eds.. Handbuch 1970. r a atd
Sutherland, E. W.: On the biological role of cyclic AMP.
der Mikroskopisches Anatomie des Menschen. Vol.
J.A.M.A. 274:1281, 1970.
6, Part 5. Berlin, Springer-Verlag, 1954.
PARACRINE SECRETION
PROTEIN SECRETION
Feyrter, F.: Uber die peripheren endokrinen (parakri-
Amsterdam, A., I. Ohad, and M. Schramm: Dynamic
nen) Drusen des Menschen. 2nd ed. Vienna, Wil¬
changes in the ultrastructure of the acinar cell ol
helm Mudrich, 1953.
the rat parotid gland during the secretory cycle. J.
Fujita, T.: Concept of paraneurons. Arch. Histol. Jpn.
Cell Biol. 41:753, 1969.
49(Suppl):l, 1977.
Blobel, G., P. Walter, C. N. Chang, B. M. Goldman, A.
Fujita, T., T. Iwanaga, Y. Kusumato, and S. Yoshie:
H. Erickson, and V. R. Lingappa: Translocation of
Paraneurons and neurosecretion. In Farner, D. S.,
protein across membranes. The signal hypothesis.
and K. Lederis, eds.: Neurosecretion: Molecules,
In Secretory Mechanisms. Symp. Soc. Exp. Biol.
Cells, Systems. New York, Plenum Publishing Corp.,
Vol. 33. Cambridge, Cambridge University Press,
1982.
1979. Pearse, A. C. E.: The diffuse neuroendocrine system
Castle, J. D., J. D. Jamieson, and G. E. Palade: Radioau-
arid the APUD concept: related endocrine peptides
tographic analysis of the secretory process in the
in brain, intestine, pituitary, placenta, and anuran
parotid acinar cell of the rabbit. J. Cell Biol. 55.290,
cutaneous glands. Med. Biol. 55:115, 1977.
1972.
 BLOOD

Blood is a fluid tissue consisting of erythro¬ it soon became evident that only the eryth¬
cytes (red blood cells) and leukocytes (white rocytes and platelets function entirely within
blood cells) suspended in blood plasma. It the confines of the vascular system. The sev¬
circulates in the vascular system, transporting eral types of leukocytes are only very tran¬
oxygen from the lungs and nutrients from siently in the blood and are constantly mi¬
the digestive tract to other tissues throughout grating through the walls of capillaries and
the body, and carrying carbon dioxide to the venules to become free cells of the connective
lungs and nitrogenous waste products to the tissues. It is there that they carry out their
kidneys for excretion. Blood also plays an functions, complete their short life span, and
essential role in the integrative function of degenerate. The blood is simply the vehicle
the endocrine system by distributing hor¬ for transport of the leukocytes from the bone
mones from their sites of production to their marrow, where they are generated, to the
distant target organs. tissues to carry out their appointed tasks.
All the connective tissues of the body are The volume of blood in humans is approx¬
composed of cells distributed in an abundant imately 5 liters, accounting for 7 per cent of
extracellular matrix. Cartilage, for example, body weight. Erythrocytes make up about 45
consists largely of intercellular material that per cent of this volume, the leukocytes and
is a firm gel, and in bone the extracellular platelets make up 1 per cent, and the re¬
matrix is a highly organized scaffolding of mainder is blood plasma, the transparent
mineralized fibers. Blood has traditionally yellow liquid that constitutes the extracellular
been classified as a connective tissue in which matrix of this tissue. When blood is drawn
the intercellular substance is fluid. Obviously from the circulation, it rapidly clots to a deep
it is not a “connective tissue” in the sense of red, jelly-like mass, but if clotting is pre¬
binding together and preserving the struc¬ vented by an anticoagulant the cellular ele¬
tural integrity of the organism, but only in ments can be centrifuged to the bottom of
the sense that it maintains logistic support the tube, providing a useful measure of the
and communication between the tissues and packed cell volume and a clear view of the
organs of the body. plasma.
The boundaries of the several kinds of A thorough knowledge of the normal his¬
tissues are not always clearly defined. The tology of blood is of great importance in
fibers of connective tissue proper extend into medical and veterinary practice, for no tissue
the extracellular matrix of cartilage and is examined more often for diagnostic pur¬
bone. Similarly the circulating blood is not poses. Study of stained blood smears under
functionally separable from the connective the microscope not only yields information
tissue around the blood vessels, for fluid about diseases that primarily affect the blood
constituents of the blood are constantly filter¬ and blood-forming organs, but also may pro¬
ing through the capillary walls to contribute vide evidence of viral, bacterial, and parasitic
to the fluid phase of the connective tissue infections. It enables the physician to identify
matrix. Concurrently, metabolites are diffus¬ the nature of the disease; to follow its course;
ing from the tissue fluid back into the blood¬ and to evaluate the effectiveness of his treat¬
stream. This continual exchange of sub¬ ment.
stances between the blood plasma and tissue
fluid is essential for survival of the cells in all
the organs.
It was formerly thought that all the blood ERYTHROCYTES
cells carried out their principal functions in
the bloodstream, but when it became possible The erythrocytes are the minute corpuscles
to radiolabel cells and trace their migrations, that impart the red color to the blood. They
111
112 • BLOOD

is well adapted to its function, for in this

develop in the bone marrow as true cells but,
form it presents a surface area 20 to 30 per
before entering the circulation, they extrude
cent greater in relation to its volume than it
their nucleus, losing the capacity for DNA-
would if it were spherical. This increased
directed protein synthesis. Their mitochon¬
surface favors the immediate saturation of its
dria and other membrane-limited organs are
hemoglobin with oxygen as the erythrocyte
also lost and they are thus reduced to plas-
passes through the pulmonary capillanes.
tids* whose cytoplasm consists mainly of
The total surface area of the erythrocytes in
hemoglobin. In this structural simplification
an average human is 3800 square meters
the erythrocytes become specialized for the
some 2000 times the total body surface. The
primary function of transporting oxygen
enormous surface area of the erythrocytes
from the lungs to the tissues and carbon
results in great efficiency in oxygen and car¬
dioxide from the tissues to the lungs. Anu-
cleate erythrocytes are characteristic of mam¬ bon dioxide transport.
The uniformity of shape of erythrocytes m
mals. In birds, reptiles, amphibia, and fish,
blood smears is not entirely representative of
the erythrocytes retain a nucleus but its chro¬
their form in the peripheral ciiculation. They
matin is functionally inert.
are very pliable and may be deformed to bell¬
The normal number of erythrocytes per
like or paraboloid shape when flowing
cubic millimeter of blood is about 5.4 million
through capillaries. This deformation is de¬
in men and 4.8 million in women. These
pendent on the velocity of blood flow and is
numbers are increased somewhat by resi¬
a consequence of hydrodynamic force and
dence at high altitude. The mammalian
viscous drag. Since the altered configuration
erythrocyte has a highly consistent and chai-
results in a further increase in surface area,
acteristic shape (Figs. 4—1B, 4—3). It is a
this transient shape change may have some
biconcave disc about 7.5 fxm in diametei, 1.9
significance in gas exchange.
|xm in maximal thickness, and has a surface
When fresh blood is examined in a thick
area of approximately 140 square microme¬
smear under the microscope, the erythro
ters. The biconcave shape of the erythrocyte
cytes are often observed to associate in stacks
called rouleaux (Fig. 4—1A). This phenome¬
*Since they lack the defining organelles of true cells, non does not occur when blood is ciiculatmg
the terms “red blood cell ” and “erythrocyte” are inap¬ but only when it is still or stagnant. It is often
propriate. Erythroplastid would be a more accurate de¬ considered a surface tension effect but its
scription, but erythrocyte is so widely used it is not likely
physical basis is in fact poorly understood.
to be replaced.

lure 4-1 Photomicrographs of fresh blood viewed by Nomarski optics. A, Erythrocytes in vitro often aggregate like
cked^coins; th"Jed rouleau formation. B, Normally, erythrocytes are biconcave discs with an obvious cen ral
nression and a thicker rim. C, Erythrocytes may assume a spiny configuration with 10 to 30 spicules evenly distributed
*r the surface and are then referred to as echinocytes. This change in form is often called crenauon, and usually
curs during specimen preparation but may occur in pathological conditions in vivo. (Micrographs courtesy of Marcel
Bessis.)
 BLOOD *113

The shape of erythrocytes is sensitive to

various factors in the surrounding medium.
In moderately hypotonic solution they swell
and may become uniconcave. In strongly
hypotonic solution the membrane is stretched
and becomes leaky or ruptures, allowing the
hemoglobin to escape, leaving behind an
empty membrane called an erythrocyte ghost.
This disruption is called hemolysis. In moder¬
ately hypertonic solution erythrocytes be¬
come flattened but do not otherwise alter
their shape. Under various experimental con¬
ditions in vitro, erythrocytes assume a spiny
configuration with 10 to 30 spicules regularly
distributed over their surface, and are then
called echinocytes (Fig. 4-1C). This change of
form is referred to as crenation. It was for¬
merly attributed to hypertonicity of the me¬
dium but is now known to be due to other
factors. Formation of echinocytes can be in¬
duced in vitro by exposure to fatty acids,
lysolecithin, anionic compounds, or elevated
pH.
Unstained erythrocytes have a pale yellow
or tan color due to their content of hemoglo¬
bin, the respiratory pigment that makes up
about 33 per cent of their mass. Hemoglobin
is the most essential biochemical component Figure 4-2. Scanning electron micrograph of two normal
of the erythrocyte. It is a protein with a biconcave erythrocytes and one crenated erythrocyte.
molecular weight of 65,000 consisting of four (Micrograph courtesy of Marcel Bessis.)
polypeptide globin chains. An iron-contain¬
ing heme group is bound to each chain of pies of disease involving defects at the molec¬
the tetramer. ular level are now of great clinical interest. A
Humans are genetically capable of synthe¬ classical example is hemoglobin S (HbS) as¬
sizing and incorporating into hemoglobin sociated with sickle cell anemia. It differs from
four structurally different polypeptide chains normal only in the substitution of valine for
designated alpha (a), beta (|3), gamma (y), glutamine at one site on the beta chain, but
and delta (6). Hemoglobin of the normal this substitution is enough to make it less
adult, called hemoglobin A (HbA), contains soluble than normal in the reduced condition
two alpha and two beta chains. This form and results in formation of long tactoids that
accounts for 96 per cent of the hemoglobin deform the red ceils into bizarre sickle forms,
while 2 per cent is of a second type (HbA2), which block capillaries and are prone to he¬
consisting of two alpha and two delta chains, molysis.
and less than 2 per cent is fetal hemoglobin Normally, the erythrocytes in a dry smear
(HbF), composed of two alpha and two of peripheral blood stain deep pink or
gamma chains. HbF greatly predominates salmon color with Wright’s stain (Fig. 4-7).
over the others during fetal life but dimin¬ Some of the young red cells that have lost
ishes in amount in the postnatal period. In their nucleus shortly before entering the cir¬
certain types of anemia (thalassemia), there culation and are not yet completely mature
is a persistence of abnormally high levels of may have a bluish or greenish tinge due to
fetal hemoglobin. the basophilic staining of small numbers of
A separate genetic locus determines the residual ribosomes. These are called polychro-
structure of each of the four globin chains, matophilic erythrocytes or reticulocytes. The latter
and a variety of inherited disorders of hemo¬ term refers to the fact that when blood
globin synthesis have been discovered that smears are stained with brilliant cresyl blue,
involve relatively minor amino acid substitu¬ the remaining ribonucleoprotein is precipi¬
tions in the beta chain but nevertheless have tated by the dye into a delicate basophilic
profound physiological effects. These exam- network in the otherwise acidophilic hemo-
114 • BLOOD

rocyte membrane than about any other. The

globin-rich cytoplasm. Within 24 hours after
biconcave shape and elasticity of the red cell
entering the blood from the bone marrow,
are attributed to a complex of peripheral
the reticulocytes mature into adult erythro¬
cytes. Reticulocyte number in adult humans proteins on the inner aspect of the mem¬
brane, which are organized in a deformable
averages about 0.8 per cent of the total eryth¬
reinforcing meshwork. The principal com¬
rocytes of blood. The reticulocyte count is used
ponent of this unusual cytoskeleton is spectrin,
clinically as a rough index of the rate of
erythrocyte formation. In patients with ane¬ a large asymmetrical molecule composed of
mia, elevation of the count is a valuable sign two polypeptide subunits of 240,000 and
220,000 M.W. It normally occurs as a tetra-
of response to treatment.
A detailed description of abnormal forms mer (920,000 M.W.), a long, flexible, rodlike
of erythrocytes is more appropriate for text¬ molecule 200 nm long. The erythrocyte cy-
books of hematology or pathology, but a few toskeletal complex is believed to be made up
of the descriptive terms may be useful here. of a meshwork of spectrin rods linked to¬
Abnormal variation in size of red cells is gether by actin oligomers. This web is at¬
described as anisocytosis. Red cells that are tached to the membrane by another protein
larger than normal are macrocytes, and an called ankyrin, which binds to both spectrin
anemia characterized by such cells is a macro¬ and one of the integral membrane proteins
(Pig 4_4). Spectrin and ankyrin were
cytic anemia. Deviation from the normal shape
is described by the term poikilocytosis. The thought to occur only in erythrocytes but
concentration of hemoglobin per cell is less there is now evidence that they occur in
than normal when the rate of red cell for¬ certain other cell types and may play a i ole
mation is relatively greater than the rate of in membrane—cytoskeleton linkage.
hemoglobin synthesis. Under these circum¬ To students, these ultrastructural details of
stances, each erythrocyte contains an abnor¬ the erythrocyte cytoskeleton may seem to be
mally small quantity of hemoglobin. It ap¬ esoteric, but they have recently acquired clin¬
pears paler and is described as hypochromic, ical significance insofar as they explain the
as opposed to normal or normochromic eryth¬ cellular abnormalities observed in certain he¬
rocytes. reditary hemolytic anemias of humans, in¬
Electron micrographs of erythrocytes re¬ cluding hereditary spherocytosis and hereditary
veal a homogeneous content of considerable elliptocytosis. In these conditions there are
density that at high magnification has a finely varying degrees of membrane instability re¬
granular texture representing molecules of sulting in deformations in shape of the eryth¬
hemoglobin (Fig. 4—3). Microtubules and fil¬ rocytes and excessive fragility. Underlying
aments found in the cytoskeleton of other these abnormalities, patients may have defi¬
cells are not observed. Owing to the ready ciencies of spectrin or of ankyrin or defective
availability of erythrocytes and the ease with binding of spectrin to actin and the integral
which the plasmalemma can be purified after protein of the erythrocyte membrane. The
hemolysis, more is known about the eryth¬ study of these skeletal protein mutants sheds

Fiaure 4-3. In a micrograph of an erythrocyte in thin section, its cytoplasm is devoid of organelles and consists of a
homogeneous suspension of hemoglobin molecules that at high magnification give the interior a finely granular texture.
 BLOOD *115

glycophorin

Figure 4-4. A, Drawing of the components of the erythrocyte cytoskeleton-a meshwork beneath the plasma membrane.
The links of the network consist of dimers of spectrin joined end to end, the nodal points being a complex of spectrin,
actin, and protein 4. The cytoskeleton is bound to the integral transmembrane protein 3 by a cross-linking protein!
ankyrin. B, Drawing depicting the configuration of the cytoskeleton as it would appear in a view of the inner aspect of
the erythrocyte membrane. (From C. M. Cohen, Semin. Hematoi. 20:141, 1983 by permission of Grune & Stratton, Inc.)

light upon the relationships of the cytoske¬ Platelets are flat, biconvex discs 2 to 3 pm
leton to the membrane not only in erythro¬ in diameter, round or ovoid when viewed on
cytes but in cells generally. the flat and fusiform in profile when seen on
edge. In human blood their number ranges
from 150,000 to 350,000 per cubic millime¬
ter. In stained blood smears they exhibit two
PLATELETS concentric zones—a thin, pale-blue periph¬
eral zone called the hyalomere, and a thicker
The thromboplastids or platelets are minute, central region, the chrojnomere or granulomere
colorless, anucleate corpuscles found in the containing small azurophil granules.
blood of all mammals. They are involved in In electron micrographs, the granulomere
the clotting of blood at sites of injury to blood usually contains one or two mitochondria and
vessels and are essential in the protection of numerous small clear vesicles (Fig. 4-5). Gly¬
the organism against excessive blood loss. cogen is present in the form of scattered
Their functional equivalents in lower verte¬ particles. Varying numbers of membrane-
brates are nucleated cells called thrombocytes. bounded dense granules, 0.2 pm in diameter,
116 • blood \

Figure 4-5. Electron micrograph of circulating platelets in the lumen of a capillary.

correspond to the azurophil granules seen tracted from platelets will form in vitro thin
with the light microscope. These are com¬ filaments and thicker filaments identifiable as
monly called oi granules. In some species the actin and myosin respectively. Thus, the mech¬
platelets have a second category of very dense anism of platelet contraction is believed to
0.5-jJim granules that are believed to contain have features in common with that of muscle.
the vasoactive substance serotonin (5-hydrox- The contractile material of circulating plate¬
ytryptamine). A few tubular invaginations lets is present mainly in monomeric form,
extend from the plasmalemma into the inte¬ but platelet activation during blood clotting
rior of the platelet. These are residual ele¬ seems to initiate polymerization of actin and
ments of the system of cytoplasmic mem¬ myosin monomers into the filamentous form
branes in the megakaryocytes from which the necessary for contraction.
platelets arise (see Chapter 7). They may have Determinations of the life span of platelets
no functional significance in the mature using the isotopic label Cr1 indicate that they
survive eight to 11 days in the normal circu¬
platelet.
The hyalomere lacks membranous orga¬ lation. Despite their small size and lack of a
nelles and is characterized by a finely fila¬ nucleus, they are able to carry out many of
mentous electron-lucent cytoplasm similar in the metabolic activities of whole cells. They
texture to the peripheral ectoplasmic zone of consume oxygen, are rich in adenosine tri¬
leukocytes and other motile cells. Its most phosphate (ATP), and contain 25 or more
conspicuous structural element in equatorial enzymes and several pharmacologically active
sections is a bundle of 10 to 15 microtubules compounds.
that run circumferentially near the plasma The principal functions of platelets are to
membrane (Fig. 4—6B). When cut tians- patch small defects in the endothelial lining
versely, the microtubule bundle is repie- of blood vessels and to limit hemorrhage by
sented by a cluster of small circular profiles promoting local coagulation of the blood. In
at either end of the sectioned platelet (Fig. the circulation, platelets exhibit no tendency
4—6A ). These microtubules are a cytoskeletal to adhere to each other, to other cells, or to
specialization serving to maintain the discoid the lining of the blood vessels, but when they
form of the platelet. A similar marginal band encounter surfaces to which they are not
is found in the nucleated erythrocytes of fish, normally exposed, they rapidly adhere. This
reptiles, and birds, which are also flattened adhesive property is expressed in vitro by
biconvex discs. sticking to glass, plastic, or other solid sub¬
Platelets contain contractile material with strates. By convention, platelet adhesion is de¬
properties similar to those of actomyosin ex¬ fined as the sticking of platelets to solid
tracted from muscle. Under appropriate surfaces, and platelet aggregation is the sticking
physicochemical conditions this material ex¬ of platelets to each other.
 BLOOD • 117

At sites of vascular injury they adhere to bosis, the basis of the familiar “heart attack.”
damaged endothelium and to exposed colla¬ Similarly, when clots that have formed in
gen, forming a layer of platelets over the injured veins of the extremities break loose
denuded area. The adhering platelets are from the vessel wall, they may be carried to
activated by this contact to break down their the lungs, resulting in fatal pulmonary embo¬
ATP and release ADP onto their surface and lism.
into the surrounding medium. ADP is a po¬ Defects in platelets or in plasma factors
tent inducer of platelet aggregation and participating in the clotting mechanism are
other platelets stick to those initially depos¬ responsible for a number of human and
ited. These in turn are activated and induce animal diseases. Clotting defects attributable
further aggregation. The mass of platelets to platelets may be due to quantitative defi¬
on the vessel wall thus continues to enlarge, ciency in their production, thrombocytopenia,
producing a platelet thrombus and finally a or to qualitative abnormalities of structure
hemostatic plug. and function, thrombocytopathia. An example
Concurrently with platelet aggregation, of the latter is the condition called thrombas¬
other complex reactions of blood clotting are thenia, in which platelet numbers may be
set in motion. A substance called tissue throm¬ normal but there is deficiency of adhesion,
boplastin released from the injured tissue of poor aggregation, and diminished clot re¬
the vessel wall initiates a series of reactions traction. In some inherited diseases such as
in blood plasma that convert prothrombin to thrombocytopenic purpura, a defect in platelet
thrombin. Thrombin catalyzes the conversion production is linked to abnormal fragility of
of plasma fibrinogen to fibrin, which polymer¬ the small vessels, resulting in spontaneous
izes as a feltwork of cross-striated fibrils that capillary bleeding manifested by multiple
enmesh erythrocytes and platelets to form a “black-and-blue” areas over the surface of
gelatinous clot. the body.
In the process of aggregation and activa¬ Blood coagulation is an extraordinarily
tion, the platelets undergo dramatic mor¬ complex process involving the interaction of
phological changes. They extend numerous at least 12 plasma factors in addition to the
slender processes, release the content of their platelets. Inherited single deficiencies of
granules, and ultimately coalesce into a co¬ many of these factors have been described.
herent viscous mass. Associated with their The best known of these disorders is the
degranulation, phospholipid is released, bleeding disease hemophilia.
which reacts with other plasma components
to produce platelet thromboplastin. This in turn
acts to promote progression of the clotting
LEUKOCYTES
process initiated by tissue thromboplastin.
Within an hour or so after its formation,
the blood clot shrinks to about half its original In addition to the red cells, the blood of
volume. This is attributed to polymerization all mammals contains a number of types of
of actin and myosin filaments during the colorless cells, the leukocytes or white blood
viscous metamorphosis of the platelets trig¬ corpuscles. They are true cells with a nucleus
gered by thrombin, and their interaction to and cytoplasm and all are spherical in the
produce contraction of the clot. The hemo¬ blood but more or less ameboid in the tissues
stasis achieved by occluding the lumen is or on a solid substrate. There are five kinds
supplemented by active constriction of the of leukocytes in the blood (Figs. 4-6, 4-8).
injured vessel. This is in part a direct conse¬ They are categorized according to the pres¬
quence of mechanical stimulation of the ves¬ ence or absence of specific cytoplasmic gran¬
sel wall at the time of injury, but there is ules {granular and nongranular) and according
evidence that diffusible substances released to the shape of their nucleus (mononuclear*
from the platelet mass also play a role. The or polymorphonuclear). The granular leuko¬
serotonin of the platelets may be involved cytes are further classified according to the
and proteolytic enzymes activated in the clot¬
ting process may result in production of *This misleading term would seem to imply possession
bradykinin and other vasoactive peptides. of a single nucleus. All the leukocytes have one nucleus.
The clotting process is essential for limita¬ Mononuclear (referring to lymphocytes and monocytes)
unfortunately has come into general use as a contraction
tion of hemorrhage but can also be life-
of monomorphonuclear (a simple unlobulated nucleus) in
threatening when it is initiated on the walls contrast to polymorphonuclear (a complex lobulated or
of coronary arteries—causing coronary throm¬ segmented nucleus).
118 • BLOOD
v

Figure 4-6. A, Micrograph of a platelet sectioned through its narrow dimension, showing its dense secretory granules
and an aggregation of glycogen particles. At arrows are cross sections of the bundle of microtubules that runs
circumferentially in the rim of the platelet. B, A platelet sectioned parallel to the broad dimension. Here the bundle of
microtubules that helps to maintain the discoid shape of the platelet is seen in longitudinal section. (Micrographs
courtesy of O. Behnke.)
 BLOOD ‘119

Figure 4-7. Human blood cells form a smear after Wright’s stain. A and D, Neutrophilic leukocytes. B and E, Eosinophilic
leukocytes. C, Basophilic leukocyte. F, Plasma cell; this is not a normal constituent of the peripheral blood but is
included here for comparison with the nongranular leukocytes. G and H, Small lymphocytes. I, Medium lymphocytes. J,
K, and L, Monocytes.
120 • BLOOD

Neutrophils
("Polys")

Eosinophils

Basophils

Lymphocytes

Monocytes
Finure 4-8 Classification of leukocytes. The granular leukocytes, especially the neutrophils, are polymorphonuclear
S '1?onaranul£Ss w often referred to as “mononuclear” leukocytes, an unfortunate term that implies that
the others are multinuclear The point of contrast is shape, not number of nuclei. Monomorphonuciear would more
accurately describe the morphological distinction between the nongranular leukocytes and the polymorphonuclear
granulocytes.

staining affinities of their granules (neutro nucleus has a simple elongate shape. Such
cells are often described as “band forms." A
phils, eosinophils, basophils).
The number of circulating leukocytes is constriction subsequently develops, resulting
normally in the range of 5000 to 9000 per in a bilobed nucleus, and the process of
mm3 of blood. The number is subject to some elongation and constriction continues with
variation with age and even at different times time until, in older neutrophils, there may
of day in the same individual. Thus, minor be five or more segments or lobes. The pro¬
variations are of little clinical significance, but portion of band forms or young cells in the
in the presence of acute infections (appen¬ differential count is a useful index of the
dicitis, pneumonia, and so forth) the white rate of entry of new neutrophils into the
blood count may rise to 20,000 or even 40,000 circulation. The normal life span of these
cells is about eight days, but a major part of
per mm3.
The relative proportions of the various this is spent in reserve in the bone marrow.
types of leukocytes are normally fairly con¬ The variability of nuclear shape is the basis
stant: neutrophils, 55 to 60 per cent; eosino¬ for the other name applied to this cell type
phils, 1 to 3 per cent; basophils, 0 to 0.7 per polymorphonuclear leukocyte. In clinical par¬
cent; lymphocytes, 25 to 33 per cent; mono¬ lance this is often abbreviated so that neutro¬
cytes, 3 to 7 per cent. Because different phils are also referred to as “polys.”
disease processes may affect the numbers of The nuclear chromatin occurs in deeply
one cell type more than others, the differential staining clumps, and a nucleolus cannot be
leukocyte count is diagnostically valuable. identified. Since these cells are fully differ¬
entiated, with no synthetic capacity, they no
longer need a nucleolus for assembly of ri-
NEUTROPHILIC LEUKOCYTES bosomal RNA. In a small proportion of the
(NEUTROPHILS) neutrophils of women, the chromatin repre¬
senting the condensed X chromosomes forms
Neutrophils are the most abundant of the a minute separate lobule—often described as
leukocytes, constituting 55 to 65 per cent of the “drumstick” because of its characteristic
the total count. In absolute numbers, there shape (Fig. 4-9). Thus, it is possible to deter¬
are 3000 to 6000 per mm3 or 20 to 30 billion mine from a blood smear the genetic sex of
in the circulation at any one time. They are the individual by examining a large number
7 pm in diameter in the circulating blood of neutrophils for the presence of this nu¬
and 10 to 12 |xm in diameter in dry smears, clear appendage.
and are easily recognized by their highly The cytoplasm of the neutrophil, when
characteristic nucleus consisting of two or properly stained, is stippled with very small
more lobules connected by narrow strands granules that have little affinity for the dyes.
(Pig 4_7A). The number of nuclear lobes These are the so-called specific granules (Fig.
depends in part on the age of the cell. When 4-7A, D). In addition to these, there are
thevJ are first released into the blood, the larger, reddish-purple azurophil granules. The
 BLOOD • 121

A B
Figure 4-9. A, Erythrocytes and a neutrophilic leukocyte from a woman showing the “drumstick” appendage
characteristic of the female. B, A comparable field from the blood smear of a man. (Photomicrograph courtesy of M.
Barr.)

granules are often quite inconspicuous in granules contain histochemically demonstra¬

routinely stained smears and they are there¬ ble acid phosphatase, (3-glucuronidase, and a
fore studied to better advantage in living number of other hydrolytic enzymes for
leukocytes viewed by phase-contrast micros¬ which no staining method exists. They are
copy or in electron micrographs. In the therefore regarded as primary lysosomes.
guinea pig and rabbit, the granules are more The specific granules, which greatly outnum¬
conspicuous than in the human, and are ber the azurophils, lack lysosomal enzymes
stainable with either acid or basic dyes but but contain alkaline phosphatase and a vari¬
show a predilection for eosin. Therefore, in ety of poorly characterized basic proteins,
these species these cells are sometimes called called phagocytins, which have significant an¬
pseudoeosinophils. tibacterial activity.
In electron micrographs of human leuko¬ Neutrophils are in the first line of defense
cytes the neutrophil granules may be found of the body against invasion by pathogenic
almost anywhere in the cell, but they tend to bacteria. At sites of inflammation, they ad¬
be absent from a thin peripheral zone of here to the walls of postcapillary venules and
cytoplasm, which is rich in fine filaments that their ameboid motility enables them to insin¬
seem to be concerned with cell motility (Fig. uate themselves between the endothelial cells
4—10). Centrally situated in the cell adjacent and into the connective tissues to attack bac¬
to the nucleus is a small Golgi complex and teria. The neutrophils are avidly phagocytic;
a pair of centrioles. The specific granules are i.e., they have the capacity to extend pseu¬
round or elongate like rice grains. In some dopods around bacteria, take them into their
species the azurophil granules are more cytoplasm in membrane-bounded vacuoles
spherical and distinctly larger, but in neutro¬ (Fig. 4—14), and destroy them with hydrolytic
phils of humans it is difficult to distinguish enzymes. In the presence of bacterial infec¬
the granule types on morphological criteria tion, a message is somehow transmitted to
alone. However, the enzyme myeloperoxi¬ the bone marrow that stimulates increased
dase is localized exclusively in the azurophil production and release of neutrophils. Thus,
granules (Fig, 4—11) and can be used as a the number of circulating polymorphonu¬
cytochemical marker to identify this granule clear leukocytes increases and the percentage
type. In addition to peroxidase, the azurophil of young band forms is elevated.
 Lobes of
nucleus

Ectoplasmic
layer

Specific
granules

Fiqur#1 4-10 Electron micrograph of a guinea pig polymorphonuclear leukocyte. The nucleus has several lobes that
appear separate in this plane of section. The cytoplasm is filled with specific granules of varying shape. A thin
ectoplasmic zone of cytoplasm rich in actin is important in pseudopod formation and ameboid locomotion 01 the cell.

Figure 4-11. The azurophil and specific granules of neutrophils are not always easily distinguished in routine electron
micrographs, but in this neutrophil stained by the cytochemical reaction for peroxidase, only the azurophil granules
(primary lysosomes) are stained. (From Bainton, D., J. Ullyot, and M. Farquhar. J. Exp. Med. 134:907, 1971.)

122
 BLOOD • 123

Although neutrophils are able to take in The neutrophil responds to attachment of

other types of particles by nonspecific phago¬ the particle by local invagination and by ex¬
cytosis, they are especially effective against tension of pseudopods (Fig. 4-12). As the
bacteria. Their efficiency in this defensive enveloping pseudopods bring more of the
role is enhanced if the body has developed cell surface into contact with the surface of
specific antibodies as a result of previous the bacterium, the ligand-receptor binding
exposure to the same type of bacteria. Under spreads laterally from the point of initial
these circumstances, blood-borne antibody contact, tightly zippering up the cell mem¬
(IgG) binds to the surface antigen of the brane to the surface of the bacterium around
bacterium and a derivative of the comple¬ its entire circumference, so that it becomes
ment system of the plasma (C3b) binds to the enclosed in a deep recess in the leukocyte
antigen-antibody complex. IgG and C3b act surface. When the enveloping cell processes
as ligands binding the bacterium to specific meet beyond the bacterium, their membranes
receptors for these molecules present in the fuse. The invaginated membrane and its en¬
plasma membrane of neutrophils. Attach¬ closed bacterium separate from the plasma-
ment and interiorization of the bacterium is lemma and move into the cytoplasm as a
thereby facilitated. Plasma components such phagocytic vacuole (Fig. 4-13). The membrane
as IgG and C3b that coat bacteria are collec¬ of one or more lysosomal granules then co¬
tively called opsonins. Phagocytosis of opson¬ alesces with the membrane of the vacuole,
ized particles is described as immune-phagocy¬ discharging into it their hydrolytic enzymes
tosis to distinguish it from nonspecific uptake (Fig. 4—14) to digest the bacterium.
of particles that have not been coated with Another mechanism associated with phag¬
ligands. ocytosis by neutrophils is the generation of

Azurophil
granules

Actin-rich
ectoplasmic
zone

^ ^ Substrate

Figure 4-12. The highly motile, avidly phagocytic neutrophils accumulating at sites of inflammation are the first line of
defense against invading microorganisms. Shown here is a neutrophil beginning to phagocytize a yeast in vitro. Notice
the thickening of the ectoplasmic zone (double arrows) and extension of lamellipodia in the region of contact with the
organism. (Micrograph courtesy of D. Bainton.)
 Ingested bacteria

Nucleus

Glycogen

Specific
granules phagosome

Figure 4-13 Micrograph of a neutrophil that has ingested several bacteria. The bacteria in the large phagocytic vacuole
show signs of inciptent degeneration resulting from this bacteriostatic and hydrolytic environment. (Micrograph courtesy
of D. Bainton.)

Ingested bacterium

Phagocytic vacuole

Glycogen

Specific
granule

Azurophil
granule

Figure 4-14. Portion of a neutrophil that has recently phagocytized a bacterium. Arrows (★) show the continuity of the
membrane of the phagosome with that of a granule about to discharge its contents into the phagosome. (Micrograph
from D. Bainton. In Williams, R. C. and H. H. Fudenberg, eds. Phagocytic Mechanisms in Health and Disease. New
York, Intercontinental Medical Book Corporation, 1972, p. 130.)

124
 BLOOD • 125

superoxide anions by an oxidase present on tissues. They are 9 pm in diameter in sus¬

their surface membrane. Some of the super¬ pension and about 12 pm when flattened out
oxide is converted to hydrogen peroxide and in dried blood smears. Eosinophils are easily
both are carried into the phagocytosis vacu¬ recognized by their relatively coarse specific
oles. There, in the presence of myeloperoxi¬ granules, which stain pink with Wright’s
dase contributed by coalescing lysosomes, hy¬ blood stain (Fig. 4—7B, E). The nucleus is less
drogen peroxide reacts with chloride ions to segmented than that of neutrophils and usu¬
form hypochlorite—a potent bactericide. ally appears bilobed. In rats and mice, the
Much has been learned about the role of nucleus has a ring form. The chromatin
the lysosomal azurophil granules, but much pattern is less coarse than it is in neutrophils.
less is known about how the specific granules There is a granule-free cell center around a
exert their antibacterial effect. However, it is pair of centrioles, a small Golgi apparatus,
clear that these cells have evolved multiple and a few mitochondria. The endoplasmic
complementary bactericidal strategies. The reticulum is not extensive.
pus that accumulates in boils and abscesses The most conspicuous feature of this cell
consists of millions of dead and dying neu¬ is its specific granules. These display rather
trophilic leukocytes that have carried out striking interspecific variations in their ultra¬
their mission. structure. In laboratory rodents, each gran¬
The azurophil and specific granules of the ule contains a single, equatorial, discoid crys¬
polymorphonuclear leukocytes function pri¬ tal (Fig. 4—15). In humans the crystals may
marily in the intracellular destruction of for¬ be single or multiple and quite variable in
eign invaders. These cells are not considered form. In cats, eosinophil granules have a
to be secretory in the usual sense. However, dense cylindrical inclusion with a concentric
under certain circumstances the hydrolytic lamellar substructure. In all species, the crys¬
enzymes that are normally confined within talline inclusions are embedded in an amor¬
the membranes of granules or within phago¬ phous or finely granular matrix and enclosed
cytosis vacuoles may escape into the sur¬ bv a membrane.
rounding tissue. Fusion of lysosomes may Upon isolation and analysis, eosinophil
occur before closure of the phagocytosis vac¬ granules contain several of the lysosomal en¬
uole, resulting in leakage of degradative en¬ zymes but are unusually rich in peroxidase.
zymes into the extracellular space. This is Fysozyme and phagocytin, the antibacterial
especially apt to occur if the ratio of particles agents present in the specific granules of
to phagocytes is high or when the foreign neutrophils, are lacking in eosinophil gran¬
particle size is too large to be completely ules. In addition to the specific granules,
ingested. Among the enzymes escaping are eosinophils contain a few azurophil granules
collagenase and elastase, which can break and thus have two classes of primary lyso¬
down collagenous and elastic fibers, and somes. The eosinophil granules also contain
other proteases that may attack cellular com¬ a protein of 11,000 M.W. rich in the amino
ponents of the connective tissue. Thus, the acid arginine. This major basic protein (MBP)
neutrophils may damage, to some extent, the present in the crystalloids is thought to be
tissues they are defending. Such effects may responsible for the strong affinity of eosino¬
contribute to the pain and swelling at sites of phil granules for acid dyes. The granules also
inflammation. contain myeloperoxidase, arylsulfatase, (T
glucuronidase, acid phosphatase, ribonu-
clease, and cathepsin.
EOSINOPHILIC LEUKOCYTES The precise role of eosinophils in the
(EOSINOPHILS) body’s defense mechanisms is not well de¬
fined. They are greatly increased in number
Eosinophils arise from precursors in the in parasitic diseases and various forms of
bone marrow. After three or four days of allergy. Repeated injections of foreign pro¬
maturation the eosinophil is released and tein are attended by local and general in¬
probably spends only three or four hours in creases in eosinophils mobilized from large
the bloodstream en route to the connective reserves in the bone marrow. They do not
tissues, where it remains for the remainder normally phagocytize bacteria, but they do
of its eight- to 12-day life span. Eosinophils selectively ingest and destroy antigen-anti¬
represent 1 to 3 per cent of the total popu¬ body complexes. Eosinophils appear to be
lation of blood leukocytes, but for every one attracted to sites where basophils and mast
in the blood there are about 300 in the cells abound. They are believed to respond
126 • BLOOD

Figure 4-15. Micrograph of a guinea pig eosinophilic leukocyte. The specific granules contain one or more dense
polyhedral crystals.
cent of the total leukocyte count. They are
to eosinophil chemotactic factors released by ba¬
slightly smaller than neutrophils, measuring
sophils and mast cells. Just how they interact
about 10 pm in diameter in stained blood
with these cells is not known but they may
smears. The elongated nucleus is often bent
counteract the inflammatory effects pro¬
into a U or J shape and may appear bilobed,
duced by the histamine and other mediators
but segmentation is less obvious than in other
that these cells secrete. Although eosinophils
granulocytes. The specific granules are
are found in connective tissue throughout
larger, fewer, and less closely packed than
the body, they are especially numerous be¬
those of eosinophils (Fig. 4-7C). They are
neath the epithelia of the alimentary and
metachromatic, staining purple with tolui-
respiratory tracts where penetration of for¬
dine blue or alcoholic thionine. They are
eign antigens is likely to occur.
The factors regulating release of eosino¬ water soluble in the human and with aqueous
stains may be distorted and partially dis¬
phils from the marrow are poorly understood
but there is evidence suggesting that a dif¬ solved, but in dried smears, or in tissue sec¬
fusible substance formed at sites of antigen- tions after alcoholic fixation, they are usually
antibody interaction in inflamed tissue is car¬ preserved. Their properties vary in different
ried back to the marrow, causing discharge species. The large oval granules of guinea
of more eosinophils into the circulation. In¬ pig basophils are insoluble in water but stain
jections of adrenocortical hormone or hydro¬ only faintly. In the dog, the granules are
cortisone result in a marked reduction in quite small and are assembled in a compact
circulating eosinophils. It is believed that group. Basophilic leukocytes seem to be ab¬
these hormones exert this effect by interfer¬ sent from the circulation in cat, rat, and
ing with the mobilization of eosinophil re¬ mouse.
In electron micrographs, basophils have a
serves from the marrow.
small Golgi apparatus, a few mitochondria,
and occasional strands of endoplasmic retic¬
BASOPHILIC LEUKOCYTES ulum. The cytoplasmic matrix contains vary¬
(BASOPHILS) ing amounts of glycogen. The specific gran¬
Basophils are the least numerous of the ules are round, oval, or angular and about
granulocytes, accounting for only 0.5 pei 1.2 pm in diameter (Fig. 4—16). They have a
 BLOOD • 127

substructure composed of 12- to 26-nm dense although these'two cell types have different
particles embedded in a less dense matrix. In origins, their functions are very similar. The
some other mammalian species, the dense tissue mast cells are relatively sessile whereas
subunits of the basophil granules are ar¬ the circulating basophils can be rapidly mo¬
ranged in a highly ordered lattice. Basophil bilized at sites where they are needed to
granules are peroxidase positive and contain supplement the resident population of mast
histamine and the sulfated mucopolysacchar¬ cells.
ide heparin, which is responsible for their An important property of basophils and
metachromasia. There is no evidence at pres¬ mast cells is the presence in their surface
ent that they contain lysosomal enzymes. membrane of specific receptors for immu¬
The basophils of the blood have a number noglobulin E (IgE). IgE is a minor compo¬
of properties in common with the mast cells nent of normal blood serum but may be
of connective tissues. Both have metachro- increased up to 20-fold in persons suffering
matic granules containing histamine and hep¬ from hay fever, asthma, or allergic dermati¬
arin. Both are degranulated by histamine- tis. It is the reaginic antibody (i.e., antibody
liberating drugs and when exposed to anti¬ against allergens) that results in immediate
gen in sensitized individuals. Many investi¬ hypersensitivity, as opposed to delayed, cell-
gators formerly considered the basophil to mediated hypersensitivity (see Chapter 13). Hu¬
be a circulating form of the mast cell, but it man basophils carry from 10,000 to 40,000
is now generally agreed that they are from molecules of IgE bound to their surface re¬
distinct cell lines. The basophils arise in the ceptors. When an allergen is introduced into
bone marrow; the mast cells probably origi¬ the tissues, it combines with IgE on the ba¬
nate from precursors in the connective tissue, sophils and mast cells. This triggers their
but this is debated. The basophil is much rapid degranulation and release of histamine
smaller and has a polymorphous nucleus and and other mediators. The histamine released
fewer granules. It is the current view that is in part responsible for itching and in-

Figure 4-16. Micrograph of a basophil leukocyte. The large granules are somewhat irregular in outline and vary in their
density.
128 • BLOOD

The lymphocytes are the cells principally

creased permeability of postcapillary venules
involved in the immune responses of the
that results in tissue edema and local swelling.
body, and their functions will be discussed in
Basophils that have migrated from the
greater detail in a later section on the im¬
blood into the dermis and subcutaneous con¬
mune system (Chapter 13). It suffices here
nective tissue are now recognized as major
to describe briefly what those responses are.
participants in the form of cell-mediated im¬
When foreign substances in the environment,
munity called cutaneous basophil hypevsensitivity.
usually proteins or polysaccharides, get into
Although in the blood there are only 30 to
the body, they stimulate formation of immune
40 basophils per mm3, they are found in
globulins that circulate in the blood plasma
concentrations of 300 to 600 per linear mil¬
and are capable of combining with and neu¬
limeter in sections of skin from previously
sensitized guinea pigs reacting to a challeng¬ tralizing the harmful effects of the inducing
ing dose of parenterally administered anti¬ substance. An animal that has become pro¬
tected in this way is said to have developed
gen.
an immunity. The substance that induces an
immune response is called an antigen and the
specific globulin molecules that appear in the
LYMPHOCYTES
blood to counteract it are called antibodies.
When an antigen is introduced into the
Lymphocytes are the second most numerous
class of leukocytes in the blood, constituting body for the first time, there is a conditioning
of the antibody-producing cells or their pre¬
20 to 35 per cent of the circulating white
blood cells. In blood smears, they are small cursors that extends over a number of weeks
spherical cells with an intensely staining, and may result in low levels of circulating
slightly indented nucleus, and a thin rim of antibody. The events taking place in this
clear blue cytoplasm (Fig. 4-7G, H). They latent period are described as the primary
are 7 to 9 \xm in diameter, which is only response. If a second exposure to the same
slightly larger than the erythrocytes. They antigen occurs weeks or months afterward,
contain no specific granules. In electron mi¬ there is a dramatic and rapid rise in synthesis
crographs they have a small Golgi complex, of antibody globulin, resulting in a titer of
a pair of centrioles, and a few mitochondria. circulating antibody 10 to 100 times the pre¬
Endoplasmic reticulum is usually lacking but vious level. This is called the secondary re¬
sponse. If a number of injections of antigen
the cytoplasm is rich in ribosomes. Occasion¬
ally observed are small, dense lysosomes that are given, the elevated level of circulating
correspond to the azurophil granules re¬ antibody may be maintained for years. This
component of the bodily defenses, which
ported by hematologists.
Histologists traditionally categorized lym¬ depends on blood-borne antibodies, is called
phocytes as large, medium, and small on the the humoral immune response. The lymphocytes
basis of their diameter and relative amount are the cells that recognize an antigen as
of cytoplasm. These were thought to repre¬ foreign to the body and respond to an initial
sent successive stages in their evolution from encounter by undergoing certain changes
a larger precursor, the lymphoblast. The that may have no effect upon their appear¬
small lymphocytes were long considered to ance but which endow them with a specific
be an end stage capable of surviving only a “memory” for that antigen, which conditions
few days, after which they were believed to their behavior when exposed at a later time
degenerate or to be eliminated from the body to the same antigen.
by migration into the lumen of the intestine. If lymphocytes are placed in tissue culture
When methods were developed for radiola¬ and stimulated by phytohemagglutinin or
beling cells, following their migrations and other mitogens, they enlarge, acquire in¬
measuring their longevity, the traditional creased numbers of ribosomes, and take on
interpretation of lymphocytes proved to be the appearance of large lymphoblasts (Fig.
erroneous. It is now known that there are 4—18). After this initial period of hypertro¬
two functionally distinct categories of small phy, they begin to divide. This unexplained
lymphocytes designated B lymphocytes and T stimulation by phytohemagglutinin is of in¬
lymphocytes. These differ in their develop¬ terest because it duplicates in vitro the nor¬
mental background, life span, and functions. mal response of lymphocytes upon exposure
They are not distinguishable morphologi¬ to antigens in vivo. The repeated division of
cally, but distinctive surface markers can be lymphoblastic cells gives rise to clones of
detected by immunofluorescence. “memory cells,” each of which expresses on
 BLOOD • 129

hi lie granules

Mitochondria Centriole

Figure 4-17. Micrograph of a guinea pig lymphocyte. The nucleus is indented and has a coarse pattern of
heterochromatin. A centriole and several mitochondria are seen in the cytoplasm. Although it is a nongranular leukocyte,
the lymphocyte may contain a few small azurophil granules.

its surface specific combining sites or recep¬ regulation of antibody production. They
tors for the inducing antigen. Some of these have the capacity to seek out antigenic for¬
lymphocytes go on to differentiate into plasma eign cells and to interact with macrophages
cells, which synthesize specific antibody so as to promote destruction of the foreign
against that antigen. The proliferative re¬ cells by cytotoxic and phagocytic mechanisms.
sponse of lymphocytes in response to antigen Protective reactions of this kind are distin¬
thus serves to amplify the production of guished from the humoral immune response
specific antibody. Although we refer to B by the term cell-mediated immunity.
lymphocytes as a single class, there are, in an The mechanisms of the interaction of lym¬
immunological sense, as many kinds of mem¬ phocytes and macrophages in immunological
ory lymphocytes as the number of different responses are still poorly understood. In in
antigens that have, at one time or another, vitro studies, T-cell functions, such as anti¬
invaded the body. gen-dependent proliferation and develop¬
The T and B lymphocytes do not function ment of “helper” capacity, have been shown
independently in the immune response. For to require a contribution from macrophages.
many antigens, subpopulations of T cells are Conversely, several macrophage functions
required to provide an additional stimulus to are modified by interaction with immune T
the B cells for the production of antibody to cells and appropriate antigen. The biologi¬
occur. These are designated “helper” T cells. cally active molecules produced by lympho¬
Other subpopulations of T cells are capable cytes in their immunoregulatory functions
of depressing production of antibody by B are called lymphokines. An increasing number
cells. These are called “suppressor” T cells. The of these are being identified. One that has
mechanism of T-cell regulation of B-cell attracted intense interest is interferon, which
function is unclear. It is not evident whether has the capacity to inhibit replication of cer¬
direct contact between the interacting cells is tain viruses and suppress the multiplication
required in vivo or whether diffusible prod¬ of some intracellular parasites. It also en¬
ucts of the T cells are involved. hances several functions of macrophages in¬
T lymphocytes participate in a number of cluding phagocytosis. In addition, interferon
immunological processes in addition to the is reported to inhibit proliferation of normal
Fiaure 4-18. A Bovine lymphocytes freshly isolated from peripheral blood and placed in culture medium containing a
lectin concanavalin A. B, At the same magnification, after 72 hours in culture many of the lymphocytes have transformed
into metabolically active large lymphoblasts with prominent nucleoli and little heterochromatin. This in vitro transformation
mimics that occurring in vivo when lymphocytes are exposed to antigen.
 BLOOD • 131

and malignant cells in vitro, and its potential should have been maintained. Thus, it was
for suppressing the growth of cancers is now shown that the lymphocytes entering the
being explored. blood from the thoracic duct are not all newly
The lymphocytes were formerly believed formed in the nodes, as had been thought.
to arise almost exclusively in the lymph nodes A substantial proportion enter the nodes
and other lymphoid organs, while other leu¬ from the blood, remain for a while, and then
kocytes were formed in the bone marrow. leave via the lymph. When tritiated thymi¬
The principal basis for this assumption was dine is infused into the afferent lymphatic to
the observation that if the thoracic duct, a node, it is found that fewer than 5 per cent
which drains lymph and lymphocytes from of the emerging cells are labeled. The vast
the lymph nodes back to the bloodstream, majority have traversed the node from the
was severed and exteriorized, the number of blood.
lymphocytes in the blood rapidly fell. Since It is now universally accepted that lympho¬
the number entering the blood daily from cytes are continually migrating between
this source was very large, it seemed reason¬ lymph nodes, spleen, and connective tissues,
able to infer that their life span must be very using the blood as a common path. The
short, for if a comparable number were not lymphocytes found in the blood, despite their
leaving the blood daily, the lymphocyte count uniform appearance, consist of different
would rise rapidly to astronomical numbers. populations. The majority are part of the
Neither of the conclusions drawn from these recirculating pool of long-lived (months to
observations has proved to be correct. years) lymphocytes, capable of participating
It is now known that large numbers of B in cell-mediated immunity. A smaller pro¬
lymphocytes are generated in the bone mar¬ portion are short-lived (weeks or months) but
row. Approximately one quarter of all the are capable of transformation to antibody-
nucleated marrow cells are lymphocytes. Al¬ producing cells on antigenic stimulation (see
though some of these no doubt belong to the Chapter 13).
pool of recirculating long-lived B and T lym¬
phocytes, many of them arise from stem cells
in the marrow, and continuously emigrate to MONOCYTES
the peripheral lymphoid organs as virgin B
lymphocytes capable of mediating primary The typical monocyte measures 9 to 12 pm
humoral immune responses. Primary B lym¬ in diameter, but in blood smears, where
phocytes are constantly turning over in the monocytes are greatly flattened in the drying
peripheral lymphoid tissues. Labeled young process, they may be up to 17 pm in diame¬
lymphocytes persist in spleen and lymph ter. They constitute 3 to 8 per cent of the
nodes for only about a week. The large-scale leukocytes of the circulating blood. Their
genesis and emigration of B lymphocytes in enumeration is subject to some error because
the marrow suggests that the population of they cannot always be sharply differentiated
rapidly renewing primary B lymphocytes in from large lymphocytes. The cytoplasm is
the spleen is maintained almost exclusively more abundant than in lymphocytes, and
by the continuous influx of cells from the instead of a clear, pale blue, it tends to have
marrow. Most of these studies have been a grayish-blue tint with scattered small azur¬
carried out on mice but it seems likely that ophil granules (Fig. 4-7 J, K, L). In older
the same events occur in humans, although monocytes, the nucleus is eccentric in posi¬
the time course may differ. tion and oval or reniform. Its chromatin is
The first indication that the lymphocytes more finely granular and more uniformly
in lymphoid organs were transient residents dispersed in the nucleoplasm, so that the
arising elsewhere came from experiments nucleus as a whole appears less intensely
that antedated application of radiolabeling stained. One or more nucleoli are present
with tritiated thymidine. It was observed that but are seldom seen in routine blood smears.
a thoracic duct fistula resulted not only in a In electron micrographs the nucleoli are
decrease in the number of blood lympho¬ obvious, and the cytoplasm contains a con¬
cytes, but also in a gradual diminution in the spicuous Golgi complex, a few cisternal pro¬
number of lymphocytes emerging from the files of rough endoplasmic reticulum, a mod¬
duct. If this outflow depended solely on new erate number of free ribosomes, and some
formation of lymphocytes in the lymph glycogen granules. In addition, there are in
nodes, the output from the thoracic duct each section 15 to 20 granules with a dense
132 • BLOOD

homogeneous content (Fig. 4—19). These cor¬ tissues. They seem to perform no function
respond to the azurophil granules seen in while in the blood but survive for months in
stained smears. They exhibit cytochemical the tissues, where they are a mobile reserve
staining reactions for acid phosphatase, ar- of scavengers that play a valuable defensive
ylsulfatase, and peroxidase, and hence are role by phagocytosis and intracellular diges¬
considered to be primary lysosomes. tion of invading microorganisms. They may
The origin of monocytes and their relation also be essential for the processing of anti¬
to lymphocytes were long obscured by the gens prior to the development of antibodies
contradictory ■ theories of blood formation. by immunocompetent lymphoid cells associ¬
More recently, experiments involving radio¬ ated with them in the tissues.
labeling of cells of the marrow and lymphoid
organs and their transfusion into x-irradiated
host animals have now resolved much of this
OTHER COMPONENTS
controversy. It has been convincingly dem¬
onstrated that monocytes originate in the
OF BLOOD
bone marrow from precursors called promon¬
ocytes. After a developmental period of one Since blood is a tissue composed of cells
to three days they enter the circulation. suspended in a fluid extracellular matrix, the
Those found in the peripheral blood repre¬ blood plasma, it may be appropriate to com¬
sent cells in transit from the marrow to their ment briefly on some of the more important
ultimate destination in the tissues. They constituents of this matrix. There are three
spend only about a day and a half in the major types of plasma proteins—albumin,
blood, then migrate into the connective tissue globulin, and fibrinogen.
of various organs throughout the body, Albumin is the most abundant and smallest
where they differentiate into tissue macro¬ of the plasma proteins with M.W. of about
phages. They are capable of mitotic division 59,000. It is synthesized by the liver and its
and of continued enzyme synthesis in the principal function is to maintain the colloid

Figure 4-19. Micrograph of a typical rabbit monocyte showing its irregularly shaped nucleus, occasional cisternal
profiles of rough endoplasmic reticulum, and a small cluster of azurophil granules (lysosomes) near the Golgi complex.
(Micrograph courtesy of D. Bainton.)
 BLOOD • 133

Marginated
leukocyte

Eiastica
interna

Figure 4-20. At one time or another, a large number of leukocytes in the blood are not circulating but are temporarily
adherent to the endothelium of small blood vessels as shown here. This population is referred to as the “marginated
pool” of leukocytes.

osmotic pressure within the blood capillaries, port protein in the plasma. In liver disease
which prevents excessive loss of fluid to the or pernicious anemia, the concentration of
tissues. In addition, a variety of substances transferrin is reduced. A protein of rather
that are relatively insoluble in water dissolve similar function, ceruloplasmin, contains
more readily in the presence of blood pro¬ nearly all the copper of the blood and is
teins. Plasma albumin plays an important believed to regulate utilization of copper by
part in the transport of these metabolic prod¬ reversibly binding and releasing it at various
ucts. sites in the body. In the rare inherited dis¬
The globulins include proteins of a wide order of copper metabolism, Wilson’s disease,
range of M.W., 80,000 to several million. ceruloplasmin is markedly reduced.
They are divided into several fractions. The Serum lipoproteins, globulins involved in
greatest interest is centered on the gamma lipid transport, attain such a large size that
globulins because this fraction includes the they can be visualized electron microscopi¬
immune globulins or antibodies, which are the cally as spherical particulates of varying size.
basis of the immunological defenses of the They can therefore be included among the
body against bacteria, toxins, and other for¬ microscopic formed elements of the blood.
eign proteins. These important globulins are These serum lipoproteins can be separated
synthesized in the cells of the lymphoid or¬ from plasma by physical techniques of ultra¬
gans. The beta globulins function in the trans¬ centrifugal flotation or electrophoresis, and
port of hormones, metal ions, and lipid. are divided into four major groups on the
The beta globulin called transferrin com¬ basis of size. (1) The chylomicrons, ranging in
bines with iron, copper, and zinc. Its princi¬ size from 100 nm to 500 nm are the largest
pal function is to transport iron. In condi¬ and are detectable by dark-field light micros¬
tions in which there is increased need for copy. (2) The very-low-density lipoproteins
iron for hemoglobin synthesis, as in nutri¬ (VLDL) generally fall within a size range of
tional deficiency of iron or in pregnancy, 25 to 70 nm and are not visible by light
there is a compensatory increase in this trans¬ microscopy. (3) The low-density or beta lipopro-
134 • BLOOD

terns (LDL) are considerably smaller. In ad¬ cytoskeletal proteins on human erythrocyte mem¬
brane. Cell 24: 24, 1981.
dition to their characteristic sizes, the lipo¬
Cohen, C. M.: The molecular organization of the red¬
proteins are distinguishable biochemically by cell membrane skeleton. Sem. Hematol. 2d: 141,
their differing proportions of various classes 1983.
of lipids associated with specific apoproteins. Harrison, P. R.: Analysis of erythropoiesis at the molec¬
In the nutrition of mammals, the lipopro¬ ular level. Nature 262:353, 1976.
Ingram, V.: The Hemoglobins in Genetics and Evolu¬
teins are the form in which lipids absorbed tion. New York, Columbia University Press, 1963.
in the gastrointestinal tract are transported Patek, J., and S. E. Lux: Red Cell membrane skeletal
in the blood to the liver. The chylomicrons defects in hereditary and acquired hemolytic ane¬
formed in the absorptive epithelium of the mias. Sem. Hematol. 2d: 189, 1983.
Perutz, M. F.: Hemoglobin structure and respiratory
intestine are transiently increased in the
transport. Sci. Am. 239:92, 1978.
blood after a fat-rich meal. The VLDL are
the form in which lipid is transported from PLATELETS
the liver to the adipose tissue. Biggs, R., and R. G. Macfarlane: Human Blood Coagu¬
Other classes of lipoproteins are persis¬ lation and Its Disorders. 3rd ed. Philadelphia, F. A.
Davis Co., 1962.
tently elevated in certain disease conditions. Fox, J. E. B., and D. R. Phillips: Polymerization and
The continued presence of high circulating organization of actin filaments within platelets. Sem.
levels of lipoprotein is thought to predispose Hematol. 2d:243, 1983.
to atherosclerosis, or hardening of the arter¬ Johnson, S. A., R. W. Monto, J. W. Rebuck, and R. C.
Horn, Jr., eds.: Blood Platelets (A Symposium).
ies. The normal lipoproteins are spherical
Boston, Little, Brown & Co., 1961.
droplets or particles but, interestingly Marcus, A. J., and M. B. Zucker: The Physiology of
enough, in diseases involving obstruction of Blood Platelets. New York, Grune & Stratton, 1965.
the bile duct, abnormal lipoproteins, which Shattil, S. J., and J. S. Bennett: Platelets and their
have an unusual discoid shape, may appear membranes in hemostasis: physiology and patho¬
physiology. Ann. Intern. Med. 94:108, 1981.
in the blood. These easily associate in rou¬
Weiss, H. J.: Platelet physiology and abnormalities of
leaux under the usual conditions of isolation platelet function. N. Engl. J. Med. 203:531, 580,
and negative staining. 1975.
Plasma clotting factors are the major de¬ White, J. G., and C. C. Clauson: Biostructure of platelets.
Ultrastruct. Pathol. 7:533, 1980.
fense of the body against serious blood loss.
An essential component of this mechanism is NEUTROPHILS
fibrinogen, a protein synthesized in the liver Anderson, D. R.: Ultrastructure of normal and leukemic
and circulating in the blood plasma as long, leukocytes in human peripheral blood. J. Ultrastr.
asymmetrical molecules of 330,000 M.W. Res. (Suppl.):5, 1966.
Athens, J. W.: Granulocyte kinetics in health and disease.
When prothrombin, also present in plasma, is
Natl. Cancer Inst. Monogr. 30:135, 1969.
activated by thromboplastin of tissue or platelet Bainton, D. F.: Primary lysosomes of blood leukocytes.
origin, enzymatically active thrombin is pro¬ In Dingle, J. T., and R. T. Dean, eds.: Lysosomes
duced, resulting in polymerization of fibrin in Biology and Pathology. Amsterdam, North Hol¬
to form the fibrous meshwork of a clot. land Publishing Co., 1976.
Cohn, Z. A., and S. I. Morse: Functional and metabolic
Various hemorrhagic disorders may result properties of polymorphonuclear leukocytes. I. Ob¬
from impaired synthesis of fibrinogen or de¬ servations on the requirements and consequences
ficiency of prothrombin. of particle ingestion. J. Exp. Med. 777:667, 1960.
Daems, W.: On the fine structure of human neutrophilic
leukocyte granules. J. Ultrastruct. Res. 24:343, 1968.
Hirsch, J. G., and Z. A. Cohn: Degranulation of poly¬
REFERENCES morphonuclear leukocytes following phagocytosis of
microorganisms. J. Exp. Med. 775:1005, 1960.
Klebanoff, S. J.: Antimicrobial mechanisms in neutro¬
ERYTHROCYTES philic polymorphonuclear leukocytes. Sem. Hema¬
Bessis, M., and J. P. Thiery: Les cellules du sang vues tol. 72:117, 1975.
au microscope a interferences (systeme Nomarski). Lisiewicz, J.: Human Neutrophils. Bowie, MD, Charles
Rev. d’Hematol. 72:518, 1957. Press Publishers, 1980.
Bennett, V., and D. Branton: Selective association of Murphy, P.: The Neutrophil. New York, Plenum Pub¬
spectrin with the cytoplasmic surface of human lishing Co., 1976.
erythrocyte plasma membranes. J. Biol. Chem. Spitznagel, J. K., F. G. Dalldorf, and M. S. Liffell:
252:2753, 1977. Characterization of azurophil and specific granules
Bennett, V., and P. J. Stenbuck: Human erythrocyte purified from human polymorphonuclear leuko¬
ankyrin. Purification and properties. J. Biol. Chem. cytes. Lab. Invest. 30:774, 1978.
255:2540, 1980. Weissmann, G., R. B. Zurier, P. J. Spieler, and I. M.
Bishop, C., and D. M. Surgenor, eds.: The Red Blood Goldstein: Mechanisms of lysosomal enzyme release
Cells. New York, Academic Press, 1964. from leukocytes exposed to immune complexes and
Branton, D., C. M. Cohen, and J. Tyler: Interaction of other particles. J. Exp. Med. 754:149, 1971.
 BLOOD • 135
EOSINOPHILS
and cellular immunity in animals and man. Hum.
Beeson, P. B., and D. A. Bass: The Eosinophil. Phila¬ Pathol. 3:454, 1972.
delphia, W. B. Saunders Co., 1977. Ishizaka, T., and K. Ishizaka: Biology of immunoglob¬
Hudson, G.: Quantitadve study of eosinophil granulo¬ ulin E. Molecular basis of reaginic hypersensitivity.
cytes. Sem. Hematol. 5:166, 1968. Prog. Allergy 19:60, 1975.
Litt, M.: Eosinophils and antigen-antibody reactions. Terry, R. W., D. F. Bainton, and M. G. Farquhar:
Ann. N.Y. Acad. Sci. 776:964, 1964. Formation and structure of specific granules in
basophilic leukocytes of the guinea pig. Fab. Invest.
BASOPHILS 21:65, 1969.
Ackerman, G. A.: Cytochemical properties of the blood Wolf-Jiirgensen, P.: The basophilic leukocyte. Ser. Hae¬
basophilic granulocyte. Ann. N.Y. Acad. Sci matol. 7:45, 1968.
103:376, 1963. Zucker-Franklin, D.: Electron microscopic study of hu¬
Askenase, P. W.: Role of basophils, mast cells and man basophils. Blood 29:878, 1967.
vasoamines in hypersensitivity reactions with a de¬
layed time course. Prog. Allergy 23:199, 1977. LYMPHOCYTES
Dvorak, H. F., and A. M. Dvorak: Basophilic leucocytes: See References, Chapter 7 and 13.
structure, function and role in disease. Clin. Hae¬
matol. 4:651, 1975. MONOCYTES
Dvorak, H. F., and A. M. Dvorak: Basophils, mast cells, See References, Chapter 5.
 5
CONNECTIVE TISSUE
PROPER

Connective tissue consists of cells and ex¬ surrounds connective tissue cells and under¬
tracellular fibers embedded in a gel-like lies epithelia.
ground substance or matrix rich in tissue There are several cell types in connective
fluid. Traditionally the fibers have been con¬ tissue. These can be. categorized as fixed cells,
sidered to be of three kinds: collagen fibers, a more or less permanent resident popula¬
reticular fibers, and elastic fibers. The collagen¬ tion, and wandering cells, which are transient
ous and reticular fibers have now been found emigrants from the bloodstream.
to be different morphological forms of the The relative abundance of the various
same fibrous protein, collagen. Nevertheless, kinds of fibers, cells, and matrix varies greatly
it is useful for descriptive purposes to retain in connective tissue from one region of the
the term reticular fiber for bundles composed body to another, depending on the local
of fibrils of collagen 50 nm or less in diameter, structural requirements. For convenience in
and the term collagen fiber for larger bundles description and communication, histologists
consisting of unit fibrils 50 to 150 nm in have attempted to classify connective tissues.
diameter. But classification is often difficult and inexact
In recent years, there has been rapid prog¬ and should not be interpreted too rigidly,
ress in characterizing the macromolecules of since various types grade into one another
the extracellular matrix of connective tissue. through transitional forms and one type may
These have been found to occur in much be transformed into another if the local con¬
greater variety than previously assumed. ditions change.
Many of these molecules belong to a class of Categorical terms are usually assigned ac¬
compounds called proteoglycans. This term cording to whether the fibers are loosely
describes complex molecules containing a interwoven or densely packed. Thus, we dis¬
core of protein to which glycosaminoglycans tinguish loose connective tissue and dense con¬
are covalently bound. The latter in turn con¬ nective tissue. Within the latter category, it is
sist of linear polymers of repeating disac¬ useful to add modifiers to indicate whether
charides. The proteoglycans may involve dif¬ the fibers have an ordered or a disordered
ferent core proteins and several classes of arrangement. Thus, in dense irregular connec¬
glycosaminoglycans linked to the core in tive tissue, the fibers are closely interwoven in
varying numbers. This results in great diver¬ random orientation, whereas in dense regular
sity at the molecular level, which endows connective tissue, the fibers are arranged in
proteoglycans with considerable versatility in parallel bundles as in tendons; or in flat
their structural and organizational functions. sheets, as in aponeuroses. In addition to these
The time-honored concept of an “amorphous designations, several kinds of specialized con¬
ground substance” has become obsolete as nective tissue are named to indicate their
more has been learned about the configura¬ predominant component, or identifying fea¬
tion and interactions of its macromolecular ture: e.g., mucous connective tissue, reticular
constituents. The general term extracellular tissue, adipose tissue, pigment tissue, and so
matrix (ECM), now widely used, embraces the on. These examples and the lamina propria of
proteoglycans, the collagen, elastin, and the the gastrointestinal mucosa are all variants of
more recently discovered structural proteins loose connective tissue, which is a common
fibronectin, chondronectin, and laminin. Collec¬ form that can be discussed as the prototype
tively these form the stable material that of connective tissues in general.

136
 CONNECTIVE TISSUE PROPER • 137

LOOSE CONNECTIVE TISSUE matrix is the medium through which all nu¬
trients and waste products must pass in
Loose connective tissue develops from the transit between the blood and the cells of the
mesenchyme that remains after the other body. The consistency, composition, and state
tissues of the embryo have been formed. An of hydration of the matrix can exert an
interlacing fabric of very fine collagenous important influence upon this vital exchange.
fibers is deposited in the meshes of the retic¬
ulum of stellate mesenchymal cells. These Collagen Fibers
fine fibrils formed early, adsorb metallic sil¬
ver when treated with alkaline solutions of Collagen fibers are present in varying
reducible silver salts, and are blackened. Be¬ abundance in all types of connective tissues.
cause of this staining property, they were In unstained preparations of loose connective
formerly considered to be distinct from col¬ tissue, they appear as colorless strands 0.5 to
lagen and were called reticular fibers and their 20 pm in diameter and of indefinite length.
substance designated reticulin. They are now They run in all directions and if not under
known to be simply one of the several molec¬ tension they tend to have a slightly wavy
ular species of collagen. As development pro¬ course (Fig. 5—1). At high magnification, a
gresses, coarser fibers are formed that asso¬ longitudinal striation can be detected in the
ciate in bundles and have the staining larger collagen bundles, indicating that these
properties of the form of collagen that pre¬ consist of smaller parallel fibers. With polar¬
dominates in the connective tissues of the ization microscopy, even the smallest collagen
adult organism. Although “reticulin” is no fibers visible with the light microscope exhibit
longer regarded as a distinct chemical entity, form birefringence attesting to the presence
the terms reticulum and reticular fibers continue of elongated submicroscopic units oriented
to be useful to designate fibrous elements in the direction of the fiber axis. Seen in
whose size is smaller than, and arrangement stained histological sections, collagen fibers
different from, those of the more prevalent are acidophilic, staining pink with eosin, blue
collagenous fibers in the adult. with Mallory’s trichrome, and green with
As development progresses the mesenchy¬ Masson’s trichrome stain.
mal cells gradually change their character, When examination of collagen is pursued
elongating and stretching out along the sur¬ down to the electron microscopic level, the
face of the collagen fiber bundles to become smaller fibers detectable by light microscopy
fibroblasts, the principal cells of connective are found to be composed of parallel fibrils
tissue. Other cell types, either differentiating 20 to 100 nm in diameter. These are the
from mesenchymal cells or emigrating from oriented subunits responsible for the form
the blood, take up residence in the interstices birefringence observed when collagen is ex¬
of the fabric of interwoven fibers. Like a amined in polarized light. In electron micro¬
collapsed sponge, this tissue contains innu¬ graphs, these unit fibrils are cross-striated with
merable potential spaces, normally occupied transverse bands repeating every 64 to 67
by a small amount of matrix but capable of nm along their length. When stained with
becoming enlarged and distended with fluid. phosphotungstic acid and viewed at high
These extracellular interstitial spaces are the magnification, several additional bands can
minute chambers or compartments seen by be resolved within each 67-nm period (Fig.
the early histologists, who are responsible for 5-2). In addition to the typical fibrils with
the term areolar tissue (from areola, a small 67-nm cross-banding, collagen fibrils with a
space), a common synonym for loose connec¬ periodicity of about 240 nm occur rarely in
tive tissue. certain tissues.
Collagen fibers are flexible but offer great
resistance to a pulling force. The breaking
point of collagen fibers in human tendon is
EXTRACELLULAR MATRIX reached with a force of several hundred
kilograms per square centimeter, and at this
The supportive functions of the connective tension they have elongated only a few per¬
tissue depend largely on the properties of its cent of their original length. When collagen
extracellular matrix. The fibers are respon¬ is denatured by boiling, it yields the familiar
sible for its tensile strength and resilience substance gelatin. With more gentle treat¬
and form a scaffolding on which its fixed ment, it is possible to extract from rapidly
cells are deployed. The aqueous phase of the developing or repairing connective tissues
Figure 5-1. Photomicrograph of collagenous fibers in a thin spread of rat mesentery. Notice the variation in fiber
diameter and the wavy course of the larger fibers. The preparation was stained by a silver method and the photograph
printed as a negative to simulate more closely the appearance of the fibers in fresh material. (From Fawcett, D. W. in
Greep, R. O., ed.: Histology. Philadelphia, Blakeston Co., 1953. Reproduced by permission of McGraw-Hill Book Co.)

Figure 5-2. Electron micrograph of the unit fibrils of collagen showing their characteristic pattern of cross striations.
(Micrograph courtesy of D. Friend.)
138
 CONNECTIVE TISSUE PROPER • 139

several collagen fractions with different sol¬ lagen is reconstituted. When a solution of
ubility properties. Extracting at neutral pH collagen and serum a-glycoprotein is dialyzed
yields a solution containing the tropocollagen against water, the fibers that are formed have
molecules, which had not yet polymerized or a periodicity of 240 nm instead of the 67 nm
had just begun to form protofilaments. This characteristic of native collagen. This is called
is the neutral-soluble fraction. If the tissue is fibrous long-spacing (FLS) collagen (Fig. 5—5).
further extracted with citrate at pH 3, the Precipitation of collagen from acid solution
acid-soluble fraction is obtained. The remain¬ by addition of adenosine triphosphate yields
ing collagen, which can only be extracted short segments about 300 nm long instead of
with very drastic procedures, is designated fibers. This form is called segment long-spacing
the insoluble fraction. These three fractions (SLS) collagen. In both of the long-spacing
are believed to represent successive stages in forms, the molecules come together side to
the extracellular polymerization of collagen side and in register with no overlap. The
molecules to form fibers. length of the period or segment is therefore
The tropocollagen molecule is about 300 approximately the same as the length of the
nm in length and 1.4 nm in diameter, and is collagen molecule. Both of these, as well as
made up of three polypeptide chains, called the native collagen fibers, can be redissolved
a chains, each having a M.W. of approxi¬ and polymerized in either of the other forms
mately 100,000. These have a left-handed by controlling the physiochemical conditions.
helical configuration and are entwined and Knowledge of these unnatural forms of col¬
cross-linked to form a right-handed triple lagen is of limited value to students of histol¬
helix with each complete turn spanning a ogy or pathology, but it has contributed sig¬
distance of 8.6 nm (Fig. 5—3). nificantly to our understanding of the
When the cold neutral-soluble fraction of molecular organization of collagen and the
collagen is warmed to body temperature, the mechanisms involved in fiber formation.
tropocollagen molecules spontaneously poly¬ Collagen was formerly considered to be a
merize to form cross-striated fibrils with the unique protein whose size, helical structure,
67-nm periodicity of native collagen. In this and amino acid composition had been highly
process, the 300-nm molecules are oriented conserved in the course of evolution. In re¬
parallel, overlapping one another by about cent years, however, improved analytical
one quarter of their length, and leaving a methods have led to the discovery of differ¬
short empty space or lacuna between the NH2 ences in amino acid sequence in a chains of
terminal of one molecule and the COOH collagen extracted from different tissues in
terminal of the next (Fig. 5-4). The stag¬ the body. Instead of a single protein, collagen
gered array of molecular and hole zones is is now regarded as a family of closely related
responsible for the cross striation of the fibers but genetically distinct molecules. At least
seen in negatively stained preparations. The seven types of collagen have been described
accumulation of the contrast medium in the to date.
hole zones results in dark bands. The light The most ubiquitous is Type I collagen
bands where no stain penetrates are the re¬ found in abundance in skin, bone, tendon,
gions in which molecular overlapping is com¬ and cornea. It occurs in striated fibrils 20 to
plete. 100 nm in diameter that aggregate to form
Banding patterns that do not normally larger collagen fibers. Its molecular subunits
occur in nature can be produced in vitro by are made up of a chains of two kinds differ¬
varying the conditions under which the col¬ ing slightly in amino acid composition and

280 nm >-

*<-8.6 nm->

Figure 5-3. In Type I collagen each molecule (tropocollagen) is composed of two alpha-1 chains (shown here unshaded)
and one alpha-2 (shaded) polypeptide chains entwined in a helical configuration. Each gyre of the helix spans a distance
of 8.6 nm. (From Junqueira, L. C., and J. Carneiro. Basic Histology. 3rd ed. Los Altos, CA, Lanqe Medical Publications
1980.)
140 • CONNECTIVE TISSUE PROPER

Figure 5-4. Schematic presentation of the biosynthetic events and organelle participation in the formation of collagen.
On left, successive intracellular events are defined; on right, the steps leading to extracellular assembly of cross-striated
collagen fibrils. (Modified after Junqueira, L. C., and J. Carneiro. Basic Histology. 3rd ed. Los Altos, CA, Lange Medical
Publications, 1980.)
 CONNECTIVE TISSUE PROPER • 141

cT?U|re 5 *Ele*ron micro9raPh of the different forms that can be produced upon reconstitution of collagen in vitro /A
Fibrils with the 64-nm period of native collagen—precipitated from solution by dialysis against 1 per cent NaCI B
Fibrous long-spacing (FLS) collagen produced from a mixture of a, acid glycoprotein of serum and collagen solution
dlaiyTDd/f?a'nS WuGr' Se9ment long-spacing (SLS) collagen precipitated from acid solution of collagen by addition
of ATP. (Micrographs courtesy of Gross, J., F. O. Schmitt, and J. H. Highberger.) y

sequence, one being designated the a 1(1) and Collagen Types I, II, and III, which form
the other the a2(I) chain. In the complex connective tissue fibers, are referred to as
nomenclature that has evolved, the complete interstitial collagen. The basal lamina under¬
molecule of Type I collagen is abbreviated lying epithelia contains collagen that does not
[ol 1 (I)2a2(I)]. polymerize into fibrils but instead forms a
Type II collagen is characteristic of cartilage mat of randomly oriented molecules in asso¬
but also occurs in embryonic cornea and ciation with proteoglycans and the structural
notochord, the nucleus pulposus, the vitreous proteins laminin and hbronectin. The colla¬
body of the eye, and elsewhere. In cartilage, gens of the basal lamina are difficult to ex¬
it forms thin 10- to 20-nm fibrils, but in other tract in pure form and have been less thor¬
microenvironments it may form larger fibrils oughly studied than the interstitial collagens.
morphologically indistinguishable from those The best characterized is Type IV collagen,
of Type I collagen. It consists of a chains of which consists of three identical a chains—
a single kind, designated al(II), and since ctl(IV)—and the molecule is designated
there are three of these, the molecule is [al(IV)]3. Chains distinctly different from
described [al(II)]3. other known a chains have been extracted
Type III collagen is abundant in loose con¬ from basal laminae and are collectively re¬
nective tissue, blood vessel walls, dermis of ferred to as Type V collagen, but their associ¬
the skin, and stroma of various glands. It ation ratio remains to be clarified.
appears to be a major constituent of the The functional significance of the molecu¬
slender 50-nm fibers that have traditionally lar heterogeneity and differing distribution
been called reticular fibers (see below). It is of collagen types is still obscure. Statements
composed of a single kind of a chain, al(III), as to distribution of the several molecular
and the molecule is abbreviated [al(III)]3. types of collagen are based largely on bio-
142 • CONNECTIVE TISSUE PROPER
\ v

chemical extraction from various organs and nm in diameter); slender nonstriated fibrils
tissues. The types specified in Table 5-1 (—10 nm); intermediate-sized striated fibrils
predominate in the locations given but it (—25 nm); and larger striated fibrils (—50
would be misleading to suggest that those are nm). Only Type I collagen, such as occurs in
the only collagens present. In most locations tendons, appears to form very large fibrils
the connective tissues include more than one up to 190 nm in diameter, but in this tissue,
type of collagen. Biochemical identification too, fibrils in the 50-nm range mingle with
of the predominant type in a given organ the large fibrils (Fig. 5-28). It seems clear
does not provide information on the range that the widely distributed 50-nm striated
of fiber sizes, their orientation, their degree fibrils with 67-nm periods may be composed
of aggregation into fiber bundles, or their of Types I, II, or III collagen and fibrils of
relationship to the proteoglycan constituents this size are not confined to reticular connec¬
of the extracellular matrix. Information of tive tissue, as has been widely assumed. The
this kind must be obtained by microscopic size and architectural pattern of the fibrils
methods such as immunohistochemical local¬ formed in different organs does not appear
ization with type-specific antibodies, and vis¬ to depend solely on genetically determined
ual analysis at high resolution with the elec¬ differences in collagen molecules, but is influ¬
tron microscope. enced by their post-translational modifica¬
When the several molecular types of col¬ tion; by their relationship to glycosaminogly-
lagen were discovered, it was tempting to cans in the matrix; and by other factors in
speculate that each might polymerize in a the regional microenvironment. Apart from
characteristic fibril size and that the different their intrinsic interest, analysis of all these
categories of fibrils were deployed in distinc¬ factors is relevant to human and veterinary
tive patterns. However, evidence to date in¬ medicine since, as discussed in a later section,
dicates that fiber diameter is not closely cor¬ a number of collagen diseases can be ex¬
related with collagen type. When examined plained by failures at specific steps in the
with the electron microscope, connective tis¬ complex process of synthesis, secretion, post-
sues that are relatively rich in Types I, II, or translational modification, and extracellular
III collagen by biochemical analysis all con¬ assembly of collagen molecules.
tain varying proportions of microfibrils (—4 Although the electron microscope is the

Table 5-1. CHARACTERISTICS OF DIFFERENT COLLAGEN TYPES

Collagen Molecular Tissue Optical Site of Interaction with
Type Formula Distribution Microscopy Ultrastructure Synthesis Glycosaminoglycans Function

I [al(I)]2a2 Dermis, bone, Closely packed, Densely packed, Fibroblast, Low level of Resistance to
tendon, thick, non- thick fibrils with osteoblast, interaction, mainly tension
dentin, fascias, argyrophilic, marked odontoblast, with dermatan sulfate
sclera, organ strongly bire- variation in chondroblast
capsules, fringent yellow diameter
fibrous or red fibers.
cartilage Collagen fibers.

II [<*1(II)]3 Hyaline and Loose, No fibers; very Chondroblast High level of Resistance to
elastic collagenous thin fibrils interaction, mainly intermittent
cartilages network visible embedded in with chondroitin pressut'e
only with abundant sulfates
picrosirius stain ground
and polarization substance
microscopy

III [al(IH)]s Smooth Loose network Loosely packed Smooth muscle, Intermediate level of Structural
muscle, endo- of thin, thin fibrils with fibroblast, interaction, mainly maintenance
neurium, argyrophilic, more uniform reticular cells, with heparan sulfate in expansible
arteries, weakly diameters Schwann cells, organs
uterus, liver, birefringent hepatocyte
spleen, greenish fibers.
kidney, lung Reticular fibers.

IV [al(IV)]3 Epithelial and Thin, Neither fibers Endothelial and Insufficient data Support and
endothelial amorphous, nor fibrils epithelial cells filtration
basal laminae weakly are detected
and basement birefringent
membranes membrane

Modified from Junqueira, L. C., and Carneiro, J. Basic Histology. 4th ed. Los Altos, CA, Lange Medical Publishers,
1983.
 CONNECTIVE TISSUE PROPER • 143

instrument of choice for identification of the

subunit structure of collagen fiber bundles,
it does not lend itself to study of the orien¬
tation and pattern of bbers with respect to
whole organs and their functional subunits.
For this purpose the light microscope is more
suitable because of its larger held and the
greater thickness of the sections that can be
examined. Its usefulness is enhanced if the
tissue is stained with the picrosirius method
and viewed with polarization optics. In react¬
ing with the basic groups of collagen, the
long molecules of the dye, Sirius red, orient
parallel to the molecules of collagen, enhanc¬
ing the form birefringence of the fibers.
Under the polarizing microscope, one can
take advantage of differential coloration and
relative enhancement of birefringence to as¬
sess the distribution of collagen Types I and
III. Thus, in dermis, tendon, fibrous carti¬
lage, bone, dentin, sclera, and cornea where
Type I is abundant, the collagen appears as Figure 5-6. Drawing comparing Type IV collagen (base¬
ment lamina), Type III (reticular fibers) and Type I (ten¬
strongly birefringent yellow or red fibers. In
don). Fibers characteristic of tendon are a mixture of fibrils
uterine smooth muscle, in the walls of arter¬ 30 to 150 nm in diameter, whereas in reticular fibers the
ies, in liver, spleen, and the investment of fibrils average 50 to 55 nm in diameter. The small dots
nerves—tissues all rich in Type III colla¬ associated with these fibrils represent granules of proto-
gen—one finds thin, green, birefringent fi¬ glyan. (From Hay, E. D., et al. In Gastpar, H., K. Kuhn,
and R. Marx, eds.: Collagen-Platelet Interactions. Stutt¬
bers corresponding in pattern to the silver- gart, Schattauer, 1978, pp. 129-151.)
impregnated reticular fibers of classical his¬
tology. Type II collagen, which is character¬
istic of cartilage, is poorly or inconsistently however, in delicate networks surrounding
stained with this method. Immunofluores- adipose cells; smooth muscle cells; the sar-
cent localization of collagen types using sen¬ colemma of striated muscle; the endoneu-
sitive specific antibodies is in general agree¬ rium of nerves; and the stroma of many
ment with the distribution observed in glandular organs. They are also found in
preparations stained with the picrosirius close association with the basal lamina of most
method. epithelia and they form the supporting tissue
of lymphoid and blood-forming organs.
With the introduction of the electron mi¬
Reticular Fibers croscope, reticular fibers were found to be
made up of unit fibrils having the 67-nm
Reticular fibers were identified as a com¬ periodicity typical of collagen. It is now
ponent of connective tissue over a century thought that the difference in argyrophilia
ago and were long regarded as a fibrous of reticular and collagen fibers is not due to
element distinct from collagen. These very a chemical difference in their fibrous pro¬
slender fibrils (0.5 to 2.0 |xm) tend to form teins, but depends on the size of the fibrils
delicate networks rather than coarse bundles and their relationship to interhbrillar proteo¬
(Fig. 5-6). They are not apparent in ordinary glycans that bind them together. It is now
histological sections but can be selectively generally accepted that collagenous and re¬
stained by reason of their property of ad¬ ticular fibers are essentially identical bio¬
sorbing metallic silver when treated with al¬ chemically. However, the terms “reticulum”
kaline solutions of reducible silver salts (Fig. and “reticular fibers,” are still widely used to
5-7). Fibers of this nature are the first to designate small fibrous elements of connec¬
appear in the differentiation of embryonic tive tissue whose size and pattern are differ¬
mesenchyme into loose connective tissue, but ent from those of more typical collagen fi¬
they gradually give way to increasing num¬ bers. Recent studies indicate that the
bers of typical collagen fibers in the connec¬ collagenous component of reticular fibers is
tive tissue of adults. Reticular fibers persist, principally Type III collagen.
144 • CONNECTIVE TISSUE PROPER

Figure 5-7. Reticular fibers are distinguished from other collagenous fibers by their smaller size, their reticular pattern,
and the fact that they blacken with silver stains. The pattern of reticular fibers in different examples is presented here.
A, Reticulum of the spleen. B, Reticular fibers of the adrenal cortex.

Elastic Fibers In unstained spreads of loose connective

tissue, the slender, refractile elastic fibers 0.2
Some organs, in their normal function, to 1.0 pm in diameter are not plentiful, but
must yield transiently to externally or inter¬ they can be distinguished from the more
nally applied force. Their connective tissue abundant collagen fibers by their small di¬
therefore must have sufficient resilience to ameter and their tendency to branch and
restore the original state. For example, the anastomose to form loose networks (Fig.
aorta, the large vessel conducting blood from 5—8). They usually are not identifiable in
the heart, is distended by the outflow from histological sections stained with hematoxylin
each contraction of the ventricle. Its elastic and eosin, but can be clearly demonstrated
recoil to resting diameter between beats is when selectively stained by Verhoeff’s stain,
essential for maintenance of continuous flow Weigert’s resorcin-fuchsin, or Halmi’s alde-
from the intermittent pumping action of the hyde-fuchsin method.
heart. Similarly, the lungs are expanded dur¬ When present in sufficient numbers, elastic
ing inspiration and return passively to initial fibers impart a yellowish color to the fresh
volume during expiration. The aorta and tissue. Certain elastic ligaments, such as the
lungs are especially rich in elastic fibers, but ligamenta flava of the human vertebral col¬
connective tissues throughout the body con¬ umn and the ligamentum nuchae of rumi¬
tain similar fibers that have the ability to be nants, are distinctly yellow. The latter is com¬
stretched by a small force and to return to posed of exceptionally coarse parallel elastic
their original dimensions when the force is fibers up to 4 or 5 pm in diameter. Elastin is
removed. Elastic fibers can be stretched to not always in the form of fibers. It takes the
their breaking point at about 150 per cent of form of fenestrated sheets or lamellae in the
their original length with a force of only 20 elastica interna and elastica externa in the walls
to 30 kg/cm2. When a fiber breaks, its ends of arteries.
quickly retract and coil up. Elastic fibers are not made up of fibrillar
 CONNECTIVE TISSUE PROPER • 145

Figure 5-8. Elastic fibers in a spread of rat mesentery, stained with resorcin-fuchsin; the photograph is printed as a
negative image. Notice that the fibers are more slender than the collagen fibers in Figure 5-1, and they branch and
anastomose to form a network. (From Fawcett, D. W. In Greep, R. O., ed. Histology. Philadelphia, Blakiston Co., 1953.
Reproduced by permission of McGraw-Hill Book Co.)

subunits that are visible with the light micro¬ gen intact. About 30 per cent of the amino
scope and therefore appear homogeneous. acid residues of elastin are glycine and about
Under polarizing optics, they exhibit a weak 11 per cent are proline. Unlike collagen,
birefringence that increases when the fibers elastin is composed mainly of nonpolar hy¬
are stretched. In electron micrographs, elas¬ drophobic amino acids and contains little
tic fibers have two components: microfibrils hydroxyproline and no hydroxylysine. It is
approximately 10 nm in diameter aggregated relatively rich in valine and contains two
in small bundles that are associated with, or unique amino acids, desmosine and isodesmo-
embedded in, a seemingly amorphous com¬ sine, which serve to cross-link the polypeptide
ponent, elastin (Fig. 5-9). chains.
Although elastin is usually described as The microfibrillar component of elastic fi¬
amorphous, a fibrillar substructure has re¬ bers differs greatly from elastin in amino
cently been reported in electron micrographs acid composition. It contains less glycine;
of purified elastin after prolonged extraction contains no hydroxyproline, desmosine, or
with osmium tetroxide or periodate. It is isodesmosine; and is composed mainly of
speculated that these treatments may dissove hydrophilic amino acids. About 5 per cent of
sugar moieties of elastin, unmasking its fi¬ the weight of microfibrillar protein consists
brillar structure. The fibrils are reported to of neutral sugars.
have a diameter of 2 to 4 nm and to be The role of the microfibrils in hbrogenesis
closely packed in parallel array. is not clear, but it is thought that they may
Elastin is resistant to boiling and to hy¬ impose a fibrous form on the polymerizing
drolysis by dilute acid or alkali under condi¬ elastin. During development, the bundles of
tions that destroy other connective tissue con¬ microfibrils are laid down first in close prox¬
stituents. It resists digestion by trypsin but imity to the surface of fibroblasts, smooth
when treated with the enzyme elastase, pre¬ muscle cells, or other mesenchymal deriva¬
pared from pancreas, it is selectively digested tives. The elastin appears later and ultimately
from tissue sections, leaving cells and colla¬ makes up the bulk of the fiber, with the
146 • CONNECTIVE TISSUE PROPER

Figure 5-9. Electron micrograph of a preparation of elastic fibers purified from ligamentum nuchae of a feta! calf. A,
Stained microfibrils are visible around and between the pale amorphous appearing areas of elastin. B, The microfibrils
have been extracted by chemical treatment that breaks disulfide bonds of protein, leaving behind only the amorphous
component of elastic fibers. Inset shows an early stage of elastic fiber formation in cross section. 11-nm microfibrils are
seen around a pale core of newly formed elastin. (Micrographs from Ross, R., and P. Bornstein. Sci Am 224:44, 1971.)

microfibrils forming a layer around its pe¬ the extracellular matrix by their higher pro¬
riphery and occurring in small fascicles in its portion of protein and by characteristic dif¬
interior. A precursor molecule, proelastin or ferences in the nature of their polysaccharide
tropoelastin, is believed to be synthesized and side chains. They usually contain hexos-
released at the cell surface. Extracellularly amine, galactose, or other sugars with sialic
the enzyme lysyl oxidase catalyzes the for¬ acid in a terminal position on the heterosac¬
mation of aldehyde groups on certain lysines. charide chain.
Three of these aldehydic residues condense Fibronectin. The term fibronectin describes
with a fourth lysine to form the ring structure a group of structurally and immunologically
of the desmosines that cross-link the elastin related glycoproteins found in connective tis¬
chains. sue, in basal laminae of epithelia, and on the
In the disease lathyrism, which occurs in surfaces of many cells. As the name implies
animals that eat the plant Lathyrus odoratus, (fibra, fiber; nectere, to bind), this protein
and in a comparable condition induced ex¬ binds to other fibrous proteins such as colla¬
perimentally by administration of (3-amino- gen and fibrin. It seems to be involved in
propionitrile, the action of lysyl oxidase is adhesion of fibroblasts and other cell types
inhibited and both collagen and elastin are to their natural substrates in the tissues, and
incompletely cross-linked. in the adhesion, spreading, and locomotion
of cells on artificial substrates in vitro. The
protein has a M.W. of about 440,000 and is
Other Structural Proteins
composed of two 220,000-dalton polypeptide
In addition to the collagenous and elastic chains. It is not confined to the tissues but
fibers long recognized by light microscopists, occurs also in the blood plasma where it was
there are at least two structural glycopro¬ first detected and called cold-insoluble globulin,
teins—-fibronectin and laminin. Glycoproteins a term that has now been discarded in favor
are distinguished from the proteoglycans of of plasma fibronectin. The circulating hbronec-
 CONNECTIVE TISSUE PROPER • 147

tin is believed to be synthesized by the en¬ exhibits no structural organization visible

dothelium lining the vascular system. The with the light microscope. It is commonly
fibronectin of the peripheral tissues is re¬ referred to, therefore, as the aiffioYphous
ported to be synthesized by fibroblasts, myo¬ ground substance. It has the physical properties
blasts, chondrocytes, Schwann cells, astroglial of a viscous solution or thin gel. It is extracted
cells, and by a number of epithelia, and the by the aqueous fixatives commonly used and
list is increasing. It is not visible in routine is not visible in histological sections. It can be
histological preparations, and what is known preserved to some extent if frozen sections
of its distribution is based mainly on immu- of fresh tissue are fixed in ether-formol va¬
nocytochemical localization with fluorescent por. In such preparations, it stains with the
antibody. periodic acid-Schiff reaction for carbohy¬
Fibronectin is an important component of drates and stains metachromatically with to-
the connective tissue matrix and is present in luidine blue. These tinctorial properties are
the basal lamina of epithelia, and in the attributable to the presence of several types
external lamina around smooth and striated of proteoglycans—complex macromolecules
muscle cells. In tissue cultures, where it has consisting of a core protein of varying length
been most extensively studied, it appears as to which glycosaminoglycan (GAG) chains are
a fine fibrillar matrix situated on cells, be¬ covalently linked. Glycosaminoglycans are
tween cells, and between them and the glass linear polymers of repeating disaccharide
or plastic substrate. There is some evidence units of which one is always a hexosamine.
suggesting transmembrane association of in¬ The GAG chains are linked to the core pro¬
tracellular actin with extracellular fibronec¬ tein at one end and radiate from it in a
tin, which would account for the observed bottle-brush configuration (Fig. 5—10). The
effects of added fibronectin on cell shape and principal glycosaminoglycans of the extracel¬
improved adhesion to substrate. lular matrix are hyaluronic acid, chondroitin
Fibronectin is found in the a granules of sulfates, dermatan sulfate, keratan sulfate, and
blood platelets, and is released onto their heparan sulfate (Table 5-2). These differ in
surface and into the environment when plate¬ molecular weight, length of chain, and the
lets are activated by thrombin or by contact nature of their disaccharide units. With the
with collagen. It has been suggested that possible exception of hyaluronic acid, these
surface fibronectin may play a role in platelet glycosaminoglycans do not exist in tissues in
aggregation and adhesion to fibrin and col¬ the free state, but are always bound to core
lagen in the formation of blood clots. proteins as components of proteoglycans.
The fibronectin molecule has along its Hyaluronic acid is a major constituent of
length a set of highly specific binding do¬ vitreous humor and synovial fluid but occurs
mains for macromolecules that occur on cell widely in extracellular matrices elsewhere. It
surfaces and in the extracellular matrix. is a very long molecule of high molecular
These binding sites enable fibronectin to me¬ weight, which if straightened out would be
diate adhesion of cells to other cells, to col¬ up to 2.5 pm in length. One of its most
lagen, and to hyaluronic acid and other gly- important properties is its very high viscosity
cosaminoglycans of the extracellular matrix. in aqueous solution, which is largely respon¬
Thus, it may play a role in organizing extra¬ sible for the consistency of the ground sub¬
cellular macromolecules as well as in binding stance. If fluid is injected into connective
cells to their substrate. tissue, it does not immediately diffuse away
Laminin. Laminin is more restricted in its from the site but remains localized for a while
distribution than fibronectin. It is a large in a discrete bleb, as though walled off by
glycoprotein composed of at least two poly¬ the viscous interstitial substance. This prop¬
peptide chains of M.W. 220,000 and 440,000 erty of the matrix is believed to serve as a
joined by disulfide bonds. Laminin is local¬ barrier to the spread of bacteria that may
ized mainly in basal laminae and seems to be gain access to the tissues. It is of interest that
involved in attachment of epithelia to their the most invasive pathogenic bacteria pro¬
supporting layer of Type IV collagen. duce an enzyme hyaluronidase, which enables
them to depolymerize the hyaluronidase in
Ground Substance the extracellular matrix. The viscosity of hy¬
aluronic acid in the synovial fluid of joints
The cells and fibers of connective tissue makes it well suited for its lubricating func¬
are surrounded by a translucent matrix that tion in this site.
148 • CONNECTIVE TISSUE PROPER

Proteoglycan
subunits

Figure 5-10. Schematic representation of the organization of the extracellular matrix of cartilage. It is composed of
collagen fibrils with intertwining proteoglycan aggregates occupying the interstices. The extracellular matrix of connective
tissues in general is believed to be similar but with differences in proportions of the several glycosaminoglycans.

Chondroitin sulfate is the predominant atan sulfate is found in cornea, cartilage, and
glycosaminoglycan in the proteoglycans of nucleus pulposus. Heparan sulfate occurs in
cartilage, bone, and large blood vessels but is liver, aorta, and lung and will no doubt be
found in other tissues as well. Dermatan detected elsewhere as more connective tissues
sulfate is most abundant in skin but also are analyzed.
occurs in lung, tendon, and elsewhere. Ker- The proteoglycans of cartilage matrix have

Table 5-2. CONSTITUTION AND DISTRIBUTION OF GLYCOSAMINOGLYCANS IN CONNECTIVE

TISSUE AND THEIR INTERACTION WITH COLLAGEN FIBERS
Electrostatic
Glycosamino¬ Interaction with
glycan Disaccharide Unit Distribution Collagen

Hyaluronic acid D-glucuronic acid + N- Umbilical cord, synovial fluid,

acetyl-O-glucosamine vitreous, cartilage

Dermatan sulfate L-iduronic acid + A-acetyl-D- Structures formed by collagen Low levels of
(chondroitin sulfate galactosamine-6-sulfate fibers: e.g., dermis, tendon, interaction, mainly
B) ligaments, heart valves, organ with collagen
capsules, sclera, fibrous cartilage, Type I
arteries (adventitial layer), nerves
(epineurium)

Chondroitin 4- or D-glucuronic acid + N- Hyaline and elastic cartilage, High levels of

6-sulfate* acetyl-D-galactosamine 4- or arterial medial layer, nucleus interaction, mainly
(chondroitin sulfate 6-sulfate pulposus of intervertebral discs with collagen
A or C) Type II

Heparan sulfate D-glucuronic acid + iduronic Structures rich in reticular fibers: Intermediate levels of
acid + A-acetyl-D- e.g., smooth muscle, liver, spleen, interaction, mainly
glucosamine + glucosamine nerves, endoneurium with collagen
Type III

*Chondroitin sulfate can present its sulfate groups in positions 4 or 6. Both of these compounds exist in cartilage
and in smaller amounts in other tissues. The biological significance of sulfation in position 4 or 6 is unknown.
From Junqueira, L. C., and Carneiro, J. Basic Histology. 4th ed. Los Altos, CA, Lange Medical Publishers, 1983.
 CONNECTIVE TISSUE PROPER • 149

been most extensively studied and can be the proteoglycans may play a role in main¬
regarded as representative of the structure taining the structural organization of the
of this class of macromolecules. The typical basal lamina.
cartilage proteoglycan has about 80 side
chains of chondroitin sulfate, each with a
M.W. of 20,000, attached to a core protein Origin of Connective Tissue Fibers
together with a smaller number of chains of The sequence of morphological events in
keratan sulfate. At one end of the core pro¬ formation of collagen is similar whether stud¬
tein is a region with few or no attached GAG ied in the embryo, in young scar tissue, or in
chains but having instead a number of cova¬ tissue cultures. Delicate networks of branch¬
lently bound oligosaccharides. This segment ing and anastomosing argyrophilic fibrils ap¬
of the polypeptide, which is referred to as pear among the hbroblasts (Fig. 5-11). The
the HA-binding region, includes an active fibrils may follow the outlines of the cells and
site that binds with high specificity to hyalu¬ their processes but they also extend far into
ronic acid. The binding of proteoglycan mon¬ the intercellular substance. When studied in
omers to hyaluronic acid results in formation electron micrographs, the finest of the devel¬
of proteoglycan aggregates of very large size. oping fibrils are extracellular and have cross
The sulfate and carboxyl groups on the re¬ striations. As the fibrils increase in number,
peating disaccharide units of the GAG chains they rearrange into parallel wavy bundles of
result in a high concentration of anionic appreciable thickness. These lose their ability
charges. Thus, the proteoglycans are poly¬ to be blackened with silver and instead accept
anions and can be stained in tissue sections stains for collagen.
with cationic substances such as colloidal iron The constant association of hbroblasts with
or Alcian blue, which have a high affinity for developing collagenous fibers both in vivo
the anionic groups on the macromolecule. and in vitro early suggested that these cells
In their native state, the long straight poly¬ were involved in hbrogenesis, but their exact
saccharide chains are extended and occupy a role remained a subject of debate. The area
large volume in relation to their molecular of controversy has been narrowed in recent
weight. It is likely that in the connective tissue years. It is now widely accepted that collagen¬
the extended proteoglycan molecules occupy ous fibers arise extracellularly by polymeri¬
essentially all the interstitial space in the ma¬ zation of molecular collagen secreted into the
trix, and the proteoglycan aggregates are extracellular matrix by hbroblasts. Consistent
intertwined among the collagen and elastic with this view is the electron microscopic
fibers (Fig. 5-10). observation that hbroblasts of growing con¬
As tissues are routinely prepared for elec¬ nective tissue have the extensive endoplasmic
tron microscopy, the polysaccharides are sel¬ reticulum and well-developed Golgi complex
dom well preserved. Better results are ob¬ that we have come to expect of cells actively
tained when the polycationic dye ruthenium engaged in protein synthesis. Moreover, if
red is included in the fixative. This appears 14C-labeled proline is given to animals in
to preserve the GAG by interaction with its which formation of connective tissue has
anionic groups, but during dehydration the been induced, labeled collagen can be de¬
chains collapse onto the protein core. Thus, tected in the microsome fraction isolated
in electron micrographs the proteoglycans from connective tissue cells. Incorporation of
are represented only by dense matrix gran¬ labeled amino acid in hbroblasts can also
ules 10 to 20 nm in diameter. These regret¬ be followed autoradiographically in animals
tably provide little information as to the con¬ with healing wounds. At early time intervals
figuration of proteoglycans in their native after administration of tritiated proline, the
state, or the relationships of the extended silver grains, betraying the location of the
molecules to each other and to the fibrous labeled precursor, are over the endoplasmic
constituents of the extracellular matrix. In reticulum; later they are seen over the Golgi
some tissues, there is a suggestion of a highly region and still later outside the cell, over
ordered interaction between the proteogly¬ newly formed collagen hbers.
cans and specific sites along the cross-banded The evidence thus points to a synthetic
collagen fibrils. A row of proteoglycan gran¬ pathway for collagen, similar to that de¬
ules forming a regular array with a spacing scribed for other proteins (Fig. 5—4). On the
of about 60 nm can be demonstrated along ribosomes of the endoplasmic reticulum, ac¬
the basal laminae of many epithelia (Fig. 5— tivated amino acids are assembled into poly¬
6). Their ordered arrangement suggests that peptide alpha chains, each composed of
150 • CONNECTIVE TISSUE PROPER
\

Mesenchymal
cell //

Fibroblast

Figure 5-11. Electron micrograph of fibroblasts in developing connective tissue. Fibroblasts actively synthesizing
collagen have a well-developed rough endoplasmic reticulum, often with distended cisternae. The relatively undifferen¬
tiated cell at upper left has more heterochromatin and little differentiation of its cytoplasm. The newly formed collagen
fibrils are slender and randomly oriented.

about 1000 amino acids. Hydroxylation of attributed to the presence of an additional

the prolyl and lysyl residues takes place while sequence of amino acids on the N-terminal
the nascent alpha chains are still being syn¬ end of the molecule. This is the original form
thesized. While the alpha chains are still as¬ in which collagen is synthesized in the cell.
sociated with the ribosomes or immediately Procollagen molecules are unable to poly¬
after their release into the lumen of the merize into collagen fibers. However, an en¬
endoplasmic reticulum, they associate in hel¬ zyme, procollagen peptidase, has been identi¬
ical configuration to form procollagen mole¬ fied, which is believed to be located in the
cules with a M.W. of 336,000. The exact cell or at its surface. This cleaves off the
location of the other post-translational events telopeptide converting procollagen to colla¬
is uncertain, but the procollagen molecules gen at the time of its release from the cell.
appear to be transported through the lumen The resulting collagen molecules are then
of the endoplasmic reticulum, and packaged free to polymerize extracellularly into cross-
in the Golgi complex. Molecular collagen striated fibrils.
segregated in Golgi vacuoles is released from A remaining point of uncertainty concerns
the fibroblast when these vacuoles move to the possible role of the cells in determining
the cell surface and discharge their contents the arrangement of the fibers. Most histolo¬
into the surrounding ground substance (Fig. gists assume that the cells simply maintain
5-4). the appropriate physicochemical conditions
A relatively recent development in our in the surrounding ground substance to per¬
understanding of collagen synthesis has been mit collagen fibers to form extracellularly by
the identification of procollagen, an anteced¬ a process of spontaneous polymerization sim¬
ent of the definitive collagen molecule. Pro¬ ilar to that observed in vitro. Such a process
collagen subunits corresponding to oq and a2 could presumably take place at some distance
chains have a M.W. of 112,000—distinctly from the fibroblast. The subsequent orienta¬
larger than the alpha chains of collagen. tion of the fibrils would be in response to
Their greater length and higher M.W. is mechanical stresses in the tissue and would
 CONNECTIVE TISSUE PROPER • 151

not be influenced by the cells. A few investi¬ along bundles of collagen fibers and appear
gators believe that new fibrils arise only in in sections as fusiform elements with long
very close relation to the cell surface, and tapering processes. In other situations they
that the cells exercise a direct control over may be flattened, stellate cells with several
their orientation in the connective tissue. It slender processes. Their cytoplasm is usually
is contended that the orthogonal patterns eosinophilic like the neighboring collagen.
and other precisely ordered arrangements of The outlines of the cell bodies are therefore
collagen fibers in the body are difficult to difficult to make out. They are more easily
explain if collagen deposition is a completely visualized after staining with iron-hematoxy-
independent extracellular phenomenon, lin.
whereas, if collagen is deposited in protofi¬ These cells have been extensively studied
brils that are oriented by fibroblasts, a mech¬ in tissue culture, where they can be better
anism is provided for ordering of collagen observed isolated from the interlacing fabric
fibrils by the cells rather than by purely of fibers in which they reside in vivo. In this
mechanical forces. Further study is needed environment the cells migrate out from the
to resolve this problem. explant into the surrounding medium, with
their processes adhering to form a cellular
network (Fig. 5-12). It is likely that the fibro¬
CELLS OF CONNECTIVE TISSUE blasts in the body also maintain tenuous con¬
tacts with one another, but for technical rea¬
It is convenient to think of the cells of loose sons this is difficult to demonstrate.
connective tissue in two categories: (1) a rel¬ The elliptical nucleus is usually smoothly
atively stable population of fixed cells—the contoured but may sometimes be slightly
fibroblasts (responsible for production and folded. There are one or two nucleoli, and
maintenance of the extracellular compo¬ the chromatin is sparse and distributed in
nents) and adipose cells (for storage of re¬ small karyosomes. A pair of centrioles and a
serve fuel); and (2) a population of mobile small Golgi apparatus are situated near the
wandering cells—macrophages, eosinophils, nucleus. Long, slender mitochondria are
mast cells, lymphocytes, and plasma cells found mainly in the perinuclear cytoplasm,
(cells concerned mainly with the shorter-term but may also occur in the processes. Under
events involved in tissue reaction to injury). conditions of stimulation, as in wound heal¬
ing, when fibroblasts are dividing and actively
Fibroblast or Fibrocyte synthesizing extracellular components, they
enlarge and their cytoplasm becomes mod¬
These are the principal cells of the connec¬ erately basophilic. In electron micrographs,
tive tissue that elaborate the precursors of quiescent fibroblasts contain a small Golgi
the extracellular matrix components.* Their complex and only a few cisternal profiles of
shape depends to some extent on their phys¬ granular endoplasmic reticulum, but in grow¬
ical substrate. They are usually deployed ing or repairing connective tissue, the Golgi
complex becomes very prominent and the
endoplasmic reticulum is much more exten¬
*The student should be aware of troublesome incon¬
sive. The cytoplasm usually contains few in¬
sistency in terminology with respect to this cell type. The
suffix -blast (Greek blastos, germ) is often used in naming
clusions except for occasional small fat drop¬
the formative stages of various cell types. Thus, an lets. Granules staining with the periodic acid-
erythroblast is an early developmental stage of the fully Schiff reaction become numerous under
differentiated cell called an erythrocyte. Some authors, some conditions and may represent intracel¬
therefore, use the term fibroblast to designate a relatively
lular precursors of the glycosaminoglycans of
immature cell actively proliferating and producing com¬
ponents of the extracellular substance, and they apply the ground substance.
fibrocyte to the relatively quiescent cells of adult connec¬ Majority opinion holds that fibroblasts are
tive tissue. This interpretation loses sight of the fact that fully differentiated cells that ordinarily do
the term “fibroblast” was originally intended to describe
not give rise to other types of cells in the
a “fiber-forming cell” and not to name an immature
form of a cell called a fibrocyte. Moreover, because most
connective tissue. There is suggestive evi¬
histologists recognize mesenchymal cells, persisting in post¬ dence, however, that in pathological states
natal life, as the undifferentiated progenitors of the and under certain experimental conditions,
connective tissue cells, the use of “fibroblast” in this they can transform into bone cells. It is also
sense makes an unnecessary distinction and introduces
widely accepted that fibroblasts may accu¬
a redundant term. The term fibroblast, therefore, is
properly used to describe the differentiated cell of adult mulate lipid and become adipose cells, but in
connective tissue, and it can be considered synonymous both these instances it is difficult to establish
with fibrocyte (as used by other authors). with certainty whether it is actually the fibro-
152 • CONNECTIVE TISSUE PROPER

Figure 5-12. Photomicrograph of fibroblasts in cell culture illustrating their spindle shape. This form is also common in
vivo but the processes may be branched, resulting in more stellate forms.

blasts or their undifferentiated mesenchymal pluripotential cells in the perivascular con¬

progenitors that undergo these transforma¬ nective tissue.
tions.
Adipose Cells
Mesenchymal Cells
Among the sessile cells in loose connective
It is widely accepted that a population of tissue are some that are specialized for the
cells that retain the developmental potential¬ synthesis and storage of lipid. These adipose
ities of embryonic mesenchymal cells persists cells or fat cells accumulate lipid to such an
in the adult organism. They are somewhat extent that the nucleus is flattened and dis¬
smaller than fibroblasts, but otherwise have placed to one side, and the cytoplasm be¬
much the same appearance and cannot easily comes so thinned out that it is resolved only
be distinguished from them in ordinary his¬ as a thin line around the rim of the single
tological sections. In loose connective tissue large lipid droplet (Figs. 5—13, 5—14). So
they are usually deployed along the blood inconspicuous are the nucleus and cytoplasm
vessels, especially along capillaries. In elec¬ that the fat cells in fresh connective tissue
tron micrographs (Fig. 5-11), these small have the appearance of large glistening drops
cells have a somewhat coarser chromatin pat¬ of oil. They may occur singly in the connec¬
tern, few mitochondria, and little or no rough tive tissue but are more often found in
endoplasmic reticulum. These characteristics groups. There is a marked tendency for them
serve to distinguish them from synthetically to be located along the course of small blood
active fibroblasts. vessels. When they accumulate in such large
Many investigators regard these cells, numbers that they become the predominant
rather than fibroblasts, as the precursors of component, crowding out other cell types,
adipose cells. When capillaries are obliged to the resulting tissue is called adipose tissue or
grow and change the character of their walls fat (see Chapter 6). All intermediate grades
in response to altered hemodynamic condi¬ between loose connective tissue and typical
tions, the smooth muscle cells required seem adipose tissue can be found.
to be recruited by differentiation of these In the preparation of the usual histological
 Developing fat cells

Macrophage

Figure 5-13. Drawing of cells in subcuta¬

neous loose connective tissue of a rat. The
osmium tetroxibe of the fixative preserved and
blackened the lipid droplets in several adipose
cells in different stages of development. (After
A. A. Maximov.)

Fibroblast

Developing fat cell

Fat cell

Mast cell Eosinophilic leukocyte

Figure 5-14. Electron micrograph of portions of two adipose cells and the intervening collagenous fibers. Relative to
the mast cell and the fibroblast at upper right, the adipose cells are enormous. Only a small portion of each is included
in this field, but enough to show their thin peripheral layer of cytoplasm and the homogeneous lipid content, which
usually is not stained black by osmium after primary glutaraldehyde fixation.
153
154 • CONNECTIVE TISSUE PROPER

section the lipid droplets of the adipose cells believed to stem from monocytes emigrating
are dissolved out during dehydration, and from the blood. The latter were thought to
there remains only the thin layer of cyto¬ originate in the connective tissue by differ¬
plasm, slightly thickened in one area to ac¬ entiation from persisting mesenchymal cells.
commodate the nucleus. Despite the extreme The development of methods for tracing
thinness of the rim of cytoplasm, a juxtanu- cell lineages by labeling with radioisotopes;
clear Golgi apparatus can be demonstrated, the use of chromosome markers; and im¬
and filamentous mitochondria are distributed munological techniques for detecting specific
around the entire circumference of the lipid receptors on the surface of ceils have all
droplet. The individual fat cells are sur¬ provided compelling evidence that the mono¬
rounded by a delicate network of reticular nuclear phagocytes all originate from precur¬
fibers. sors in the bone marrow; are transported in
Adipose cells develop from fusiform cells the blood as monocytes; migrate through the
that resemble fibroblasts, but, as indicated endothelium of postcapillary venules; and
above, these are generally believed to be take up residence in the tissues. To some
mesenchymal cells that persist into postnatal extent, their morphology is dependent on
life in the adventitia of the small blood vessels the organ in which they are found and on
of the connective tissue. During their early the degree of their.activity. It is now widely
development they may contain multiple small accepted that the free and fixed macrophages
lipid droplets, but these ultimately coalesce of the connective tissue are different func¬
into a single drop (Fig. 5-13). The fully tional phases in the life history of cells of the
formed fat cell is incapable of mitotic divi¬ same lineage.
sion. Any new areas of adipose cells that The circulating monocytes of the blood are
develop in adult life must therefore arise a reserve of potential phagocytes that can be
from undifferentiated precursors. rapidly mobilized at sites where they are
needed. In inflammatory reactions, additions
are made to the resident population of phag¬
Macrophages
ocytes by emigration of monocytes from the
The connective tissues contain mononu¬ blood and their transformation to free mac¬
clear cells of varying appearance that have in rophages. A monocyte that has recently ar¬
common the ability to take particulate mate¬ rived in the tissues is a spherical cell 10 to 14
rial into their cytoplasm and to degrade the jxm in diameter with an eccentric, reniform
ingested substances with hydrolytic enzymes. nucleus and a faintly basophilic cytoplasm.
This activity is called phagocytosis (see Chapter In electron micrographs it has a relatively
1) and any cells that exhibit it are described smooth surface, a sparse endoplasmic retic¬
as phagocytes. The principal phagocytes in ulum, and few cytoplasmic inclusions. After
normal connective tissue are the macrophages. its activation and transformation to a mac¬
Because of their remarkable capacity to de¬ rophage, the surface is highly irregular with
stroy bacteria, dead cells, and debris resulting many microvilli and lamellipodia, and the
from injury, such cells are important in main¬ cytoplasm contains numerous phagocytic vac¬
tenance and repair of tissues and in the uoles, secondary lysosomes, and residual bod¬
body’s defense against invading microorga¬ ies (Fig. 5—15). These cells have an ameboid
nisms. locomotion. Therefore, the shape they pre¬
Traditionally, histologists were obliged to sent in sections depends on their configura¬
rely exclusively upon the shapes of cells, their tion at the moment of their immobilization
staining properties, and their relationship to by the fixative.
other tissue components for their identifica¬ A large proportion of the macrophages in
tion. Using these morphological criteria, two normal connective tissue are relatively inac¬
main categories of phagocytes were distin¬ tive and are deployed along fibrous compo¬
guished—-free macrophages and fixed macro¬ nents of the tissue as stellate or fusiform cells
phages; one motile and wandering through that are difficult to distinguish from fibro¬
the ground substance of the connective tis¬ blasts. Their nucleus is somewhat smaller and
sue, the other sessile and stretched out along more deeply stained. The cytoplasm is more
the bundles of collagen fibers. These two heterogeneous, often containing granules
forms of tissue phagocytes were considered and vacuoles of varying size. In this config¬
to be distinct in their origins and, to some uration, the cells correspond to the fixed
extent, in their functions. The former were macrophages or histiocytes of the classical lit-
 CONNECTIVE TISSUE PROPER • 155

Lysosomes
Pinocytotic
vacuoles

Heterophagic
vacuoles

Figure 5-15. Micrograph of a free macrophage in edematous loose connective tissue showing the typical heterogeneity
of the cytoplasm containing numerous lysosomes, vesicles, and heterophagic vacuoles. Thin ruffles or lamellipodia were
fixed in various phases of their pinocytotic activity.

erature on histology and histopathology. transform to macrophages that eliminate by

When appropriately stimulated, these sessile phagocytosis the dead and dying leukocytes
macrophages detach from the fibers, with¬ of other types. Thus, in a few days, pure
draw their processes, and migrate as free cultures of macrophages are obtained (Fig.
macrophages to sites of tissue injury or bac¬ 5—16). The cells flatten out uniformly spaced
terial invasion. After the acute inflammatory on the glass substrate or, if sufficiently
reaction subsides, the survivors presumably crowded, may assume an epithelioid appear¬
disperse and revert to their sessile, resting ance. The culture vessel acts as a foreign
form. body and, after prolonged cultivation, mul¬
In certain forms of chronic inflammation, tinucleate giant cells are formed.
aggregated macrophages may take on a po¬ Monocytes and their derivatives have the
lygonal shape due to close packing and mu¬ unique property of rapid adherence to glass
tual deformation. In this form, they are de¬ or plastic surfaces. When mixed populations
scribed as epithelioid cells. Around splinters or of cells from blood, peritoneal exudates, or
other foreign bodies in the tissues that are spleen are introduced in suspension into a
too large to be engulfed and destroyed by culture flask, monocytes and mature macro¬
intracellular digestion, macrophages may co¬ phages settle onto the floor and soon become
alesce to form huge multinucleate masses firmly adherent. Other cell types remain un¬
called foreign-body giant cells. The morpho¬ attached and after a few hours can be washed
logical transformations of mononuclear away. By this device, pure cultures can be
phagocytes that occur in inflamed connective obtained for experimental purposes without
tissue in vivo can also be observed when waiting for the contaminating cell types to be
leukocytes from blood are cultivated in vitro. eliminated by phagocytosis. Thus, adherence
In such cultures, the only leukocytes capable to glass has become one of the defining
of survival are the monocytes. These rapidly characteristics of mononuclear phagocytes.
156 • CONNECTIVE TISSUE PROPER

it binds to antigen on the surface of bacteria.

A component of complement (C3) in turn binds
to the antigen-antibody complex and serves
as a ligand promoting phagocytosis by facili¬
tating attachment of the bacterium to specific
receptors (Fc) in the membrane of the mac¬
rophage. The phagocytic and cytotoxic ca¬
pabilities of macrophages are also enhanced
by products of other cells in the connective
tissue, notably the lymphocytes.
Conversely, products of macrophages in¬
fluence lymphocytes. Their role in immunity
will be discussed in more appropriate context
in Chapter 13. It will suffice here to note
that the interaction of macrophages with lym¬
phocytes is an essential step in the initiation
and regulation of an immune response. An¬
tibody production, both in vivo and in vitro,
is greatly impaired in the absence of macro¬
phages. It is postulated that they process
antigen to make it more immunogenic, but
to date the exact mechanisms by which mac¬
rophages participate in antibody production
have not been clearly defined. They are also
required to generate cytotoxic cells in cell-
mediated immune responses.

Figure 5-16. Photomicrograph of a culture of macro¬ FREE CELLS OF CONNECTIVE TISSUE

phages that have differentiated in vitro from peripheral
blood monocytes.
In addition to the fixed or sessile cell types
of connective tissue, there are several types
The ability to follow in vitro the same of migratory cells that are emigrants from
sequence of cellular transformation that oc¬ the blood—lymphocytes, monocytes, eosinophils,
curs in inflamed tissues has made it possible basophils, and neutrophils. Because of their
to study in considerable detail the cytological capacity for ameboid locomotion and their
and cytochemical changes associated with the tendency to congregate at sites where their
transition of monocytes to macrophages and services are needed, their numbers in the
epithelioid and giant cells. Accompanying the connective tissue are highly variable, depend¬
acquisition of phagocytic properties, there is ing on the local conditions. They were for¬
an increase in cell volume and a striking merly regarded as blood cells carrying out
enlargement of the Golgi apparatus, which essential functions in the circulation and only
becomes the site of active formation of nu¬ secondarily leaving the blood to take up res¬
merous lysosomes. During the intracellular idence in the connective tissue. The devel¬
digestion of ingested material, the lysosomes opment of ingenious methods for tagging
discharge their content of hydrolytic enzymes these cells, determining their life span, and
into the phagocytotic vacuoles. Thus, the following their movements in the body has
number of lysosomes diminishes during ac¬ brought about a fundamental change in our
tive phagocytosis. They accumulate again in interpretation of their mission. It is now
abundance in the epithelioid and giant cells known that the primary functions of these
that develop after cellular debris and other cells are performed extravascularly in the
material available for phagocytosis has been connective tissue.
eliminated.
In their defensive role against pathogenic
Monocytes and the Mononuclear
bacteria, macrophages do not function inde¬
Phagocyte System
pendently. They are assisted by antibodies
(immunoglobulins) and complement present in The structure of monocytes was described
blood serum. If specific antibody is present, and illustrated in Chapter 4, and their capac-
 CONNECTIVE TISSUE PROPER • 157

ity to transform into macrophages, giant cells, essary but an insufficient criterion for inclu¬
and epithelioid cells has been discussed in sion in the system.
the foregoing section. It remains to consider The development of methods for marking
them in the context of a broader spectrum cells with radioisotopes and for recognizing
of phagocytic cell types that are widely dis¬ stable surface antigens made it possible to
tributed in the body. In addition to those trace cell lineages and to carry out cytokinetic
already mentioned, the alveolar phagocytes studies that have significantly changed our
of the lung, the microglia of the brain, the concept of this system. Van Furth (1969)
Kupffer cells of the liver, and certain stromal proposed the term mononuclear phagocyte sys¬
cells of the bone marrow and lymphoid or¬ tem to include all highly phagocytic cells and
gans all share the property of avid phagocy¬ their precursors. This includes a number of
tosis. Although these were formerly believed the cell types previously assigned to the reti¬
to arise from different precursors, histolo¬ culoendothelial system and several additional
gists early recognized the desirability of ones (Fig. 5—18). It excludes fibroblasts,
grouping them together on the basis of their endothelial cells, reticulum cells, dendritic
shared structural and behavioral features. cells, and others that ingest vital dyes at very
Metchnikoff (1892) considered them as a low rates. The great appeal of the unifying
diffuse cell system, which he termed the concept of a mononuclear phagocyte system
macrophage system. Relying on the uptake and is that labeling and cytokinetic studies show
storage of vital dyes as a measure of phago¬ that all of its members belong to the same
cytic capacity, Aschoff (1924) broadened the cell lineage, originating in the bone marrow
concept to include the specialized endothe¬ and being transported in the blood to their
lium lining the sinusoids of spleen and bone functional sites in the tissues. Wherever
marrow and proposed the inclusive term re¬ found they exhibit a positive histochemical
ticuloendothelial system (Fig. 5-17), which is still reaction for esterase, contain peroxidase-pos¬
widely used and strongly defended by its itive granules, and have receptors for IgG
proponents. and complement on their surface.
The term and its definition are now criti¬ Since the cells of the mononuclear phago¬
cized on several grounds. The vital dye try¬ cyte system pass through several develop¬
pan blue not only accumulates in highly mental stages and express different degrees
phagocytic cells, but when used in high con¬ of functional activity, there is need for de¬
centration may be taken up in smaller scriptive terms for these states. Macrophages
amount by fluid-phase pinocytosis in unspe¬ that are present at any given site in the
cialized endothelium, in fibroblasts, and even absence of an exogenous stimulus are called
in adipose cells—cells not truly phagocytic. resident macrophages. Those that have acquired
The uptake of vital dyes is therefore a nec¬ enhanced functional activity in response to

RETICULOENDOTHELIAL SYSTEM

Microglia Reticulum cells of

Macrophages lymphatic tissue
lining sinuses

Lymph Tissue Blood

Blood
sinuses sinuses macrophages macrophages
(monocyte)

Liver Spleen Bone Adrenal Anterior

(Kupffer cells) marrow cortex pituitary
Figure 5-17. The reticuloendothelial system as traditionally defined—a diffuse system of macrophages and phagocytic
endothelial cells lining blood sinuses in various organs. Although widely separated, they were grouped together because
they shared similar phagocytic properties. The sinusoids of the pituitary and adrenal were originally included but are
shown here in interrupted lines because electron microscopy has revealed that it is not the endothelium, but perivascular
macrophages, that are phagocytic in these organs. The alleged phagocytic activity of splenic sinusoids is now seriously
questioned, and in the liver these properties reside in the Kupffer cells and not in the endothelium proper. Thus, the
endothelial component of the system has been largely eliminated by application of modern research methods.
158 • CONNECTIVE TISSUE PROPER

Mononuclear Phagocyte System found in small numbers in normal connective

tissue throughout the body but are greatly
Stem cell increased at sites of chronic inflammation.
I They are normally abundant in the lamina
Committed stem cell propria of the respiratory tract and are es¬
I Bone marrow pecially concentrated in the loose connective
Monoblasts >
I tissue that underlies the epithelium of the
Promonocytes gastrointestinal tract, where they are involved
I in protective immunosurveillance against the
Monocytes
bacterial flora and antigenic foreign sub¬
1 stances present in the gut lumen.
Monocytes Peripheral blood
I
Macrophages Tissues
Plasma Cells
%

Plasma cells are ovoid cells with an eccentric

nucleus and an intensely basophilic cyto¬
Normal State
connective tissue (histiocyte) plasm. They vary in size but usually fall
liver (Kupffer cell) within the range of 10 to 20 pm. The nucleus
lung (alveolar macrophage) is spherical, and its abundant heterochro¬
lymph nodes (free and fixed macrophages; matin occurs in unusually coarse clumps that
interdigitating cell?)
spleen (free and fixed macrophages)
tend to be evenly spaced around the periph¬
bone marrow (fixed macrophage) ery in a radial pattern that is a useful iden¬
serous cavities (pleural and peritoneal tifying characteristic of this cell type. A prom¬
macrophages) inent, lightly staining area adjacent to the
bone (osteoclasts)
central nervous system (CSF macrophages; brain
macrophages)
skin (histiocyte; Langerhans cell?)
synovia (type A cell)
other organs (tissue macrophage)

Inflammation
exudate macrophage
exudate-resident macrophage
epithelioid cell
multinucleated giant cell (Langhans type
and foreign-body type)
Figure 5-18. The concept of a mononuclear phagocyte
system has replaced the reticuloendothelial system. This
is a group of widely disseminated cell types that not only
share similar morphology and phagocytic potential but
also have a common origin from the monocytes of the
blood, included in this system are all the cell types
specified in parenthesis after the organ or tissue in which
thdy are located. (After von Furth, R. Immunobiology
767:178, 1982.)

an environmental stimulus are called activated

macrophages. Macrophages of varying degrees
of activity attracted to the same site by a
given substance are referred to as elicited
macrophages. These terms find their greatest
use in studies on the inflammatory response
of tissues.

Lymphocytes
The smallest of the free cells of the con¬ Figure 5-19. Plasma cells from connective tissue near
nective tissue are lymphocytes that have em¬ the human tonsil. Several transitional forms between
lymphocytes and plasma cells are drawn. The mature
igrated from the blood. Their structural and plasma cells are intensely basophilic, and a conspicuous
functional role in immune responses have paler region adjacent to the nucleus is the site of the
been discussed in Chapter 4. They may be large Golgi complex. (After A. A. Maximov.)
 CONNECTIVE TISSUE PROPER • 159

nucleus is the site of its large Golgi apparatus. be found anywhere in the connective tissue
The remainder of the cytoplasm is strongly where lymphocytes abound. Thus, they are
basophilic.
common among the population of free cells
In electron micrographs the coarse chro¬ in the lamina propria of the intestinal tract.
matin pattern is equally striking. There is a The B lymphocytes that are the immediate
well-developed juxtanuclear Golgi complex precursors of plasma cells make up only a
and a pair of centrioles. The most conspicu¬ small percentage of the circulating lympho¬
ous ultrastructural feature of the cytoplasm cytes. They carry in their surface membrane
is an extensive system of cisternae of the molecules of immunoglobulin that they have
rough endoplasmic reticulum (Fig. 5-20). produced in response to an initial encounter
The cisternae are usually narrow and in close with an antigen earlier in their relatively long
parallel array, but in some cells they may be life span. These serve as specific receptors
less highly ordered and their lumen may be for that antigen in a subsequent exposure
distended with a flocculent material of low anywhere in the body. When antigen binds
electron density. The plasma cell thus has to these surface receptors, the antigen-anti-
the ultrastructural features of a highly active body complexes aggregate and are taken into
protein-secreting cell. The varying appear¬ the cell by endocytosis. These events trigger
ance of its endoplasmic reticulum probably transformation of the lymphocyte into a lym¬
reflects differing degrees of activity and ac¬ phoblast that divides repeatedly, giving rise
cumulation of secretory product. to a clone of B lymphocytes all bearing the
Plasma cells are the principal producers of same antigenic specificity. Some members of
antibodies, the immunoglobulins that partic¬ this clone differentiate into plasma cells that
ipate in the humoral immune responses of synthesize and release large amounts of spe¬
the body (Fig. 5—21). These cells arise extra- cific antibody. The remainder are added to
vascularly by further differentiation of B the population of recirculating B lympho¬
lymphocytes that have migrated into the tis¬ cytes with specificity for that particular anti¬
sues from the blood. They are most numer¬ gen. This enables the individual to mount a
ous in the lymph nodes and spleen but may more vigorous and sustained immune re-

Figure 5 20. Electron micrograph of a plasma cell illustrating the coarse pattern of heterochromatin and the extensive
rough endoplasmic reticulum. The large Golgi complex is not included in this plane of section.
160 • CONNECTIVE TISSUE PROPER

of their plasma cells. It was later found that

humans with hyperglobulinemia, an excess of
circulating antibodies, had a high concentra¬
tion of these cells in their tissues, whereas
persons with congenital agammaglobulinemia
developed no plasma cells at sites of antigenic
stimulation and had a complete failure of
antibody synthesis. Compelling direct evi¬
dence finally resulted from the demon¬
stration of antibody production in vitro by
individual plasma cells isolated from the tis¬
sues by microsurgery. Immunocytochemical
methods localized antibody by light micros¬
copy within plasma cells (Fig. 5—21) and elec¬
tron microscopy localized antibody within the
Figure 5-21. Fluorescence photomicrograph of a human cisternae of the endoplasmic reticulum.
plasma cell reacted with fluorescein-conjugated antibody
against gamma globulin. The strong fluorescence of the
cytoplasm identifies the cell as a site of immunoglobulin
Eosinophils
synthesis. (From Mellors, R., and L. Korngold. J. Exp.
The structure and functions of eosinophils
Med. 118:387, 1963.)
have been presented in Chapter 4 and need
not be reviewed here. Their function in the
sponse to reintroduction of the antigen to normal connective tissue is yet to be eluci¬
the body at a later time. The plasma cells dated. What little is known has been deduced
formed have a limited life span in the lymph¬ from observations of their behavior at sites
oid organs and connective tissue, and degen¬ of inflammation or parasitic invasion.
erate after a few weeks of active antibody Eosinophils are often found in close prox¬
synthesis. imity to basophils and mast cells, and it is
Plasma cells are unusual among protein- possible that they inactivate histamine and
secreting cells in that their product is not other mediators released by these cells in
normally packaged and stored in conspicuous inflammation. The most dramatic eosinophil
cytoplasmic granules. Instead, it appears to responses are seen in parasitic diseases such
be transported from the Golgi complex to as schistosomiasis, trichinosis, and ascariasis
the cell surface in small vesicles and is re¬ in which circulating eosinophils may rise to
leased at the same rate as it is produced. 90 per cent of the leukocyte count. In hu¬
Occasional plasma cells contain spherical mans or experimental animals that have pre¬
inclusions called Russell bodies. These stain viously mounted an immune response to the
with histochemical reactions for both protein parasite, a second invasion is rapidly followed
and carbohydrate and also to some degree by accumulation of very large numbers of
with fluorescein-conjugated antibody against eosinophils around the parasite larvae. De¬
immunoglobulin. In electron micrographs granulation and release of MBP and other
they are found within distended cisternae of substances not yet identified results in killing
the endoplasmic reticulum. Although the gly¬ and destruction of the larvae. This killing
coprotein Russell bodies contain immuno¬ effect of the eosinophils is dependent on the
globulin, they are not regarded as normal presence of antiparasite antibody. In animals
secretory product but probably represent an lacking such antibodies, this rapid mobiliza¬
aberrant state of the cell. Whether their pres¬ tion and degranulation of eosinophils does
ence is indicative of a defect in synthesis or not take place, and dissemination of parasites
transport is not known. It is speculated that and severe disease may occur before immu¬
they may be accumulations of light chains nity develops.
not used in the assembly of complete immu¬
noglobulin molecules. Mast Cells
The train of evidence implicating plasma
cells in antibody production extends back The mast cells of connective tissue have
over 30 years and includes contributions several of the cytological and functional char¬
from several disciplines. It was first observed acteristics previously described for basophilic
that intensive immunization of animals was leukocytes (Chapter 4), but they are a distinct
attended by a marked increase in the number cell type. The basophils develop in the mar-
 CONNECTIVE TISSUE PROPER • 161

row, circulate in the blood, and migrate arise from preexisting mast cells by mitotic
through the endothelium of small venules division.
into the connective tissue. The mast cells Mast cells have a round or oval nucleus
originate in the connective tissue. They are and their cytoplasm is filled with several
especially abundant near the small blood ves¬ hundred granules (Fig. 5-22), which stain
sels and have long been thought to develop metachromatically with toluidine blue or
from persisting perivascular mesenchymal thionine. This property of changing the color
cells. However, recent experimental evidence of blue dyes to purple is due to their content
indicates that they differentiate from hemo¬ of heparin, a sulfated acid mucopolysaccha¬
poietic stem cells. Thus, it now seems likely ride.
that circulating stem cells enter the connec¬ In electron micrographs, mast cells are
tive tissues from the blood and there differ¬ observed to have numerous villous projec¬
entiate into mast cells. Mast cells can also tions and undulant surface folds (Fig. 5-23).

Macrophages

Eosinophilic leukocytes

Fibroblast Mast cells

Macrophages

Eosinophilic
Fibroblast leukocytes
Collagenous
fibers

Mast cells

Macro¬ Fibroblast
phage

Mast cel

V
Macrophage

Macro- Elastic fiber

phages

Figure 5-22. Mast cells in loose connective tissue of the rat. Upper figure was drawn from a preparation fixed and
stained with hematoxylin-eosin-azure II. Lower figure depicts a fresh preparation stained supravitally with neutral red
The mast cell granules have bound the dye. Other elements of the tissue are largely unstained. (After A. A. Maximov.)
162 • CONNECTIVE TISSUE PROPER

Figure 5-23. Electron micrograph of a mast cell from loose connective tissue of the rat.

The Golgi complex is well developed; the substances that serve to recruit other cell
endoplasmic reticulum is sparse and the mi¬ types to participate in local defense mecha¬
tochondria relatively few. The dense gran¬ nisms.
ules are bounded by a membrane and display Degranulation of mast cells can be experi¬
considerable variability in their substructure mentally induced by a number of nonspecific
from species to species. Their content is finely stimuli such as compound 48/80, polylysine,
granular in rodents, but they are quite het¬ polymyxin B, certain snake venoms, and by
erogeneous in humans, with coarse subunits lowering of the pH or osmolarity of the
consisting of scrolls of thin lamellae of dif¬ surrounding fluid. However, the stimulus
fering size and orientation. In some species most commonly invoking this response in
the granules are soluble in aqueous fixatives. vivo is invasion of the tissues by any foreign
It has long been known that mast cell substance (antigen) to which the individual is
granules contain heparin, an anticoagulant; sensitive (allergic). Thus, the activation and
histamine, which increases vascular permea¬ degranulation of mast cells is intimately re¬
bility; and serotonin, which causes contraction lated to the body’s immune system (Chapter
of smooth muscle. The conditions under i3).
which these substances are released and the In addition to the major class of antibodies
way in which they function in the connective (IgG) involved in humoral immunity, the
tissues have become better understood in blood serum contains a minor immunoglob¬
recent years. Mast cells are now recognized ulin component, designated IgE, which is
as sensitive sentinels detecting foreign sub¬ implicated in pathogenesis of allergies. The
stances and initiating a local inflammatory mast cell membrane contains from 50,000 to
response in the tissues. Located near mucosal 300,000 receptors that bind IgE molecules to
surfaces and along blood vessels, they are the cell surface (Fig. 5-24). When exposed
strategically situated to detect a variety of to specific antigen, the antigen molecules
threats to the host. Their activation results in form complexes with the surface IgE that
prompt release of the preformed chemical alter the permeability of the mast cell mem¬
mediators stored in their granules and in brane, permitting influx of calcium. The
slower generation of other biologically active membrane of the mast cell granules then
 CONNECTIVE TISSUE PROPER • 163

fuses with the plasmalemma, and the granule Mast cell granules also contain the enzymes
contents are discharged into the surrounding P -glucuronidase hexosaminidase, arylsulfatase,
connective tissue matrix. This whole process and chymase. These too are released by im-
of degranulation or explosive secretion is munologically activated mast cells. Their ef¬
completed in two to three minutes. The his¬ fects in the extracellular environment are
tamine released causes transudation of fluid poorly understood, but they may degrade
from dilated small venules, resulting in the glycosaminoglycans of the connective tis¬
edema and local swelling. If the antigen is sue ground substance.
inhaled, histamine released by mast cells on Much remains to be learned about the
the surface of the airways and in the sub¬ functions of mast cells. It is well established
mucosal connective tissue activates smooth that in allergic individuals their immunolog-
muscle, constricting bronchi and bronchioles, ically induced degranulation is responsible
thereby precipitating an asthma attack in for an important train of events in the path¬
susceptible individuals. The actions of hista¬ ogenesis of both immediate and more persis¬
mine are supplemented by similar effects of tent inflammatory processes. Recent investi¬
another mediator generated after granule gations have focused on these dramatically
release called slow-reacting substance of anaphy¬ rapid responses in sensitized animals and
laxis (SRS-A). In addition to these vasoactive allergic humans, but it is likely that under
and bronchoconstrictive mediators, discharge of normal conditions they secrete more or less
mast cell granules is attended by liberation continuously, at a slower rate. The possible
of chemotactic mediators—substances that at¬ significance of this activity for the homeosta¬
tract leukocytes. The best characterized of sis of the connective tissues is largely un¬
these is eosinophil chemotactic factor of anaphy¬ known.
laxis (ECF-A), which induces emigration of The secretory activity of mast cells in in¬
eosinophils from the blood and their aggre¬ flammatory reactions is widely accepted but
gation in the vicinity of activated mast cells. it is possible that they possess other nonse-

Plasma cells

Target tissues and cells

SMOOTH MUSCLE
MUCOUS GLANDS
SMALL BLOOD VESSELS
SENSORY NERVE ENDINGS
EOSINOPHIL LEUKOCYTES
BLOOD PLATELETS

mediators
Figure 5-24. Schematic representation of the role of mast cells in allergic reactions. IgE produced by mast cells binds
to IgE receptors in the mast cell membrane. On a second exposure, antigen binds to IgE and this event triggers mast
cell degranulation with release of histamine and other chemical mediators that affect various target tissues and organs.
164 • CONNECTIVE TISSUE PROPER

cretory functions in normal connective tissue. The omentum in man extends downward
They take up thorium dioxide, used as an from the greater curvature of the stomach
undigestible cell marker, and incorporate it like a loose curtain or veil over the intestines
in the granules. Marked cells are still detect¬ and is of great clinical importance in the
able in rats after ten months indicating, that limitation of disease processes in the abdom¬
under normal conditions mast cells are very inal cavity. When patients with a recently
long-lived, and there is less turnover of their perforated ulcer are operated upon, it is
granules than would be expected if they were usually found that the highly mobile omen¬
continuously secreting by degranulation. tum has already become locally adherent to
the wall of the gut in an effort of nature to
close the opening. Similarly, the omentum
SEROUS MEMBRANES adheres at sites of inflammation and tends to
\ wall off the process so that a local abscess will
The serous membranes, peritoneum, pleura, form instead of a generalized and often fatal
and pericardium, are thin layers of loose con¬ peritonitis. In addition to the protection af¬
nective tissue covered by a layer of meso- forded by the adhesion of the omentum, the
thelium. When the membranes are folded, free cells of its connective tissue constitute an
forming the omentum or the mesentery, both important mobile .reserve to combat infec¬
free surfaces are covered with mesothelium. tions in the peritoneal cavity.
The cavities lined by serous membranes al¬
ways contain a small amount of liquid, the Free Cells of the Serous Exudate
peritoneal fluid or pleural fluid. The cells in this
exudate originate from the serous mem¬ Normally, the amount of serous exudate
brane. in the body cavities is small, but in patholog¬
All the elements of the loose connective ical conditions it may increase enormously. It
tissue previously described are found in ser¬ contains a variety of freely floating cells,
ous membranes, such as the mesentery. Be¬ including (1) macrophages that originate in
cause they are very thin and require no the milky spots of the omentum and migrate
sectioning, the mesenteries have been favor¬ into the cavity; (2) desquamated mesothelial
ite sites for the microscopic study of loose cells that keep their squamous form or round
connective tissue. A mesentery contains a up; (3) small lymphocytes, the vast majority
loose network of collagenous and elastic fi¬ of which have migrated from the blood ves¬
bers, scattered fibroblasts, macrophages, mast sels of the omentum; (4) eosinophilic leuko¬
cells, and a varying number of fat cells. cytes of hematogenous origin; (5) free mast
Physiologically the most important and his¬ cells, which are especially abundant in serous
tologically the most interesting of the serous exudates of rats and mice; and (6) in patho¬
membranes in mammals is the omentum. The logical inflammatory exudates, enormous
membrane is pierced by innumerable holes numbers of neutrophilic leukocytes.
and is thus reduced to a fme lacelike net
formed by collagenous bundles covered by
mesothelial cells. Such thin, fenestrated areas
DENSE CONNECTIVE TISSUE
have few or no vessels. In the thicker areas
where the omentum is a continuous sheet,
macrophages are numerous. There are also Dense connective tissue differs from the
many small lymphocytes and plasma cells loose form mainly in the great preponder¬
and, occasionally, esoinophils and mast cells ance of the fibers over the cellular and matrix
(Fig. 5—25). The number of lymphocytes and components. Where the fiber bundles are
plasma cells varies considerably in different randomly oriented, the tissue is described as
animal species. dense irregular connective tissue. Where the fi¬
In certain areas, the macrophages and bers are oriented parallel to one another or
other free cells accumulate in especially dense in some other consistent pattern, it is called
masses. Such macroscopically visible areas are dense regular connective tissue.
often arranged along the blood vessels as
round or oval patches called milky spots. These Dense Irregular Connective Tissue
are sometimes found in the thin netlike part
of the omentum. They are especially char¬ This tissue is found in the dermis; in the
acteristic of the omentum of the rabbit. capsules of many organs; sheaths of tendons
 CONNECTIVE TISSUE PROPER • 165

Small
Mesothelial Mast lympho-
Small lymphocyte cell nucleus cell cyte Capillary

Elastic
Un differ¬
fiber
entiated
periuascu-
Macro¬ tar cell
phage
Capillary

Erythro-
cyte

Endothe¬
lium

Mast cell

Mesothe¬ Elastic
lial cell fiber
nucleus

Undiffer¬
entiated
perivascu¬ Collage¬
lar cell nous fibers

Fibroblast
Fibroblasts

Elastic fiber Fat cell Macrophages Hole Mesothelial cell nucleus

Figure 5-25. Drawing of a spread preparation of human omentum stained with hernatoxylin-eosin-azure II (After A A
Maximov.)
166 • CONNECTIVE TISSUE PROPER

Fiqure 5-26. Photomicrographs illustrating connective tissues with differing amounts of collagen. A, Loose connective
tissue from an 8-month-old fetus, showing relatively sparse, slender collagen fibers. B, Moderately dense irregular
connective tissue with coarse, irregularly oriented bundles of collagen. C, Dense connective tissue with very abundant
collagen in parallel wavy bundles.

and nerves; beneath the epithelium in parts plan, and the specific arrangement reflects
of the urinary tract; and in many other sites the mechanical requirements of the particu¬
in the body. Its structure in the dermis of lar tissue. Macroscopically, the tissue has a
the skin can be taken as typical. The elements distinctly fibrous structure and a character¬
are the same as in the loose connective tissue istic shining white appearance.
but the collagenous bundles are thicker and Tendon consists predominantly of Type I
are woven into a compact feltwork. They are collagen. It is composed of thick, closely
accompanied by extensive elastic networks. packed bundles of collagen fibrils oriented
The fibers from the dermis continue directly parallel to the long axis of the tendon. Deli¬
into those of the subcutaneous tissue, but cate networks of elastin have been described
there the fiber bundles are thinner and their between the bundles of collagen fibrils, but
arrangement is correspondingly looser. these are inconspicuous and detectable only
There is less ground substance in the dense after specific staining of elastin.
connective tissue. Among the densely packed In transverse sections examined with the
collagenous and elastic fibers are the cells, electron microscope, the collagen fibrils are
but these are much more difficult to identify of two sizes, one population averaging 60 nm
than in the loose tissue. The macrophages in diameter and the other 175 nm (Fig.
are recognizable only by vital staining. Along 5—28A). The relative numbers of the larger
the small vessels there are always many in¬ and smaller fibrils vary from tendon to ten¬
conspicuous fusiform cells, which are undif¬ don and in different areas of the same ten¬
ferentiated mesenchymal cells. don. Delicate cross-bridging strands of un¬
known nature extend laterally from fibril to
fibril (Fig. 5-285).
Dense Regular Connective Tissue
The fibroblasts, which are the only cells
The collagen bundles of regular connective present, are arranged in long parallel rows
tissue are arranged according to a precise in the spaces between the parallel collagenous
 CONNECTIVE TISSUE PROPER • 167

bundles. The cell bodies are rectangular, tri¬ ers may derive as much as 50 per cent of
angular, or trapezoidal when seen in surface their locomotor energy from this source. An¬
view, and rod-shaped when seen in profile. imals, such as the kangaroo, that have
Their cytoplasm stains darkly with basic dyes evolved a hopping gait make maximal use of
and contains a clear centrosome adjacent to this property of tendons. A kangaroo chang¬
the single flattened nucleus. Although the ing from walking to running actually de¬
limits between the successive cells in a row creases its oxygen consumption and at 30
are distinct, the lateral limits of the cells are miles per hour utilizes no more than an
indistinct. In a stained cross section of a animal half its size running on all four feet
tendon, the cells appear as dark star-shaped at the same speed. The tendons are thus an
figures between the collagenous bundles. A important source of “free” energy for loco¬
tendon consists of a varying number of small motion.
tendon bundles bound by loose connective Ligaments are similar to tendons, except
tissue into larger bundles (Fig. 5—27). that the elements are somewhat less regularly
In the tendon the fibers form a tissue that arranged. In other examples of dense regular
is flexible but offers great resistance to pull¬ connective tissue, such as the fasciae and apo¬
ing force. In certain forms of locomotion, neuroses, the collagenous bundles and fibro¬
the tendons of limb muscles are stretched blasts are arranged regularly in multiple
when the foot strikes the ground, acting like sheets or lamellae. In each layer the fibers
a tension spring. The subsequent recoil of follow a parallel and often slightly wavy
the tendons to their original length helps course. In the different layers the direction
push the foot off the ground in the next may be the same or it may change. The fibers
stride. This mechanical or elastic energy re¬ often pass from one layer into another.
quires no metabolic input such as that in¬ Therefore, a clear isolation of the sheets is
volved in muscle contraction. Human sprint- seldom possible. The cells that correspond to
the tendon cells adapt their shape to the
spaces between the collagenous bundles.
In dense connective tissue layers with
somewhat less regularly arranged elements,
such as the periosteum, sclera, and the like,
a section perpendicular to the surface shows
successive layers of collagenous bundles cut
in the longitudinal, oblique, or transverse
direction, and cells that are irregular, flat, or
fusiform. In these tissues there are always
gradual transitions to neighboring areas,
where the elements have a quite irregular,
dense arrangement. There is also no sharp
distinction between them and the surround¬
ing loose connective tissue.
The substantia propria of the cornea pro¬
vides an example of dense regular connective
tissue made up of successive layers of colla¬
gen with the fibrils of one layer oriented at
approximately 90 degrees to those in the next
layer (Fig. 5—29).

CONNECTIVE TISSUE WITH

SPECIAL PROPERTIES

Mucous Connective Tissue

Figure 5-27. Cross section of human tendon. Massive This tissue is found in many parts of the
bundles of collagen are separated by darker-staining
embryo and is a form of loose connective
loose connective tissue. Dots in the paler areas of colla¬
gen are nuclei of modified fibroblasts in narrow clefts tissue. The classic example of this type of
among the collagen fibers. connective tissue is Whartons jelly of the um-
Figure 5-28. Electron micrographs of human tendon. A, Transverse section of Achilles tendon showing variations in
the distribution and ratio of larger 175-nm fibrils to smaller 60-nm fibrils. B, High magnification of a small area of
plantaris tendon illustrating linear densities extending from fibril to fibril. (Micrographs from Dyer, R. F. Cell Tissue Res.
168:247, 1976.)

Figure 5-29. Electron micrograph of dense regular connective tissue of the cornea. The collagen is arranged in lamellae
with the fibrils in alternate layers oriented at right angles to those in adjacent lamellae. (Micrograph courtesy of M.
Jakus.)
168
 CONNECTIVE TISSUE PROPER • 169

bilical cord. The cells are large, stellate fibro¬ Elastic tissue forms layers in the walls of
blasts whose processes often are in contact hollow organs upon which a changing pres¬
with those of neighboring cells. A few mac¬ sure acts from within, as in the largest arteries
rophages and lymphoid wandering cells are and in the trachea and bronchi. In the large
also present. The intercellular substance is elastic arteries, the elastic tissue takes the
very abundant, soft, jelly-like, and homoge¬ form of a fenestrated membrane, a sheet of
neous in the fresh condition. When fixed, elastin of variable thickness provided with
much of it is extracted, and the residue many irregular openings. The fenestrated
contains granular and fibrillar precipitates. It membranes are arranged in multiple layers
has the staining reactions of mucin and con¬ concentric with the lun^en of the vessel and
tains thin, collagenous fibers that increase in are connected with one another by oblique
number with the age of the fetus. ribbon-like branches. The spaces between the
Examples of mucous connective tissue in lamellae contain a mucoid ground substance
adult animals are limited to the cockscomb and smooth muscle cells of irregular outline.
and the dermis and hypodermis of the sex Fibrous elastic networks, as well as fenes¬
skin of monkeys. In both of these the ground trated elastic membranes, exist in the walls
substance is extraordinarily abundant, is of of these vessels and it is difficult to distinguish
firm consistency, and is influenced by the sex clearly between the two in sections.
hormones.

Reticular Connective Tissue

Elastic Connective Tissue
Most of the fibrous elements around the
In the dense connective tissue of a few sinusoids of the liver and in the stroma of
stuctures in the body, elastic fibers predomi¬ lymphatic organs, hemopoietic tissue, and the
nate, and the tissue has a yellow color on spleen are blackened by silver stains and are
inspection with the naked eye. It may appear thus identified as reticular fibers. The pat¬
in the form of strands or sheets of coarse terns of the fibers in these examples of retic¬
parallel fibers, as in the ligamenta flava of ular connective tissue are also distinctive. Small
the vertebral column, in the vocal cords, in bundles of thin collagenous fibrils form com¬
the ligamentum stylohyoideum, and in the plex three-dimensional networks, whose in¬
ligamentum suspensorium penis. In these terstices are occupied by large numbers of
situations, the elastic fibers are thick and free cells. Stellate cells of mesenchymal origin
either round or flattened in cross section. are also associated with the argyrophilic re¬
They branch frequently and rejoin with one ticulum in these organs.
another at acute angles, as in a stretched
fishing net. In cross section the angular or
round areas representing the fibers form
small groups. The spaces between the elastic HISTOPHYSIOLOGY OF
fibers are filled with a delicate feltwork of CONNECTIVE TISSUE
collagenous fibrils and a few fibroblasts.
The classical example of dense regular elastic
Normal Functions
connective tissue is found in the massive liga¬
mentum nuchae of ruminants. It consists of Connective tissue functions in mechanical
coarse elastic fibers 10 to 15 pm in diameter, support, exchange of metabolites between blood
closely associated in parallel bundles. Its and the tissues, storage of energy reserves in
stretch and elastic recoil permits grazing an¬ adipose cells, protection against infection, and
imals to cut off grass with their lower incisors repair after injury.
with relatively little expenditure of energy. For its mechanical role the fibrous com¬
Scarpa’s fascia of the human anterior ab¬ ponents are most important, and their abun¬
dominal wall, which aids in the support of dance and distribution are adapted to the
the viscera, also consists largely of elastic local structural requirements. Delicate net¬
fibers. The corresponding layer in the large works of reticular fibers support the base¬
quadrupeds, called the tunica abdominalis, is ment lamina of epithelia, surround the cap¬
a thick yellow sheet of dense elastic tissue illaries and sinusoids, and envelop individual
several millimeters in thickness, which plays muscle fibers and the groups of parenchymal
an important role in supporting the viscera cells that form the functional units of organs.
while permitting some degree of abdominal The coarser collagenous fibers abound where
distention. greater tensile strength is required. They
170 • CONNECTIVE TISSUE PROPER

form the tendons and the aponeuroses, the which increases the caliber and rate of flow
septa and fibrous capsules of organs. Elastic through the small vessels, resulting in redness
fibers give the tissues their suppleness and and local elevation of tissue temperature.
their ability to spring back to their normal Increased permeability of the walls of post¬
relations after stretching. They are especially capillary venules results in escape of fluid,
abundant in hollow organs subject to periodic fibrinogen, and immunoglobulins into the
distention. Loose connective tissue, with its extravascular compartment, causing edema
abundant, highly hydrated ground sub¬ and associated swelling.
stance, is commonly found beneath the integ¬ Other chemical mediators induce selective
ument, between muscles, and in other sites margination of blood leukocytes, diapedesis,
where mobility of the parts is advantageous. and attraction to the site of injury. The poly¬
On the other hand, where strength is more morphonuclear leukocytes are the earliest
important than mobility, dense connective and most abundant emigrants, followed after
tissue is formed, and its bundles of collagen a day or so by monocytes that transform into
fibers tend to be oriented so as to resist most macrophages, which phagocytose bacteria
efficiently the local mechanical stresses. and tissue debris and eliminate them by in¬
Connective tissue plays a significant role in tracellular digestion. If the invading bacteria
nutrition of the other tissues that it surrounds are of species resistant to destruction, chronic
and permeates. It is evident that all sub¬ inflammation ensues with a change in the nature
stances reaching the cells of these other tis¬ of the cellular infiltration, with macrophages,
sues from the blood, and all the waste prod¬ lymphocytes, and plasma cells predominat¬
ucts of their metabolism that are returned to ing. If the tissue invasion is not successfully
the blood and lymph, must pass through a controlled by the mobilization of phagocytic
layer of connective tissue. These metabolites cells, the process is contained by proliferation
are believed to diffuse through the aqueous of fibroblasts and formation of a dense wall
phase of the gelatinous ground substance or of collagenous fibers.
along thin films of fluid coating the fibers. The mobilization of neutrophils and mon¬
The exchange of materials is probably influ¬ ocytes at the site of injury is the result of
enced by the viscous properties of the ground chemotaxis—a vectorial motility of these ceils
substance. The polyelectrolyte properties of toward substances that are generated by cer¬
its glycosaminoglycans suggest that the con¬ tain bacteria; by activation of serum comple¬
nective tissue extracellular matrix may also ment; and by activation of Factor XII of the
participate in maintaining water and electro¬ blood clotting\system, which leads to forma¬
lyte balance. In addition to the storage of tion of the chemotactic agents kallikrein and
energy in adipose cells, it is noteworthy that plasminogen activator. The bacteria are ren¬
approximately half of the circulating proteins dered more susceptible to phagocytosis by
of the body are in the interstitial spaces and, being coated with serum complement and
because the proportions of albumin and glob¬ specific antibody that gain access to them as
ulin there differ from those in plasma, it is a consequence of the increased permeability
speculated that the connective tissue may of the blood vessels. This enhancement of
exercise some selectivity in its depot function. phagocytosis is called opsonization. Some en¬
capsulated bacteria are resistant to destruc¬
tion by phagocytes because they appear to
Inflammation
have the capacity to inhibit fusion of lyso-
Of great importance in the defense against somes with the phagocytic vacuoles.
disease is the reaction of the connective tis¬
sue, called inflammation. A detailed considera¬ Repair
tion of this process falls into the sphere of
general pathology, but some awareness of its The regenerative capacity of fibroblasts
mechanisms is essential to an understanding and the fact that they respond so readily to
of the functions of the cellular elements of injury by proliferation and fibrogenesis make
the blood and connective tissues. them the principal agents of repair. They are
Bacteria or other injurious agents breach¬ involved in the healing of defects, not only
ing the mechanical barrier provided by epi- in connective tissue proper, but also in other
thelia induce an intense reaction in the con¬ tissues that have little or no regenerative
nective tissue, resulting in local redness, capacity of their own. For example, the heart
swelling, heat, and pain. Tissue damage muscle that degenerates following a heart
causes release of histamine from mast cells, attack is replaced by a connective tissue scar.
 CONNECTIVE TISSUE PROPER • 171

Although much has been learned about ized by bone deformities has been traced to
the production of collagen in the histioge- the ingestion of the sweet pea Lathyrus odor-
nesis and repair of connective tissue, it must atus. All the abnormalities of this disease,
be realized that there are also mechanisms called lathyrism, can be reproduced by admin¬
foi the removal of collagen. This is of great istration of (3-aminopropionitrile, the toxic
importance in the growth and remodeling of agent extracted from the sweet pea plant.
bone and involves local production and re¬ Tropocollagen appears to be synthesized nor¬
lease of the enzyme collagenase. Collagenase mally in such animals but is defective in its
is also detected in mammalian tissues subject ability to form stable collagen fibrils. When
to cyclic regression, such as the uterine en¬ tropocollagen is extracted at 0° C from normal
dometrium.
animals and induced to form fibers by an
increase in the temperature to 37° C, the
Hormonal Effects longer it is held at that temperature the less
soluble it becomes at 0° C. This time-depend¬
The adrenocorticotropic hormone of the ent loss of solubility is believed to be due to
pituitary and cortisone of the adrenal cortex formation of increased numbers of bonds
both tend to lower the glycosaminoglycan between the tropocollagen units. In lathyritic
content of the ground substance. They also animals there is a marked increase in the
diminish the intensity of the cellular response extractability of tropocollagen from connec¬
in inflammation. The response of connective tive tissue at 0° C, and although the extracted
tissues to sex hormones varies greatly with tropocollagen is capable of forming typical
the species, the sex, and the site in the body. cross-striated fibers on warming, these fibers
The most dramatic effects are on the sex skin retain the ability to redissolve upon cooling
of monkeys, where estrogens greatly increase even after prolonged periods at 37° C. This
the glycosaminoglycans of the extracellular and other evidence suggests that ingestion of
matrix and in the cockscomb, where testos¬ lathyritic agents results in synthesis of abnor¬
terone stimulates accumulation of hyaluronic mal chains that are not able to cross-link
acid and formation of collagen. Less dramatic adequately.
hormonal effects are detected in the human A rare inherited disease of humans, Ehlers-
female in the periodic increase in hydration Danlos syndrome, is characterized by short stat¬
of the tissues during certain phases of the ure, unusually stretchable skin, hypermobil-
menstrual cycle. ity of joints, and a tendency for joint dislo¬
cations. There is a related disease of cattle
Disturbances of Collagen Metabolism and sheep called dermatosparaxis in which the
skin is fragile and very easily torn. Recent
Collagen metabolism is affected by age and evidence indicates that the defect may reside
nutrition and may be rather specifically dis¬ in an abnormally low activity of the enzyme
turbed in a number of disease states. In some procollagen peptidase, which normally converts
of these specific disorders, recently acquired procollagen to collagen by cleaving off a
knowledge of the mechanisms of hbrogenesis peptide from the amino-terminal ends of the
has provided a partial explanation of the cq and a2 chains of procollagen.
defect or has made it possible to identify the
step in the process where the normal biosyn¬
thetic mechanism fails. It has long been
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Griffin, F. M., J. A. Griffin, J. E. Leider, and S. C.
Gotte, L., M. G. Giro, D. Volpin, and R. W. Horne: The
ultrastructural organization of elastin. J. Ultrastruct. Silverstein: Studies on the mechanism of phagocy¬
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Ross, R., and P. Bornstein: Elastic fibers in the body. Uananue, R. R., and J. Cerotti: The function of mac¬
rophages in the immune response. Semin. Hematol.
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Ross, R., and P. Bornstein: The elastic fiber, j. Cell Biol. 7:225, 1970.
Weiss, L. P., and D. W. Fawcett: Cytochemical observa¬
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tions on chicken monocytes, macrophages and giant
Sandberg, L. B., N. T. Soskel, and J. G. Leslie: Elastin
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GROUND SUBSTANCE MAST CELLS

Balacz, E. D.: Glycosaminoglycans and proteoglycans. In Dvorak, H. F., and A. M. Dvorak: Basophils, mast cells
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1970. Fawcett, D. W.: An experimental study of mast cell
Compar, W. D., and T. C. Laurent: Physiological func¬ degranulation and regeneration. Anat. Rec. 727 :29,
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Rev. 58:255, 1978. Ishizaka, H., H. Tomioka, and T. Ishizaka: Mechanism
Hascall, V. C., and G. K. Hascall: Proteoglycans. In Hay, of passive sensitization. I. Presence of IgE and IgG
E. D., ed.: Cell Biology of the Extracellular Matrix. molecules on human leukocytes. J. Immunol.
New York, Plenum Press, 1981. 765:1459, 1970.
Lindahl, U., and M. Hook: Glycosaminoglycans and Ishizaka, T.: Functions and development of cell recep¬
their binding to biological macromolecules. Annu. tors for IgE. In Johannsen, Strandberg, and Uvnas,
Rev. Biochem. 47:385, 1978. eds.: Molecular and Biological Aspects of Acute
Rosenberg, L., W. Heilman, and A. K. Kleinschmidt: Allergic Reactions. New York, Plenum Publishing
Electron microscopic studies of proteoglycan aggre¬ Co., 1976.
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Timpl, R., H. Rohde, P. G. Robey, S. I. Rennard, J. M. Invest. Dermatol. 77:81, 1978.
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 ADIPOSE TISSUE

Adipose tissue was long considered to be a is less abundant and occurs only in certain
metabolicaily inert tissue that passively stored specific areas. There are marked species dif¬
fat, provided insulation against heat loss, and ferences in the relative amounts of the two
functioned in mechanical support in certain types of fat. Brown adipose tissue is most
regions of the body. The allocation of a abundant in hibernating species. Although it
separate chapter to it in recent textbooks of is present in primates, including man, it is
histology is a consequence of its belated rec¬ relatively inconspicuous and probably does
ognition as a diffuse organ of primary met¬ not assume great importance in the economy
abolic importance. of these species.
Most animals feed intermittently but con¬ The peripheral parts of lobules of brown
sume energy continuously; there must there¬ adipose tissue often have an appearance
fore be provision for temporary storage of strongly suggestive of a transition from one
fuel. Lipid is the most favorable substance form of fat to the other. This has fostered
for this purpose because it weighs less and the widespread belief that brown fat is simply
occupies less volume per calorie of stored an immature or transitional form of ordinary
chemical energy than either carbohydrate or adipose tissue. For this reason it is sometimes
protein. Although many tissues contain small referred to in the literature of pathology as
amounts of carbohydrate and fat, the adipose fetal fat. The term is not appropriate how¬
tissue serves as the body’s most capacious ever, for in those species in which it is best
reservoir of energy. About 10 per cent of the developed, brown fat persists throughout
total body weight of an average human is fat, adult life and is morphologically and meta-
representing approximately a 40-day reserve bolically sufficiently different to warrant its
of energy. In obese individuals this may in¬ designation as a distinct type of adipose tis¬
crease to the equivalent of a year or more of sue. We will return to this point in discussing
normal metabolism. By accumulating lipid in the histogenesis of the adipose tissues.
periods of excess food intake and releasing
fatty acids in periods of fasting, adipose tissue
plays an important role in maintaining a
Unilocular Adipose Tissue
stable supply of fuel. Far from being inert, Fat varies in color from white to deep
the cells of this tissue are capable of actively yellow, depending in part on the abundance
synthesizing fat from carbohydrate and are of carotinoids in the diet. The color resides
highly responsive to hormonal and nervous mainly in the stored lipid. The cells are very
stimulation. large, ranging up to 120 pm in diameter.
They are typically spherical but may assume
polyhedral shapes because of mutual defor¬
HISTOLOGICAL mation (Fig. 6-1). A single droplet of lipid
occupies most of the volume of the cell.
CHARACTERISTICS OF THE Therefore, adipose cells of this kind are de¬
ADIPOSE TISSUES scribed as unilocular to distinguish them from
brown fat cells, which contain multiple small
In most mammals there are two distinct droplets and are described as multilocular.
types of adipose tissue, which differ in their The nucleus is displaced to one side by the
color, distribution, vascularity, and metabolic accumulated lipid and the cytoplasm is re¬
activity. One is the familiar yellow or white duced to a thin rim constituting only about
adipose tissue, which composes the bulk of one fortieth of the total volume of the cell.
body fat. The other, called brown adipose tissue, The lipid is usually extracted during prepa-
174
 ADIPOSE TISSUE • 175

of endoplasmic reticulum, and a moderate

number of free ribosomes. The thin layer of
cytoplasm surrounding the lipid droplet also
contains a few mitochondria, 10-nm fila¬
ments, and minute vesicles, which may rep¬
resent the agranular reticulum. Not infre¬
quently there also are small droplets of newly
formed lipid that has not yet coalesced with
the principal lipid drop (Figs. 6-3 and 6-4).
The lipid droplet is not bounded by a mem¬
brane but its interface with the cytoplasm
may stain more intensely, giving a specious
appearance of a limiting membrane. A reg¬
ular network of orthogonally arranged 6-nm
filaments is often observed around the lipid
droplet (Fig. 6-6). Similar filaments occur¬
ring singly or in small bundles are found
randomly oriented elsewhere in the cyto¬
plasm. Each adipose cell is invested by a layer
of glycoprotein resembling the basal lamina
of epithelia. The plasma membrane shows
numerous minute invaginations of the kind
that are usually interpreted as evidence for a

Figure 6-1. Common unilocular adipose tissue prepared

by routine methods. The lipid droplet of the cells has been
extracted during dehydration and only a thin rim of cyto¬
plasm remains.

ration of histological sections, so that only

the plasmalemma and a thin shell of cyto¬
plasm remain. With silver stains each cell is
found to be surrounded by delicate reticular
fibers. In the angular spaces between the cells
are cross sections of capillaries that form a
loose plexus throughout the tissue. If well
preserved, adipose tissue appears in section
as a delicate network with large polygonal
meshes (Figs. 6-1 and 6-2), but the cell rims
often collapse to varying degrees during
preparation, giving the cells an irregular out¬
line.
Adipose tissue is often subdivided into
small lobules by connective tissue septa. This
compartmentation, visible with the naked
eye, is most obvious in regions where the fat
is subjected to pressure and has a cushioning
or shock-absorbing effect. In other regions,
the connective tissue septa are thinner and
the lobular organization of the tissue is less Figure 6-2. A thin spread of adipose tissue of mesentery
apparent. stained with Sudan black without previous dehydration.
Examined with the electron microscope, Here the lipid droplet has been retained and is stained by
the cytoplasm near the nucleus is found to the fat-soluble dye, while the surrounding rim of cytoplasm
is unstained. (From Fawcett, D. W. In Greep, R. O., ed.:
contain a small Golgi complex, a few filamen¬ Histology. Philadelphia, Blakiston Co., 1953. Reproduced
tous mitochondria, occasional short profiles by permission of McGraw-Hill Book Co.)
176 • ADIPOSE TISSUE
v

Figure 6-3. Electron micrograph of portions of two unilocular adipose cells from the epididymal fat pad of the rat. Notice
the relative sizes of the neutrophil leukocyte and the very large fat cell. The cell at upper right has several small lipid
droplets that have not yet coalesced with the large lipid droplets.

Figure 6-4. Micrographs of the rim of cytoplasm of an adipose cell. A few of the numerous small lipid droplets appear
to be discharging their contents into the main lipid droplet at the top of the figure.
 ADIPOSE TISSUE • 177

Figure 6-5. Micrograph of a portion of a developing adipose cell. After glutaraldehyde fixation the lipid often is only
slightly stained by osmium, and the interface between the lipid and the cytoplasm (at arrows) can be seen more clearly.
Notice the absence of a membrane around the lipid. (Micrograph courtesy of E. Wood.)
178 • ADIPOSE TISSUE
\

nal lamina no longer conforms to the cell

surface but becomes folded. The cells never
revert to simple fusiform elements resem¬
bling fibroblasts.

Distribution of Unilocular Adipose

Tissue (White Fat)
This type of fat is widely distributed in the
subcutaneous tissue but exhibits regional dif¬
ferences, which are influenced by age and
sex. In infants and young children there is a
continuous subcutaneous layer of fat, the
panniculus adiposus, over the whole body. In
adults it thins out in some regions but persists
and grows thicker in certain sites of predilec¬
tion. These sites differ in the two sexes and
are largely responsible for the characteristic
differences in body form of males and fe¬
males. In the male, the principal areas are
the nape of the neck, the subcutaneous area
overlying the deltoid and triceps muscles, the
lumbosacral region, and the buttocks. In the
female, subcutaneous fat is most abundant in
the breasts, the buttocks, the epitrochanteric
region, and the lateral and anterior aspects
of the thighs.
Figure 6-6. Micrograph of portions of two lipid droplets
and the intervening cytoplasm in a developing adipose
cell. The lipid-cytoplasm interface is sectioned obliquely,
revealing (at arrows) an ordered array of 10-nm filaments
at the boundary. (Micrograph courtesy of E. Wood.)

submicroscopic form of pinocytosis. The sig¬

nificance of these vesicles has been the subject
of considerable debate. It has been suggested
that they may be involved in uptake of ma¬
terials used by the cell in lipid synthesis.
However, their number is reported to in¬
crease greatly during prolonged starvation
and after norepinephrine administration.
This has led to the speculation that fatty acid
and glycerol formed during lipolysis may be
transported in vesicles to the cell surface for
release. Since it is not easy to determine from
fixed images the direction of vesicular trans¬
port, the role of these vesicles in the economy
of the adipose cell remains unresolved.
In prolonged fasting or in the emaciation
associated with chronic illness, adipose tissue
may give up much of its stored lipid and
revert to a highly vascular tissue containing
aggregations of ovoid or polygonal cells with
multiple small lipid droplets. In electron mi¬
crographs of fat cells decreasing in size dur¬
ing fasting, the cell surface becomes highly Figure 6-7. Photomicrograph of typical multilocular adi¬
pose tissue (brown fat). The polygonal cells contain more
irregular in outline with numerous pseudo¬ cytoplasm than in white fat and have multiple lipid droplets
pod-like processes, and the redundant exter¬ of varying size.
 ADIPOSE TISSUE • 179

In addition to these superficial fat deposits, the cell. A small Golgi apparatus is present,
there are extensive accumulations in both as well as numerous large spherical mito¬
sexes in the omentum, mesenteries, and ret¬ chondria. In electron micrographs the mito¬
roperitoneal areas. All these areas readily chondria occupy a large part of the cytoplasm
give up their stored lipid during fasting. and have numerous cristae that may extend
There are other areas of adipose tissue, how¬ across the full width of the organelle (Fig.
ever, that do not give up their stored fuel so 6-8). The endoplasmic reticulum is not well
readily. For example, the adipose tissue in developed, and only a few profiles of the
the orbit, in the major joints, and on the smooth-surfaced form can be found. The
palms of the hands and soles of the feet does lipid droplets do not appear to develop
not seem to be grist for the metabolic mill within the reticulum but are free in the
but instead has the mechanical function of cytoplasm. Scattered ribosomes and variable
support. These areas diminish in size only amounts of glycogen are also present in the
after very prolonged starvation. cytoplasmic matrix.
The connective tissue stroma of brown
Muitilocular Adipose Tissue (Brown adipose tissue is very sparse and the blood
Fat) supply exceedingly rich (Fig. 6-9). The cells
are therefore in more intimate association
The color of this form of adipose tissue with one another and with the capillaries than
ranges from tan to a rich reddish brown. Its is the case in unilocular fat. Numerous small
cells are smaller than those of white fat and unmyelinated nerve fibers can be demon¬
are polygonal in section. The cytoplasm is strated among the brown fat cells by silver
more abundant and there are multiple lipid staining methods and in electron micro¬
droplets of varying size (Fig. 6-7). The spher¬ graphs. Naked axons are frequently encoun¬
ical nucleus is somewhat eccentric in position tered in close apposition to the surface of the
but is seldom displaced to the periphery of adipose cells.

Figure 6-8. Electron micrograph of the nucleus and adjacent cytoplasm of a brown adipose cell from a bat recently
aroused from hibernation. Typical of this tissue is a great abundance of very large mitochondria with cristae traversing
the entire width of the organelle.
180 • ADIPOSE TISSUE

no new areas develop after birth. This is in

contrast to ordinary fat cells, which may de¬
velop in almost any area of loose connective
tissue, and new adipose cells may appear
at any time in postnatal life. Brown adipose
tissue may not occur in all mammals, but its
presence has been established in representa¬
tives of at least seven of the orders, including
primates. It is prominent in the newborn of
all the species in which it occurs and is a
distinct and conspicuous tissue in the adults
of hibernating species. In some nonhibernat¬
ing species, including man, the multilocular
condition of the lipid in its cells gradually
diminishes postnatally by coalescence of the
droplets, so that the cells may gradually come
to resemble those of unilocular adipose tis¬
sue. For this reason, there has been some
debate as to whether or not there are two
physiologically distinct types of adipose tissue
in well-nourished human adults. The bulk of
the evidence now indicates that two types do
exist, even though they may be difficult to
distinguish morphologically in well-nour¬
ished adults. Brown fat is well differentiated

Figure 6-9. Photomicrograph of a thick section of brown

and white adipose tissue in which the blood vessels have
been injected with India ink. The vascular network of the
brown adipose tissue (above) is extraordinarily rich and
has a glandlike pattern, while that of the white fat (below)
is relatively sparse. (From Fawcett, D. W. J. Morphol.
90:363, 1952.)

The histological organization of brown fat

is always distinctly lobular, and the pattern
of distribution of the blood vessels within
lobes and lobules resembles that found in
glands. In animals subjected to prolonged
fasting the brown fat gradually becomes
more deeply colored and reverts to a com¬
pact, glandlike mass of epithelioid cells bear¬
ing no resemblance to a connective tissue
(Fig. 6-10). The depletion of lipid in brown
adipose tissue is more rapid in animals sub¬
jected to a cold environment.
The brown color of the tissue is in large
part attributable to the high concentration of
cytochromes in its extraordinarily abundant
mitochondria. The relative oxidative capacity
of brown adipose tissue, based on cytochrome
oxidase, is greater than that of cardiac mus¬
cle.

Distribution of Multilocular Adipose

Tissue Figure 6-10. Brown adipose tissue that has been de¬
pleted of lipid after prolonged fasting, or after hypophy-
Brown or multilocular adipose tissue arises sectomy, takes on the appearance of a compact glandular
epithelium. Its cells bear no resemblance to fibroblasts.
in embryonic life in certain specific sites, and
 ADIPOSE TISSUE • 181

as early as the twenty-eighth week in the lar adipose cells differentiate from special
human fetus, and in the newborn constitutes formative cells, called lipoblasts, that arise
about 2 to 5 per cent of total body weight. from mesenchymal cells in late fetal and early
In adults, all the fat may appear to be of the postnatal life. According to this interpreta¬
unilocular variety, but in the elderly, in per¬ tion, the characteristic pattern of fat deposits
sons with chronic wasting diseases, or in star¬ in the adult would reflect the distribution
vation, glandlike masses of multilocular fat and relative abundance of the lipoblasts
cells reappear in the same regions where they formed in the fetal and neonatal period.
are found in the newborn. Moreover, two These hypothetical lipoblasts are fusiform or
types of lipomas (fatty tumors) occur in stellate cells with no cytological features that
man—one resembling unilocular adipose tis¬ clearly distinguish them from other relatively
sue and the other resembling brown adipose undifferentiated connective tissue cells. Some
tissue. These observations lend support to histologists therefore consider it unnecessary
the view that both types of adipose tissue are to postulate a separate category of committed
represented in man throughout life. stem cells but prefer to interpret the imme¬
In the common laboratory rodents, brown diate precursors of adipose cells as persisting
fat occurs in two symmetrical interscapular pluripotential mesenchymal cells.
fat bodies, in thin lobules between muscles The view prevailing now is that there are
around the shoulder girdle, and in the axil¬ two processes of adipose tissue formation in
lae. It fills the costovertebral angle and forms mammals (Fig. 6—11). In one taking place in
long slender lobules on either side of the fetal life and called primary fat formation, spe¬
thoracic aorta. Smaller lobules are also found cial epithelioid precursor cells are laid down
in the anterior mediastinum, along the great in lobular, glandlike arrangements. These
vessels in the neck, and in the hilus of the accumulate multiple lipid droplets and be¬
kidney. Brown adipose tissue is less extensive come the multilocular adipose tissue found
in primates, but in young macaques and in in most mammalian species. In addition, adi¬
newborn humans, sizable masses can be pose cells can arise in late fetal and early
found in the axillae and in the posterior postnatal life by accumulation of lipid in
triangles of the neck. Smaller lobules are relatively undifferentiated cells of connective
found near the thyroid, along the carotid tissue without these cells becoming organized
sheath, and in the hilus of the kidney. into epithelioid glandlike lobules. This secon¬
dary fat formation results in the disseminated
unilocular adipose tissue of the adult.
HISTOGENESIS OF ADIPOSE These two distinct modes of histogenesis
of fat are identifiable in the human. In post¬
TISSUE natal life, however, the multiple lipid droplets
in cells of areas of primary fat formation
Histogenesis of adipose tissue has long tend to coalesce, resulting in a tissue closely
been controversial and unanimity has yet to resembling unilocular adipose tissue. Thus,
be achieved. Histologists of the last century in the well-nourished adult there appears to
considered adipose tissue to be merely loose be only one morphological type of adipose
connective tissue in which lipid had accu¬ tissue, but there are in fact two, differing in
mulated in many of the fibroblasts. Accord¬ their ontogeny and physiology.
ing to this interpretation, any and all connec¬ In affluent developed countries, obesity is
tive tissue could become adipose tissue when a major health problem. Excess adipose tissue
dietary intake exceeded energy expenditure. puts an added strain on the circulatory sys¬
Connective tissue is ubiquitous; however, in tem and increases the risk for those predis¬
obesity, adipose cells do not become univer¬ posed to myocardial infarction and hyper¬
sally and evenly distributed but develop pref¬ tension. Obesity developing in adult life com¬
erentially in certain sites while others remain monly results from accumulation of excess
unchanged. For example, the eyelids, nose, lipid in a normal number of unilocular adi¬
ears, scrotum and genitalia, and the back of pose cells (hypertrophic obesity). In severe obe¬
the hands and feet rarely accumulate fat. sity, however, there may also be a greater than
This would be hard to explain if adipose cells normal number of adipose cells (hypercellular
could arise from fibroblasts wherever they obesity). The precursor cells are formed in the
occurred. early postnatal period and adipose cells are
Later histologists maintained that unilocu¬ incapable of proliferation later in life. There
182 • ADIPOSE TISSUE

Multilocular

Figure 6-11. Schematic representation of histogenesis of the adipose tissues. Mesenchymal cells differentiate into
fibroblasts and two kinds of lipoblasts. One forms glandlike aggregations of epithelioid cells that accumulate multiple
lipid droplets and develop into multilocular adipose tissue. In primates, including humans, the lipid droplets may coalesce
to varying degrees in well-nourished adults, so that the tissue comes to resemble unilocular adipose tissue. The other
kind of lipoblast, which is fusiform and more widely dispersed in the embryonic connective tissues accumulates lipid
that coalesces into a single large droplet, forming typical unilocular adipose tissue. In prolonged fasting, when the lipid
is depleted, the two types of adipose tissue revert to tissues of different appearance. Neither reverts to a spindle-shaped
cell that could be mistaken for a fibroblast.

is now experimental and clinical evidence HISTOPHYSIOLOGY OF

that overfeeding in the early weeks of life ADIPOSE TISSUE
can induce formation of greater numbers of
adipose cell precursors, resulting in a greater
risk of hypercellular obesity in adult life. Since the introduction of isotopic tracers
Infants achieving an early body weight .over for use in studying metabolism, it has been
the 97th percentile are reported to be three clearly shown that the lipid in fat depots is
times as likely to become obese adults as are not an inert energy reserve drawn upon only
other infants. Conversely, infants who were in periods of inanition. On the contrary, the
born in a period of famine in Europe near lipid is continuously being mobilized and
the end of World War II had roughly one renewed even in an individual in caloric bal¬
third the incidence of adult obesity compared ance. The half life of depot lipid in the rat is
with a similar group born in a period of about eight days, which means that almost 10
plenty in the first summer of peace. per cent of the fatty acid stored in adipose
Thus, it seems clear that the level of nutri¬ cells is replaced each day by new fatty acid.
tion in the early weeks of life can influence The same kind of continual renewal occurs
the number of adipose cell precursors. While in man, but the quantities and time course
adult hypertrophic obesity may occur from of these events are not known with the same
dietary excess in anyone, severe hypercellular precision as in laboratory animals.
obesity is more likely to occur in those who The histophysiology of adipose tissue can
were overnourished as infants. best be understood by analogy to deposits
 ADIPOSE TISSUE • 183

and withdrawals from a metabolic reserve increases up to 100-fold. Comparable in¬

bank or revolving fund. The “deposits” may creases are measured by incorporation of
be in the form of (1) fatty acids from the exogenous fatty acid into triglyceride. The
chylomicrons formed from dietary lipid, (2) activity of lipoprotein lipase also increases
fatty acids synthesized from glucose in the two orders of magnitude during in vitro
liver and transported to the adipose tissue in adipose conversion.
the form of serum lipoprotein, or (3) triglyc¬
eride synthesized from carbohydrate in the
adipose cells themselves. “Withdrawals” are
Hormonal Influences
made by enzymatic hydrolysis of triglyceride In many tissues under endocrine control,
and release of free fatty acids into the blood. the hormones act by stimulating the enzyme
With a continuous supply of glucose, lipolysis adenyl cyclase in the cell membrane, which
and release of free fatty acids are negligible. generates from ATP, the intracellular “sec¬
With alternations of fasting and feeding, ond-messenger” cyclic AMP. The differing
which is the usual feeding pattern, lipolysis responses of various target organs to this
is increased severalfold during periods of common messenger depend on its activation
fasting. The normal balance is greatly af¬ of organ-specific protein kinases. Several hor¬
fected by hormones and by the action of the mones affect the release of fatty acid from
nervous system. adipose tissue—the hypophyseal hormones
In the deposition of fat from circulating adrenocorticotropin (ACTH), thyrotropin
chylomicrons and very-low-density lipopro¬ (TSH), and luteinizing hormone (LH) and
teins, the triglycerides of these blood-borne the adrenomedullary hormone epinephrine.
particles are hydrolyzed in the adipose tissue The fat cell membrane contains specific re¬
capillaries by an enzyme, lipoprotein lipase, ceptors for each of these. The receptors reg¬
which is believed to be synthesized by adipose ulate access of the hormones to adenyl cy¬
cells and subsequently localized in membrane clase. The cyclic AMP generated within the
at the luminal surface of the endothelium in adipose cell activates lipase that degrades
the neighboring capillaries. The fatty acid stored triglyceride to fatty acids and glycerol,
resulting from lipoprotein hydrolysis tra¬ which are released into the bloodstream. The
verses the endothelium, diffuses across the importance of this process is evident when
narrow extracellular space, and enters the one considers that in humans, it results in
adipose cells. The role of micropinocytotic release of nearly 200 g of free fatty acid a
vesicles in fatty acid transport and uptake is day, the oxidation of which in other tissues
moot. In the thin rim of cytoplasm, fatty accounts for about 80 per cent of the basal
acids are combined with a glycerophosphate, oxygen consumption.
an intermediate product of glucose metabo¬ The characteristic differences in adipose
lism, to form triglyceride (neutral fat), which tissue distribution in males and females has
is added to the lipid droplet. The smooth already been alluded to. Because hormones
endoplasmic reticulum is the organelle prin¬ circulate freely in the blood and presumably
cipally involved in the reesterihcation of the reach all tissues in approximately equal con¬
fatty acids to form the triglycerides. centrations, these differences in fatty distri¬
Much has been learned in recent years bution imply either that there are genetically
about the morphological and biochemical dif¬ determined differences in distribution of
ferentiation of adipose cells by studying a cell cells having the capacity to develop into fat
line, designated 3T3, which originated from cells, or that there are regional differences
disaggregated cells of a mouse fetus at the in sensitivity of the cells to circulating sex
stage when adipose cell precursors were first hormones. This regional difference in sensi¬
appearing. When growth is arrested in cul¬ tivity does not seem to be restricted to the
tures of these 3T3 preadipose cells, they effects of sex hormones, for an excess of
exhibit in vitro the same morphological and adrenocortical hormone results in a characteris¬
biochemical changes that occur in adipose tic distribution of fat, of which a prominent
tissue development in vivo. Lipid droplets feature is an accumulation of fat over the
accumulate and the cells progressively as¬ lower cervical region, producing a deformity
sume the spherical shape of fat cells. Accom¬ referred to as “buffalo hump.”
panying triglyceride accumulation, the rate The hormone insulin is the main physiolog¬
of their endogenous fatty acid synthesis, ical factor controlling the uptake of glucose
measured by incorporation of labeled acetate, by adipose tissue and the synthesis of fat
184 • ADIPOSE TISSUE \

from carbohydrate. Whether given in vivo or of insulin, the required energy is derived
added to the medium of adipose tissue in¬ principally from fat.
cubated in vitro, it appears to stimulate the
transport of glucose into the cell and to Influence of the Autonomic Nervous
accelerate its metabolism along all of the System
paths open to it. It has an effect on the rate
at which glucose is converted to glycogen and Adipose tissue is rather richly innervated,
also acts on fatty acid synthetase. Oxygen especially the brown fat. The function of the
consumption is stimulated because of the nerves can be demonstrated experimentally
accelerated conversion of glucose to fatty by cutting the nerves of the interscapular fat
acid. The effects of insulin in promoting body on one side of the midline, leaving the
glycogen deposition can be demonstrated nerve supply to the other side intact. Within
morphologically and are very much more the first few postoperative days it becomes
pronounced in brown than in white fat (Figs. apparent that the fat cells on the denervated
6-12, 6-13). side are larger than those on the normal side.
In the absence of insulin, as in diabetes, The differences in the two sides are more
there is a rise in blood glucose, a diminished dramatic if the animal is then deprived of
utilization of glucose, an increase in unester- food and placed in a cold environment.
ihed fatty acids in the plasma, and an increase These environmental conditions ordinarily
in blood lipoproteins. Carbohydrates are nor¬ lead to a rapid mobilization of lipid from the
mally used preferentially as an energy source, fat depots. In animals unilaterally dener¬
but in the diabetic, in whom carbohydrates vated, the fat cells on the side with the nerves
cannot be utilized because of the deficiency intact are rapidly depleted of lipid, while the

Figure 6-12. A, White adipose tissue of a rat refeeding after a period of fasting. The dark granular deposits in the rim
of cytoplasm are glycogen, which subsequently disappears as the carbohydrate is used in synthesis of triglycerides. B,
Under the same experimental conditions, considerably more glycogen is deposited in multilocular adipose tissue, but
not all cells accumulate carbohydrate to the same degree. Some, therefore, stain relatively little with the periodic
acid-Schiff reaction used here. A similar deposition of glycogen can be induced in unfasted animals by administration
of excess insulin. (From Fawcett, D. W. J. Morphol. 90:363, 1952.)
 ADIPOSE TISSUE • 185

Very low density lipoprotein

/ Chylomicron

Figure 6-13. Schematic representation of lipid transport between the blood and adipose cells. An enzyme, lipoprotein
lipase, probably synthesized by adipose cells, is localized in the endothelium of the adjacent capillaries. Lipid is carried
in the bloodstream to the adipose cells in the form of chylomicrons from the intestine and very-low-density lipoproteins
from the liver. In the capillaries, these are hydrolyzed by lipoprotein lipase to fatty acids and glycerol, which diffuse to
the cytoplasm of the adipose cells where they are reesterified to triglyceride, which is added to the stored lipid. Glucose
diffusing to the fat cells may also be used in the synthesis of triglycerides from carbohydrate. Upon neural or hormonal
stimulation, elevated cyclic AMP activates*^ cytoplasmic lipase, which hydrolyzes triglyceride from the periphery of the
droplet, and the resulting fatty acids and glycerol diffuse back to the capillaries and are carried in the blood to tissues
throughout the body.

fat cells on the denervated side retain an Brown Adipose Tissue as a Heat
almost normal content of lipid. It is thus Generator
demonstrated that the presence of nerves is
necessary for normal mobilization of lipid It has long been noted that brown adipose
from adipose tissue. tissue is more abundant in animal species
The chemical mediator norepinephrine is that hibernate, and it was assumed to have a
present in abundance in innervated adipose function related to winter dormancy. There
tissue but is low or absent after denervation. is now evidence that one of its functions is to
It is through release of norepinephrine from serve as a “chemical furnace”—an oil burner
the nerve endings that the nerves control the to heat the animal during arousal from hi¬
mobilization of fatty acids from adipose tis¬ bernation.
sue. Injection of small amounts of exogenous Homeothermic animals maintain their
norepinephrine inhibits the action of insulin body temperature within a rather narrow
on fat cells and approximately doubles the range. When exposed to an unfavorable cold
amount of free fatty acid in the blood plasma. environment, they decrease their heat loss by
The norepinephrine brings about the acti¬ constricting peripheral blood flow and in¬
vation of adipose tissue lipase, increasing the crease their metabolism to generate more
rate of hydrolysis of triglycerides. In patients heat. The basal metabolic rate is the heat pro¬
who have an adrenomedullary tumor (pheo- duction or oxygen consumption in a resting,
chromocytoma) that secretes excessive fasting animal in a thermoneutral environ¬
amounts of norepinephrine, the plasma con¬ ment. In a cold environment, extra heat can
centration of fatty acids may be several be produced by shivering— a form of invol¬
hundred times the normal level. untary isometric exercise. In addition to shiv-
186 • ADIPOSE TISSUE

ering thermo gene sis, many species are capable from brown adipose tissue. A deficiency of
of nonshivering thermogenesis—an increase in oxidative phosphorylation in mitochondria
heat production that occurs without any in¬ would be a surprising finding in any other
crease in electrical activity in skeletal muscle. normal tissue, but it is consistent with the
Brown adipose tissue has been found to be needs of a system concerned mainly with heat
the most important site of nonshivering ther¬ production.
mogenesis. Upon stimulation, it is capable of The heat generation by brown fat can be
the highest oxygen consumption recorded demonstrated visually by the new technique
for any organ. Its maximal consumption may of thermography (Fig. 6--14). The thermo¬
represent a rate of heat production hundreds graph scans across the body, detecting the
of times the average heat production of other infrared radiation from surfaces, and regis¬
organs, and the blood flow through this tissue ters the temperature-dependent intensity of
can increase to seven times its own volume radiation on a photographic plate. When a
per minute. bat arousing from hibernation is scanned,
When a hibernating animal begins to the thin wing membranes rapidly equilibrate
arouse, there is a marked increase in oxygen with the ambient temperature, and most of
consumption and generation of heat without the body is still relatively cool. However, a
shivering. Nerve impulses to the brown adi¬ sharply delineated “Tot area” is found on the
pose tissue release norepinephrine at the thermograph, coinciding with the location of
nerve endings, which leads to activation of the interscapular brown fat. Thus, in hiber¬
lipase in the fat cells and breakdown of tri¬ nating species brown adipose tissue performs
glyceride to fatty acid and glycerol. Oxidation two important roles during arousal from dor¬
of fatty acid and reesterification of some of mancy: oxidation of lipid to produce heat
the fatty acid then occurs, with consumption within the brown fat, and release into the
of oxygen and generation of heat that serves circulation of oxidizable substrates for utili¬
to warm the blood flowing through the fat, zation by other tissues.
secondarily raising the temperature of the When newborns of nonhibernating species
animal as a whole. such as the rat and rabbit are exposed to the
Correlated with this function are some un¬ cold or when they are infused with physio¬
usual features of the mitochondria. Oxidative logical amounts of norepinephrine, there is
phosphorylation is difficult to demonstrate a substantial increase in oxygen consumption,
in mitochondrial fractions from brown adi¬ and thermocouples embedded in their inter¬
pose tissue. The elementary particles or inner scapular brown fat register a local production
membrane subunits that are usually present of heat.
on the mitochondrial cristae are thought to It has now been shown that the human
be the sites of phosphorylation enzymes. infant makes use of the same mechanism for
These have not been demonstrated in normal heat generation. An infant placed in an en¬
numbers in negatively stained mitochondria vironment at 23°C immediately after birth

Figure 6-14. Thermograph of a bat during arousal from hibernation. Scanning the animal for detection of infrared
radiation reveals a “hot spot” in the area corresponding to the interscapular brown adipose tissue. During arousal this
tissue acts as a chemical “furnace,” producing heat carried in the bloodstream to warm the rest of the body.
(Thermograph courtesy of J. Hayward.)
 ADIPOSE TISSUE • 187

will have approximately double the metabolic Napolitano, L., and H. T. Gagne: Lipid depleted white
rate of an infant at 33°C and will accomplish adipose cells. Anat. Rec. 747:273, 1963.
this without shivering. There is an associated Renold, A. E., and G. F. Cahill, eds.: Adipose Tissue
increase in the level of glycerol in the blood, Handbook of Physiology. Sec. 5. Washington, D.C.,
American Physiological Society, 1965.
resulting from lipolysis of triglycerides. Un¬ Sheldon, H.: The fine structure of the fat cell. In Rodahl,
der these conditions, thermography reveals K., and B. Issekutz, eds.: Fat As A Tissue. Baltimore,
areas of elevated skin temperature over the McGraw-Hill Book Co., 1964.
sites of brown adipose tissue at the nape of Slavin, B. G.: The cytophysiology of mammalian adipose
tissue. Int. Rev. Cytol. 55:297, 1972.
the neck and in the axillae. It is possible that
this adipose tissue may also have some slight MULTILOCULAR ADIPOSE TISSUE
thermogenic function in adults, but this is yet Cannon, B., and B. W. Johansson: Non-shivering ther¬
to be demonstrated. mogenesis in the newborn. Mol. Aspects Med. 5:119,
1980.
Fawcett, D. W.: A comparison of the histological orga¬
nization and histochemical reactions of brown fat
REFERENCES and ordinary adipose tissue. J. Morphol. 90:363,
1952.
UNILOCULAR ADIPOSE TISSUE Foster, D. O., and M. L. Frydman: Tissue distribution
of cold-induced thermogenesis in conscious warm-
Cushman, S. W.: Structure-function relationships in the or cold-acclimated rats reevaluated from changes in
adipose cell. I. Ultrastructure of the isolated adipose tissue blood flow: the dominant role of brown adi¬
cell. II. Pinocytosis and factors influencing its activity pose tissue in the replacement of shivering by non¬
in the isolated adipose cell. J. Cell Biol. 46:326, 342 shivering thermogenesis. Can. J. Physiol. Pharma¬
1970. col. 57:257, 1979.
Green, H.: The adipose conversion of 3T3 cells. In Himms-Hagen, J.: Cellular thermogenesis. Annu. Rev.
Ahmad, F., T. R. Russell, J. Schultz, and R. Werner, Physiol. 38:315, 1976.
eds.: Miami Winter Symp. Differentiation and De¬ Lindberg, O., ed.: Brown Adipose Tissue. New York,
velopment. Vol. 15. New York, Academic Press, American Elsevier Publishing Co., 1970.
1978. Lindberg, O., J. de Pierre, E. Rylander, et ah: Studies
Greenwood, M. R. C., and J. Hirsch: Postnatal devel¬ of the mitochondrial energy-transfer system of
opment of adipocyte cellularity in the normal rat. J. brown adipose tissue. J. Cell Biol. 34:293, 1967.
Lipid Res. 75:474, 1979. Merklin, R. J.: Growth and distribution of human fetal
Heindel, J. J., L. Orci, and B. Jeanrenaud: Fat mobili¬ brown fat. Anat. Rec. 178:637, 1974.
zation and its regulation by hormones and drugs in Nedergaard, J., and O. Lindberg: The brown fat cell.
white adipose tissue. In Peters, C. (ed.): Interna¬ Int. Rev. Cytol. 74:310, 1982.
tional Encyclopedia of Pharmacology and Therapy. Nicholls, D. G.: Brown adipose tissue mitochondria.
Oxford, Pergamon Press, 7:175-373, 1975. Biochim. Biophys. Acta 549:1, 1979.
Hirsch, J., and B. Batchelor: Adipose tissue cellularity Sidman, R. L., and D. W. Fawcett: The effect of periph¬
in human obesity. Clin. Endocrinol. Metab. 5:299, eral nerve section on some metabolic responses of
1976. brown adipose tissue. Anat. Rec. 118:487, 1954.
Jeanrenaud, B.: Dynamic aspects of adipose tissue me¬ Sidman, R. L., M. Perkins, and N. Weiner: Noradrena¬
tabolism. A review. Metab. Clin. Exp. 10:535, 1961. line and adrenaline content of adipose tissues. Na¬
Napolitano, L.: The differentiation of white adipose ture 193:36, 1962.
cells: an electron microscope study. J. Cell Biol. Smith, R. E., and B. A. Horwitz: Brown fat and ther¬
18:663, 1963. mogenesis. Physiol. Rev. 49:330, 1969.
 CARTILAGE

Cartilage is a specialized form of connec¬ understood from a consideration of its mode

tive tissue consisting of cells, called chondro¬ of development.
%

cytes, and extracellular fibers embedded in a

gel-like matrix. The intercellular components Histogenesis of Cartilage
predominate over the cells, which are isolated
in small cavities within the matrix. Unlike At sites of future cartilage formation in the
other connective tissues, cartilage has no embryo, the mesenchymal cells first withdraw
nerves or blood vessels of its own. The col¬ their processes and become crowded together
loidal properties of its matrix are therefore in dense aggregations called protochondral tis¬
important to the nutrition of its cells and are sue or centers of chondrification. The nuclei of
in large measure responsible for its firmness the cells are very close together and the cell
and resilience. The capacity of cartilage for boundaries are indistinct (Fig. 7-2A,B). As
rapid growth while maintaining a considera¬ the cells enlarge and differentiate, they se¬
ble degree of stiffness makes it a particularly crete around themselves a metachromatic ex¬
favorable skeletal material for the embryo. tracellular matrix (Fig. 7—2C). Tropocollagen
Most of the axial and appendicular skeleton is secreted at the same time, but the fibrils
is first formed in cartilage models, which are that form extracellularly tend to be masked
later replaced by bone. by the hyaline matrix in which they are
Cartilage is of more restricted occurrence embedded. As the amount of interstitial ma¬
in postnatal life, but it continues to play an terial increases, the cells become isolated in
indispensable role as the long bones grow in separate compartments or lacunae and grad¬
length in the immature individual, and it ually take on the cytological characteristics of
persists in the adult on the articular surfaces mature cartilage cells or chondrocytes (Fig.
of the long bones. Except where it is exposed 7-3).
to the synovial fluid in joints, cartilage is The continuing growth of cartilage takes
invariably enclosed in a dense fibrous con¬ place by two different mechanisms. Mitosis is
nective tissue called the perichondrium. observed among the cells for a rather long
Three kinds of cartilage, hyaline, elastic, and period. After the constriction of the cyto¬
fibrocartilage, are distinguished on the basis of plasm in such a division, a new partition of
the amount of extracellular matrix and the interstitial substance quickly develops and
relative abundance of the collagenous and separates the two daughter cells. These in
elastic fibers embedded in it. Hyaline carti¬ turn may divide, giving rise to clusters of
lage is the most common and most character¬ four, and so on. The mitotic division of the
istic type, and the others can be regarded as chondrocytes and the secretion of new matrix
modifications of it (Fig. 7-1). between the daughter cells lead to an expan¬
sion of the cartilage from within, which is
referred to as interstitial growth.
The mesenchyme surrounding the carti¬
HYALINE CARTILAGE lage primordium condenses to form a special
layer, the perichondrium, which merges with
In the adult, hyaline cartilage is found on the cartilage on one side and with the adja¬
the ventral ends of the ribs, in the tracheal cent connective tissue on the other (Fig. 7—4).
rings and larynx, and on the joint surfaces Throughout embryonic life the cells on the
of bones. It is a somewhat elastic, semitrans¬ inner or chondrogenic layer of the perichon¬
parent tissue with an opalescent bluish gray drium constantly differentiate into chondro¬
tint. Its histological appearance is most easily cytes, secrete matrix around themselves, and

188
 CARTILAGE • 189

Figure 7-1. Hyaline cartilage from the trachea of a guinea pig. Notice the more intense staining of the capsular or
territorial matrix immediately surrounding the groups of isogenous cells. The cells immediately beneath the perichondrium
(top) recently added in appositional growth are single and elongated.

in this way contribute new cells and matrix eny of a single chondrocyte that underwent
to the surface of the mass of cartilage. This a few mitotic divisions in the course of the
process is called appositional growth. The abil¬ interstitial growth of the cartilage. In the
ity of the perichondrium to form cartilage cartilage of the epiphyseal plates of long
persists but remains latent in the adult. bones, cell division in a consistent plane re¬
sults in an arrangement of the cartilage cells
The Chondrocytes in long columns that are later invaded by
advancing bone (Fig. 7-6).
In the layers of cartilage immediately be¬ The nucleus of the chondrocyte is round
neath the perichondrium or under the free or oval and contains from one to several
surface of articular cartilage, the lacunae are nucleoli, depending on the species. There is
elliptical in section, with the long axis parallel a juxtanuclear cell center with a pair of cen-
to the surface, while deeper in the cartilage trioles and a well-developed Golgi apparatus.
they are semicircular or angular. The cells in The surrounding cytoplasm contains elon¬
living cartilage usually conform to the shape gated mitchondria, occasional lipid droplets,
of the lacunae that they occupy, but fixation and variable amounts of glycogen. When new
and dehydration may result in their retrac¬ matrix is being formed in growing or regen¬
tion from the wall of the lacuna, so that they erating cartilage, the cytoplasm becomes
may appear stellate. Mature cartilage cells in more basophilic and the Golgi region be¬
higher vertebrates rarely if ever have proc¬ comes unusually large. Under these condi¬
esses visible with the light microscope, but in tions of active growth, electron micrographs
electron micrographs their surface is quite show a well-developed granular endoplasmic
irregular. The cells tend to be clustered in reticulum with moderately distended cister-
small groups (Fig. 7-1). Each group is said nae. The saccules of the Golgi complex tend
to be isogenous because it represents the prog¬ to be dilated, and there are numerous asso-
190 * CARTILAGE

Figure 7-3. Development of cartilage from mesenchyme

in a 15-mm guinea pig embryo. The mesenchyme (below)
gradually merges into the protochondral tissue with inter¬
stitial substance (above). (After A. A. Maximow.)

ciated vacuoles of varying size that sometimes

contain filaments or granules. Similar vacu¬
oles are also seen at the cell surface, where
m they appear to be discharging their contents
I
•L® m into the surrounding matrix. In cartilage that
is not actively growing, the endoplasmic re¬
* j&!k ?*T‘
'- Sh*^' " ticulum is less extensive and the Golgi com¬
T ^ £ mr plex not as prominent.
rSi,

jN*|L
Z M*
0R}^
m
jlg^fllip ^0»’>*tmk #w '
Cartilage Matrix
W^ m ip
mii^^* «J
Pgs
In fresh hyaline cartilage the matrix ap¬
*
^>-^1 pears homogeneous. This is due in part to
•Tjgr && * Wm •*.. i ♦# |IJi the fact that the ground substance and the
-<ss
- . •*> 4w^»e collagen embedded within it have approxi¬
Figure 7-2. In the histogenesis of cartilage, mesenchymal mately the same refractive index and in part
cells (A) withdraw their processes and become crowded to the small size of the collagen fibrils. The
together to form an area of precartilage (B). In newly matrix is deeply colored with the periodic
formed embryonic cartilage (C), the densely aggregated acid-Schiff reaction for carbohydrates. It
cells of the precartilaginous stage have been moved apart
by deposition of clear hyaline matrix between them. The
also has a marked affinity for basic dyes and
cells then become angular (D) and isolated in clearly stains metachromatically with toluidine blue.
demarcated lacunae. The principal constituents of the extracellu-
 CARTILAGE • 191

plex carbohydrates called glycosaminogly-

cans. These radiate from the core protein in
a bottle-brush configuration. The principal
glycosaminoglycans of cartilage matrix are
chondroitin sulfate and keratan sulfate. There
are typically about 80 chondroitin sulfate
chains and about 100 keratan sulfate chains
associated with each core protein molecule.
At one end of the core protein is a polypep¬
tide segment relatively free of glycosamino-
glycan side chains, the so-called hyaluronic
acid-binding region. The proteoglycan mol¬
ecules are bound at this end, via a link pro¬
tein, to very long hyaluronic acid molecules
spaced at intervals of about 30 nm. The
proteoglycan aggregates so formed occupy the
interstices of the meshwork of collagen fibrils.
In the hydrated state in vivo the extended
proteoglycan molecules occupy very large
volumes relative to their molecular weight.
The entwining of the proteoglycan aggre¬
gates with the collagen fibrils is depicted
schematically in Figure 5-10, Chapter 5.
These relationships cannot be directly visu¬
alized by routine methods, for when tissues
are processed for electron microscopy, the
glycosaminoglycan chains collapse onto the
core protein during dehydration. The pro¬
teoglycan monomers thus appear in sections
of ruthenium red-stained cartilage as dense
granules of irregular shape dispersed in the
Figure 7-4. Hyaline cartilage from xiphoid process of rat.
A, Transition layer adjacent to perichondrium. B, Contin¬ meshes of the collagenous framework. The
uation of collagenous fibers from perichondrium into in¬ molecular organization of the extracellular
terstitial substance of cartilage. C, Columns of isogenous matrix in the hydrated state is ideally suited
groups of cartilage cells, some of which have fallen out to its function on the weight-bearing articular
of the lacunae in processing. (After A. A. Maximow.)
surfaces of bones. The fibrillar scaffolding of
collagen determines and maintains tissue
shape and resists tensile forces, while the
proteoglycan aggregates occupying its inter¬
lar matrix of hyaline cartilage are Type II stices provide a hydrated viscous gel that
collagen and proteoglycans. The collagen fibrils absorbs compressive forces.
are generally quite thin (10-20 nm) and The matrix immediately surrounding each
unlike other collagens they usually lack a group of isogenous cells usually stains more
distinct cross-banding. They are assembled deeply than elsewhere (Fig. 7-1). This deeply
from molecules composed of a single type of basophilic rim is called the capsular or territo¬
a chain designated a 1(11). The fibrils are not rial matrix, while the less basophilic areas
organized in bundles but form a loose mesh- between cell groups are called the intercapsu-
work throughout the matrix. lar or interterritorial matrix. The deeper stain¬
The proteoglycans of cartilage have been ing of the territorial matrix suggests that the
thoroughly studied and have a more compli¬ concentration of chondroitin sulfate is higher
cated structure than many other members of in the immediate vicinity of the cells.
this class of macromolecules. The core pro¬
tein forming the backbone of the proteogly¬
Secretion of Matrix Components
can is about 300 nm in length and has a
molecular weight of about 250,000. The re¬ The chondrocytes synthesize and secrete
mainder of the molecule consists of the com¬ the collagen and proteoglycans of the sur-
192 CARTILAGE

Figure 7-5. Electron micrograph of chondrocytes and matrix of mouse trachea illustrating the irregular outlines of the
cells, their well-developed granular endoplasmic reticulum, and other organelles. (Micrograph from Seegmiller R C
Ferguson, and H. Sheldon. J. Ultrastruct. Res. 38:288, 1972.)
 CARTILAGE • 193

Figure 7-6. Hyaline cartilage of the epiphyseal plate of rabbit tibia. Here the cartilage cells are arranged in lonq parallel
columns. From above downward, zones of cartilage cell proliferation, maturation, hypertrophy, and degeneration can be

rounding matrix. After injection of tritiated granular material can be found in the same
proline into experimental animals that are vacuole, suggesting that collagen and proteo¬
actively forming cartilage, this amino acid glycans are synthesized concurrently and
precursor of collagen can be localized by packaged together for exocytosis. Secretion
radioautography over the endoplasmic retic¬ of hyaluronic acid and linking proteins prob¬
ulum of the chondrocytes in 10 minutes; over ably take place similarly. However, the pro-
the Golgi complex in 30 minutes; over secre¬ teoglycans are secreted as monomers, and
tory vacuoles at the cell periphery at three their assembly into aggregates occurs extra-
hours; and at longer time intervals over the cellularly.
extracellular matrix. Thus, the synthesis of In hyaline cartilage of adults the collagen
collagen follows the same intracellular path network in the matrix does not seem to be
as the secretion of protein by glandular cells. renewed, but the proteoglycans slowly turn
The same train of events can be demon¬ over and are replaced by molecules newly
strated in the elaboration of matrix proteo¬ synthesized by the chondrocytes. These cells
glycans. The synthesis of the core protein are thus involved in maintenance of the nor¬
and the initial steps of oligosaccharide addi¬ mal structure of the matrix, and are capable
tion occur on the ribosomes of the endo¬ of responding to altered conditions of weight
plasmic reticulum. After accumulation of the bearing and attrition by altering the rate of
initial product in the cisternae and transport turnover of the proteoglycans. Some of the
to the Golgi, chondroitin sulfate chains are structural changes observed in the cartilage
rapidly added to complete assembly of pro¬ of aging osteoarthritic individuals are prob¬
teoglycans. The completed molecules, pack¬ ably a consequence of a declining ability of
aged in secretory vacuoles, are moved to the chondrocytes to replace proteoglycans at a
cell surface and released by exocytosis. In rate sufficient to keep pace with wear and
electron micrographs both filamentous and tear. In the inflammatory reaction associated
194 • CARTILAGE

Figure 7-7. Diagram of the intracellular pathway for

synthesis of matrix components. Amino acids are incor¬
porated into protein at the ribosomes. Sugar and sulfates
are believed to be incorporated into polysaccharide in the
Golgi region and combined to form the protein-polysac¬
charide mucopolysaccharide released into the surround¬
ing matrix. (Courtesy of E. Hay and J. P. Revel.)

with rheumatoid arthritic disorders, pro¬ velops, containing fibroblasts and fibrillar
teases released may rapidly degrade the core bundles that do not give reactions character¬
proteins of proteoglycans and impair their istic of either collagen or elastin. These in¬
normal function in the matrix. different fibers later acquire typical staining
properties for elastin. The cells secrete ma¬
trix around themselves and become recogniz¬
able as chondrocytes. As in hyaline cartilage,
ELASTIC CARTILAGE a perichondrium is formed and initiates ap-
positional growth.
In mammals this variety of cartilage is
found in the external ear, the walls of the
external auditory and eustachian canals, the
epiglottis, and in parts of the corniculate and
FIBROCARTILAGE
cuneiform cartilages. It differs from hyaline
cartilage macroscopically in its yellowish color Fibrocartilage occurs in a few regions of
and its greater opacity and elasticity. dense connective tissue in the body. In small
Its cells are similar to those of hyaline areas with poorly defined limits, typical car¬
cartilage; they are of the same rounded tilage cells and a small amount of matrix are
shape, are also surrounded by capsules, and found among the abundant fibrous elements.
are scattered singly or in isogenous groups It occurs in the intervertebral discs; in certain
of two or four cells. The interstitial substance articular cartilages; in the symphysis pubis;
differs from that of hyaline cartilage by being in the ligamentum teres femoris; and in the
permeated by frequently branching fibers, sites of attachment of certain tendons to
which are positive in all staining methods for bones. The encapsulated cartilage cells lie
elastin (Fig. 7—8). They form a network that singly or in pairs, or are sometimes aligned
is often so dense that the ground substance in rows between bundles of collagen fibers
is obscured. In the layers beneath the peri¬ (Fig. 7-9). The matrix is quite inconspicuous
chondrium, the feltwork of the elastic fibers except in the immediate vicinity of the cells,
is looser. The elastic fibers of the cartilage where its presence can be inferred from the
continue into the perichondrium. characteristic form of the lacunae.
In the histogenesis of elastic cartilage in Fibrocartilage is closely associated with the
the embryo, a primitive connective tissue de¬ connective tissue of the capsules and liga-
 CARTILAGE • 195

Figure 7-8. Elastic cartilage of the epiglottis of a child. Notice the dark-staining elastic fiber bundles in the matrix
between cell groups. (From Fawcett, D. W., In Greep, R. O., ed.: Histology. Philadelphia, Blakiston Co 1953
Reproduced by permission of McGraw-Hill Book Co.)

ments of joints. It is a transitional form be¬ comes infiltrated only slightly, with cartilagi¬
tween cartilage and dense connective tissue, nous ground substance.
and the gradual transition from one to the
other can be observed in the adult, as well as
during histogenesis in the embryo. In the OTHER VARIETIES OF
intervertebral discs, for example, the hyaline
cartilage connected with the vertebrae shows
CARTILAGE AND
distinct collagenous fibers in its matrix. These CHONDROID TISSUE
then become associated into thick bundles,
which almost entirely displace the ground There is a transitory phase in the embry¬
substance while the cartilage cells retain their onic development of hyaline cartilage when
typical form and their capsular matrix. Fi¬ it is composed of closely packed cells having
nally, this typical hbrocartilage merges into thin capsules and collagen fibers in its inter¬
connective tissue, the fusiform cells of which cellular substance. In this underdeveloped
have tapering processes and are not enclosed condition the cartilage may remain through¬
in lacunae. out life in certain sites in the bodies of higher
Fibrocartilage develops in much the same organisms. It occurs often in lower verte¬
way as ordinary connective tissue. In the brates (fishes and amphibians) and is still
beginning there are typical fibroblasts sepa¬ more common in invertebrates. Such tissue
rated by a large amount of collagen. Then has been variously called pseudocartilage, fi-
these cells become rounded, are transformed brohyaline tissue, vesicular supporting tissue, and
into cartilage cells, and surround themselves chondroid tissue. This tissue serves as a local
with a thin layer of capsular matrix. The mechanical support for the surrounding tis¬
abundant fibrous interstitial substance be¬ sue.
196 • CARTILAGE

in the embryo. After a wound or excision of

a portion of cartilage in adult mammals, no
such independent regeneration takes place.
Instead, one sees at first in the injured area
only necrotic and atrophic changes. The de¬
fect is then filled by connective tissue, which
grows in from the perichondrium or from
fascia in the vicinity of the injured area. The
fibroblasts of this ingrowing granulation tis¬
sue may then round up, produce capsules
around themselves, and become transformed
into new cartilage cells. Thus, if cartilage is
replaced in the adult mammal, this takes
place mainly by metaplasia of loose connec¬
tive tissue.
Such cartilaginous metaplasia sometimes
takes place in connective tissue under the
influence of simple mechanical forces acting
from the outside, such as pressure, particu¬
larly when combined with friction. It is
claimed that the presence of cartilage on the
joint surfaces of bones is related to the con¬
stant mechanical influences to which a nor¬
mal joint is subjected while functioning.
When these mechanical conditions disappear,
as happens in dislocation of bones, the carti¬
lage often undergoes dedifferentiation. On
the other hand, cartilage is laid down in the
primordia of the joint surfaces in the embryo
at a time when there are probably no me¬
chanical forces acting on the joint.
Although cartilage has only limited rege¬
nerative capacity, it has been shown that
components of the matrix can be rapidly re¬
formed if the cells remain intact. Injection of
Figure 7-9. Low-power drawing of fibrocartilage at inser¬ a crude preparation of papain into young
tion of tendon into the tibia of a rat. Note the direct rabbits results in a collapse of their ears. This
transformation of rows of tendon celis (top) into cartilage is attended by a loss of basophilia of the
cells surrounded by deeply staining matrix. (Drawn by
cartilage matrix and by a loss of its proteo¬
Miss A. Nixon.)
glycans and elastic components, demonstra¬
ble in electron micrographs. After 48 hours
The tissue composing the notochord of ver¬ the regeneration of matrix components is
tebrate embryos has a similar structure. Here already far advanced and the ears are largely
there is a cylindrical shaft of variable thick¬ restored to their normal erect position.
ness, which consists of large, closely packed
cells distended with fluid. The notochordal
tissue has a different embryologic origin REGRESSIVE CHANGES IN
from that of the cartilage and of the other
CARTILAGE
connective tissues.

The most important regressive change in

cartilage, calcification, normally precedes the
REGENERATION OF type of bone formation called endochondral
CARTILAGE ossification (Fig. 7-10). The cartilage cells in
a center of ossification undergo a regular
In amphibians, cartilage is regenerated in sequence of cytological changes accompanied
a manner resembling histogenesis of cartilage by characteristic changes in the neighboring
 CARTILAGE • 197

in the zone of hypertrophic cartilage cells. In

the early stages of calcification of the hyaline
matrix, minute crystals of hydroxyapatite are
seen within and at the surface of the matrix
vesicles. It is believed that the vesicles may
have the capacity to bind and concentrate
calcium, resulting in precipitation of hydrox¬
yapatite in their immediate vicinity. As the
nests of apatite crystals enlarge and merge,
the cartilage becomes opaque, hard, and brit¬
tle. Because of these changes in the matrix,
the zone of cartilage cell hypertrophy is also
known as the zone of provisional calcification.
The relation of these events to the process
of ossification will be discussed in greater
detail in Chapter 8.
In man, ossification of certain cartilages
also occurs as a normal age change and may
take place in some parts of the larynx as early
as 20 years of age.

HISTOPHYSIOLOGY OF
CARTILAGE

Cartilage in joints has the remarkable

property of sustaining great weight and at
Membrane bone the same time allowing the bones, which carry
Figure 7-10. Two stages in calcification of the cartilage this weight, to move easily and smoothly
model of the calcaneus in rats. A, Two days after birth; against one another. In other places, such as
B, four days after birth. The calcium salts appear black
because of the silver nitrate stain. Undecalcified prepa¬
the ear and the respiratory passages, cartilage
rations stained with von Kossa’s method. (After W. Bloom serves as a pliable yet resistant framework
and M. A. Bloom.) that prevents the collapse of the tubular or¬
gan it surrounds. Finally, cartilage in many
bones makes possible their growth in length
and is important in determining their size
matrix. In the epiphyseal plate of long bones and shape.
where the cells become arranged in parallel Far from being an inert tissue, cartilage,
columns, these changes are observed in suc¬ through its participation in the growth of
cessive zones along the length of the column. bones, is a fairly delicate indicator of certain
Distinct zones of proliferation, maturation, car¬ metabolic disturbances. It reflects nutritional
tilage cell hypertrophy, and cell degeneration can deficiencies, especially of protein, minerals, or
be recognized (Fig. 7—6). In the zone of vitamins. For example, the thickness of the
hypertrophic cartilage cells, the matrix epiphyseal cartilage plate diminishes rapidly
undergoes calcification. when a young rat is placed on a protein-
At sites where cartilage matrix undergoes deficient diet or on one lacking in vitamin A.
calcification, small membrane-limited struc¬ When vitamin C is withheld from guinea
tures are found, called matrix vesicles. Because pigs, producing scurvy, cessation of matrix
they are limited by a typical unit membrane formation may be accompanied by changes
and sometimes contain ribosomes or qther in the cells and by distortion of their colum¬
recognizable cytoplasmic components, it is nar arrangement. Absence of vitamin D is
assumed that they arise by being budded attended by a deficient absorption of calcium
off from the chondrocytes. The vesicles and phosphorus from the diet and leads to
have been isolated in centrifugal fractions rickets, in which the epiphyseal cartilages con¬
and found to contain acid phosphatase and tinue to proliferate but fail to calcify, and the
ATPase activity. They are found at all levels growing bones become deformed by weight
in the epiphyseal plate but are concentrated bearing.
198 • CARTILAGE

The participation of cartilage in the growth Christner, J. E., M. L. Brown, and D. D. Dziewiatkowski:
Interactions of cartilage proteoglycans with hyaluro-
in length of bones is in part under control of
nate. J. Biol. Chem. 254:4624, 1979.
several hormones, of which the most impor¬ Comper, W. D., and T. C. Laurent: Physiological func¬
tant is the pituitary growth hormone. Hypophy- tion of connective tissue polysaccharides. Physiol.
sectomy in young rats leads to a thinning of Rev. 55:255, 1978.
the epiphyseal plate of long bones, with ces¬ Gliicksmann, A.: Studies on bone mechanics in vitro. II.
The role of tension and pressure in chondrogenesis.
sation of mitosis and a decrease in the num¬
Anat. Rec. 75:39, 1939.
ber and especially in the size of its cells. After Goldman, G. C., and N. Lane: On the site of sulfation
a short time the cartilage fails to be eroded in the chondrocyte. J. Cell Biol. 27:353, 1964.
and growth of the bone ceases. When growth Goldman, G. C., and K. R. Porter: Chondrogenesis,
studies with the electron microscope. J. Biophys.
hormone is injected into such animals, the
Biochem. Cytol. 5:719, 1960.
cartilage undergoes a striking metamorphosis Elascall, G. K.: Cartilage proteoglycans: comparison of
and within a few days resembles that of a sectioned and spread whole molecules. J. Ultra-
normal young growing animal, and the bone struct. Res. 76:369, 1980.
resumes its growth. The response of the Elascall, V. C.: Interaction of cartilage proteoglycans
with hyaluronic acid. J. Supramol. Struct. 7:101,
cartilage varies with the dose level and has
1977.
been used to assay extracts containing the Lane, J. M., and C. Weiss: Review of articular cartilage
hormone. Long-continued administration of collagen research. Arthritis Rheum. 75:553, 1975.
the hormone produces giant rats, this being McCluskey, R. T., and 6L. Thomas: The removal of
cartilage matrix in vivo by papain. I. Exp. Med.
made possible in part by growth of cartilage
765:371, 1958.
after it would normally have ceased growing. Minns, R. J., and F. S. Stevens: The collagen fibril
Further, the injection of the hormone into organization in human articular cartilage. J. Anat.
older rats, in which cartilage proliferation 725:437, 1977.
has stopped, can to some extent reactivate its Revel, J. P., and E. D. Hay: An autoradiographic and
electron microscopic study of collagen synthesis in
growth, with subsequent increase in the size differentiating cartilage. Zeitschr. f. Zellforsch.
of its bones. 67:110, 1963.
Seegmiller, R., C. C. Ferguson, and H. Sheldon: Studies
on cartilage. VI. A genetically determined defect in
tracheal cartilage. J. Ultrastruct. Res. 55:288, 1972.
REFERENCES Seegmiller, R., E. C. Fraser, and H. Sheldon: A new
chondrodystrophic mutant in mice. Electron mi¬
Ali, S. Y., S. W. Sajdera, and H. C. Anderson: Isolation croscopy of normal and abnormal chondrogenesis.
and characterization of calcifying matrix vesicles J. Cell Biol. 45:580, 1971.
from epiphyseal cartilage. Proc. Natl. Acad. Sci USA Sheldon, H., and F. B. Kimball: Studies on cartilage.
67:1513, 1970. III. The occurrence of collagen within vacuoles of
Anderson, H. C.: Vesicles associated with calcification of the Golgi apparatus. J. Cell Biol. 72:599, 1962.
the matrix of epiphyseal cartilage. J. Cell Biol. 47:59, Sheldon, H., and R. A. Robinson: Studies on cartilage.
1969. I. Electron microscope observations on normal rab¬
Anderson, D. R.: The ultrastructure of elastic and hya¬ bit ear cartilage. II. Electron microscopic observa¬
line cartilage in the rat. Am. J. Anat. 774:403, 1964. tions on rabbit ear cartilage following the adminis¬
Anderson, H. C., T. Matsuzwa, S. W. Sajdera, and S. Y. tration of papain. J. Biophys. Biochem. Cytol. 4:401,
Ali: Membranous particles in calcifying matrix. 1958; 8:151, 1960.
Trans. N.Y. Acad. Sci. 52:619, 1970. Silberberg, R., M. Hasler, and M. Silberberg: Submicro-
Anderson, H. C. and S. W. Sajdera, 1971. The fine scopic response of articular cartilage of mice treated
structure of bovine nasal cartilage. Extraction as a with estrogenic hormone. Am. J. Pathol. 46:289,
technique to study proteoglycans and collagen in 1965.
cartilage matrix. J. Cell Biol. 49:650 Silberberg, R., M. Silberberg, and D. Feir: Life cycle of
Becks, H., C. W. Asling, M. E. Simpson, C. H. Li, and articular cartilage cells: an electron microscope
H. M. Evans: The growth of hypophysectomized study of the hip joint of the mouse. Am. I. Anat.
female rats following chronic treatment with pure 774:17, 1964.
pituitary growth hormone. III. Skeletal changes— Thomas, L.: Reversible collapse of rabbit ears after
tibia, metacarpal, costochondral junction and caudal intravenous papain and prevention of recovery by
vertebrae. Growth 75:175, 1949. cortisone. J. Exp. Med. 764:245, 1956.
Bonucci, E.: Fine structure of early cartilage calcification. Wolbach, S. B., and C. L. Maddock: Vitamin-A acceler¬
J. Ultrastr. Res. 20:33, 1967. ation of bone growth sequences in hypophysecto¬
Bonucci, E.: Fine structure and histochemistry of calci¬ mized rats. Arch. Pathol. 55:273, 1952.
fying globules in epiphyseal cartilage. Zeitschr. f. Zambrano, N. Z., Montes, G. S., Shigihara, K. M.,
Mikroskop. Anat. 765:192, 1970. Sanchez, E. M., and Junqueira, L. C.: Collagen
Cameron, D. A., and R. A. Robinson: Electron micros¬ arrangement in cartilages. Acta Anat. (Basel)
copy of epiphyseal and articular cartilage matrix in 775:26,1982.
the femur of the newborn infant. J. Bone Joint
Surg. 46:163, 1958.
 BONE

Bone, in common with other connective able, compact (substantia compacta) and spongy
tissues, consists of cells, fibers, and ground
(substantia spongiosa), also called cancellous
substance, but unlike the others its extracel¬ bone. The latter consists of a three-dimen¬
lular components are calcified, making it a sional lattice of branching bony spicules or
hard, unyielding substance ideally suited for trabeculae delimiting a labyrinthine system of
its supportive and protective function in the intercommunicating spaces that are occupied
skeleton. It provides for the internal support by bone marrow. Compact bone appears as a
of the body and for the attachment of the solid continuous mass in which spaces can be
muscles and tendons essential for locomo¬ seen only with the aid of the microscope. The
tion. It protects the vital organs of the cranial two forms of bone grade into one another
and thoracic cavities, and it encloses the without a sharp boundary (Fig. 8-3).
bloodforming elements of the bone marrow. In typical long bones, such as the femur or
In addition to these mechanical functions, it the humerus, the diaphysis (shaft) consists of
plays an important metabolic role as a mobi- a thick-walled hollow cylinder of compact
lizable store of calcium, which can be drawn bone with a voluminous central medullary cav¬
upon as needed in the homeostatic regulation ity (marrow cavity) occupied by the bone
of the concentration of calcium in the blood marrow. The ends of long bones consist
and other fluids of the body. mainly of spongy bone covered by a thin
Bone has a remarkable combination of peripheral cortex of compact bone (Figs. 8-2,
physical properties—high tensile and com¬ 8-3). The intercommunicating spaces among
pressive strength while at the same time hav¬ the trabeculae of this spongy bone, in the
ing some elasticity and being a relatively adult, are directly continuous with the mar¬
lightweight material. At all levels of the or¬ row cavity of the shaft. In the growing animal
ganization of bones, from their gross form the ends of long bones, called the epiphyses,
to their submicroscopic structure, their con¬ arise from separate centers of ossification and
struction ensures the greatest strength with are separated from the diaphysis by a carti¬
great economy of material and minimal laginous epiphyseal plate (Fig. 8-1), which is
weight. Despite its strength and hardness, united to the diaphysis by columns of spongy
bone is a dynamic living material, constantly bone in a transitional region called the meta-
being renewed and reconstructed through¬ physis. The epiphyseal cartilage and the ad¬
out the lifetime of the individual. Owing jacent spongy bone of the metaphysis consti¬
to its continual internal reconstruction and tute a growth zone, in which all increment in
its responsiveness to external mechanical length of the growing bone occurs. On the
stimuli, it can be modified by the surgical articular surfaces at the ends of long bones,
procedures and appliances of the orthopedic the thin cortical layer of compact bone is
surgeon or the orthodontist. It is also sur¬ covered by a layer of hyaline cartilage, the
prisingly responsive to metabolic, nutritional, articular cartilage.
and endocrine influences. Disuse is followed With few exceptions, bones are invested by
by atrophy with loss of substance; increased periosteum, a layer of specialized connective
use is accompanied by hypertrophy, with an tissue, which is endowed with osteogenic po¬
increase in the mass of bone. tency. That is to say, it has the ability to form
bone. A covering of periosteum is lacking on
those areas at the ends of long bones that are
MACROSCOPIC STRUCTURE covered with articular cartilage. It is also
OF BONES absent at the sites where tendons and liga¬
ments are inserted and on the surfaces of the
cJpon inspection with the naked eye or patella and other sesamoid bones that are
hand lens, two forms of bone are distinguish¬ formed within tendons. It is also lacking on
199
200 • BONE

their structure or osteogenic potency from

the periosteum and endosteum of long
bones. However, defects in the calvaria re¬
sulting from injury often do not heal com¬
pletely in adults.

MICROSCOPIC STRUCTURE
OF BONES
If a thin ground section of the shaft of a
long bone is examined with the microscope,
it is apparent that the contribution of the
cellular elements of bone to its total mass is
small. Compact bone is largely composed of
the mineralized interstitial substance, bone
matrix, deposited in layers or lamellae 3 to 7
pm thick (Figs. 8-4, 8-5). Rather uniformly
spaced throughout the interstitial substance
of bone are lenticular cavities, called lacunae,
each completely filled by a bone cell or osteo-
cyte. Radiating in all directions from each
lacuna are exceedingly slender, branching
tubular passages, the canaliculi, that penetrate
the interstitial substance of the lamellae, an¬
astomosing with the canaliculi of neighboring
lacunae (Figs. 8-6, 8-7). Thus, although the
lacunae are spaced some distance apart, they
form a continuous system of cavities inter¬
connected by an extensive network of minute
canals. These slender passages are believed
Figure 8-1. Photograph of the upper half of the tibia of a to be essential to the nutrition of the bone
young girl showing the proximal bony epiphysis, the cells. Whereas cartilage can be sustained by
cartilaginous epiphyseal plate, and the shaft or diaphysis. diffusion through the aqueous phase of the
gel-like hyaline matrix, the deposition of cal¬
cium salts in the interstitial substance of bone
the subcapsular areas of the neck of the evidently reduces its permeability. However,
femur and of the astragalus. Where func¬ the maintenance of a system of intercom¬
tional periosteum is absent, the connective municating canaliculi provides avenues for
tissue in contact with the surfaces of bone exchange of metabolites between the cells
lacks osteogenic potency and does not con¬ and the nearest perivascular space.
tribute to the healing of fractures. The mar¬ The lamellae of compact bone are disposed
row cavity of the diaphysis and the cavities in three common patterns. (1) The great
of spongy bone are lined by the endosteum, a majority are arranged concentrically around
thin cellular layer that also possesses osteo¬ longitudinal vascular channels within the
genic properties. bone to form cylindrical units of structure
In the flat bones of the skull, the substantia called haversian systems or osteons. These vary
compacta forms, on both surfaces, relatively in size, being made up of four to 20 lamellae.
thick layers that are often referred to as the In cross section, the haversian systems appear
outer and inner tables. Between them is a layer as concentric rings around a circular opening
of spongy bone of varying thickness called (Figs. 8—6, 8—8A). In longitudinal section,
the diploe. The periosteum on the outer sur¬ they are seen as closely spaced bands parallel
face of the skull is called the pericranium, to the vascular channels (Figs. 8—5, 8-9). (2)
while the inner surface is lined by the dura Between the haversian systems are angular
mater. Although different terms are applied fragments of lamellar bone of varying size
to these connective tissue coverings of the and irregular shape. These are the interstitial
flat bones, they do not differ significantly in systems (Fig. 8—4). The limits of the haversian
 Figure 8-2. Photograph of a sagittal section of the proximal end of the humerus in relation to the glenoid fossa of the
scapula at the shoulder joint. These are dry bones, and the cartilaginous articular surfaces of the joint are not present.
he figure is presented here to illustrate the appearance and distribution of spongy and compact bone. (After A
Feininger, from Anatomy of Nature. Crown Publishers. With permission of Time, Inc.)

Figure 8-3. A thick ground section of the tibia illustrating the cortical compact bone and the lattice of trabeculae of the
cancellous bone.

201
 interstitial system
Outer basic lamellae Sharpey’s
Sharpey’s fiber fiber Area being
absorbed

Haversian
system Cement
Hauer si an ne
canal

Interstitial
system

Cement line
Ca a! of Volkmann
Canal of Volhmann Inner basic lamellae
Figure 8-4. Portion of a ground cross section of a human metacarpal bone. Stained with fuchsin, mounted in Canada
balsam. (After Schaffer.)

Haversian canal

Interstitial system

Cement line
Lacuna

Lamella of haver si an t;..,

system

Hauersian canal

Cement line

Figure 8-5. Portion of a longitudinal, ground section of the ulna of man; stained with fuchsin. (After Schaffer.)
202
 BONE • 203

systems and interstitial systems are sharply communicate with the free surface and with
demarcated by refractile lines called cement the marrow cavity via transverse and oblique
lines. In cross section, the interior of compact channels called Volkmann’s canals. These can
bone thus appears as a mosaic of round and be distinguished from the haversian canals in
angular pieces cemented together (Fig. sections by the fact that they are not sur¬
8-8A). (3) At the external surface of the rounded by concentrically arranged lamellae
cortical bone, immediately beneath the peri¬ but traverse the bone in a direction perpen¬
osteum, and on the internal surface, subja¬ dicular or oblique to the lamellae. The blood
cent to the endosteum, there may be several vessels from the endosteum and, to a lesser
lamellae that extend uninterruptedly around extent, from the periosteum communicate
much of the circumference of the shaft. with those of the haversian systems via Volk-
These are the outee and innev ciYcumfeYential mann s canals, d he vessels are often larger
lamellae (Fig. 8—9). than those in the osteons.
Two categories of vascular channels are Although it is generally correct, the tradi¬
distinguished in compact bone on the basis tional description of haversian canals as being
of their orientation and their relation to the longitudinal and Volkmann’s canals as being
lamellar structure of the surrounding bone. oblique or transverse is an oversimplification.
The longitudinal channels in the centers of Reconstruction of osteons from serial sections
the haversian systems are called haversian has shown that they are not always simple
canals. They are 22 to 110 pm in diameter cylindrical units but may branch and anasto¬
and contain one or two blood vessels en- mose and have a rather complex three-di¬
sheathed in loose connective tissue. The ves¬ mensional configuration. Thus, obliquely ori¬
sels are, for the most part, capillaries and ented vascular channels surrounded by
postcapillary venules, but occasional arte¬ concentric lamellae may be encountered. De¬
rioles may also be found. The haversian ca¬ spite their atypical orientation, these are
nals are connected with one another, and clearly cross-connecting haversian canals.
204 • BONE

Fiqure 8-7. Sector of a cross section of a haversian system of a macerated human hip bone. The cavities and canaiiculi
are filled with a dye: e, connection of canaiiculi of the haversian system with those of an intermediate system. .After A.
A. Maximow.)

Figure 8-8. Section of bone from the midshaft of the human fibula as revealed by four different optical methods. A,
Ground section photographed through the ordinary bright field microscope. The lacunae, haversian systems, and
interstitial lamellae are clearly shown. B, The same section photographed through the polarizing microscope shows the
alternating bright and dark concentric layers in the haversian systems that result from the differing orientation of collagen
fibers in the successive lamellae. C, In a historadiogram of the same section, the differing shades of gray in the scale
from nearly white to nearly black reflect the differing concentrations of calcium. In the haversian canals, there has been
no absorption of the x-rays and the film is therefore black. The most recently deposited haversian systems are
incompletely calcified and appear dark gray, whereas older ones containing higher concentrations of calcium are lighter.
The old interstitial lamellae, being fully calcified, are most highly absorptive and therefore appear white. D, The 14-year-
old girl from whom this specimen was taken had been given a daily dose of the antibiotic Achromycin (tetracycline) for
15 consecutive days at one period of her illness. Amputation of the leg was carried out 230 days later. Achromycin is
incorporated into the matrix of bone being deposited at the time of its administration and imparts a fluorescence to the
newly formed bone, in the section shown here, transilluminated with ultraviolet light in a fluorescence microscope, the
white areas represent areas of bone laid down during the 15-day Achromycin treatment. The nonfluorescent central
portions of the same haversian systems represent bone deposited after cessation of the treatment. (Courtesy of R.
Amprino.)
 BONE • 205

mIM

Figure 8-8. See opposite page for legend.

206 • BONE

Outer circumferential
lamellae

Interstitial
lamellae

Inner
circumferential -Haversian systems
lamellae (osteons)

Periosteum

Trabeculae
of cancellous
Blood vessels
bone

'Sharpey’s
fibers

Endosteum-
Haversian
canals

Volkmann’s'
canals
Figure 8-9. Diagram of a sector of the shaft of a long bone, illustrating the disposition of the lamellae in the osteons or
haversian systems, the interstitial lamellae, and the outer and inner circumferential lamellae. (After A. Benninghoff.
Lehrbuch der Anatomie des Menschen. Berlin, Urban & Schwarzenberg, 1949.)

Cancellous bone is also composed of la¬ vessels traverse the deeper layer and enter
mellae, but its trabeculae are relatively thin Volkmann’s canals, through which they com¬
and usually are not penetrated by blood ves¬ municate with the vessels of the haversian
sels. Therefore, there generally are no hav¬ canals. These numerous small vessels enter¬
ersian systems but merely a mosaic of angular ing Volkmann’s canals from the periosteum
pieces of lamellar bone. The bone cells are may contribute to maintaining its attachment
nourished by diffusion from the endosteal to the underlying bone. In addition, coarse
surface via the minute canaliculi that inter¬ bundles of collagenous fibers from the outer
connect lacunae and extend to the surface. layer of the periosteum turn inward, pene¬
The periosteum is subject to considerable trating the outer circumferential lamellae
variation in its microscopic appearance, de¬ .and interstitial systems of the bone. These
pending on its functional state. During em¬ are called Sharpey’s fibers or perforating fibers
bryonic and postnatal growth there is an (Figs. 8-9, 8-10). They arise during growth
inner layer of bone-forming cells, osteoblasts, of the bone when thick collagenous bundles
in direct contact with the bone. In the adult, become incarcerated in the bone matrix de¬
the osteoblasts revert to a resting form (osteo- posited during the subperiosteal formation
progenitor cells) and are indistinguishable from of new lamellae. When the perforating fibers
other spindle-shaped connective tissue cells. are uncalcified, they occupy irregular canals
If a bone is injured, however, the bone form¬ penetrating the compact bone from the peri¬
ing potentiality of these cells is reactivated; osteal surface in a direction perpendicular or
they take on the appearance of typical osteo¬ oblique to the lamellae. When calcified, they
blasts and participate in the formation of new appear as irregular radial streaks in the outer
bone. The outer layer of the periosteum is a portion of the cortical bone. They serve to
relatively acellular dense connective tissue anchor the periosteum firmly to the under¬
containing blood vessels. Branches of these lying bone. They vary greatly in number in
 BONE • 207

about 50 per cent of its dry weight. The

oi ganic matrix consists of collagenous fibers
embedded in a ground substance. In adult
mammals, about 95 per cent of the organic
matrix is collagen.

Ground Substance
The chemical composition of the extracel¬
lular substance of bone has not been studied
as thoroughly as that of cartilage, owing in
part to the fact that it represents a relatively
small fraction of the organic matrix of bone.
The positive periodic acid—Schiff reaction,
the faint metachromasia of some areas of
bone matrix, and the autoradiographic dem¬
onstration of incorporation of 35S all provide
indirect histochemical evidence for the pres¬
ence of glycosaminoglycans. Analyses of ex¬
tracts of whole bones have identified at least
three such amino sugar-containing macro-
molecular components—chondroitin sulfate,
keratan sulfate, and hyaluronic acid. How¬
ever, it is evident that the concentration of
sulfated glycosaminoglycans in the ground
Figure 8-10. Portion of a cross section of a human fibula. substance of bone is much less than in carti¬
SF, Sharpey’s fibers. (After Schaffer.) lage, for bone matrix is usually acidophilic in
its staining properties.

Collagen
different regions, being particularly numer¬
ous in some bones of the skull and at sites of The collagen of bone, like that of common
attachment of muscles and tendons to the connective tissue, occurs in the form of cross-
periosteum of long bones. In addition to striated fibers 50 to 70 nm in diameter, with
Sharpey’s fibers, some elastic fibers penetrate a 67 nm repeating period. Collagen of bone
the cortical bone from the periosteum, either is predominantly Type I, but differs in some
together with or independent of the collage¬ of its physical properties. It fails to swell in
nous bundles. dilute acid and is insoluble in solvents used
The endosteum is a thin layer of squamous successfully to extract collagens from other
cells lining the walls of those cavities in the tissues. Thus, it seems to have a greater
bone that house the bone marrow. It is the degree of intermolecular bonding.
peripheral layer of the stroma of the bone In mature lamellar bone, the collagen fi¬
marrow where it is in contact with bone, and bers are highly ordered in their arrangement.
it resembles the periosteum in its osteogenic Those within each lamella of a haversian
potencies, but is much thinner—usually a system are parallel in their orientation, but
single layer of cells without associated con¬ the direction of the fibers in the successive
nective tissue fibers. All the cavities of bone, lamellae changes. This change in orientation
including the haversian canals and the mar¬ of the fibers is responsible for the alternation
row spaces within spongy bone, are lined by of bright and dark layers in haversian systems
endosteum. viewed with polarizing optics (Fig. 8-8B).
Some disagreement persists as to the precise
arrangement of the fibers. In decalcified
preparations viewed at high magnification,
BONE MATRIX
refractile lamellae with a fine circumferential
striation alternate with less refractile layers
The interstitial substance of bone is com¬ having a stippled or punctate aspect. This
posed of two major components, an organic appearance was originally interpreted as in¬
matrix and inorganic salts, each comprising dicating a regular alternation of lamellae with
208 • BONE

circularly and with longitudinally oriented fission of uranium in nuclear reactors or of

fibrils. This was apparently an oversimplifi¬ uranium or plutonium in the detonation of
cation. Some investigators have insisted that nuclear weapons, a large number of radio¬
collagen-rich lamellae alternate with colla¬ active elements are liberated. Some of these,
gen-poor lamellae and that this difference is on gaining access to the body, are incorpo¬
as important as the direction of the fibers in rated in bone. The most hazardous of these
accounting for the microscopic appearance bone-seeking isotopes is 90Sr. As a result of their
of the haversian systems. Others have sug- radioactivity, isotopes may cause severe dam¬
ested that the fibers within a given collagen- age to bone and to the blood-forming cells
rich lamella are not parallel but form two in the marrow. A few of these bone-seeking
sets of fibers intersecting in a lattice-like pat¬ isotopes, including 239Pu, do not enter the
tern. Most histologists, however, seem to be¬ bone mineral but have instead a special affin¬
lieve that the fibrils in all the lamellae run ity for the organic constituents of bone. Stud¬
helically with respect to the axis of the hav¬ ies of the rate of turnover of the inorganic
ersian canal, but that the pitch of the helix substances in bone have been greatly aided
changes sufficiently from one lamella to the by the use of bone-seeking isotopes.
next to account for the differences observed During development and growth, the
under bright-held and polarization micro¬ amount of organic material per unit volume
scopes. remains relatively constant, but the amount
of water decreases and the proportion of
bone mineral increases, attaining a maximum
Bone Mineral of about 65 per cent of the fat-free dry weight
The inorganic matter of bone consists of of the tissue in the adults. In the poorly
submicroscopic deposits of a form of calcium calcified bone of individuals suffering from
phosphate, very similar, but not identical, to rickets or osteomalacia, the mineral content may
the mineral hydroxyapatite (Ca10 [P04]6 be as low as 35 per cent.
[OH]2). Bone mineral is probably deposited If bone is exposed to a weak acid or a
initially as amorphous calcium phosphate and chelating agent, the inorganic salts are re¬
subsequently reordered to form crystalline moved. The bone thus demineralized loses
hydroxyapatite. In its final phase, the calcium most of its hardness but is still very tough
phosphate is present as thin plates or slender and flexible. It retains its gross form and a
rodlike crystals 1.5 to 3 nm in thickness and nearly normal microscopic appearance. On
about 40 nm long. These are situated on and the other hand, if the organic constituents
within the substance of collagen fibers in the are extracted from a bone, the remaining
matrix. The crystals are not randomly dis¬ inorganic constituents retain the gross form
tributed but recur regularly at intervals of 60 of the bone and, to a certain extent, its
to 70 nm along the length of the fibers. microscopic topography, but the bone has
Bone mineral contains significant amounts lost much of its tensile strength and is as
of the citrate ion C6H5Ovs and the carbonate brittle as porcelain. Thus, it is clear that the
ion COs=. Citrate is considered to be in a hardness of bone depends on its inorganic
separate phase, located on the surfaces of constituents, while its great toughness and
crystals. The site of carbonate is still a matter resilience reside in its organic matrix, partic¬
of debate; it may be located on the surface ularly the collagen. Without either one, bone
of crystals, or it may substitute for P04~ in would be a poor skeletal material, but with
the crystal structure, or both. Substitution of both, it is a highly ordered, remarkably re¬
the fluoride ion F~ for OH- in the apatite sistant tissue, superbly adapted, at all levels
crystal is common; its amount depends of its organization, for its chemical and me¬
mainly on the fluoride content of the drink¬ chanical functions.
ing water. Magnesium and sodium, which
are normal constituents of body fluids, are
also present in bone mineral, which to some
extent serves as a storage depot for these
THE CELLS OF BONE
elements. The isotopes 45Ca and 32P can, of
course, substitute for the stable 40Ca and 31P In actively growing bones four kinds of
in the hydroxyapatite crystal. Foreign cations, bone cells are distinguishable: osteoprogenitor
such as Pb+ + , Sr+ + , and Ra++ (22bRa), if in¬ cells, osteoblasts, osteocytes, and osteoclasts. Al¬
gested, may also substitute for Ca+ + . In the though the first three are usually described
 BONE • 209
as distinct cell types, there is clear evidence
meantime, a majority of contemporary inves¬
of ti ansformation from one to the other, and
tigators of bone prefer the more limited
it is evidently more reasonable to regard
implications of the term osteoprogenitor cell.
them as different functional states of the
same cell type. Such reversible changes in
appearance are examples of cell modulation, Osteoblasts
in contrast to differentiation, which is the term
The osteoblasts are responsible for forma¬
reserved for progressive and apparently ir¬ tion of bone matrix and are invariably found
reversible specialization in structure and on the advancing surfaces of developing or
function. The osteoclasts have a separate or¬ growing bone. During active deposition of
igin from monocytes formed in the bone
new matrix, they are arranged in an epithe¬
marrow and circulating in the blood.
lioid layer of cuboidal or low columnar cells
connected to one another by short slender
Osteoprogenitor Cells processes. The nucleus with its single prom¬
inent nucleolus is often at the end of the cell
Like other connective tissues, bone devel¬ farthest from the bony surface. The cells are
ops from embryonic mesenchyme. It retains clearly polarized toward the underlying bone,
in postnatal life a population of relatively with a well-developed Golgi apparatus situ¬
undifferentiated cells that have the capacity ated between the nucleus and the cell base.
for mitosis and for further structural and The mitochondria are elongated and fairly
functional specialization. These osteoprogenitor numerous. The cytoplasm is intensely baso¬
cells have pale-staining, oval or elongate nu¬ philic, owing to its large content of rough
clei and inconspicuous acidophilic or faintly endoplasmic reticulum.
basophilic cytoplasm. They are found on or The osteoblasts give a strong histochemical
near all the free surfaces of bone: in the reaction for alkaline phosphatase, and the
endosteum; in the innermost layer of the periodic acid-Schiff reaction reveals in cyto¬
periosteum; lining the haversian canals; and plasmic vacuoles small pink-staining granules
on the trabeculae of cartilage matrix at the that are believed to represent precursors of
metaphysis of growing bones. the bone matrix. When active new formation
The osteoprogenitor cells are active during of bone ceases and the osteoblasts revert to
the normal growth of bones and may be spindle form, these granules disappear from
activated in adult life during internal reor¬ the cytoplasm, and the phosphatase reaction
ganization of bone or in the healing of frac¬ of the cells rapidly declines.
tures and repair of other forms of injury. In electron micrographs, osteoblasts are
Under any of these conditions, they undergo seen to have the structure expected of cells
division and transform into the bone-forming actively engaged in protein synthesis (Fig.
osteoblasts. 8-11). The endoplasmic reticulum is exten¬
After administration of tritiated thymidine, sive and its cisternae are often in parallel
the osteoprogenitor cells are the only cells array. Their membranes are studded with
found to be labeled in autoradiographs at ribosomes, and these are also present in great
early time intervals. At later times, silver numbers in the cytoplasmic matrix. The
grains can also be found over the nuclei of Golgi membranes are well developed and
osteoblasts, indicating that some of the osteo¬ have numerous associated vacuoles. Sizable
progenitor cells have become transformed vesicles containing an amorphous or floccu-
into bone-forming cells. The osteoblasts can lent material of appreciable density appar¬
also revert to osteoprogenitor cells when os¬ ently correspond to the PAS-staining gran¬
teogenesis subsides. ules observed with the light microscope.
Many authors refer to these potentially Small lipid droplets and membrane-limited
osteogenic cells as mesenchymal cells, but this dense bodies, interpreted as lysosomes, are
term implies that they have a broader range also encountered occasionally.
of latent developmental potentialities than
has been demonstrated. It may yet be shown
Osteocytes
that these cells can also develop into adipose
cells, into fibroblasts, and into hemopoietic The principal cells of fully formed bone
cells of the bone marrow. If this should be are the osteocytes, which reside in lacunae
true, ’’mesenchymal cell” would indeed be within the calcified interstitial substance. The
the more appropriate designation. In the cell body is flattened, conforming to the
210 • BONE

Figure 8-11. Edge of a resorption canal being filled in by lamellar bone. At upper left is a portion of an osteoblast
containing a prominent Golgi zone and abundant granular reticulum. Subjacent to it are the collagen fibrils of two
unmineralized lamellae, and at lower right is the dense mineralized bone. (From Cooper, et al. J. Bone Joint Surg.
48A:1239, 1966.)

shape of the lenticular cavity that it occupies, The nuclear and cytoplasmic characteris¬
but there are numerous slender processes tics of osteocytes at the light microscopic level
that extend for some distance into canaliculi » are similar to those of osteoblasts except that
in the surrounding matrix. How far they the Golgi region is less conspicuous and the
penetrate into the canaliculi of adult mam¬ cytoplasm exhibits less affinity for basic dyes.
malian bone could not be ascertained by light In electron micrographs of osteocytes that
microscopy. However, electron microscopic have only recently been incorporated into
studies have shown that the processes of bone, the Golgi apparatus is still rather large
neighboring osteocytes are in contact at their and the endoplasmic reticulum is quite exten¬
ends. Moreover, their apposed membranes sive (Fig. 8-13). In osteocytes situated deeper
are specialized to form gap junctions or nex¬ in bone matrix, these organelles have under¬
uses at their sites of contact (Fig. 8-12). Thus, gone some regression (Fig. 8—14). Although
the bone cells are not completely isolated in these cells appear less active in protein syn¬
their lacunae but appear to be in communi¬ thesis, they are by no means metabolically
cation with one another and ultimately with inert.
the cells at the surface via a series of cell-to- In its development, an osteocyte is essen¬
cell junctions of low electrical resistance, per¬ tially an osteoblast that has become sur¬
mitting flow of ions and possibly of small rounded by bone matrix. Isolated within its
molecules. This finding may explain how cells lacuna, it undergoes some cytological dedif¬
deep within the calcified matrix of bone can ferentiation, but remains active. Considerable
respond to hormonal stimuli that would seem indirect evidence has accumulated indicating
to have direct access only to cells in the that the osteocyte exerts an important influ¬
immediate vicinity of blood vessels. ence on the surrounding osseous matrix. The
 BONE • 211

tion, it may revert to a quiescent osteopro-

genitor cell.

Osteoclasts
Closely associated with areas of bone re¬
sorption are the osteoclasts—giant cells 20 to
100 p.m in diameter and containing as many
as 50 nuclei. They were first described a
century ago and believed to be the active
agents in bone resorption. Although this view
has been a subject of continuing debate, the
great bulk of recent evidence supports this
interpretation. Osteoclasts are frequently
found in shallow concavities in the surface of
bone, called Howship’s lacunae. It was this
relationship that first suggested to early in¬
vestigators of the histology of bone that these
lacunae were formed by an erosive action of
osteoclasts. In the turnover and remodeling
of bones that occurs in growing animals,
osteoclasts are always most abundant in those
areas known to be undergoing resorption.
No one questioned the close topographical
relation of these cells to sites of resorption,
but some argued that the osteoclasts arose by
coalescence of osteocytes liberated from sur¬
Figure 8-12. Electron micrograph of portions of cell
rounding matrix in the course of bone re¬
processes from two neighboring osteocytes traversing the
zone of unmineralized matrix lining a lacuna. Note the sorption, and that they therefore should be
nexus or gap junction where the processes overlap. (From regarded as products rather than as agents
Holtrop, M. E., and M. J. Weinger. Proc. Fourth Parathy¬ of bone resorption. This interpretation now
roid Conference, Internat. Congress Series No. 243. Am¬ has few adherents, as the evidence for an
sterdam, Excerpta Medica, 1971.)
active role of osteoclasts in bone resorption
has become compelling. Osteoclasts actively
engaged in bone resorption show an obvious
phenomenon of osteolysis is an active physio¬ polarity. The nuclei tend to congregate near
logical process whereby the bone matrix im¬ the outer surface, which is smooth contoured,
mediately surrounding osteocytes is modified while the side adjacent to the bone exhibits a
and bone salt is resorbed. It is the current radial striation that was long interpreted as a
belief that the osteocytes play an active role “brush border” but which has now been
in the release of calcium from bone to blood, shown by electron microscopy to have an
and hence participate in the homeostatic reg¬ infolded structure that makes the term “ruf¬
ulation of its concentration in the fluids of fled border” more appropriate. There are
the body. deep infoldings of the plasma membrane,
Parathyroid hormone is the principal reg¬ which delimit a large number of closely
ulator of the blood calcium level. Its admin¬ packed clavate or foliate processes of highly
istration has a microscopically visible effect variable size and shape, separated by narrow
on the osteocytes and on the staining reac¬ extracellular clefts (Fig. 8-16). Small crystals
tions of the adjacent bone matrix. Because of bone mineral liberated from the bone
the response of the blood calcium level to matrix may be found deep in these clefts.
parathyroid hormone is too rapid to be ac¬ The plasma membrane itself is specialized
counted for by osteoclastic erosion of bone, in the region of the ruffled border. It bears
the primary action of the hormone may be on its inner or cytoplasmic surface a nap of
to stimulate osteocytic osteolysis. exceedingly small, bristle-like appendages 15
The osteocyte is believed to be capable of to 20 nm in length and spaced about 20 nm
modulation to other cell types. When re¬ apart. This bristle coat makes the membrane
leased from its lacuna during bone resorp¬ in this elaborately infolded region appear
Figure 8-13. Electron micrograph of an osteocyte. The plane of section does not include the nucleus. Notice that the
Golgi complex is still quite well developed, and numerous cisternal profiles of endoplasmic reticulum are present. At the
left, a cell process is seen extending into a canaliculus. An area similar to that in the rectangle is seen at higher
magnification in Figure 8-15. (Micrograph courtesy of M. Holtrop.)

Figure 8-14. Electron micrograph of an osteocyte. Notice that it completely fills its lacuna. The clear area around the
cell is in fact occupied by unmineralized matrix in which the collagen fibers are faintly visible. The mineralized matrix is
black owing to the electron scattering of the apatite crystals. (Micrograph courtesy of M. Holtrop.)
212
 BONE • 213

Figure 8-15. Osteocyte process extending from cell body

(above) into a canaliculus (beiow). Notice the high con¬
centration of cytoplasmic filaments in the cell process.
(Micrograph courtesy of M. Holtrop.)

thicker than the unspecialized unit mem¬ dicative of their lysosomal nature. In electron
brane elsewhere on the cell surface. In con¬ micrographs, the granules are for the most
trast to a typical brush border, which is a part dense, spherical, 0.2 to 0.5 pim in di¬
very well-ordered and stable differentiation, ameter, and membrane limited, but larger
the ruffled border of the osteoclast seems to vesicular structures 0.5 to 3 pm in diameter
be highly active and constantly changing its may also be secondary lysosomes.
configuration. Cinematographic studies of Despite the evidence of activity of the os¬
these cells in vitro have shown that processes teoclast surface where it is in contact with
are continually being extended and retracted bone, there is little evidence that these cells
and changing their position. are mechanically erosive or even highly phag¬
The nuclei of osteoclasts resemble those of ocytic. The exact mechanisms by which they
osteoblasts and are in no way unusual. The accomplish the simultaneous degradation of
cytoplasm is slightly basophilic when stained the organic matrix and the dissolution of
with basic dyes at controlled pH, but in rou¬ bone mineral still elude us, but there are
tine histological sections it is usually eosino¬ rather clear indications that they secrete hy¬
philic and highly vacuolated. There are mul¬ drolytic enzymes, including collagenase,
tiple Golgi complexes distributed among the which are responsible for digestion of matrix
nuclei, and a number of centriole pairs cor¬ components.
responding to the number of nuclei. The The experiments upon which this conclu¬
centrioles may gather together in a centro- sion is based depend on the fact that main¬
some region. The rod-shaped or short fila¬ tenance of normal levels of blood calcium in
mentous mitochondria of osteoclasts are very the intact animal depend on the actions of
numerous and tend to congregate near the two hormones that act antagonistically on
ruffled border. In contrast to other bone bone. Administration of parathyroid hormone
cells, the cytoplasmic vacuoles of osteoclasts causes mobilization of calcium by promoting
stain supravitally with neutral red—a prop¬ bone resorption, while calcitonin acts to sup¬
erty that can be used to advantage in locating press mobilization of calcium from bone.
these multinucleate cells in fresh bone for Both hormones are also effective when ap¬
experimental manipulation. Many of the vac¬ plied to isolated bone fragments maintained
uoles and granules also give a positive histo- in vitro. Thus, addition of parathyroid hor¬
chemical reaction for acid phosphatase, in¬ mone to organ cultures results in appearance
214 • BONE

Mitochondria

SH Nucleus

Ruffled

Resorbing
bone
matri x

Figure 8-16. Electron micrograph of a portion of an osteoclast, including the nucleus above, several Golgi elements,
and the ruffled border closely applied to an area of resorbing bone matrix at bottom of figure. (Micrograph courtesy of
P. Garrant.)
 BONE • 215

of resorption cavities in the bone and mark¬ posed that lysosomal acid hydrolases of os¬
edly accelerated release of calcium and phos¬ teoblasts are active in resorption of the or¬
phate into the medium as a result of solubi¬ ganic matrix of bone and that the stimulated
lization of bone mineral. There is also an local acid production solubilizes bone mineral
increase in liberation of hydroxyproline from and at the same time creates a pH favorable
degradation of collagen of the bone matrix. to the action of acid hydrolases.
Concomitant with these evidences of bone The surface membrane of osteoclasts
resorption, appreciable quantities of several around the periphery of the active ruffled
lysosomal acid hydrolases are also released border is very smooth and closely applied to
into the medium, whereas there is no detect¬ the underlying bone (Fig. 8—17). This obser¬
able release of nonlysosomal enzymes from vation has led to the speculation that the
the cells of the culture. It is suggested, there¬ close application of the cell to its substrate in
fore, that the lysosomal enzymes are secreted the clear zone around the periphery of the
by osteoclasts. Parathyroid hormone also ruffled area might help to seal off the active
greatly increases the rate of production of portion of the cell and enable it to maintain
lactate and citrate by the bone explants, caus¬ a microenvironment conducive to solubiliza¬
ing them to acidify their medium much more tion of mineral and to optimal activity of
rapidly than other cells in culture. From hydrolytic enzymes.
consideration of such experiments, it is pro¬ There is additional physiological evidence

Figure 8-17. Electron micrograph of a portion of an osteoclast around the end of a spicule of bone. Notice the ruffled
border at the end of the bone, where it is undergoing resorption. A smooth contoured portion of the osteoclast surface
with a subjacent clear zone of ectoplasm is closely applied to the bone around the periphery of the ruffled border.
(Micrograph courtesy of M. Holtrop.)
216 • BONE

for a direct action of hormones on the osteo¬ modes of osteogenesis are recognized in em¬
clast. Administration of parathyroid hor¬ bryos! When bone formation occurs directly
mone is reported to cause depolarization of in primitive connective tissue, it is called
the osteoclast membrane and an increase in intramembranous ossification. When it takes
the rate of RNA synthesis, while calcitonin place in preexisting cartilage, it is called in-
polarizes the cell membrane and inhibits the tracartilaginous or endochondral ossification. In
effect of parathyroid hormone on RNA syn¬ endochondral ossification the bulk of the
thesis. Calcitonin also causes disappearance cartilage must be removed before bone dep¬
of the ruffled border, a change in the char¬ osition begins, and the distinctive features of
acter of the membrane, and separation of this mode of ossification are more concerned
osteoclasts from the bone surface. with the resorption of cartilage than with
It is generally accepted that the multinu- deposition of bone. The actual deposition of
cleate osteoclasts arise by coalescence of uni¬ bony tissue is essentially the same in the two
nucleate cells, but a consensus as to the iden¬ modes of ossification. Bone is first laid down
tity of the precursor cell has been reached as a network of trabeculae, the primary spon¬
only recently. Osteoprogenitor cells, osteo¬ giosa, which is subsequently converted to
blasts, and osteocytes liberated from the ma¬ more compact bone by a filling in of the
trix have all been proposed and strongly interstices between trabeculae. Occasionally,
defended. However, some 30 years ago, in under pathological ‘ conditions, bone may
experiments that involved labeling of hemo¬ arise in tissues not belonging to the osseous
poietic cells with tritiated thymidine, it was system, and in connective tissues not ordinar¬
observed in radioautographs that some osteo¬ ily manifesting osteogenic properties. This is
clasts contained labeled nuclei. Since the nu¬ called ectopic bone formation.
clei of such cells do not divide and hence
would not have incorporated thymidine dur¬
Intramembranous Ossification
ing DNA replication, it was concluded that
the labeled osteoclasts arose by fusion of Certain flat bones of the skull—the frontal,
mononuclear cells of bone marrow origin. parietal, occipital, and temporal bones and
Although this interpretation was slow to gain part of the mandible—develop by intramem¬
acceptance, confirmatory observations with branous ossification and are referred to as
labeling experiments of other design have membrane bones. The mesenchyme condenses
accumulated. Perhaps the most compelling into a richly vascularized layer of connective
evidence for the origin of osteoclasts from tissue, in which the cells are in contact with
cells circulating in the blood has been pro¬ one another by long tapering processes, and
vided by ingenious experiments with mice of the intercellular spaces are occupied by ran¬
a mutant strain that have osteopetrosis—an domly oriented delicate bundles of collagen
excessive accumulation of spongiosa in their fibrils embedded in a thin, gel-like extracel¬
long bones due to impaired osteoclastic re¬ lular matrix. The first sign of bone formation
sorption of bone during development. When is the appearance of thin strands or bars of
such osteopetrotic mice are surgically joined a denser eosinophilic matrix (Fig. 8—18).
to normal littermates in parabiosis, the excess These strands of bone matrix tend to be
spongiosa disappears from their bones within deposited approximately equidistantly from
six weeks, and they remain cured even after neighboring blood vessels, and since the ves¬
separation from their normal parabiont. The sels form a network, the earliest trabeculae
conclusion is inescapable that the mononu¬ of bone matrix also develop in a branching
clear progenitors of the osteoclasts originated and anastomosing pattern (Fig. 8-19). Si¬
from the blood of the normal mouse while multaneously with the appearance of those
their circulatory systems were in continuity. first strands of eosinophilic extracellular ma¬
Osteoclasts are now believed to arise by fu¬ trix, there are changes in the neighboring
sion of monocytes, and although they are not primitive connective tissue cells. They en¬
phagocytic in the usual sense, osteoclasts are large and gather in increasing numbers on
considered members of the diffuse mono¬ the surface of the trabeculae, assuming a
nuclear phagocyte system. cuboidal or columnar form while still remain¬
ing adherent to one another via shortened
processes. Concurrently with the changes in
HISTOGENESIS OF BONE their size and shape, the cells become in¬
tensely basophilic and are thereafter identi¬
Bone always develops by replacement of a fied as osteoblasts. Through their synthetic
preexisting connective tissue. Two different and secretory activity, additional bone matrix
 BONE • 217

Figure 8-19. Photomicrograph of the pattern of trabecu¬

lae in the primary spongiosa of intramembranous bone
formation.

bone in which the collagen fibers run in all

directions is often called woven bone, to distin¬
guish it from the lamellar bone formed in
subsequent remodeling, which contains col¬
lagen in highly ordered parallel arrays.
Woven bone is permeated by relatively large
tortuous channels occupied by blood vessels
and connective tissue. The osteocytes are
distributed uniformly but oriented at ran¬
dom. In lamellar bone, on the other hand,
the osteocytes are arranged in regular con¬
centric order around relatively straight ves¬
sels in slender haversian canals (Fig. 8-20).
At a very early stage in the replacement of
the interstitial substances of primitive con¬
Figure 8-18. Beginning of intramembranous bone for¬ nective tissue by bone matrix, the latter be¬
mation in the skull of a 5.5-cm cat embryo. B, Homoge¬ comes a site of deposition of calcium phos¬
neous thickened collagenous fibers, which become the
interstitial bone substance. C, Collagenous interstitial sub¬
phate. All of the matrix subsequently
stance. F, Connective tissue cells. O, Connective tissue secreted by the osteoblasts calcifies after a
cells, with processes, which become osteoblasts and later very short lag. Thus, in electron micro¬
bone cells. Eosin-azure stain. (After A. A. Maximow.) graphs, there is only a narrow zone of osteoid
between the bases of the osteoblasts and the
(osteoid) is deposited, and the trabeculae be¬ heavily mineralized matrix of the underlying
come longer and thicker. trabeculae (Fig. 8-11). As the trabeculae
Collagen molecules are secreted together thicken by accretion, some of the osteoblasts
with the proteoglycans of the matrix, and at their surface become incarcerated in the
they polymerize extracellularly to form great newly deposited matrix, and one by one they
numbers of randomly interwoven fibrils of are buried within its substance to become
collagen throughout the trabeculae of os¬ bone cells—osteocytes. The osteocytes thus
seous matrix. This early intramembranous sequestered in lacunae within the newly de-
218 • BONE

Figure 8-20 Three-dimensional diagrammatic representation of the differences in architecture of woven bone (A) and
lamellar bone (B). (From Hancox, N. M. Biology of Bone. Cambridge, England, Cambridge University Press, 1972.)

posited matrix nevertheless remain con¬ osteoblasts that have remained on the surface
nected to osteoblasts at the surface by slender of the bone during its development revert to
processes. The canaliculi of bone are formed the fibroblast-like appearance as growth
by deposition of matrix around these cell ceases and persist as the quiescent osteopro-
processes. As rapidly as the ranks of osteo¬ genitor cells of the endosteum or of the
blasts on the surface of the trabeculae are periosteum. If they are again called upon to
depleted by their incorporation into the form bone, their osteogenic potentialities are
bone, their numbers are restored by differ¬ reactivated and they again take on the mor¬
entiation of new osteoblasts from primitive phological characteristics of osteoblasts.
cells of the surrounding connective tissue.
Mitotic division is frequent in these progeni¬
tor cells but is rarely if ever observed in Endochondral Ossification
osteoblasts.
In areas of the primary spongiosa that are Bones at the base of the skull, in the
destined to become compact bone, the tra¬ vertebral column, in the pelvis, and in the
beculae continue to thicken at the expense of extremities are called cartilage bones because
the intervening connective tissue until the they are first formed of hyaline cartilage,
spaces around the blood vessels are largely which is then replaced with bone in the
obliterated. The collagen fibrils in the layers process called endochondral ossification. This
of bone that are deposited on the trabeculae can best be studied in one of the long bones
in this progressive encroachment upon the of an extremity. The first indication of the
perivascular spaces gradually become more establishment of a center of ossification is a
regularly arranged and come to resemble striking enlargement of the chondrocytes at
lamellar bone. Although the irregularly con¬ the middle of the shaft of the hyaline carti¬
centric layers formed may bear a superficial lage model (Fig. 8—21). The cells in this
resemblance to haversian systems, they are region hypertrophy, glycogen accumulates
not true lamellar bone because their collagen within them, and their cytoplasm becomes
is randomly oriented. highly vacuolated. As the chondrocytes hy¬
In those areas where spongy bone will pertrophy there is an enlargement of their
persist, the thickening of the trabeculae lacunae at the expense of the intervening
ceases and the intervening vascular connec¬ cartilage matrix, which is gradually reduced
tive tissue is gradually transformed into hem¬ to thin fenestrated septa and irregularly
opoietic tissue. The connective tissue sur¬ shaped spicules. The remaining hyaline ma¬
rounding the growing mass of bone persists trix in the region of hypertrophic cartilage
and condenses to form the periosteum. The cells becomes calcihable, and small granular
 BONE • 219

Endochondral Nucleus Endochondral

ossification center puiposus ossification center

Figure 8-21. Photomicrograph of the cartilaginous vertebral column of a mouse embryo, showing in the center of each
vertebra an area of hypertrophied cartilage cells that represents an early stage in the establishment of a center of
endochondral ossification.

aggregati°ns and nests of crystals of calcium tiate into osteoblasts, which congregate in an
phosphate are deposited within it (Fig. epithelioid layer on the irregular surfaces of
8-25C). Regressive changes in the hypertro¬ spicules of calcified cartilage matrix and be¬
phied cartilage cells, including swelling of gin to deposit bone matrix upon them. The
their nuclei and loss of chromatin, are fol¬ earliest bony trabeculae formed in the inte¬
lowed by their death and degeneration. rior of the cartilage model thus have a core
Concurrently with these hypertrophic and of calcified cartilage and an outer layer of
regressive changes in the chondrocytes in the bone of varying thickness. Owing to the dif¬
interior of the cartilage model, the osteogenic ferent staining affinities of calcified cartilage
potencies of cells in the perichondrium are and bone, these trabeculae have a mottled
activated, and a thin layer of bone, the per¬ heterogeneous appearance and are easily dis¬
iosteal band or collar, is deposited around the tinguished from the homogeneous trabeculae
midportion of the shaft (Figs. 8-22B, 8-25A). of woven bone formed under the periosteum
At the same time, blood vessels from the by intramembranous ossification.
investing layer of connective tissue (now It is common practice to include in the
called the periosteum) grow into the dia- term primary center of ossification all the early
physis, invading the irregular cavities in the morphological changes described above,
cartilage matrix created by the enlargement whether they occur in the interior of the
of the chondrocytes and confluence of their cartilage model or under the perichondrium.
lacunae (Figs. 8-22E,F, 8-23, 8-24). The This usage is intended only to distinguish the
thin-walled vessels branch and grow toward diaphyseal center, which appears first, from
either end of the cartilage model, forming secondary centers of ossification, which develop
capillary loops that extend into the blind ends much later in the epiphyses. Some investiga¬
of the cavities in the calcified cartilage. Prim¬ tors, however, reserve the term primary cen¬
itive pluripotential cells are carried into the ter of ossification for the subperiosteal collar
interior of the cartilage in the perivascular on the grounds that this is the first true bone
tissue of the invading blood vessels. Some of formed, even though its formation is her¬
these cells differentiate into hemopoietic ele¬ alded by earlier changes in the chondrocytes
ments of the bone marrow. Others differen¬ in the interior of the model.
220 • BONE

Figure 8-22. Diagram of the development of a typical long bone as shown in longitudinal sections (A to J) and in cross
sections A', B', C\ and D' through the centers of A, B, C, and D. Pale blue, cartilage; purple, calcified cartilage; black,
bone; red, arteries. A, Cartilage model. B, Periosteal bone collar appears before any calcification of cartilage. C,
Cartilage begins to calcify. D, Vascular mesenchyme enters the calcified cartilage matrix and divides it into two zones
of ossification (E). F, Blood vessels and mesenchyme enter upper epiphyseal cartilage and the epiphyseal ossification
center develops in it (G). A similar ossification center develops in the lower epiphyseal cartilage (H). As the bone ceases
to grow in length, the lower epiphyseal plate disappears first (I) and then the upper epiphyseal plate (J). The bone
marrow cavity then becomes continuous throughout the length of the bone, and the blood vessels of the diaphysis,
metaphyses, and epiphyses intercommunicate.
 BONE • 221

Bone Osteoblast Periosteum

Mesenchymal
cell in
mitosis

Vessel
Hemocyto-
blast

Cartilage\ Vessel
cells

Hemocyto-
blasts

Mesenchymal
bud

Bone Hemocytoblasts
Figure 8-23. Part of longitudinal section through the middle of theaiiaphysis of the femur of a 25-mm human embryo.
Mesenchyme with vessels entering calcified cartilage through an opening in the periosteal bone collar. Eosin-azure
stain. (Afler A. A. Maximow.)

Mechanism of Calcification tals when introduced into metastable solu¬

tions of inorganic calcium and phosphate.
The mechanism by which mineral is de¬ The fact that fibers of the native type, with
posited in the organic matrix of cartilage and 67-nm periodicity, are the only form of col¬
bone has been a subject of much debate and lagen capable of inducing this change led to
numerous hypotheses. One of the most speculation that nucleation of bone mineral
widely accepted of these holds that the initi¬ is dependent on, or is at least facilitated by,
ation of mineralization in bone is analogous the particular arrangement of molecules in
to the familiar induction of crystallization these fibers. This interpretation derives
from metastable solutions in vitro by adding added support from the observation that in
a seed crystal or by scratching the wall of the electron micrographs of early stages of min¬
beaker—a process called heterogeneous nuclea- eralization, the deposits of calcium phosphate
tion. The foreign matter disturbs the equilib¬ are localized on specific regions of the cross-
rium in the solution and causes a clustering banded fibers.
of molecules that results in formation of small During their formation of linear polymers,
nuclei capable of growing to form crystals. the collagen molecules overlap a short dis¬
In the case of bone, it is believed to be the tance (Fig. 8-27). When these linear poly¬
highly ordered “crystalline” collagen fibers of mers associate laterally to form fibrils, they
the matrix that act as the nucleation catalyst pack in such a way that discontinuities or
for transformation of calcium and phosphate “holes” exist within the fibers between the
in solution in the tissue fluids into the solid- tails and heads of successive molecules. In
phase mineral deposits. In support of this negatively stained collagen, the contrast me¬
theory, it has been shown that reconstituted dium penetrates the fiber and fills these
pure collagen fibers and demineralized bone holes, producing the broad, dark bands that
are able to induce formation of apatite crys¬ are characteristic of such preparations. The
222 • BONE

Osteoblast Bone Periosteum

Osteoclast

Vessel

Cartilage
cells
; •
I f
' <

Osteoblast

Mesen¬
chymal bud
Bone Vessel Osteoblast Bone

Figure 8-24. Part of longitudinal section through the middle of the diaphysis of the humerus of a human embryo of
eight weeks. The process of ossification has advanced slightly farther than in Figure 8-23. Eosin stain. (After A. A.
Maximow.)

earliest deposits of bone mineral produce a irregular pattern of crystal deposits on and
similar pattern of dense bands and therefore between collagen fibers. It may also have an
are believed to be localized in the holes within advantage over the collagen-centered nuclea-
the collagen fibers (Fig. 8-27). tion theory in helping to explain those in¬
Persuasive as the evidence is for this at¬ stances of mineralization, such as in epiphy¬
tractive theory, it does not fully explain the seal cartilage, in which the initial deposits are
localization of the process or the absence of localized without obvious topographical re¬
calcification in many other collagen-rich tis¬ lation to typical 67-nm collagen fibers.
sues. It is evident too that collagen cannot be Clearly, this chapter in the story of bone
the only initiator of calcification. The organic development is not closed. Many details of
matrix of the developing enamel of teeth, for the calcification process and its regulation
example, is composed of a very different remain to be worked out.
protein, but it is rapidly and heavily miner¬
alized. A number of investigators insist that Growth in Length of Long Bones
collagen does not act alone in vivo but that
its interaction with chondroitin sulfate or In the continuing growth in length of the
some other glycosaminoglycan in the matrix cartilage model after the appearance of the
may result in a particular stereochemical con¬ diaphyseal center of ossification, the chon¬
figuration that promotes apatite crystal for¬ drocytes in the adjacent regions of the epi¬
mation. Evidence for this hypothesis is frag¬ physes become arranged in longitudinal col¬
mentary, but it would seem to be in better umns instead of in the small groups usually
accord with morphological observations on found in hyaline cartilage (Fig. 7-6). The
early stages of mineralization that show an cells within the columns are separated by thin
 BONE 223

Cal erf i e d cart11age

First stage of calcification Intervertebral

of cartilage disk

Figure 8-25. Photomicrographs showing several stages of bone formation in developing rats. From formalin-fixed,
undecalcified sections stained with silver nitrate to show bone salt (black). A, Longitudinal section through second rib of
18-day rat embryo; calcification of the periosteal bone collar is further advanced than that of the cartilage. B, Section of
metatarsal of 4-day rat, in which ossification is proceeding toward the epiphyses; the hypertrophic cartilage is not
completely calcified. C, Three stages in calcification of vertebrae in 20-day rat embryo. (Courtesy of W. Bloom and M.
A. Bloom.)

transverse septa, while adjacent columns are division of the small flattened cells provides
separated by wider longitudinal bars of hya¬ for continual elongation of the columns. Next
line matrix. As endochondral ossification comes a zone of maturation, in which the cells
progresses from the center of the shaft to¬ that are no longer dividing gradually enlarge.
ward either end of the cartilage model, the This is followed by a zone of hypertrophy, with
chondrocytes undergo the same sequence of very large vacuolated cells. Since the matrix
changes as described for the establishment of in this latter region becomes the site of cal¬
the diaphyseal center, but the process is now cium deposition, this may also be called the
more orderly. Along the length of the epi¬ zone of provisional calcification. And finally, at
physeal cell columns are several recognizable the diaphyseal end of the columns is a zone
zones, corresponding to various stages in the wherein the chondrocytes are degenerating
cytomorphosis of the cartilage cells. At some and the open ends of their enlarged lacunae
distance from the diaphyseo-epiphyseal junc¬ are being invaded by capillary loops and
tion is a zone of proliferation, where frequent osteoprogenitor cells from the marrow spaces
224 • BONE

Proliferating
cartilage cells

Hypertrophic
cartilage cells

Provisional
^ calcification

Invasion of
“cartilage

Primary
spongiosa

Secondary
spongiosa

Figure 8-26. Endochondral ossification in longitudinal sections through the zone of epiphyseal growth of the distal end
of the radius of a puppy. A, Neutral formalin fixation; no decalcification. Von Kossa and hematoxylin-eosin stain. All
deposits of bone salt are stained black; thus, bone and calcified cartilage matrix stain alike. B, Zenker-formol fixation;
decalcified. Hematoxylin-eosin-azure II stain. Persisting cores of cartilage matrix in trabeculae of bone take a deep blue
or purple stain, whereas bone stains red. It is impossible to tell where calcium deposits had been.

of the diaphysis (Fig. 8—26). As the spaces at rickets in growing children or as osteomalacia
the lower ends of the columns are invaded, in adults.
osteoblasts differentiate and congregate on The distribution of calcified cartilage and
the surfaces of the irregularly shaped longi¬ new bone is best demonstrated in undecalci¬
tudinal bars of calcified cartilage that persist fied sections in which the bone mineral has
between them. A thin new layer of bone been stained black with silver by the von
matrix is then deposited on the surface of Kossa method (Fig 8—26A). The transitional
the cartilage. Under favorable conditions it zone where the cartilage is being replaced by
begins to calcify nearly as rapidly as it is laid advancing bone is called the metaphysis. The
down, and thus it becomes bone. Electron primary spongy bone in this zone undergoes
microscopy has shown, however, that a su¬ extensive reorganization as the growth proc¬
perficial layer of uncalcified preosseous tissue esses pass it by. As the bone grows longer,
or osteoid, 1 pm or less in thickness, is always the diaphyseal ends of the trabeculae are
present on forming bony surfaces (Fig. continually being eroded by osteoclasts at
8—11). There may be further lag in calcifica¬ about the same rate that additions are made
tion under conditions of local failure in the at the epiphyseal end, with the result that the
supply of calcium or phosphate. When such spongiosa of the metaphysis tends to remain
a failure becomes general and osteoid accu¬ relatively constant in length.
mulates in excess, the condition is known as Centers of ossification have appeared in
 BONE • 225

Length JU 4.4 units

/v Collagen in balance with the rate of their degeneration
Macromolecule
and removal at the diaphyseal end of the
Overlap zone columns. The epiphyseal plate therefore re¬
o = 0.4 units O

A A tains approximately the same thickness.

—■■ . Protofibril
Growth in length is the result of the cartilage
(linear polymer) cells continually growing away from the shaft
Stagger Vgd+h) Hole zone
s-1.0 units o h = 0.6units
AAi and being replaced by bone as they recede.
•

'!>> f.v.*4 The net effect is an increase in the length of

Fibril the shaft.
At the end of the growing period, prolif¬
eration of cartilage cells slows and finally
ceases. Continued replacement of the carti¬
lage by bone at the diaphyseal end of the
columns then results in complete removal of
the epiphyseal cartilage, and the bony trabe¬
culae of the metaphysis then become contin¬
uous with the spongiosa of the bony epi¬
Fiber physis. This process of elimination of the
epiphyseal plate is referred to as closure of the
epiphysis. When this has taken place, no fur¬
ther longitudinal growth of the bone is pos¬
sible. The times of closure and the relative
contribution of each of the two epiphyses of
a long bone to its overall growth may differ
markedly. Growth in length of the femur,
for example, takes place mainly at the distal
epiphysis; growth of the tibia, mainly at the
proximal epiphysis. Such information is of
Figure 8-27. Diagram of the overlapping staggered ar¬ clinical value in radiology and orthopedic
rangement of molecules in a collagen fiber, showing the
small discontinuities or holes that are thought to be sites
surgery.
of nucleation of apatite crystals in the mineralization of Because all increment in length of a bone
bone. (Modified from Glimcher, M. J., and S. M. Krane. is limited to its epiphyseal plates, injury to
In Gould, B. S., and G. N. Ramachandran, eds.: A Treatise this region may result in serious impairment
on Collagen. Vol. II. New York, Academic Press, 1968.)
of growth. In cases of retarded growth of
one leg attributable to general neurovascular
the diaphysis of each of the principal long disturbances, such as may occur in the limb
bones of the skeleton by the third month of of a child who has had poliomyelitis, the
fetal life. Much later, usually after birth, the orthopedic surgeon can take advantage of
epiphyses show in their interior the charac¬ existing knowledge of the normal rates of
teristic chondrocyte hypertrophy that heralds growth at the various epiphyseal plates and
the onset of endochondral ossification, and of the times of their normal closure, to select
they in turn are invaded by blood vessels and the appropriate time and site for a surgical
osteogenic tissue from the perichondrium to obliteration of an epiphysis in the normal leg.
establish secondary centers of ossification at Such a procedure, if appropriately timed,
either end of the developing long bones (Fig. may retard growth of the normal leg just
8—22C,H) These differ from the diaphyseal enough to permit the slower growing leg to
centers in that there is no associated deposi¬ catch up and thus achieve an equalization of
tion of subperichondral bone. The expansion leg length by the time that growth in stature
of these secondary centers gradually replaces of the individual ceases.
all of the epiphyseal cartilage except that
which persists as the articular cartilage and a
Growth in Diameter of Long Bones
transverse disc between the epiphysis and
diaphysis called the epiphyseal plate (Fig. The long bones of the extremities are first
8—22J). The latter contains the cartilage col¬ laid down in cartilage models, and, as indi¬
umns whose proliferative zone is responsible cated in the foregoing section, their growth
for all subsequent growth in length of long in length depends on endochondral ossifica¬
bones. Under normal conditions the rate of tion. The growth in diameter of the shaft,
multiplication of cartilage cells in this zone is however, is the result of deposition of new
226 • BONE

membrane bone beneath the periosteum. marrow cavity. The rates of external apposi¬
The compact bone forming the shaft of a tion of new bone and internal resorption are
fully developed long bone is almost entirely so adjusted that the cylindrical shaft expands
the product of subperiosteal intramembran- rapidly while the thickness of its wall in¬
ous ossification. creases more slowly.
After establishment of the primary ossifi¬ Because of the continual internal resorp¬
cation center, the ends of the cartilage model tion and reorganization of bone during de¬
continue to elongate and broaden by prolif¬ velopment, the record of the topographical
eration of chondrocytes and elaboration of distribution of endochondral and intramem-
new matrix, but such interstitial growth is no branous ossification in earlier stages of de¬
longer possible in the center of the diaphysis, velopment is continually erased. Therefore,
where the cartilage is regressing and being the extent of the contribution of the perios¬
replaced by bone. The diameter of the en¬ teum to the fully formed long bone is seldom
dochondral component in the middle of the fully appreciated. It is informative in this
diaphysis, therefore, cannot be appreciably regard to examine developing long bones of
greater than the diameter of the cartilage the manatee, an aquatic mammal in which
model in the early embryo at the time of resorption of bone to form the secondary
establishment of the primary center of ossi¬ marrow cavity does not take place. In fetal
fication. To keep pace with the rapid inter¬ bones of this species (Figs. 8—28, 8—29), the
stitial growth of the cartilage at the ends, primary spongiosa of endochondral bone has
increase in thickness of the shaft is accom¬ a characteristic hourglass distribution. The
plished by a progressive thickening of the two conical regions, with their apices meeting
periosteal band or collar formed around the at the site of the original ossification center,
middle of the cartilaginous diaphysis at the result from uniform growth in length and
onset of ossification. This results in deposi¬ breadth of the ends of the cartilage model.
tion of a lattice of trabeculae of intramem- The area between the diverging sides of the
branous woven bone, forming the wall of the two cones is filled in by a thick collar of
diaphysis. trabeculae of periosteal origin. Such bones,
Bone resorption is as important to the lacking the capacity for the resorption that
growth of bones as is bone deposition, and occurs in the histogenesis of long bones in
the deposition of new bone on the outside of other species, provide an instructive view of
the shaft is accompanied by the appearance the basic topography of the cartilaginous and
of osteoclasts that erode the inner aspect of membranous components of all long bones
the subperiosteal trabeculae to enlarge the (Fig. 8-29).

Primary spongiosa

Figure 8-28. Photomicrograph of the humerus in a fetal manatee in longitudinal section. In this species, whose bones
lack a secondary marrow cavity, the respective contributions of subperiosteal and endochondral bone to the formation
of the shaft of a long bone are more evident than in bones of other species. (After Fawcett, D. W. Am. J. Anat. 77:271,
1942.)
 BONE • 227

isotopes or the antibiotic tetracycline, both of

which are deposited preferentially in newly
forming bone.
Typical of such experiments are those lo¬
calizing the sites of osteogenesis in the grow¬
ing rat tibia. This bone supports a large
articular surface, and the epiphysis is consid¬
erably broader than the shaft. Thus, it is
possible to distinguish a cylindrical region in
the middle of the shaft and a conical region
toward the end, where it expands to the
width of the epiphysis. If a bone-seeking
isotope is given to a growing rat and autora¬
diographs are then made of longitudinal sec¬
tions of the tibia, the sites of new bone
formation are disclosed by the distribution of
silver grains in the overlying emulsion. In
the conical region of the bone, the silver
grains are aligned immediately subjacent to
the endosteum, whereas in the cylindrical
portion of the shaft they are found beneath
the periosteum (Fig. 8-31). Study of parallel
histological sections reveals numerous osteo¬
clasts beneath the periosteum of the conical
region and beneath the endosteum of the
cylindrical segment. Thus, it is clear that in
the surface remodeling of this bone, the
Figure 8-29. Diagrammatic representation of the devel¬
periosteum plays opposite roles in neighbor¬
opment of a manatee bone (above) compared with that
of a typical mammal (below). (After Fawcett, D. W. Am. ing regions on the surface of the same bone.
J. Anat. 77:271, 1942.) Subperiosteal bone deposition is occurring in
the cylindrical portion of the shaft while
subperiosteal bone absorption is taking place
Surface Remodeling of Bones in the conical region. Similarly, bone is being

Although growing bones are constantly

changing their internal organization, they
retain approximately the same external form
from an early fetal stage into adult life. It is
apparent that this would not be so if new
bone were deposited at a uniform rate at all
points beneath the periosteum. Instead, the
shape of a bone is maintained during growth
by a continual remodeling or sculpturing of
its surface, which involves bone deposition in
some areas of the periosteum and bone ab¬
sorption in other areas. That this is true was
demonstrated in the middle of the eighteenth
century by madder feeding experiments.
With this method of vital staining, the bone
deposited during a period of feeding on
madder root was stained red, while areas that
were stable or were undergoing resorption
remained unstained. It was clearly shown that
some areas of the surface of long bones
stained while others did not. The general
features of these early experiments have now Figure 8-30. Diagram to illustrate remodeling during
been confirmed and extended by means of growth of tibia and fibula of rat, viewed from anterior
newer techniques employing bone-seeking aspect and in profile. (After Wolbach.)
228 • BONE

concurrently with bone deposition on the

outer surface, but also the rates of deposition
and absorption must differ from the center
to the periphery of each cranial bone in order
to account for its flattening as the radius of
curvature of the skull vault increases. How
these local variations in function of endos¬
teum and periosteum are controlled in space
and time to mold and shape the bone con¬
stantly during its growth is a fascinating un¬
solved problem in morphogenesis.

Internal Reorganization of Bone

The conversion of the primary lattice of
trabeculae laid down in intramembranous
ossification into compact bone is attributed to
thickening of the trabeculae and a progres¬
sive encroachment‘of bone upon the perivas¬
cular spaces until these are largely obliter¬
ated. As the process advances, bone is
deposited in ill-defined layers with randomly
oriented collagen fibers but since these are
disposed more or less concentrically around
vascular channels, they come to bear a super¬
ficial resemblance to haversian systems. They
Figure 8-31. Diagram based on an autoradiograph of the are sometimes called primitive haversian sys¬
head of the tibia of a growing rat killed a few hours after tems, but they should be clearly distinguished
receiving an injection of 32P. The localization of high from the more precisely ordered lamellar
concentrations of silver grains in the autoradiograph is
depicted here in black. In addition to the new bone in the systems comprising the definitive haversian sys¬
epiphysis and at the metaphysis, bone is being deposited tems of adult bone. The latter arise only in
under the endosteum in the conical portion and beneath the course of the internal reorganization of
the periosteum in the cylindrical portion of the shaft primary compact bone that is referred to as
(arrows). (Drawing based on studies of Leblond et al. Am.
secondary bone formation.
J. Anat. 86:289, 1950. After Fawcett, D. W. In Greep, R.
O., ed.: Histology. Philadelphia, Blakiston Co., 1954. At scattered points in the compacta, usually
Reproduced by permission of McGraw-Hill Book Co.) in those areas laid down earliest, cavities
appear as a result of osteoclastic erosion of
primary bone. Such absorption cavities enlarge
formed at the endosteal surface of the cone to form long cylindrical cavities occupied by
and absorbed on the inner aspect of the blood vessels and embryonic bone marrow.
cylinder. As a consequence of these activities, When they reach a considerable length, de¬
the midportion of the shaft is expanding struction of bone ceases; the osteoclasts give
radially and its marrow cavity is being en¬ way to osteoblasts, and concentric lamellae of
larged. While the bone as a whole is elongat¬ bone are laid down on the walls of the cavity
ing by growth at the epiphyseal plate, the until it is filled in to form a typical haversian
diverging wall of the conical region of the system of lamellar bone. The lamellae of this
shaft is being straightened and is contribut¬ and subsequent generations of haversian sys¬
ing, at its lower end, to the lengthening of tems have the ordered arrangement of col¬
the cylindrical region of the shaft. lagen and the change in its orientation in
Similarly, in the skull vault, the assumption successive layers that are characteristic of
that growth of the flat bones at the sutures osteons in adult bone. In man from about
could account for enlargement of the cranial the age of 1 year onward, only lamellar bone
cavity to accommodate the growing brain is of this character is deposited within the shafts
not sufficient as an explanation. As the radius of long bones. This secondary bone eventu¬
of curvature of the growing skull vault in¬ ally replaces all the primitive haversian sys¬
creases, the bones become less convex. tems.
Therefore, not only must bone resorption The outer limits of secondary haversian
take place on the inside of the calvarium systems are defined by distinct cement lines.
 BONE • 229

Figure 8-32. Diagram of stages in formation of three generations of haversian system, H, H', and H". I, Interstitial
lamellae. (Modified from Prenant.)

These are layers of bone matrix formed be seen in a cross section (1) mature osteons,
whenever a period of resorption is followed in which all rebuilding activity has come to
by new bone formation. They are collagen an end and which form the great mass of
poor, have staining properties different from structural bone upon which the weight-bear¬
other layers of matrix, and are not traversed ing function of the skeleton depends; (2)
by canaliculi. actively forming new osteons, in which con¬
The internal bone destruction and recon¬ centric layers of preosseous tissue are being
struction do not end with the replacement of laid down and progressively calcified; and (3)
primary by secondary bone, but continue absorption cavities being hollowed out in
actively throughout life. Resorption cavities preparation for formation of new osteons.
continue to appear and to be filled in by The rate of lamellar bone formation can
third, fourth, and higher orders of haversian be determined by administration of tetracy¬
systems (Fig. 8-32). The interstitial lamellae cline at two different times and measurement
of adult bone represent persisting fragments of the thickness of bone between the two
of earlier generations of haversian systems resulting bands of labeled bone (Fig. 8-33).
largely removed in the continuing internal Such studies show that 1 pm per day is a fair
reorganization. At any one time there may average for the human and for any given

Figure 8-33. A pair of haversian systems from the midshaft of the tibia of a 9-month-old dog given two 5-day courses
of treatment with a tetracycline separated by an interval of 19 days. A, Ordinary microscopy. B, Historadiogram. C and
D, Fluorescence microscopy. The bone deposited during Ledermycin (demeclocycline) treatment fluoresces, and the
design of this experiment permits one to visualize the amount of bone deposited in each 5-day period. Of particular
interest is the fact that the inner band, corresponding to the second period of administration, is narrower than the first,
demonstrating that in this instance there is slowing down in the rate of concentric bone deposition as the formation of
the haversian system progresses. (Courtesy of R. Amprino.)
230 • BONE

haversian system the rate of deposition slows tilage and hbrocartilage then develop within
as the osteon nears completion. The forma¬ it, forming a fibrocartilaginous callus that fills
tion time for a haversian system in the adult the gap between the ends of the fragments.
is four to five weeks. Different values are The new bone, which will ultimately unite
found in young growing bone and in patho¬ the fragments, begins to form at some dis¬
logical states. The newly deposited lamellar tance from the fracture line, by activation of
bone continues to calcify over a considerable osteoprogenitor cells of the deeper layers of
period of time. A historadiogram therefore the periosteum and endosteum. A mesh-
reveals a mixture of haversian systems of work of subperiosteal trabeculae, the bony
varying age, displaying all degrees of miner¬ callus, is formed, and a similar formation of
alization (Fig. 8-8C). By this continuous turn¬ new bone of endosteal origin occurs in the
over, the organism is assured a continuing medullary cavity around the cartilaginous cal¬
supply of new bone to carry out its skeletal lus. As healing progresses, the latter is grad¬
and metabolic functions. It also provides the ually eroded, with only enough cartilage ma¬
plasticity that enables bone to alter its internal trix remaining to provide a framework for
architecture to adapt to new mechanical con¬ deposition of new bone in the area. As in
ditions. endochondral bone formation, ossification of
the fibrocartilaginous callus is accomplished
by its gradual replacement with bone. Bony
Repair of Bone
union of the fracture is complete when the
After a fracture the usual reactions of any new spongy bone from the two fragments
tissue to severe injury are seen, including meets and becomes continuous across the
hemorrhage and organization of the clot by fracture line. After this there is compaction
ordinary granulation tissue. The granulation and reorganization, with resorption of excess
tissue becomes denser connective tissue. Car¬ bone and internal reconstruction, resulting

Figure 8-34. Cross section of the anteromedial sector of the midshaft of the femur as revealed by negative
historadiography. A, At age 7. B, At age 20. C, At age 65. Notice in the child (A) there are large resorption cavities
(black) and large, irregularly shaped haversian systems. At the surface of the compacta, the thick zone of periosteal
bone is invaded by resorption cavity. Large remnants of periosteal primary bone are found in the interstices between
the secondary osteons in the middle zone of the compacta. These remnants are fewer and smaller in the older
perimedullary zone at the bottom of the figure. In the 20-year-old man (B), the compacta is much thicker. Secondary
haversian systems and remnants of primary bone persist in the subperiosteal zone. Elsewhere, the osteons are fairly
regular in outline and are separated by remnants of preexisting osteons. (Courtesy of R. Amprino.)
 BONE • 231
finally in a bridging of the gap with compact conclusion is supported by experimental pro¬
bone.
duction of bone in connective tissue after
In certain locations, where the connective ligation of the renal artery and vein and after
tissue surrounding the bone lacks osteogenic a variety of experimental manipulations, such
potency, such as the subcapsular areas of the as transplantation of bladder epithelium to
neck of the femur and of the astragalus, and the fascia of the anterior wall of the abdo¬
the surfaces of the sesamoid bones formed men, or after injection of alcoholic extracts
within tendons (e.g., patella, pisiform), heal¬ of bone into muscle.
ing of fractures occurs without a periosteal
Many attempts have been made to utilize
reaction and without a fibrocartilaginous cal¬
the osteogenic potencies of periosteum and
lus. If there is good apposition of the frag¬
bone by transplanting these tissues to areas
ments, the cancellous bone of the marrow
in which it is desired that new bone be
cavity unites, without any callus formation.
formed. The modern “bone bank,” which
If apposition is poor or nonexistent, repair
supplies fragments of bone preserved by
may occur only as a relatively weak, fibrous
freezing or by other means, is the fruit of
union.
these efforts. Transplants of fresh autoge¬
nous bone ordinarily survive and proliferate.
Ectopic Bone Formation Flomografts are antigenic and give rise to an
immune response that leads ultimately to
As already stated, intramembranous bone death of the transplanted tissue. Heterografts
forms from a connective tissue, with the usually do not survive, but if calf bone is
transformation of mesenchymal cells into refrigerated and stored, it loses some of its
osteoprogenitor cells, osteoblasts and osteo- antigenicity and may therefore be suitable
cytes. The return of these cells to fibroblast¬ for use in bone banks. Grafts of such tissue
like osteoprogenitor cells has also been favor induction of new bone formation by
described. The influences under which ordi¬ the cells of the host.
nary connective tissue gives rise to bone in Primitive connective tissue cells within the
the embryo are poorly understood, but it is orbit of advancing bone, as in the formation
clear that previously undifferentiated cellular of medullary bone in birds, assume the form
elements of primitive connective tissue are of osteoblasts before they actually participate
capable of transformation to the cells char¬ in osteogenesis. This observation, together
acteristic of bone.
with those upon the behavior of bone grafts
It would appear that, once cells have ex¬ just cited, suggests that the presence of bone
hibited osteogenic potencies, these can be itself may be an important factor in activating
readily evoked again for an indeterminate osteogenic potencies. There is thus histolog¬
period after the cells have returned to an ical evidence in favor of induction of bone
indifferent morphological state. Thus, in the formation, although attempts to isolate a spe¬
healing of fractures, cells in the deepest lay¬ cific inductor substance have so far given
ers of the periosteum and endosteum, under equivocal results.
the stimulus of trauma, reassume the form
of osteoblasts and once again are actively
engaged in osteogenesis. Moreover, cells
grown from bone in tissue culture, and hav¬
ing lost the morphological characteristics of
HISTOPHYSIOLOGY OF BONE
osteoblasts, once again form bone when im¬
planted into the anterior chamber of the eye. As the principal tissue making up the skel¬
Furthermore, under certain conditions, etal system, bone functions in support of the
bone may be formed spontaneously from soft tissues; it carries the articulations and
connective tissue that is not in association provides attachment for the muscles involved
with the skeleton. This ectopic ossification has in locomotion; and it forms a rigid covering
been described in such diverse locations as for protection of the nervous system and of
the pelvis of the kidney, the walls of arteries, the hemopoietic tissue. In addition to these
the eyes, muscles, and tendons. In the long mechanical functions, it plays an important
tendons of the legs of turkeys, bone forma¬ role as a large store of calcium and phospho¬
tion is a normal event. From these observa¬ rus that can be drawn upon to maintain the
tions it may be inferred that many types of normal levels of these elements in the blood
connective tissue have latent osteogenic po¬ and to provide for the mineral requirements
tencies that are exhibited only rarely. This of other tissues.
232 BONE

Head Acetabular
Capsule cartilage
Fovea
Trochanter

Blood
vessels

Epiphysis

Future joint cavity

Epiphysis

Figure 8-35. Section through head of the femur of a 26-cm human fetus. (From a preparation of H. Hatcher.)

Articular surface

Figure 8-36. Articular surface of head of the femur of a man. (From a preparation of H. Hatcher.)
 BONE • 233

Bone as a Store of Mobilizable calcium at the normal level of 10 mg per 100

Calcium ml is mediated by parathyroid hormone, and
involves resorption of bone mineral and or¬
It is impossible to overemphasize the im¬
ganic matrix through the action of osteo¬
portance of calcium in the vital functions of
clasts.
the body. It is essential for the activity of
The responsiveness of the skeleton to the
many enzymes. It is indispensable for main¬
metabolic needs of other organ systems is
tenance of cell cohesion and of the normal
best illustrated in those species in which there
permeability of cell membranes. It is required
are unusual periodic demands for calcium.
for contraction of muscles and for coagula¬
Perhaps the most striking example is found
tion of the blood. It is not surprising, there¬
in birds in the laying cycle, during which
fore, that the homeostatic mechanisms of the
considerable amounts of calcium are re¬
body regulate the concentration of plasma
quired in the oviduct for deposition of the
calcium with remarkable constancy, the nor¬
eggshell. To meet this need, many trabeculae
mal range being between 9 and 11 mg per
in the marrow cavities of the long bones are
100 ml. Most of the calcium in the body (99
resorbed, only to be restored after the egg is
per cent) is, of course, in the bones. There is
laid and again removed to provide the shell
a constant interchange of calcium between
for the next egg in the clutch. Less dramatic
bone and the blood, which results in main¬
examples of mobilization of calcium from the
tenance of the relatively constant calcium ion
skeleton are also observed in mammals.
concentration in the plasma. The minute
While the antlers of deer are growing, there
hydroxyapatite crystals present a surface area
is a mild rarefaction of bone throughout the
for exchange with the extracellular fluids that
skeleton, and in dairy cows producing large
is of the order of 100 to 300 square meters
amounts of rnilk there may be a detectable
per gram. It has been estimated that during
osteoporosis associated with the considerable-
each minute in the life of an adult man, one
calcium loss in the milk. Human reproduc¬
of every four calcium ions present in the
tion does not involve such unusual demands
blood exchanges with similar ions in the
for calcium. There is no doubt, however, that
bones.
during pregnancy the maternal skeleton is
A dual mechanism for homeostatic regu¬
drawn upon to some extent for calcification
lation of the blood calcium level has been
of the fetal skeleton, and during prolonged
postulated. One part, acting by diffusion and
lactation it is the source of some of the
simple equilibrium between blood and the
calcium lost in milk. In normal individuals
labile fraction of bone mineral, is adequate
there is no detectable radiological change in
to maintain a constant but low calcium level
the skeleton, but when pregnancy or lactation
of approximately 7 mg per 100 ml of blood
is superimposed upon severe nutritional de¬
plasma. Not all of the bone contributes ficiency or impaired absorption of calcium,
equally to this function. The most labile cal¬
osteomalacia results, and may become so se¬
cium apparently is located in the younger vere as to result in pathological fractures.
and incompletely calcified osteons. It is these
that are most sensitive to ionic variations in
Endocrine Effects Upon Bone
the internal environment. Therefore, the
continued remodeling of the adult skeleton The skeletal system is affected by several
has metabolic as well as mechanical signifi¬ hormones. The most important of these is
cance. It provides a pool of young osteons, parathyroid hormone. Its participation in main¬
which can rapidly respond in homeostatic tenance of the normal levels of circulating
regulation by taking up or releasing calcium. calcium was referred to earlier. The activity
As these osteons mature and become more of the parathyroid glands appears to be reg¬
heavily mineralized, they become progres¬ ulated by a negative feedback mechanism, in
sively less available to the extracellular fluids, which the blood Ca++ level itself exerts a
and these older osteons probably contribute direct effect upon parathyroid activity. Para¬
more to the mechanical function of the skel¬ thyroid hormone has multiple sites of action.
eton. They are ultimately replaced in their One of the earliest detectable effects after its
physiological function by a new generation administration is on the kidney, where it
of osteons. These two categories are some¬ causes a rapid increase in excretion of phos¬
times referred to as metabolic and structural phate in the urine. This in turn affects the
bone. The second part of the dual mechanism plasma calcium levels. The hormone appears
required to elevate and maintain the plasma to have a dual effect upon bone. Its initial
234 • BONE

where it is stored in the egg yolk as phospho-

effect is believed to be on osteocytes, stimu¬
vitellin, the major source of phosphate for
lating osteocytic osteolysis. A long-continued
growth and development of the chick em¬
increase in circulating parathyroid hormone
results in induction of osteoclast formation bryo.
and accelerated bone remodeling. Since bone Mice react to administration of estrogens
resorption under the influence of parathy¬ in a manner qualitatively similar to that of
roid hormone results in destruction of stable birds. Endosteal bone formation is enhanced,
crystals of hydroxyapatite, as well as of the but in this case does not seem to serve any
organic matrix, this mechanism makes avail¬ physiological function. Endosteal bone for¬
able, for homeostatic regulation, an otherwise mation has not been reported in rats. In this
inaccessible source of calcium. species, estrogens inhibit normal resorption
Grafts of parathyroid to bone in vivo and of the spongiosa during endochondral ossi¬
confronted cultures in vitro have demon¬ fication, resulting in a greatly elongated and
strated that the gland causes resorption by dense spongiosa in the metaphysis. The os¬
direct action on bone. In clinical hyperparathy¬ teoporosis occasionally seen in women after
roidism, bone is extensively absorbed and is the menopause is attributed by some to the
replaced by fibrous tissue containing large decline in ovarian function, but it does not
numbers of osteoclasts. This results in the respond favorably to treatment with estro¬
pathological condition described as osteitis fi¬ gens.
brosa (von Recklinghausen’s disease). The gonadal hormones in some way play
Opposing the action of parathyroid hor¬ an important part in determining the rate of
mone is the polypeptide hormone calcitonin skeletal maturation. In normal human devel¬
or thyrocalcitonin, which originates from spe¬ opment the time of appearance of the various
cial cells in the thyroid gland. This hormone ossification centers and the time of fusion of
inhibits bone resorption and thus tends to the epiphyses with their diaphyses is remark¬
lower blood calcium. It is currently hypoth¬ ably constant. The progress of these events
esized that parathyroid hormone and calci¬ at any given time during development is
tonin act together to prevent or counteract intimately related to the developmental state
any significant perturbation of the homeos¬ of the reproductive system. Thus, in preco¬
tatic regulation of plasma calcium concentra¬ cious sexual development, skeletal matura¬
tion. A fall in plasma calcium below a certain tion is accelerated and growth is stunted,
level would presumably result in increased owing to premature epiphyseal closure. On
release of parathyroid hormone and suppres¬ the other hand, in testicular hypoplasia or
sion of calcitonin release. The effect of this prepubertal castration, epiphyseal union in
would be an increased rate of bone resorp¬ the long bones is delayed, and the arms and
tion and movement of calcium from bone to legs become disproportionately long.
blood, thereby returning the plasma calcium The growth of bone is also markedly influ¬
to normal. Conversely, a supranormal blood enced by the growth hormone (somatotropin)
calcium concentration would stimulate re¬ of the anterior hypophysis. Hypophysectomy
lease of calcitonin and suppress release of results in cessation of growth at the epiphy¬
parathyroid hormone. These effects would seal plate; upon administration of growth
tend to return the elevated plasma calcium hormone, growth recommences. Growth hor¬
to normal. mone injected into rats that have been both
The effects of the gonadal hormones upon thyroidectomized and hypophysectomized
bone vary greatly with the species. In the produces skeletal growth, whereas thyroxine
example of the laying bird cited previously, produces maturation but only moderate
an entire new system of trabeculae of med¬ growth. Coordination between growth and
ullary bone is produced by stimulation of the maturation may be restored by administra¬
endosteal lining in the estrogenic phase of tion of both hormones.
the egg-laying cycle. These trabeculae serve
to accumulate calcium for later use in for¬ Nutritional Effects Upon Bone
mation of the eggshell. The same changes
can be induced by administration of exoge¬ Growth of the skeleton is quite dependent
nous estrogens. Concurrently with the storage upon nutritional factors. Deficiencies of min¬
of calcium in medullary bone, the liver forms erals or of essential vitamins are often de¬
a phosphoprotein that appears in the blood tected more easily in bone than in other
and is transported to the ovarian follicle, tissues. A gross dietary deficiency of either
 BONE • 235

calcium or phosphorus leads to rarefaction degrees of movement between the adjoining

of bone and increased liability to fractures. bones. Such structures are called joints or
Even if the intake of these elements is ade¬ articulations. These present extreme varia¬
quate, a deficiency of vitamin D may interfere tions in character, which depend primarily
with their intestinal absorption and lead to on the type of bones that are joined and the
rickets. In this condition, ossification of the varying degrees of motion permitted by the
epiphyseal cartilages is disturbed, the regular articulation. Thus, in some cases, as in the
columnar arrangement of the cartilage cells skull, the joints are immovable, and the con¬
disappears, and the metaphysis becomes a nected bones are separated only by a thin
disordered mixture of uncalcified cartilage connective tissue layer, the sutural ligament.
and poorly calcified bone matrix. Such bones Other joints are slightly movable, such as the
are easily deformed by weight bearing. intervertebral articulations. Here the suc¬
In long-standing deficiency of calcium and ceeding vertebrae are joined by dense fibrous
of vitamin D, especially when aggravated by tissue and cartilage. Still other bones are
pregnancy, the bones of adults come to con¬ freely movable upon one another, and here
tain much uncalcified osteoid tissue, a con¬ the ends of the bones are completely covered
dition known as adult rickets or osteomalacia. by cartilage, and the articular surfaces are
Although the condition is aggravated by the surrounded by fibrous capsules.
increased demands of pregnancy, the dimi¬ Joints in which there is little or no move¬
nution in calcium content in this condition is ment are called synarthroses. There are three
due mainly to failure of calcification of new types of these: if the connection between the
bone formed in the turnover of this tissue, bones is of bone, it is a synostosis; if it consists
rather than to decalcification of previously of cartilage, a synchondrosis; and if of connec¬
calcified bone. tive tissue, a syndesmosis, joints that permit
Deficiency of vitamin C leads to profound free movement of the bones are called diar¬
changes in tissues of mesenchymal origin, throses.
producing the condition known as scurvy or The articular surface of the bones in diar¬
scorbutus, in which the primary defect is an throses is covered with hyaline cartilage. Al¬
inability to produce and maintain the inter¬ though the opposing cartilages are not cov¬
cellular substance of connective tissues. In ered by connective tissue, a small area of
the case of bone, it results in deficient pro¬ perichondrium at their margins is reflected
duction of collagen and bone matrix, with backward into the lining of the joint capsule.
consequent retardation of growth and de¬ At this point, there are many cartilage cells
layed healing of fractures. extending into the synovial membrane.
Deficiency of vitamin A results in a dimi¬ The joint capsules are composed of an
nution in the rate of growth of the skeleton. external layer consisting of dense fibrous
The vitamin controls the activity, distribu¬ tissue called the fibrous layer and an inner
tion, and coordination of osteoblasts and os¬ synovial layer, which is more cellular and is
teoclasts during development. Among other thought to secrete the viscid, colorless liquid
things, resorption and remodeling fail to en¬ of the joint cavity. However, joint membranes
large the cranial cavity and spinal canal at a exhibit many variations in structure. The
rate sufficient to accommodate growth of the synovial layer is sometimes thrown into folds,
brain and spinal cord. Serious damage there¬ which may project for surprising distances
fore results to the central nervous system. In into the cavity. The larger of these folds
hypervitaminosis A, erosion of the cartilage frequently contain vessels. In other cases the
columns accelerates without a compensating two layers appear fused, or the synovial layer
increase in the rate of multiplication of cells may rest directly on muscle, fatty tissue, or
in the proliferative zone. The epiphyseal periosteum. It has been suggested that the
plates may therefore be completely obliter¬ synovia be classified according to the tissues
ated, and growth may cease prematurely. on which they lie: that is, loose connective,
dense fibrous, or adipose, tissue.
Synovial membranes that rest on loose con¬
JOINTS AND SYNOVIAL nective tissue usually cover those parts of the
joints not subjected to strain or pressure. As
MEMBRANES a rule, they have a definite surface layer,
separated from the underlying tissue of the
Bones are joined to one another by con¬ joint by loose connective tissue. The surface
nective tissue structures that permit varying layer consists of collagenous fibers inter-
236 • BONE

Villi

Synovial
membrane

Subsynovial
connective
tissue

Capsule

Artery

Figure 8-37. Section through capsuie of the knee joint of a young man, showing the villi and connective tissue
components. The area outlined is shown at higher magnification in Figure 8—38. (From a preparation of H. Hatcher.)

Fat cells
Vessel

Figure 8-38. Synovial membrane of a young adult. (Higher magnification of the area outlined in Fig. 8-37.) Note the
irregularity in concentration of cells toward the free surface of the villus and the irregular distribution of fat cells. (From
a preparation of H. Hatcher.)
 BONE • 237

spersed with fibroblasts. The collagenous fi¬ Belanger, L. F.: Osteocytic osteolysis. Calcif. Tissue Res.
bers either are irregularly arranged or may 4:1, 1969.
be oriented along the main lines of stress. In Belanger, L. F., J. Robichon, B. B. Migicovscy, D. H.
Copp, and J. Vincent: Resorption without osteo¬
addition to the fibroblasts, there are a few
clasts (osteolysis.) In Sognnaes, R. F., ed.: Mecha¬
macrophages, leukocytes, and lymphoid wan¬ nisms of Hard Tissue Destruction. Washington, DC,
dering cells. In addition to blood vessels, the American Association for the Advancement of Sci¬
loose connective tissue contains many lym¬ ence, 1963.
phatics. Bernard, G. W., and D. C. Pease: An electron micro¬
scopic study of initial intramembranous osteogene¬
The fibrous type of synovial membrane sis. Am. J. Anat., 725:271, 1969.
covers the interarticular ligaments and ten¬ Bloom, W., M. A. Bloom, and F. C. McFean: Calcifica¬
dons and lines those parts of the joints that tion and ossification: medullary bone changes in the
are subject to strain. The adipose type of reproductive cycle of female pigeons. Anat. Rec.
57:443, 1941.
synovial membrane covers the fat pads that
Bonucci, E.: The locus of initial calcification in cartilage
project into some joint cavities. The synovial and bone. Clin. Orthop. 75:108, 1971.
membrane in this case usually consists of a Cohen, J., and W. H. Harris: The three dimensional
single layer of cells resting on a thin layer of anatomy of haversian systems. 1. Bone joint Sur?.
connective tissue. 40AA10, 1958.
Cooper, R. R., J. W. Milgram, and R. A. Robinson:
Folds of the synovial membrane either may Morphology of the osteon. An electron microscopic
be transient formations, which depend on study. J. Bone Joint Surg. 48A: 1239, 1966.
the position of the joint, or permanent villi, Fawcett, D. W.: The amedullary bones of the Florida
which project into the joint cavity. Some of manatee. Am. J. Anat. 77:271, 1942.
Fishman, D. A., and E. D. Hay: Origin of osteoclasts
these villi are short and have a broad base
from mononuclear leucocytes in regenerating new
(Fig. 8-37), while others may be thin and limbs. Anat. Rec. 743:329, 1962.
long. The larger folds contain blood vessels, Gaillard, P. J.: Parathyroid gland and bone in vitro. VI.
lymphatics, and occasionally lobules of adi¬ Dev. Biol. 7:152, 1959.
pose tissue (Fig. 8—38). There is an increase Glimcher, M. J.: Molecular biology of mineralized tissues
with particular reference to bone. In Oncley, J. F.,
in the size and number of the villi with age. et al., eds.: Biophysical Science—A Study Program.
New islets of cartilage are formed in them by New York, John Wiley & Sons, 1959.
metaplasia of the synovial fibroblasts. Glimcher, M. J., A. J. Hodge, and F. O. Schmitt: Ma-
There are two plexuses of lymphatics, as a cromolecular aggregation states in relation to min¬
eralization: the collagen-hydroxyapatite system as
rule, within the synovial membranes—a su¬
studied in vitro. Proc. Natl. Acad. Sci. USA 43:860,
perficial and a deep plexus. The nerves that 1957.
accompany the blood vessels end beneath the Glimcher, M. J., and S. M. Krane: The organization and
surface in terminal arborizations or end structure of bone and the mechanism of calcifica¬
tion. In Ramachandran, G. N., and B. S. Gould,
bulbs. Pacinian corpuscles are always present
eds.: Treatise on Collagen. IIB: Biology of Collagen.
in synovial membranes. Fondon, Academic Press, 1968.
When injured, the synovial membrane Goldhaber, P.: Oxygen dependent bone resorption in
reacts, like any other connective tissue, by tissue culture. In Greep, R. O., and R. V. Talmage,
forming granulation tissue, and after some eds.: The Parathyroids. Springfield, IF, Charles C
Thomas, 1961.
weeks may be completely regenerated. The
Hancox, N. M., and B. Boothroyd: The osteoclast in
synovial fluid is normally small in amount resorption. In Sognnaes, R. F., ed.: Mechanisms of
and seems to be a dialysate of blood, to which Hard Tissue Destruction. Washington, DC, Ameri¬
have been added small amounts of hyalu¬ can Association for the Advancement of Science,
1963.
ronic acid and a very few lymphocytes, mono¬
Harris, W. H., and R. P. Heaney: Skeletal renewal and
cytes, and macrophages. metabolic diseases of bone. N. Engl. J. Med. 25:253
1969.
Heinen, J. H., G. H. Dabbs, and H. A. Mason: The
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238 • BONE

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 BONE MARROW AND
BL OOD CELL
FORMA T/ON

The cells of the blood are short-lived and mur, and in vertebrae, ribs, sternum, and
are continuously replaced from sources out¬ ilia. Fatty transformation of the marrow in
side of the circulation. The process of blood the peripheral segments of the appendicular
cell formation is called hemopoiesis and the skeleton is believed to be related to the
sites where it takes place are termed hemo¬ slightly lower temperature of these parts.
poietic organs. The principal hemopoietic or¬ Yellow marrow can revert to red in response
gans in adult mammals are the bone marrow, to elevated temperature or unusual demands
spleen, lymph nodes, and thymus. This chap¬ for blood cells.
ter is devoted to the bone marrow, which is
by far the most important site of hemopoiesis.
The role of the spleen, lymph nodes, and
thymus in the formation and maturation of PRENATAL HEMOPOIESIS
blood cells will he discussed in later chapters.
Marrow occupies the cylindrical cavities of During prenatal life, there are three suc¬
the long bones and the interstices of the cessive phases in which the principal site of
spongiosa in vertebral bodies, ribs, sternum, hemopoiesis shifts from one region of the
and the flat bones of the cranium and pelvis. embryo to another. Formation of blood is
It accounts for 4 to 6 per cent of body weight first detectable in the mesenchyme of the
and has a total volume nearly equal to that body stalk and in neighboring areas of the
of the liver. It is a soft, highly cellular tissue yolk sac in the second week of life when the
consisting of precursors of the blood cells, embryo is only a few millimeters long. Clus¬
macrophages, adipose cells, reticular cells, ters of mesenchymal cells in these areas
and reticular fibers. The relative proportions round up and differentiate into large baso¬
of these several cell types undergo changes philic cells that assemble in aggregations
with age, and vary in different regions of the called blood islands. In this mesoblastic phase of
skeleton. At birth, all the bones contain deep hemopoiesis, nearly all of the cells formed are
red hemopoietically active marrow. The red erythrocytes. The earliest basophilic cells of
color is largely attributable to the enormous the blood islands differentiate into primitive
numbers of developing erythrocytes it con¬ erythroblasts. These synthesize hemoglobin
tains. At 4 to 5 years of age, the number of and develop into erythrocytes, which differ
blood-forming cells begins to diminish and from those of postnatal life in the nature of
the number of adipose cells increases. With their hemoglobin and in the fact that they
this shift in relative abundance of hemo¬ have a nucleus (Figs. 9-1, 9-2).
poietic and adipose cells, the color of the After establishment of blood islands in the
marrow gradually changes from deep red to yolk sac, accumulations of round basophilic
yellow. The transformation of hemopoieti¬ cells also appear, at about six weeks of ges¬
cally active red marrow to relatively inactive tation, in the primordium of the liver, initi¬
yellow marrow occurs earlier and progresses ating the hepatic phase of hemopoiesis. These
further in the distal portions of the long cells resemble the erythroblasts of postnatal
bones. In adults, red marrow persists only in hemopoiesis and are called definitive erythro¬
the proximal ends of the humerus and fe¬ blasts. They ultimately give rise to anucleate
239
240 • BONE MARROW AND BLOOD CELL FORMATION

Splanchnic mesotheliurrr-*

&*

Splanchnic mesotheliunr ESTHER -30HL MAN

? ’ ■I
v-* *

Splanchnic
mesothelium

';V- o*

; w 9
- ' vf
V -f *

Splanchnic mesothelium
Figure 9-1. Two sections through folds of the wall of the yolk sac of a 24-day human embryo (HI516 Univ. Chicago
Emb. Coll.). A, Early stage of hemopoiesis, consisting of proliferating extravascular hemocytoblasts (1, V); L, lumen of
a small vessel containing a few primitive polychromatophilic erythroblasts. B, Later stage of hemopoiesis showing
transformation of hemocytoblasts (1) into primitive basophilic erythroblasts (la), primitive polychromatophilic erythroblasts
(2, 3), and primitive erythrocytes (4); 5, mesenchymal cells; 6, endothelium. Hematoxylin-eosin-azure II stain. (From
Bloom, W., and F. W. Bartelmez. Am. J. Anat. 67:21, 1940.)

erythrocytes unlike those arising from prim¬ itive bone marrow, initiating the myeloid phase
itive erythroblasts, which retain their nucleus. of hemopoiesis. Hemopoiesis in the liver and
In the second month, granular leukocytes spleen then declines, and thenceforth the
and megakaryocytes appear in small numbers bone marrow is the major blood-forming
in the sinusoids of the liver. Somewhat later organ.
the spleen, as well as the liver, becomes a site In the transition from mesoblastic to he¬
of hemopoiesis. patic, to myeloid hemopoiesis, it was formerly
In the early embryo, the skeleton consists believed that mesenchymal cells in each of
entirely of hyaline cartilage that is subse¬ these successive sites gave rise independently
quently replaced by bone. In the fourth to primitive blood-forming cells. It has now
month, blood vessels begin to penetrate into been demonstrated that various blood-cell
cavities created by programmed degenera¬ types in the adult, including pluripotential
tion of chondrocytes in the cartilage models stem cells, can migrate in the bloodstream
of the bones. The blood vessels carry with from organ to organ. Therefore, it is now
them mesenchymal cells that differentiate thought that, in the embryo, successive sites
into bone-forming osteoblasts and reticulum of hemopoiesis are probably seeded by mi¬
cells destined to form the stroma of the bone gration of stem cells from the preexisting
marrow. Concurrently with establishment of sites of blood formation.
these ossification centers in the cartilaginous The liver and spleen in the adult do not
skeleton, blood formation begins in the prim¬ normally participate in blood formation, but
 BONE MARROW AND BLOOD CELL FORMATION • 241

Figure 9 2. Section through yolk sac of a 20-mm human embryo. In addition to circulating primitive erythrocytes there
are two foci developing polychromatophilic definitive erythroblasts. 1, Hemocytoblast; 4, primitive erythrocytes- 5
mesenchymal cells; 7 and 8, early and late definitive polychromatophilic erythroblasts with one in mitosis at 7'- 9
jnTarSSzVi°’l^OP)0id wandering cell‘ Hematoxy|in-e°sin-azure II stain. (From Bloom, W., and F. W. Bartelmez. Am.

in diseases attended by marrow failure, extra¬

within the medullary cavity enter Volkmann’s
medullary hemopoiesis may be resumed in these
canals and join the vascular network within
organs.
the cortex. Very few intramedullary branches
communicate directly with the sinuses of the
marrow. Thus, most of the blood reaching
STRUCTURAL ORGANIZATION the sinuses has first entered the osseous tissue
OF BONE MARROW either from periosteal vessels or from endos¬
teal branches of the nutrient artery, and has
secondarily entered the medullary sinuses at
The bone marrow consists of closely the corticomedullary junction. The signifi¬
packed hemopoietic cells, reticular cells, and cance of this indirect transosteal routing of
adipose cells occupying all the extravascular the blood is not known, but it may be neces¬
spaces around an extensive system of thin- sary in order to maintain the optimal physi¬
walled venous sinuses. The blood cells de¬ cochemical environment for hemopoiesis.
velop extravascularly and must pass through The vascular sinuses of the marrow are 50
the walls of the sinuses to enter the circula¬ to 75 pm in diameter with a thin endothelial
tion. Some familiarity with the unusual vas¬ lining. Histologists had long assumed that
cular architecture of bones is essential to an the endothelium was discontinuous to allow
understanding of marrow histology. passage of newly formed blood cells into the
The arterial blood supply comes from two circulation. This view was sustained by early
sources. The cortical osseous tissue is pene¬ electron microscopic studies, which reported
trated from outside by branches of a network sizable openings in the sinus wall. It is now
of small vessels in the surrounding perios¬ agreed that these were artifacts, for in scan¬
teum. Some of the capillaries within the cor¬ ning and transmission electron microscopic
tex are continuous at the corticomedullary studies in which care has been taken to pre¬
boundary with an elaborate network of an¬ serve the marrow in situ and avoid mechanical
astomosing thin-walled venous sinuses within damage, the sinuses have been shown to have
the marrow. These sinuses, in turn, converge no permanent apertures that would permit
upon wider collecting sinuses radially arranged passage of blood cells. The endothelium is
around a large, longitudinally oriented central composed of flattened cells joined together
sinus. The major arterial supply to long bones by junctional complexes as in other epithelia.
is from a nutrient artery, which enters the In the extremely attenuated peripheral por¬
marrow cavity through the nutrient canal and tions of the cells, clusters of endothelial pores
bifurcates into ascending and descending may be found. A typical basal lamina is absent
branches. The great majority of the fine but there may be scattered extravascular de¬
branches resulting from their ramification posits of flocculent material of similar nature.
242 • BONE MARROW AND BLOOD CELL FORMATION

Reticular cells are deployed in a discontin¬ cover may be reduced to less than 20 per
uous adventitial layer over the abluminal sur¬ cent of the sinus surface.
face of the sinuses with their long branching The passage of mature blood cells into the
processes extending into the surrounding circulation does not take place by separating
myeloid tissue in intimate association with endothelial cells at their junctions, but is
the differentiating blood cells. They are stra¬ transcellular. The migrating cell presses the
tegically situated to participate in inductive abluminal membrane of the endothelial cell
or regulatory interactions with the hemo¬ into contact with the adluminal membrane,
poietic stem cells, but if they have such a and the two fuse to form a transient migration
function it has yet to be convincingly dem¬ pore (Fig. 9-3). This opening may be some¬
onstrated. Their principal function seems to what enlarged by the cell in transit but does
be mechanically supportive. They synthesize not exceed 4 pm in diameter, and continuity
and maintain the delicate framework of re¬ of the endothelium is rapidly restored after
ticular fibers that form the sparse stroma of the blood cell has become free in the lumen
the marrow. of the sinus. The endothelium is the major
The adventitial reticular cells in normal barrier in the path of the exiting mature
marrow are estimated to cover 40 to 60 per blood cells. It appears to be actively involved
cent of the abluminal surface of the sinuses, in the process and there is suggestive evi¬
leaving the remainder accessible to mature dence that it may exercise some selectivity in
blood cells for transmural migration into the control of the transcellular traffic.
bloodstream. In response to circulating tox¬ The organization of the extravascular hem¬
ins and possibly to hemopoietic hormones, opoietic compartment of the marrow is dif¬
the reticular cells may change their shape, ficult to study with the light microscope ow¬
exposing more endothelial surface for egress ing to the crowding and superimposition of
of cells. In experimentally induced active cells in histological sections, and the difficulty
transmura! cell transport, the adventitial of obtaining adequately fixed preparations

Sinus endothelium

Sinus lumen

Figure 9-3. Electron micrograph illustrating transmural migration of a lymphocyte through the endothelium into the
sinus lumen.
 BONE MARROW AND BLOOD CELL FORMATION • 243
Fixed Normoblast
Hemocytoblast macrophage nuclei Megakaryocyte Heterophilic leukocyte

Primitive reticular
cell
Small lymphocyte Heterophilic
myelocytes
Small lymphocyte
Primitive reticular
cell Polychromatophilic
erythroblasts
Megakaryocyte
Fixed macrophage
(lining cell)
Basophilic
Venous sinus
erythroblasts

Free macrophage

Hemocytoblast
( mitosis)
Migrating Heterophilic
macrophage myelocyte
Arteriole
Plasma cell

Small lymphocyte
Lining cells

Heterophilic
Heterophilic
leukocyte
leukocyte

Normoblast nucleus Eryth- Polychro- ^ot cells Normoblasts Eosinophilic myelocytes

rocytes matophilic
erythroblasts in mitosis

Figure 9-4. Section of bone marrow of rabbit that had injections of lithium carmine and India ink. Hematoxvlin-eosin
azure II stain. (After A. A. Maximow.)

without mechanical damage (Fig. 9-4). How¬ newed, a small number of cells have the
ever, some patterns of cell association have capacity for both self-duplication and differ¬
emerged. Erythropoiesis tends to occur in entiation. Such cells are called stem cells. If
clusters near the sinuses, while granulocytes their progeny are able to differentiate into
develop near the center of the hemopoietic several different types of mature blood cells,
spaces. Megakaryocytes are typically situated they are described as pluripotential hemopoietic
adjacent to the sinus wall with processes pro¬ stem cells (PHSC). The differentiation of such
jecting through apertures into the lumen. a cell involves first, a loss in the ability to
Macrophages and lymphocytes are scattered develop along multiple alternative paths and
throughout the marrow. Adipose cells arise later, gradual acquisition of distinctive new
by accumulation of lipid in adventitial retic¬ morphological features and functional prop¬
ular cells and hence are usually in close erties typical of more mature blood cells. The
proximity to sinuses. The spatial and func¬ immediate progeny of a pluripotential stem
tional relationships of cell types are consid¬ cell that retain the capacity for self-renewal,
ered further below. but are able to differentiate only into a single
end-cell type, are termed unipotential stem cells
or committed stem cells.
Recognition of hemopoietic stem cells
HEMOPOIETIC STEM CELLS poses an insuperable problem for the mi-
croscopist. For visual identification of differ¬
In the bone marrow, as in other organs ent cell types, he is obliged to rely on size,
where the cell population is continually re¬ shape, staining properties, nuclear conhgu-
244 • BONE MARROW AND BLOOD CELL FORMATION

PHSC
I

— CFU-E
I

Proerythroblast Megakaryocyto-
I blast

Basophilic
erythroblast Promyelocyte

Polychromatophilic
erythroblast
I

Metamyelocyte

Reticulocyte Megakaryocyte
Band form
l
<g>

Neutrophil Eosinophil Platelets

Erythrocytes Basophil

Figure 9—5. Diagram of the principal myelopoietic cell lineages. Above the dotted line are the stem cells, which are not
microscopically distinguishable but are identified by spleen assay—pluripotential hemopoietic stem cell (PHSC); colony¬
forming unit-spleen (CFU-S); colony-forming unit-burst (CFU-B); colony-forming unit-erythrocyte (CFU-E); colony¬
forming unit-granulomonocyte (CFU-GM); and colony-forming unit-megakaryocyte (CFU-M). Below the dotted line are
the morphologically recognizable cell lines that differentiate from these stem cells.
 BONE MARROW AND BLOOD CELL FORMATION • 245
ration or chromatin pattern, and presence or
sites. This may account for the occurrence of
absence of specific granules. The distinctive
extramedullary hemopoiesis in certain dis¬
features required for identification are ac¬
eases. The pluripotential stem cells are slowly
quired late in the course of differentiation
proliferating but give rise to more rapidly
and are largely lacking in the early stages of
proliferating unipotential stem cells (CFU-E
the several blood-forming cell lines. Thus,
or CFU-M), limited to production of granu¬
although histologists and hematologists can
locytes and/or cells of the monocyte-macro-
agree upon a functional definition of a stem phage lineage.
cell, they cannot identify it with any degree
Much of our understanding of the kinetics
of confidence at the light or electron micro¬
and cell lineages in the early stages of hemo¬
scope level. It is safe to assume that it is a
poiesis has been based on spleen colony as¬
small spherical cell with a rim of basophilic
says in irradiated mice, but there is every
cytoplasm around a nucleus relatively poor
reason to believe that the same principles
in condensed chromatin. Thus, it cannot be
apply to the human. Culture systems have
dependably distinguished from one of the
now been developed in which progenitor cells
several kinds of lymphocytes. Therefore,
from human marrow are stimulated with
stem cells are commonly dealt with as hypo¬
hemopoietic growth factors and give rise to
thetical or statistical entities rather than rec¬
clonal colonies in a semisolid matrix of agar,
ognized cell types.
fibrin, or methyl cellulose. These in vitro
Stem cells are detected and their several
systems promise to permit studies of kinetics
categories are distinguished by testing their
and control mechanisms of human hemo¬
developmental potential in an in vivo or in
poiesis comparable with those carried out in
vitro assay system. The most widely used the mouse.
experimental procedure consists of injecting
suspensions of hemopoietic cells into the
bloodstream of mice that have been X-irra-
diated with a dosage sufficient to destroy the ERYTHROPOIESIS
proliferative capacity of their own cells. The
injected cells “home’’ into the spleen and The erythrocytes have a life span of about
marrow of the recipient mouse. After several 120 days and are then removed from the
days, the spleen contains grossly visible small blood and destroyed in the spleen. The main¬
colonies, each having developed by prolifer¬ tenance of normal numbers in the circulation
ation of a single stem cell and differentiation requires their continuous formation in the
of its progeny. This assay detects a broad bone marrow. Each day some 2 X 1010 new
class of stem cells designated colony-forming erythrocytes enter the circulation.
units—spleen (CFU-S). The stem cells giving Although blood cell development is a con¬
rise to the individual colonies can be more tinuous process, it is customary to consider it
narrowly defined by examining the nature of as occurring in three phases: hemopoietic
their differentiated progeny. If these include stem cells, committed progenitor cells, and
two or more blood cell types, the cell of origin morphologically recognizable maturation
was a pluripotential stem cell (PHSC). If the stages. Through lack of criteria for its visual
progeny are all of the erythrocyte lineage, identification, the pluripotential stem cell giv¬
they arose from a unipotential stem cell, des¬ ing rise to all blood cells is designated the
ignated a colony-forming unit—erythroid (CFU- colony-forming unit (CFU). Some of the
E). Similarly, if they all belong to the mega¬ progeny of this slowly replicating cell lose
karyocyte line, they arose from a colony¬ then pluripotentiality and become irreversi¬
forming unit—megakaryocyte (CFU-M). Other bly committed to production of erythrocytes.
colonies contain both granulocytes and mon¬ Hematologists studying the characteristics of
ocytes and arise from a bipotential stem cell these unipotential progenitor cells in cultures
referred to as a colony-forming unit—granulo- have distinguished two successive stages:
monocyte (CFU-GM). erythroid burst-forming units (E-BFU or CFU-
Kinetic studies indicate that there are in B) and erythroid colony-forming units (E-CFU
the mouse as few as one pluripotential stem or CFU-E). The former have a higher prolif¬
cell per 10,000 nucleated marrow cells—a eration rate and require a relatively high
total of about 40,000. The spleen contains concentration of the stimulating factor eryth¬
about one twentieth this number. They have ropoietin. The latter proliferate more slowly
also been detected in small numbers in pe¬ and are responsive to erythropoietin in low
ripheral blood and are therefore capable of concentration. With further differentiation,
being transported from the marrow to other the erythroid progenitor cells (E-CFU or
246 * BONE MARROW AND BLOOD CELL FORMATION

CFU-E) become morphologically identifiable Each proerythroblast undergoes a series of

as proerythroblasts. These are round cells 14 to divisions to produce several smaller basophilic
19 pm in diameter, with a thin rim of baso¬ erythroblasts. These have a deeply basophilic
philic cytoplasm and a large nucleus, which cytoplasm, a more coarsely clumped chro¬
has a rather uniformly dispersed chromatin matin pattern, and no visible nucleoli (Fig.
pattern and two or more nucleoli. 9—6). The disappearance of nucleoli is asso-

Orthochromatfc erythroblast
Neutrophilic leukocytes
(normoblast), extruded nucleus

Eosinophilic leukocyte

Late polychromatophilic
Neutrophilic metamyelocyte
erythroblast

Eosinophilic metamyelocyte

Polychromatophilic
Neutrophilic myelocyte
erythroblast

Eosinophilic metamyelocyte

Early polychromatophilic
Early neutrophilic myelocyte
erythroblast

Eosinophilic myelocyte

Very early myelocyte Basophilic erythroblast

Free stem cell (blast) Very primitive free stem cell Early basophilic erythroblast

Figure 9—6. Photomicrographs of developing blood cells in human bone marrow, showing steps in the transformation
of stem cells into neutrophilic and eosinophilic leukocytes and into erythrocytes, as seen in dry smears stained with
Wright’s blood stain.
 BONE MARROW AND BLOOD CELL FORMATION • 247

ciated with cessation of nuclear synthesis of same time, the hemoglobin synthesized on
new ribosomal protein. In electron micro¬
the polyribosomes steadily accumulates (Fig.
graphs, the cytoplasm contains a profusion 9—7). The range of tinctorial affinities exhib¬
of free polyribosomes but few or no profiles
ited by the polychromatophilic erythroblasts
of endoplasmic reticulum. Synthesis of hemo¬ reflects the progressively changing propor¬
globin is already in progress at this stage. It tions of ribosomes (which bind the blue com¬
is recognizable in electron micrographs as ponents of the dye mixture) and of hemoglo¬
fine cytoplasmic particles of relatively low bin (which has an affinity for the eosin).
density. In stained marrow smears observed When the cells have acquired nearly their
with the light microscope, its presence is full complement of hemoglobin, their cyto¬
obscured by the intense blue staining of the plasm is distinctly eosinophilic with only a
ribonucleoprotein of the cytoplasm. slight tinge of residual blue. The nucleus is
The progeny of the division of basophilic now small, eccentric, and deeply stained.
erythroblasts are polychromatophilic erythro- Such cells (7 to 14 pm) are called orthochro-
blasts, recognizable by their smaller overall matic erythroblasts (also called normoblasts). In
size, their smaller nucleus with more con¬ electron micrographs, their heterochromatin
densed chromatin, and the characteristic is in coarse blocks with little intervening nu¬
staining reaction of their cytoplasm, which cleoplasm. The cytoplasm is devoid of organ¬
ranges from bluish-gray through gray-greens elles except for an occasional mitochondrion
of diminishing intensity (Fig. 9-6). Since no (Figs. 9—8, 9-9C). The cytoplasmic matrix
new ribosomes are formed after disappear¬ presents a rather uniform fine gray granu¬
ance of the nucleoli, there is a progressive larity, owing to the high concentration of
decrease in their concentration as a conse¬ hemoglobin. Widely scattered clusters of ri¬
quence of the succeeding divisions. At the bosomes are still present (Fig. 9-9C).

Figure 9-7. Polychromatophilic erythroblasts from guinea pig marrow showing coarse blocks of heterochromatin in the
nucleus, and cytoplasm consisting mainly of hemoglobin and polyribosomes. In inset, polyribosomes (at arrows) can be
distinguished from the smaller, less dense granular background of hemoglobin.
248 • BONE MARROW AND BLOOD CELL FORMATION

Figure 9-8. Electron micrographs of human orthochromatic erythroblasts (normoblasts). The cell at right is extruding
its nucleus. Notice that the nucleus is pinched off, enclosed in a portion of the cell membrane and a thin layer of
cytoplasm, and does not pass through a break in the membrane as was previously believed.

The eccentric nucleus is then extruded and There is a reserve of reticulocytes in the
pinched off, with a very thin him of cyto¬ marrow somewhat greater than the number
plasm enclosed by a portion of the plasma in the circulation. In the continual turnover
membrane (Fig. 9-8). The extruded nuclei of erythrocytes, about 1.9 x 1010 erythrocytes
are ingested and destroyed by macrophages. are destroyed each day and the same number
The anucleate portion, an erythrocyte, is ul¬ of new ones are produced in the marrow.
timately released into the bloodstream. Newly Old or damaged erythrocytes undergo
formed erythrocytes contain small amounts changes in their surface membrane, notably
of basophilic material dispersed in the hemo¬ a loss of sialic acid residues, and are selec¬
globin, and consequently have a slightly tively removed from the circulation in the
greenish tint instead of the clear salmon pink spleen and phagocytized by macrophages.
of more mature forms. These are described Iron released in the degradation of their
as polychromatophilic erythrocytes. However, if hemoglobin is transported by plasma trans¬
fresh blood smears are stained with cresyl ferrin back to the marrow for reutilization in
blue, the residual ribosomes are clumped and hemoglobin synthesis. The remainder of the
the aggregates formed appear as a bluish degraded hemoglobin is transformed into the
skein or network in the otherwise pink cyto¬ bile pigment bilirubin and excreted.
plasm. Under these conditions of staining,
the recently formed erythrocytes are called
reticulocytes. GRANULOPOIESIS
The percentage of reticulocytes in a blood
smear is a dependable index of the rate of In common with all other blood cells, the
formation of new red blood cells and is widely granulocytes originate from the morpholog¬
used in following the recovery of patients ically undefined pluripotential stem cell now
from blood loss or their response to treat¬ referred to as the colony-forming unit (CFU-
ment for anemia. In man, the erythrocyte S). These cells having a high capacity for self¬
generation time from stem cell to circulating renewal give rise to several types of progen¬
blood cell is about a week. itor stem cells including one committed to
 BONE MARROW AND BLOOD CELL FORMATION • 249

formation of granulocytes and monocytes

(GM-CFU). These differentiate into myelo¬
blasts, the earliest morphologically recogniz¬
able cell of the granulocyte series.
The myeloblast is a relatively small cell with
a large nucleus containing three or more
nucleoli. The cytoplasm is basophilic and
devoid of granules. In its further develop¬
ment, the cell enlarges and acquires small,
metachromatic azurophil granules. It is then
called a promyelocyte.
The promyelocyte has a reniform nucleus,
multiple nucleoli, and a dispersed chromatin
pattern. Early in its development, a small
number of azurophil granules are clustered
near the cell center (Fig. 9—6). In electron
micrographs, these are dense, somewhat ir¬
regular in outline, 0.1 to 0.25 pm in diame¬
ter, and membrane bounded. A few cisternal
profiles of rough endoplasmic reticulum are
present, and abundant free ribosomes. The
Golgi complex is situated in an indentation
of the nucleus, and numerous transitional
vesicles are observed between the cisternae
of the reticulum and the convex outer face
of the Golgi. Condensing vacuoles containing
dense cores arise at the inner concave face
of the Golgi. These dense-cored vacuoles
fuse, forming larger ones containing multiple
dense bodies that coalesce to form azurophil
granules. In the human, the content of the
condensing vacuoles is a flocculent precipi¬
tate instead of dense cores and morula forms
characteristic of these cells in the rabbit. Oth¬
erwise, the mechanism of azurophil granule
formation is not significantly different.
More advanced promyelocytes exhibit a
striking increase in size from about 16 to
about 24 pm in diameter (Figs. 9-10, 9—11,
9-12). There is a concomitant increase in
number of azurophil granules, which are now
dispersed throughout the cytoplasm. The
chromatin is more condensed and clumped
along the nuclear envelope and around the
nucleoli. The endoplasmic reticulum is more
extensive and is often somewhat distended
during this phase of rapid synthesis of gran¬
ule protein.
After one or more mitotic divisions, the
resulting late promyelocytes are considerably
smaller, the chromatin is more condensed,
and nucleoli are inconspicuous. Although
azurophil granules still occupy much of the
Figure 9-9. A series of electron micrographs illustrating cytoplasm, the peak period of their formation
the decline in number of ribosomes and progressive has passed and there is a notable decrease in
increase in hemoglobin in the cytoplasm during the differ¬ the size of the Golgi and the extent of the
entiation of the guinea pig erythrocyte. A, Basophilic reticulum.
erythroblast; B, polychromatophilic erythroblasts; C, or-
thochromatic erythroblast (normoblast); D, reticulocyte; E, From stem cell through the late promy¬
erythrocyte. elocyte stage, the development of all types of
250 • BONE MARROW AND BLOOD CELL FORMATION

granulocytes is essentially the same. The

granulocyte lineage then diverges along three
separate paths of differentiation with the
appearance in the myelocytes of specific granules
with differing tinctorial properties and dis¬
tinctive ultrastructure.

Neutrophilic Myelocytes
The neutrophilic myelocyte is distinguished
from the promyelocyte by its smaller size; by
the more variable shape of the nucleus and
greater condensation of its chromatin; by the
smaller size of its Golgi complex; and espe¬
cially by the presence in its cytoplasm of a
second type of granule that has little affinity
for stains in routine marrow smears. In elec¬
tron micrographs these are less dense than
azurophil granules ^nd often elongated, re¬
sembling rice grains. The formative stages of
the specific granules are said to be associated
with the convex surface of the Golgi complex.
Thus, in the development of neutrophils, two
populations of granules are formed at differ¬
ent times. The azurophil granules arise in
the promyelocyte stage at the concave face of
the Golgi apparatus, and the specific granules
are formed in the myelocyte stage at its
convex face. The azurophil granules contain
histochemically demonstrable peroxidase,
acid phosphatase, arylsulfatase, (3-galactosi-
dase, (3-glucuronidase, esterase, and 5'-nu¬
cleotidase and therefore are considered to be
primary lysosomes (Fig. 9—13). The specific
granules contain alkaline phosphatase and
poorly characterized bacteriostatic proteins.
From the progenitor cell through the my¬
elocyte stage, four to seven divisions occur.
Mitotic divisions then cease and subsequent
differentiation involves changes in nuclear
form, diminution in number of mitochondria
and other organelles, slight condensation of
the cytoplasm, and the appearance in it of
small amounts of glycogen.
The next stage, the metamyelocyte, is distin¬
guishable from the myelocyte mainly by the
shape of its nucleus, which is deeply indented
(Fig. 9—14). Two granule types are still pres¬
ent, but specific granules now make up over
80 per cent of the granule population. Nu¬
clear modeling continues, resulting in a cell
with a slender, straight, or curved nucleus,
referred to as a band-form. During final mat¬
uration of the neutrophil, the nucleus be¬
Figure 9-10. Phase-contrast photomicrographs of pro¬ comes constricted into multiple small lobules
myelocytes (A, B), neutrophilic myelocytes (C, D, E), and
metamyelocytes (F) from human bone marrow. (Photo¬ connected by extremely thin regions.
graphs from Ackerman, A. Zeitschr. f. Zellforsch. 121:153, The entire transit time from stem cell to
1972.) mature granulocyte is about ten days. The
 BONE MARROW AND BLOOD CELL FORMATION • 251

Azurophil

Ele<f°n m'cr°9raph of Promyelocyte from rabbit marrow. Dense-cored vacuoles associated with the Golai
ripnCPi m!?PreSen °rmatlve s?9es° azur°Phl1 granules. Enlargement and coalescence of these gives rise to the larqe
CeH BioT 28r2797ai,966S)Seen e S@Where ln the cy,°P|asm- (Electron micrograph from Bainton, D„ and M. Farquhar.9J

production rate in the human is estimated to ules are rich in peroxidase, and cytochemical
be about 1.6 X 104/kg/day, of which the great staining for this enzyme is useful in revealing
majority are neutrophils. A large reserve of the intracellular pathway for synthesis, seg¬
metamyelocytes, band-forms, and mature regation, and packaging of the enzyme into
neutrophils is maintained in the marrow, specific granules. In eosinophilic myelocytes,
perhaps as great as ten times the daily pro¬ peroxidase reaction product can be demon¬
duction. These can be mobilized to meet strated in the perinuclear cistern, throughout
unusual demands. Mature neutrophils are the endoplasmic reticulum, and in the Golgi
preferentially released, but in infections saccules and forming granules (Fig. 9-16).
many band-forms and even metamyelocytes In later stages after granule formation has
enter the circulation. ceased, the enzyme is found only in the
specific granules. A similar distribution is
Eosinophilic Myelocytes reported for acid phosphatase and arylsul-
fatase. Thus, the myelocytes appear to be
These are less numerous than neutrophilic capable of synthesizing and concentrating
myelocytes. The nucleus has a rather coarse several proteins simultaneously.
pattern of peripheral clumps of chromatin. The nucleus of eosinophil metamyelocytes
The cytoplasm is slightly basophilic and the is indented, and in mature eosinophils it is
specific granules are distinctly larger and usually bilobed. No band-form occurs and
eosinophilic. In electron micrographs there the nucleus never acquires the degree of
are many cisternal profiles of endoplasmic lobulation seen in neutrophils. In late my¬
reticulum and abundant free ribosomes. Two elocytes and metamyelocytes, the contents of
populations of granules are discernible— the specific granules begin to crystallize, and
dense azurophil granules and somewhat less one finds in micrographs a heterogeneous
dense specific granules. The eosinophil gran¬ population of granules, some remaining
Figure 9-12. Electron micrograph of a polymorphonuclear promyelocyte, the largest cell of the neutrophilic series,
treated for peroxidase. The cytoplasm is packed with peroxidase-positive azurophil granules, and a positive reaction is
seen throughout the reticulum. Specific granules are not yet present. (From Bainton, D., J. Ullyot, and M. Farquhar. J.
Exp. Med. 734:907, 1971.)

Figure 9-13. Electron micrograph of polymorphonuclear neutrophilic myelocyte stained with the peroxidase reaction.
The azurophil granules are strongly reactive but the specific granules are unstained. (From Bainton, D., J. Ullyot, and
M. Farquhar. J. Exp. Med. 734:907, 1971.)
252
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Hlncle.9~14'^ICr°9r^ph °f \ gr0Uf? 0f neutr°Phi,ic metamyelocytes and band forms from baboon bone marrow. The
dense azurophil granules are few relative to the specific granules.

Figure 9-15. Eosinophilic myelocyte from guinea pig bone marrow. There is a well-developed endoplasmic reticulum
and the specific granules are distinctly larger than those of neutrophilic myelocytes. The crystals in the granules have
not yet developed.

253
254 • BONE MARROW AND BLOOD CELL FORMATION

Fiaure 9-16 Eosinophilic myelocyte of rat, stained by the cytochemical reaction for peroxidase. Formation of specific
granules is continuing and reaction product is found in the perinuclear cisterna and throughout the endoplasmic
reticulum-also in the Golgi complex and specific granules. (Micrograph from Bainton, D., and M. Farquhar. J. Cell Biol.
45:54, 1970.)

dense and homogeneous, others having crys¬ line shares with the granulocytes a common
tals of varying form surrounded by a matrix committed stem cell, the colony-forming
of low density (Fig. 9-17). unit-granulocyte/macrophage (CFU-GM). A
monoblast is described from colonies in ceil
culture but is identified with difficulty in the
Basophilic Myelocytes marrow. Its division gives rise to promonocytes.
These are rarely observed in preparations About half of the promonocytes of the mar¬
of human marrow owing to their small num¬ row are rapidly proliferating to generate
bers and the difficulty of preserving their nonproliferating monocytes. The remainder
granules, which are soluble in aqueous media constitute a reserve of very slowly renewing
and are partially or completely extracted dur¬ progenitor cells that can be activated to meet
ing fixation and staining. They have a paler- unusual demands for tissue macrophages.
staining nucleus than other myelocytes. The The pool of promonocytes in human mar¬
cytoplasm contains relatively few specific row is estimated to be about 6 X 108/kg body
granules, which are metachromatic and vary weight and the monocyte production rate
considerably in size. The nucleus of meta¬ about 7 x 106/kg body weight per hour. The
myelocytes is deeply indented, and that of stem cell to monocyte transit time is about 55
mature basophils is constricted into two and hours and they probably remain in the cir¬
occasionally three lobes. culation no more than 16 hours before emi¬
grating to the tissues, where they differen¬
tiate into macrophages—the functional phase
of this cell line. Maintenance of the macro¬
MONOPOIESIS phage population in the tissues depends
mainly on the continuous influx of monocytes
Experimental studies of spleen colonies has from the blood. Macrophages are capable of
revealed that the monocyte-macrophage cell cell division, but under normal conditions
 BONE MARROW AND BLOOD CELL FORMATION 255

Figure 9 17 Micrograph of a pair of late eosinophilic myelocytes from rat bone marrow. Endoplasmic reticulum is less
extensive and crystals are forming in the specific granules (at arrows).

their proliferation does not contribute sub¬ The term thrombopoiesis refers to the devel¬
stantially to renewal of the population in the opmental events in hemopoietic organs lead¬
tissues. ing to formation of thrombocytes and plate¬
lets. The thrombocytes in lower vertebrates
are nucleated cells that develop by successive
THROMBOPOIESIS division and differentiation of precursor cells
called thromboblasts in a manner similar to
the derivation of erythrocytes from erythro-
Thrombocytes and platelets are cellular blasts. The anucleate platelets of mammals,
elements in the blood of vertebrates con¬ which are the functional equivalent of the
cerned with protection against blood loss by thrombocytes of amphibia, reptiles, and
promoting clotting at sites of tissue injury. birds, are formed by fragmentation of the

Marrow

Mitotic - Post-mitotic- •Blood-*

[^-Storage-

Figure 9-18. Summary diagram J-H day- -10 days - •10 hr-
of the timing of events in differ¬ ■41 xl09/kg- 17x107 kg1
entiation of polymorphonuclear
neutrophils. (From Boggs, D., and |-24xl09/kg-
M. Chernack. J. Reprod. Fertil.,
Suppl. 70:32, 1970.)
256 • BONE MARROW AND BLOOD CELL FORMATION

cytoplasm of huge polymorphonuclear cells polysomes, a few cisternae of endoplasmic

called megakaryocytes, found among the other reticulum, a juxtanuclear Golgi complex, and
hemopoietic cells in the bone marrow. The large mitochondria. If two nuclei are present,
nucleus of these giant polyploid cells contains these fuse at the next nuclear division and
from eight to 16 sets of chromosomes and the cell (2n) subsequently increases rapidly in
occasionally as many as 64. Thus, if platelets size while undergoing a series of atypical
arose by nuclear and cytoplasmic division as nuclear divisions that are not accompanied
do thrombocytes in lower vertebrates, 64 cells by division of the cytoplasm. In each of these
at most would result. But megakaryocytes divisions, the centrioles replicate and a com¬
will produce 4000 to 8000 platelets. There¬ plex multipolar spindle is formed. At meta¬
fore, in mammals a significant increase in phase, the chromosomes become arranged in
productive capacity has been gained by the several equatorial planes and give rise at
unique device of cytoplasmic fragmentation. anaphase to several chromosome groups that
The very large size of the megakaryocytes, then reconstitute a single lobulated nucleus
50 to 70 pm, makes them the most conspic¬ of larger size. After a short interval, another
uous blood-forming cell in the marrow. The similar episode of division occurs with daugh¬
cell body is roughly spherical but may have ter groups of chromosomes again reconsti¬
blunt cytoplasmic processes projecting from tuting a single large nucleus at telophase.
its surface (Fig. 9-19). The nucleus is re¬ Thus, cells with 4n, 8n, 16n, and higher
markably elaborate, with multiple lobes of orders of polyploidy are formed without di¬
varying size interconnected by constricted vision of the cytoplasm. The successive dou¬
regions. The nucleoplasm has a moderately bling of DNA content in cells of this series
coarse chromatin pattern and contains nu¬ has been verified by microspectrophotome¬
merous indistinct nucleoli. The cytoplasm try. Frequencies of 4n, 8n, 16n, 32n, and 64n
appears rather homogeneous in routine mar¬ are respectively 1.6, 10, 71.2, 17.1, and 0.1
row smears, but after appropriate fixation per cent.
and special staining a concentric zonation is Although there is a concurrent increase in
apparent. A narrow perinuclear zone is sur¬ volume of the megakaryocyte, there is little
rounded by a broad region stippled with differentiation of its cytoplasm until it has
numerous fine azurophil granules, which synthesized all of the DNA it will ultimately
may be either uniformly distributed or gath¬ possess. Since the volume attained by a cell
ered in small clusters, depending on the stage is a function of its ploidy, and not all progress
of development of the megakaryocyte. to the same degree of polyploidy, megaka¬
Groups of centrioles are found in bays among ryocytes vary considerably in size and in the
the lobules of the complex nucleus. Mito¬ number of platelets they can form.
chondria are numerous and quite small. A In the promegakaryocyte stage (30—45 |xm in
single compact Golgi apparatus is demonstra¬ diameter), a prominent cell center develops
ble in a juxtanuclear location in young mega¬ containing a number of centriole pairs cor¬
karyocytes, but in mature forms multiple responding to the degree of polyploidy. In
small Golgi bodies are widely dispersed in the subsequent increase in cytoplasmic vol¬
the cytoplasm. At the periphery is a rim of ume, its basophilia diminishes and newly
clear cytoplasm of irregular outline and vary¬ formed azurophilic granules become dis¬
ing thickness, which is devoid of granules. persed throughout.
The fully formed reserve megakaryocyte, not
yet active in platelet formation, is 50 to 70
Megakaryocyte Development fxm in diameter and possesses a very large,
The committed stem cell of mammalian highly lobulated nucleus. Fine azurophil
thrombopoiesis, designated colony-forming granules are widely disseminated in the cen¬
unit—megakaryocyte (CFU-M), gives rise to the tral region of the cytoplasm, but generally
earliest morphologically identifiable cell of absent from a narrow peripheral zone of pale
this lineage, the megakaryoblast. This is a large blue ectoplasm. This rim of clear ectoplasm
cell with round or indented nucleus, loose becomes less conspicuous in mature platelet¬
chromatin pattern, and inconspicuous nu¬ forming megakaryocytes, and the azurophil
cleoli. The cytoplasm is basophilic and largely granules become clustered in small groups
free of specific granules. In electron micro¬ separated by narrow aisles of agranular cy¬
graphs, it contains moderate numbers of toplasm.
 BONE MARROW AND BLOOD CELL FORMATION • 257

. -.—-

Figure 9-19. Micrograph of rat bone marrow including a portion of a megakaryocyte. The limits of the cell are outlined
^“re V6ry 'ar9e SiZe multilobulate nucleus can be compared with the^ smaNer hemopoiefc cells al
258 * BONE MARROW AND BLOOD CELL FORMATION

ing megakaryocytes ultimately fuse side-to-

Ultrastructural Basis of
side*and their coalescence results in a three-
Platelet Formation
dimensional lattice of paired platelet demar¬
The fragmentation of the megakaryocyte cation membranes that bound a continuous
cytoplasm to form platelets was quite accu¬ system of narrow clefts or cisternae that out¬
rately described by Wright in 1906 using line the nascent platelets (Fig. 9-20).
the light microscope, but the nature of the Since its discovery, the platelet demarca¬
cytoplasmic differentiation that makes this tion system has been variously interpreted as
possible had to await electron microscopic an unusual configuration of smooth endo¬
examination. Concurrently with the aggre¬ plasmic reticulum; a derivative of the Golgi
gation of azurophil granules into clusters, complex; a unique organelle arising from
small vesicles appear in the cytoplasm and membranogenic centers in the cytoplasm; or
become aligned in rows that pursue a mean¬ a product of invasion of the megakaryocyte
dering course between the groups of gran¬ cytoplasm by the surface membrane. There
ules. Elecron micrographs in various planes is now little reason to believe that the demar¬
of section suggest that the “vesicles” originally cation system originates from any of the
described may, in fact, be parallel tubular membranous organelles of the cytoplasm.
structures that appear to be vesicles when cut When these cells are exposed to solutions of
transversely. If it were possible to visualize ferritin or horseradish peroxidase, which do
these linear arrays of tubules in three dimen¬ not penetrate biomembranes, these extracel¬
sions, it would be obvious that they are ac¬ lular tracers readily enter the cisternae of the
tually arranged in intersecting planes that demarcation system. Such experiments
partition the cytoplasm into units 1 to 3 \xm clearly demonstrate communication with the
in diameter, each containing a cluster of extracellular space and strongly support de¬
granules and representing a future platelet. rivation of the system from the surface mem¬
The limiting membranes of the aligned tu¬ brane. The alternative possibility that the
bular profiles seen in micrographs of matur¬ system develops from cytomembranes and

Figure 9-20. Micrograph of a small area of megakaryocyte cytoplasm, showing the platelet demarcation channels
outlining future platelets.
 BONE MARROW AND BLOOD CELL FORMATION • 259

secondarily establishes open communication Their large size results in their entrapment
with the cell surface is rendered unlikely by at sites where the narrowing of the pulmo¬
the cytochemical demonstration that the de¬ nary vessels prevents their passage. Tens of
marcation membranes possess enzymatic ac¬ thousands of megakaryocytes are believed to
tivity that is characteristic of the cell surface reach the lungs every minute. Their signifi¬
and not found in membranes of the cyto¬ cant contribution to platelet production is
plasm. It is now widely accepted that the made evident by the observation that the
tubules found in the cytoplasm arise by in¬ platelet count in blood taken from the pul¬
vagination of the plasmalemma. These be¬ monary veins is higher than in that from the
come oriented with their long axes parallel pulmonary artery.
and, by side-to-side fusion of their mem¬
branes, the tubules are reorganized into flat
sheets limiting an extensive system of cister- Kinetics of Thromhopoiesis
nae. The two sheets of membrane bounding Considerable information has accumulated
any one segment of the system will form the on the kinetics of thromhopoiesis. The gen¬
membranes of adjacent nascent platelets. eration time from stem cell to platelet-pro¬
ducing megakaryocyte is estimated to be
Release of Platelets about ten days in humans. A humoral regu¬
latory mechanism seems to ensure that pro¬
Megakaryocytes are located in subendo- duction is responsive to the need for circu¬
thelial relationship to the vascular sinuses of lating platelets. Bleeding or performance of
the marrow. From this position, processes of exchange transfusions producing low levels
mature megakaryocytes penetrate through of platelets (thrombocytopenia) is followed, in
the endothelium, and large segments of their several days, by a three- to fourfold increase
cytoplasm come to project into the lumen. in megakaryocyte numbers in the marrow
These structures, sometimes called proplate¬ and a rebound in circulating platelets to 150
lets, may contain as many as 1200 platelet to 200 per cent of the initial level. A hypo¬
subunits and it is estimated that a single thetical humoral agent, thrombopoietin, is be¬
megakaryocyte may produce and cast off lieved to be responsible for this positive feed¬
about six proplatelets, ultimately giving rise back, but efforts to isolate a thromhopoiesis
to 8000 platelets. The discharge of the pro¬ stimulating factor from plasma and urine
platelets leaves behind around the polymor¬ have not been notably successful to date.
phous nucleus a thin residual layer of cyto¬
plasm bounded by an intact cell membrane.
The possibility that these residual megakaryo¬
cytes might reconstitute their cytoplasm and LYMPHOPOIESIS
produce another set of platelets has not been
excluded. It is generally assumed, however, Blood cells were traditionally divided into
that they degenerate and are replaced by two categories according to their presumed
differentiation of new megakaryocytes from sites of origin. The lymphocytes and mono¬
stem cells. About 10 per cent of megakary¬ cytes were referred to as the “lymphoid”
ocytes observed in bone marrow appear to elements of the blood reflecting the belief
be degenerating cells that have lost nearly all that they arose in the lymphoid organs
of their cytoplasm in platelet production. (lymph nodes, spleen, and thymus). The
Fragmentation of the proplatelets probably granulocytes, erythrocytes, and platelets
takes place mainly in the vascular sinuses of formed in the bone marrow were referred to
the marrow, but some enter the general cir¬ as the “myeloid” elements and their devel¬
culation and are carried to the lungs where opment was collectively termed myelopoiesis.
they break up into individual platelets. Entire This terminological distinction persists in
megakaryocytes may also enter the circula¬ common usage although some of the assump¬
tion. Their presence in the blood has been tions upon which it was based are no longer
abundantly documented. They are also valid. Monocytes have now been shown to
found occasionally in spleen, liver, and kid¬ arise in the marrow from a progenitor cell
ney, but are most abundant in the vascular that is also capable of giving rise to granulo¬
channels of the lung. These pulmonary cytes, It is therefore no longer appropriate
megakaryocytes originate in the bone mar¬ to consider it a lymphoid cell.
row and are carried in the blood to the lung. Although considerable proliferation of
260 • BONE MARROW AND BLOOD CELL FORMATION
s '
stimulated lymphocytes occurs in the periph¬ REGULATION OF
eral lymphoid organs throughout life, the HEMOPOIESIS
lymphopoietic stem cells originate in the bone
marrow. Those destined to become T lym¬
Much research has been done to discover
phocytes leave the marrow and are carried
how hemopoiesis is regulated to maintain the
in the blood to the cortex of the thymus
normal numbers of each cell type in the
where they proliferate and acquire their
circulation. We still know little about what
characteristic surface markers as they move
determines which of several alternative path¬
into the medulla, whence they are trans¬
ways of differentiation will be taken by the
ported to the spleen and there undergo fur¬
progeny of pluripotential stem cells; how¬
ther maturation before becoming members
ever, it is becoming apparent that multiple
of the long-lived recirculating population of
factors are involved. Some originate locally
small lymphocytes.
as components of the hemopoietic microenviron¬
The genesis of B lymphocytes has been
ment. Others are humoral agents that originate
more clearly defined in birds than in mam¬
elsewhere in the body and are blood-borne
mals. The stem cells migrate from the bone
to the marrow to stimulate the proliferation
marrow to an appendage of the avian cloaca
called the bursa of Fabricius in much the same of particular cell lineages.
way that T-cell precursors populate the thy¬ 6

mus. In the bursa, they differentiate into Hemopoietic Microenvironment

mature B cells and then enter the recirculat¬
ing pool of small lymphocytes. The location Although stem cells may be carried in the
of the functional equivalent of the bursa of blood throughout the body, hemopoiesis is
Fabricius in mammals has been a subject of confined to certain tissues and organs that
controversy. The genesis of B lymphocytes provide a favorable environment for their
appears to take place in multiple sites includ¬ proliferation. Appropriate local conditions
ing gut-associated lymphoid tissue, spleen, may be acquired by some organs and lost by
and bone marrow. It has become apparent, others, as exemplified in the changing loca¬
however, that the bone marrow is probably tion of the principal site of hemopoiesis dur¬
the major primary site of lymphopoiesis in ing embryonic life from yolk sac to liver and
mammals. In laboratory rodents in which it spleen, and then to the marrow. The stem¬
can be studied with radiolabeled tracers, the cell assay based on colony formation in the
marrow has a high continuous rate of lym¬ spleen of irradiated mice depends on the
phocyte production throughout fetal and existence in that organ of an environment
postnatal life. Lymphocytes constitute 30 per capable of attracting and supporting prolif¬
cent of all nuclelated marrow cells and at all eration of pluripotential cells transported in
ages exceed the number of erythroblasts. The the blood. Creation of such an environment
dividing precursors of small lymphocytes are is a function of radio-resistant cellular ele¬
somewhat larger cells with a leptochromatic ments of the stroma. Ectopic sites of hemo¬
nuclear pattern, a very thin rim of cytoplasm poiesis can be established in experimental
with few organelles. These so-called transi¬ animals by transplantation of marrow stroma.
tional cells are actively proliferating and con¬ After some initial degenerative changes, the
stitute about one fifth of all marrow lympho¬ tissue becomes revascularized and ultimately
cytes. Production rate of small lymphocytes is repopulated with hemopoietic cells. The
in mouse marrow is calculated to be of the mechanism by which migratory stem cells
order of 108 cells per day. Although ethical recognize and lodge in tissues that will sup¬
considerations exclude quantitation by simi¬ port their growth is not known. The next
lar methods in the human, proportionally step of commitment to a particular path of
high rates of marrow lymphocytes are as¬ differentiation seems to require interaction
sumed. The resulting lymphocytes progres¬ with cells of the stroma. Local factors pro¬
sively acquire the IgG surface markers typical vided by the stroma are also necessary to
of B lymphocytes and continuously emigrate sustain proliferation and maturation of the
in very large numbers to the peripheral hemopoietic cells. Whether stem cell commit¬
lymphoid tissues. ment and proliferation are accomplished by
 BONE MARROW AND BLOOD CELL FORMATION • 261

cell contact or production of diffusible stim¬ newal, the relative numbers of the several
ulatory substances is not known, but the most cell types remain remarkably constant. This
likely candidate for carrying out these func¬ implies a need for some mechanism of mon¬
tions is believed to be the dendritic reticular itoring the numbers in the circulation, and
cell, although other cells may be involved. the existence of specific humoral growth reg¬
The stroma of different organs or in dif¬ ulators acting back upon the hemopoietic
ferent regions of the same organ favor de¬ tissue to control the rate of formation and
velopment of different cell lineages. Stem release of new cells of each type. While in
cells lodging in the spleen of irradiated mice most cases the means of sensing changes in
give rise to colonies that are predominantly the circulating population remains obscure,
erythroid, while those settling in depleted considerable progress has been made in de¬
marrow produce a preponderance of gran¬ tecting and characterizing the molecules in¬
ulocyte/monocyte colonies. Stem cells rarely volved in humoral regulation of specific
form lymphoid colonies in the spleen but hemopoietic cell lineages.
those lodging in thymus or lymph nodes do The control of erythropoiesis was the first
so regularly. to be studied and is best understood. Blood
The study of long-term cultures of bone loss stimulates the marrow to increase its
marrow has provided equally persuasive evi¬ production and release of erythrocytes.
dence of the influence of microenvironment. Transfusion of excess erythrocytes is fol¬
In such cultures the cells produced are gran¬ lowed by suppression of erythropoiesis. With
ulocytes, largely neutrophils. In addition, no change in blood volume, an enhanced
there is an extensive growth of stromal ele¬ need for oxygen transport also stimulates the
ments including endothelial cells, reticular marrow. It was observed nearly a century ago
cells, adipose cells, and macrophages. A dom¬ that in the physiological adaptation to high
inant feature of cultures active in granulo¬ altitude, the body responds to hypoxia by
poiesis is the presence of clusters of adipose increasing the number of circulating eryth¬
cells. The differentiating granulocytes are rocytes. It was initially concluded that the
invariably found in very close relationship to marrow was directly responsive to the oxygen
fat cells or the processes of reticular cells. content of the blood. However, this reason¬
Granulopoietic mouse marrow cultures can able interpretation was later shown to be
be switched to erythropoiesis by addition of incorrect in ingenious experiments on para¬
serum from anemic mice to the medium. biotic rats that share a common circulation.
The changeover is accompanied by a regres¬ When only one member of the pair was
sion of the population of adipose cells and exposed to hypoxia, both members showed
emergence of a new pattern of cell associa¬ enhanced erythropoiesis. This and other ex¬
tion. All stages of the erythroid series are periments clearly demonstrated that the hy¬
found clustered around large mononuclear poxic stimulus to the marrow was mediated
phagocytes in a configuration closely resem¬ by a blood-borne humoral agent. This was
bling the “erythroblastic islets” commonly ob¬ initially named erythropoiesis stimulating factor
served in marrow in vivo. but is now called erythropoietin. Erythropoietin
Thus, it is evident from experimental stud¬ is a glycoprotein of 70,000 molecular weight.
ies both in vivo and in vitro that the microen¬ It can be detected in plasma and urine of
vironment plays a very important role in the hypoxic animals and in humans with diseases
regulation of hemopoiesis. The stromal cells attended by oxygen deficiency. Erythropoie¬
are intimately involved, and the specific cell tin synthesis is inversely related to the oxygen
associations favoring granulopoiesis differ tension prevailing in the tissues. The site of
from those favoring erythropoiesis. its synthesis is mainly in the kidney.
Maintenance of normal numbers of circu¬
Humoral Regulation of Hemopoiesis lating erythrocytes depends on (1) continuing
stimulation of the marrow by erythropoietin,
Because blood cells are short-lived and may (2) a marrow capable of responding, and (3)
spend only a small portion of their life span an adequate supply of iron to meet the needs
in the circulation, they must be continually of the marrow for synthesis of hemoglobin.
replaced by formation of large numbers of A normal marrow not only provides for con¬
new cells. In this process of continual re¬ tinual replacement of erythrocytes, but is also
262 • BONE MARROW AND BLOOD CELL FORMATION

capable of rapid increase in output to four less, as infection continues, there is an accel¬
or five times the basal rate. The body contains eration of granulopoiesis. A humoral agent,
only limited reserves of iron in the form of variously termed leukopoietin or granulopoietin,
ferritin, a protein capable of accumulating is believed responsible for carrying to the
2000 or more atoms of iron. It is found in marrow the signal to produce more neutro¬
various cells throughout the body but notably phils. Convincing experimental evidence for
in the liver. Iron is transported in the plasma such a substance has been difficult to obtain
by an iron-binding globulin, transferrin, and in vivo, but systems have now been developed
is taken up by specific transferrin receptors for cloning populations of hemopoietic cells
on the surface of erythropoietic cells and in semisolid culture medium in vitro. This
interiorized for use in hemoglobin synthesis. has made it possible to detect specific mac¬
The amount of available iron may limit the romolecules that control proliferation and
response of the marrow to erythropoietic maturation of the progeny of single colony-
stimulation, resulting in iron deficiency anemia. forming cells, which are the earliest commit¬
To account for the regulation of circulating ted progenitors of the various hemopoietic
granulocytes, hematologists have long pos¬ cell lines. Cell proliferation within each col¬
tulated a specific humoral factor comparable ony is dependent on the presence in the
with erythropoietin. Validation of the exist¬ medium of a specific colony-stimulating factor
ence of such a substance requires demonstra¬ (CSF). Committed cells that give rise to gran¬
tion that it is capable of stimulating new ulocytes are bipotential. The colonies formed
granulocyte production in vivo. This has may contain only granulocytes, only mono¬
been difficult to achieve because factors other cyte-macrophages, or mixtures of the two.
than formation of new cells influence the These cells are therefore called granulocyte-
granulocyte count. In the presence of a bac¬ macrophage colony-forming cells (GM-CFC). The
terial infection, there is a rapid and large colony-stimulating factor controlling devel¬
increase in the number of circulating neutro¬ opment of these cell types is designated GM-
phils. The rapidity of this response suggests CSF. It is a glycoprotein with a molecular
that it is due to mobilization of reserves weight of about 45,000 and biochemical
rather than to the production of new cells. properties resembling those of erythropoie¬
The population of neutrophils within the tin. It can be found in various tissues but its
vascular system customarily falls into two major source seems to be the mononuclear
categories: (1) those that are flowing with the phagocyte system. Bacterial endotoxins in¬
blood—the circulating pool of leukocytes and (2) crease the release of this colony-stimulating
those that are temporarily adherent to the factor by macrophages in vitro, suggesting a
vascular endothelium in various parts of the mechanism whereby the presence of bacterial
circulatory system—the marginated pool of leu¬ products in infections can increase granulo¬
kocytes. Normally the neutrophils are about cyte production by bone marrow. For this
equally divided between these two pools, but colony-stimulating factor to qualify as a true
in response to exercise, epinephrine admin¬ granulopoietin, it will have to be demon¬
istration, and other stimuli there may be a strated that its repetitive or continuous
massive movement of marginated cells into administration will result in a sustained in¬
the bloodstream, doubling the number in the crease in new circulating granulocytes in vivo.
circulating pool without increased granulo¬ This is yet to be accomplished.
poiesis in the marrow. Perturbations of the numbers of circulating
Under normal circumstances, eight to ten platelets are followed by compensatory ad¬
days elapse after the last myelocyte division justments in thrombopoiesis. Blood plasma
before the myelocytes complete their matu¬ from thrombocytopenic patients is reported
ration and enter the blood. Thus, the greater to stimulate platelet production when in¬
part of the life span of these cells is spent in jected into laboratory animals. The stimula¬
the marrow where there are very large re¬ tory effect of thrombocytopenic plasma can
serves of band-forms and mature neutrophils be detected not only by platelet counts, but
capable of entering the circulation on de¬ by measuring increased incorporation of ra¬
mand—approximately 15 times as many as dioisotope into platelets after administration
there are in the blood. Very marked increases of 75Se-selenomethionine or 35C-sodium sul¬
in circulating neutrophils can thus be fate. Thus, there is strong indirect evidence
achieved by their release without an imme¬ for feedback regulation of platelet numbers
diate change in production rate. Neverthe¬ by a thrombopoietin, but little is known about
 BONE MARROW AND BLOOD CELL FORMATION • 263

the sensing mechanism responsible for acti¬ Dexter, T. M., T. D. Allen, L. G. Lajtha, L. G. Krizra,
vating its synthesis and release. N. Testa, and M. A. S. Moore: In vitro analysis of
There is no persuasive evidence for a feed¬ self-renewal and commitment of hemopoietic stem
cells. In Hematopoietic Mechanisms. Cold Spring
back control of B-lymphocyte production Harbor Symp. Quant. Biol. 42:63, 1978.
comparable with the poietins that regulate Lajtha, L. G.: Haemopoietic stem cells: Concepts and
production of other blood cells. By analogy dehnitions. Blood Cells 5:447, 1979.
with effects of bleeding or hemolysis on Quesenberry, P., and L. Levitt: Haematopoietic stem
cells. N. Engl. J. Med. 30:155, 819, 1979.
erythropoiesis, one might expect a marrow
Till, J. E., and E. A. McCulloch: Hemopoietic stem cell
response to deletion of circulating lympho¬ differentiation. Biochim. Biophys. Acta 665:431
cytes. immunological suppression of circulat¬ 1980.
ing lymphocytes has no significant effect ERYTHROPOIESIS
upon lymphocyte production in the marrow.
Bessis, M.: Life Cycle of the Erythrocyte. Sandoz Mon¬
On the other hand, mice raised under germ- ographs. Basle, Sondoz Ltd., 1966.
free conditions have a markedly reduced rate Campbell, F. R.: Nuclear elimination from the normo¬
of lymphocyte production. It is concluded blast of fetal guina pig liver as studied with electron
that a normal basal level of marrow lympho¬ microscopy and serial section techniques. Anat. Rec
766:539, 1968.
poiesis is regulated by local microenviron¬
Harrison, P. R.: Analysis of erythropoiesis at the molec¬
mental factors, but may be augmented by ular level. Review article. Nature (Lond.) 262:353
exogenous antigenic stimulation. 1976.
Hillman, R. S., and C. A. Finch: Erythropoiesis normal
and abnormal. Semin. Hematol. 4:427, 1967.

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PRENATAL HEMOPOIESIS in the polymorphonuclear leukocytes. Two types
derived from opposite faces of the Golgi complex
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De Bruyn, P. P. H., P. C. Breen, and T. B. Thomas: Meuret, G.: Origin, ontogeny and kinetics of mononu¬
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HEMOPOIETIC STEM CELLS THR OMB OPOIE SIS
Dexter, T. M.: Hemopoiesis in long-term bone marrow Becher, R. P., and P. P. H. DeBruyn: The transmural
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Behnke, O.: An electron microscope study of the me¬ origin, structure, and developmental potential. Int.
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 w
MUSCULAR TISSUE

Contractility is one of the fundamental trachea to the alveolar ducts, and in the
properties of protoplasm and is exhibited in urinary and genital ducts. The walls of the
varying degree by nearly all cell types. In arteries, veins, and larger lymphatic trunks
muscular tissue the ability of the cells to contain smooth muscle. In the skin it forms
convert chemical energy into mechanical minute muscles called arrectores pilorum,
work through contraction has become highly responsible for elevation of hairs. In the
developed. areola of the mammary gland it participates
Two broad categories of muscle are distin¬ in the erection of the nipple, and in the
guished—smooth muscle and striated muscle. subcutaneous tissue of the scrotum it causes
Smooth muscle is composed of fusiform, uni¬ the wrinkling of the skin that accompanies
nucleate cells. It contracts in response to elevation of the testes. In the eye it forms the
stimulation by the autonomic nervous system musculature of the iris and of the ciliary
and thus is not subject to voluntary control. body, which is concerned with accommoda¬
Striated muscle associated with the skeleton tion and with constriction and dilation of the
is responsible for locomotion and other vol¬ pupil.
untary movements of vertebrates and is inner¬
vated by the cerebrospinal nervous system. It
The Smooth Muscle Fibers
consists of very long cylindrical units, which
are multinucleate syncytia containing large Smooth muscle fibers are long, spindle-
numbers of closely packed cytoplasmic fila¬ shaped cells. Where they are closely associ¬
ments in a highly ordered arrangement that ated in bundles or sheets, their boundaries
results in a distinctive pattern of cross stria- are difficult to resolve with the light micro¬
tions along the length of the cellular ele¬ scope, but by special maceration techniques
ments, traditionally called muscle fibers. the fibers can be isolated, and their long
The muscular wall of the heart is a unique fusiform shape is then evident. They vary
form of striated muscle whose rhythmic con¬ greatly in length in different organs. In the
traction is involuntary. Cardiac muscle differs pregnant human uterus, they may reach 0.5
from skeletal muscle in that its branching fibers mm in length. Their average length in the
are not syncytial but are formed of individual musculature of the human intestine is about
cellular units joined end to end. 0.2 mm. The smallest smooth muscle cells in
the walls of small blood vessels may be only
20 pm long.
In longitudinal sections, a single elongated
SMOOTH MUSCLE
nucleus is found to occupy the thickest part
of the fiber, about midway along its length.
Smooth muscle forms the contractile por¬ The long nuclear profile is rounded at the
tion of the wall of the digestive tract from (ends (Fig. 10-1). The chromatin usually
the middle of the esophagus to the internal forms a delicate pattern uniformly dispersed
sphincter of the anus. It provides the motive in the nucleoplasm, but in the smooth muscle
power for mixing the ingested food with }of some organs it tends to be aggregated
digestive juices and for its propulsion along the inner surface of the nuclear enve¬
through the absorptive and excretory por¬ lope. There are two or more nucleoli, de¬
tions of the tract. Smooth muscle is also pending on the species. In smooth muscle
found in the walls of the ducts in the glands fixed in contraction, the passively distorted
associated with the alimentary tract, in the nuclei may be deeply indented along their
walls of the respiratory passages from the margins or may take on a helical form.

265
266 * MUSCULAR TISSUE

Figure 10-1 Figure 10-2

Figure 10-1. Photomicrograph of a longitudinal section of smooth muscle from the tunica muscularis of intestine.
Figure 10-2. Photomicrograph of a transverse section c)f smooth muscle from the tunica muscularis of the human
stomach. The section was stained with the periodic acid- Schiff reaction, which stains the glycoprotein external lamina
of the muscle cells, accentuating their outline.

The cells of smooth muscle are offset with anisotropic transverse bands that are char¬
respect to one another, so that the thick acteristic of the myofibrils of striated muscle.
middle portion of one is juxtaposed to the After appropriate cytological staining
thin ends of adjacent cells. In transverse methods, mitochondria can be seen through¬
sections, smooth muscle therefore presents a out the sarcoplasm, but they tend to congre¬
mosaic of rounded or irregularly polygonal gate near the poles of the elongated nucleus.
profiles varying from less than 1 pm to sev¬ A pair of centrioles and a small Golgi appa¬
eral micrometers across (Fig. 10—2). Only the ratus can also be demonstrated. In some
largest profiles, those representing sections organs the sarcoplasm of smooth muscle may
through the thick middle portion of the fi¬ contain glycogen.
bers, contain a centrally placed cross section In the development of mammary and
of the nucleus. No nucleus is found in the sweat glands, certain cells of ectodermal ori¬
smaller profiles, which represent sections at gin become specialized for contraction. The
various levels in the tapering ends of the cell body of these myoepithelial cells has some
fusiform cells. of the characteristics of epithelial cells, but
The cytoplasm of muscle cells is called their base is drawn out into several radiating
sarcoplasm. In smooth muscle it appears quite processes that contain myofilaments. These
homogeneous in the living state and is almost portions of the cell have an appearance
equally devoid of structure after routine fix¬ closely resembling the sarcoplasm of smooth
ation and staining. However, after use of muscle cells.
special stains, or after gentle maceration in
acid, fine longitudinal striations can be dem¬ Mode of Association of Smooth
onstrated, running the full length of the cell. Muscle Fibers
These are the myofibrils, the contractile ma¬
terial of smooth muscle. They are birefrin- Smooth muscle cells may occur singly or in
gent under the polarizing microscope, but small groups in ordinary connective tissue, as
show no sign of the alternating isotropic and in the lamina propria of the intestinal villi,
 MUSCULAR TISSUE • 267

where their contraction shortens the villi and and numerous polyribosomes. A small Golgi
helps expel lymph from the lacteals. In the complex is located near one pole of the
walls of blood vessels, where smooth muscle nucleus. The bulk of the sarcoplasm is occu¬
fibers serve only to change the caliber of the pied by exceedingly thin, parallel myofila¬
lumen, they are oriented circumferentially, ments associated in bundles that correspond
occurring as isolated fibers in the smallest to the myofibrils seen with the light micro¬
arterioles and as a continuous layer in vessels scope. These are oriented, for the most part,
of larger size. In the wall of the intestine, parallel to the long axis of the muscle cell.
smooth muscle is arranged in separate lon¬ Interspersed among the bundles of myofila¬
gitudinal and circumferential layers. The co¬ ments are mitochondria, occurring singly or
ordinated action of these layers forms con¬ in small clusters, and having a prevailing
strictions that move along the intestine as longitudinal orientation. Scattered through
peristaltic waves, propelling the contents the contractile substance of the cell are oval
through the lumen. In other hollow organs, or fusiform dense areas (Figs. 10-3 and
such as the bladder or uterus, the smooth 10-4). At high magnification these appear to
muscle forms poorly defined layers of elab¬ be traversed by myofilaments embedded in a
orately interlacing coarse bundles oriented in dense amorphous matrix. Similar densities
different directions. The connective tissue are distributed at intervals along the inner
fibers outside the muscle layer continue into aspect of the plasmalemma. The exact nature
the spaces between the cells and bind them of these dense areas in the sarcoplasm is not
into bundles. Between the thicker bundles or known. Their hne structural resemblance to
layers of smooth muscle cells is a small the dense regions found at desmosomes, at
amount of loose connective tissue containing zonulae adherentes of epithelia, and the Z
fibroblasts, collagenous and elastic fibers, and bands of striated muscle suggests that the
a network of capillaries and nerves. Connec¬ dense component of all of these may be
tive tissue cells are seldom found within similar and may have a cohesive function.
smooth muscle bundles, but the clefts be¬ The occurrence of dense bodies in smooth
tween muscle cells are nevertheless occupied muscle at nodal points where myofilaments
by thin reticular fibers, which branch irreg¬ seem to be bonded together laterally, and
ularly and form a delicate network investing also where they are attached to the cell sur¬
the individual smooth muscle cells. This re¬ face, is consistent with this speculation. The
ticulum can be stained with Mallory’s aniline plasmalemma between the specialized sites of
blue method and still more distinctly with the myofilament attachment is characteristically
Bielschowsky silver impregnation method. studded with small vesicular inpocketings or
The reticular fibers are embedded in a layer caveoli like those seen in endothelial cells and
of intercellular material that appears in sec¬ commonly interpreted as evidence of micro-
tions stained with the periodic acid—Schiff pinocytosis (Fig. 10-6). However, these vesi¬
(PAS) reaction as a continuous pattern of cles do not appear to dissociate from the
magenta lines outlining every muscle cell membrane and move into the sarcoplasm as
(Fig. 10-2). one would expect if they were involved in
The pull of each contracting cell in smooth endocytosis. It has been suggested that they
muscle is transmitted to the surrounding may sequester and release calcium and may
sheath of reticular fibers, which are continu¬ thus play a role in control of contraction and
ous with those of neighboring cells and ulti¬ relaxation comparable with that of the sar¬
mately with those of the surrounding con¬ coplasmic reticulum of striated muscle. But
nective tissue. This arrangement permits the the exact physiological significance of these
force of contraction of the entire layer of the membrane invaginations remains unclear.
smooth muscle to be uniformly transmitted In the early electron microscopic studies
to the surrounding parts. all vertebrate striated muscles and many un-
striated muscles of invertebrates were found
The Fine Structure of Smooth Muscle to contain two distinct sets of parallel fila¬
ments. This led to the expectation that all
In electron micrographs the elongated nu¬ muscle cells would be found to have a con¬
cleus of the extended smooth muscle cell is tractile apparatus consisting of two kinds of
smoothly contoured and rounded at the sarcoplasmic filaments. This expectation was
ends. The juxtanuclear sarcoplasm contains not immediately borne out in studies of mam¬
long slender mitochondria, a few tubular malian smooth muscle. The filaments were
elements of granular endoplasmic reticulum, thinner and less well ordered than in striated
Figure 10-3. Electron micrograph of smooth muscle in transverse section. The cells are separated by wide intercellular
clefts occupied by glycoprotein cell coats and small bundles of collagen fibrils. Scattered through the cytoplasm and at
the periphery of the cell (at arrows) are densities that are sites of lateral bonding of filaments of the cytoskeleton and
sites of attachment of myofibrils to the cell membrane.

muscle, and thicker filaments were only grouped together in bundles associated with
rarely seen. The inconsistency in results of myosin filaments in a ratio of 12:1 or higher,
electron microscopy of smooth muscle compared with 6:1 in striated muscle. Infor¬
proved to be a technical problem. The rou¬ mation on the three-dimensional arrange¬
tine procedure of posthxation with osmium ment of the filaments is still incomplete, but
solutions distorts or depolymerizes actin fila¬ they appear to be grouped together in con¬
ments and is probably destructive to myosin tractile units attached at each end to the cell
filaments in this tissue. The corresponding membrane. The organization of the myosin
filaments in striated muscle are much more molecules and the structural polarity of the
stable. In more recent studies using improved filaments formed differs from that of myosin
methods of preservation, it has been possible in striated muscle. Thick filaments isolated
to distinguish punctate profiles of two sizes from smooth muscle are quite variable in
that are interpreted as cross sections of two length (3-8 pm) and they possess projecting
kinds of filaments comparable with the cross bridges at regular intervals along their
myosin and actin filaments of striated muscle entire length. Thus, the molecules do not
(Fig. 10—5). The existence of a two-filament have the antiparallel arrangement that results
contractile system in smooth muscle is now in a central bare area on myosin filaments of
generally accepted, and the morphological striated muscle (Fig. 10-255). The molecular
evidence is strongly supported by biochemi¬ organization of the thin filament is very sim¬
cal isolation of both actin and myosin and ilar to filamentous actin of striated muscle.
their interaction in vitro to form a complex How the shearing action between the thick
capable of contraction upon addition of and thin filaments takes place during con¬
adenosine triphosphate. traction is still poorly understood. The integ¬
Actin is relatively much more abundant rity of the contractile system is strongly
than in striated muscle and the filaments are dependent on the extracellular calcium con-
 MUSCULAR TISSUE • 269

centration. If smooth muscle is perfused with

mammalian Krebs-Ringer solution contain¬
ing a physiological concentration of calcium
ions, the myofilaments are present in normal
abundance and distribution in electron mi¬
crographs (Fig. 10-6T). If it is then perfused
with the same solution containing the chelat¬
ing agent EDTA, myofilaments are largely
absent (Fig. 10—65). The filaments are re¬
stored and contractility regained upon sub¬
sequent perfusion with the original solution.
In addition to the contractile filaments,
smooth muscle cells contain a well-developed
cytoskeleton consisting of 10-nm hlaments,
which form structural attachments with the
sarcoplasmic dense bodies and the plasma
membrane. The cytoskeletal elements are not
easily identified in routine micrographs, but
the hlaments and their association with the
dense bodies are clearly visualized in prepa¬
rations in which the myosin and actin hla¬
ments have been extracted (Fig. 10—65).
Opinion is divided as to whether the dense
bodies are at nodal points in the cytoskeletal
network of 10-nm hlaments or whether they
are also sites of attachment of the actin hla¬
ments.

Cell-to-Cell Relations in
Smooth Muscle
In electron micrographs the surface of
each smooth muscle cell is invested by a thick
extracellular coating that corresponds in its
hne structure to the basal lamina of epithelia
and to the external lamina of other mesenchy¬
mal derivatives. This is clearly the component
responsible for the PAS reaction of the inter¬
cellular spaces of smooth muscle (Fig. 10-2).
Small bundles of collagen hbrils, which cor¬
respond to the argyrophilic reticulum, are
lodged in clefts between or within the glyco¬
protein surface coatings of adjacent smooth
muscle cells.
Owing to the presence of this thick extra¬
cellular layer, adjacent smooth muscle cells
are separated by a distance of 40 to 80 nm.
Typical desmosomes are not found. How¬
ever, the specialized dense areas in which the
myofilaments terminate at the surface of ad¬
jacent cells are too often opposite one an¬
other for their distribution to be entirely
Figure 10-4. Micrograph of a central segment of a smooth random. An intermediate dense line may be
muscle cell in longitudinal section. The uniform gray region found in the intercellular material between
at the periphery is occupied by myofilaments that are not two such opposing dense regions. Thus, in
resolved at this magnification. A conical region of sarco¬
spite of the considerable distance that sepa¬
plasm extending from the pole of the elongate nucleus
contains numerous mitochondria and a few profiles of rates the cells, there is a complementarity to
rough endoplasmic reticulum. the specialized areas of their surfaces that
270 • MUSCULAR TISSUE

Figure 10-5. Micrograph of a vascular smooth muscle cell in cross section, clearly showing thick and thin filaments.
Inset shows the same at higher magnification. (Micrograph from Somlyo, A. P., C. E. Devine, and A. V. Somlyo. In
Vascular Smooth Muscle. Heidelberg, Springer-Verlag, 1972.)

suggests cell-to-cell cohesion at these sites, as tance, permitting free movement of ions and
at the desmosomes of epithelia. Contraction spread of excitation from one cellular unit to
results in force applied at many points of another throughout the muscle mass.
insertion of myofilaments on the periphery Although smooth muscle cells are special¬
of the cell. The contracted cell becomes el¬ ized for their primary contractile function,
lipsoid and may exhibit numerous invagina¬ they also synthesize and release components
tions of its surface at points of attachment of of the extracellular matrix. Cultures of
myofilament bundles. The force is probably smooth muscle cells have been shown to pro¬
transmitted to neighboring cells, mainly duce collagen and elastin. Thus, they are not
through the reticular connective tissue dependent on fibroblasts for formation of
sheath, but long-range forces of attraction their extracellular matrix.
acting at multiple dense areas of specializa¬
tion on the opposing cell surfaces may also Physiological Properties and
be involved. Contractile Mechanism of
In certain limited areas of the surface of Smooth Muscle
visceral smooth muscle, the intercellular sub¬
stance is lacking and the membranes of Smooth muscle is distinguished from
neighboring cells come into very close asso¬ striated muscle not only by its histological
ciation. At these sites, the intercellular space and cytological appearance but also by its
is narrowed to 2 nm or less, and the opposing physiological and pharmacological proper¬
plasmalemmae, in freeze-cleaved prepara¬ ties. Its contractions are slower than those of
tions, exhibit closely packed 9-nm particles in other types of muscle, but it is able to sustain
hexagonal array within the plane of the mem¬ forceful contraction for long periods with
brane. These junctional specializations are relatively little expenditure of energy. De¬
therefore typical gap junctions or nexuses, and pending on the site, contraction may be ini¬
are believed to be sites of low electrical resis¬ tiated by nerve impulses, hormonal stimula-
 MUSCULAR TISSUE • 271

Figure 10-6. A, Micrograph of smooth muscle cell perfused with Krebs-Ringer solution, showing bundles of longitudinal
filaments. B, Muscle perfused with Krebs-Ringer solution containing the calcium chelating agent EDTA. The myofilaments
are no longer present. The 10-nm cytoskeletal filaments and dense bodies remain. Upon reperfusion with the original
solution the filaments reappear and contractility is restored. (Micrographs from Cooke, P. Eur. J. Cell Biol. 27:55, 1982.)

tion, or local changes arising within the being conducted from cell to cell via the gap
muscle itself. One of the more important junctions.
local stimuli initiating contraction is stretch¬ Two forms of contraction are recognized
ing of the muscle fibers, which can change in visceral smooth muscle: rhythmic contraction
the membrane potential and initiate a wave and tonic contraction. In the former, periodic
of contraction. The ability of smooth muscle spontaneous impulses are generated and
to respond to stretch is particularly important spread through the muscle, accompanied by
in the physiology of the bladder, gastrointes¬ a wave of contraction. In addition, smooth
tinal tract, and other hollow viscera, whose muscle maintains a continuous state of partial
contents are evacuated by contractions. contraction called muscle tone or tonus. The
Although usually treated by morphologists cause of the tonic contraction is no better
as a single type of muscle, smooth muscle is understood than the genesis of the rhythmic
adapted to a variety of functions and differs contractions. The degree of tonic contraction
markedly in its physiological properties in may change greatly, without any change in
different organs. Vascular smooth muscle, in the frequency of the rhythmic contraction, and
the blood vessels, behaves rather like skeletal vice versa. The two forms of contraction thus
muscle in that its activity is usually initiated appear to be independent.
by nerve fibers (vasomotor nerves), and there There are several other physiological and
is little evidence of conduction between cel¬ pharmacological differences in smooth mus¬
lular units. Visceral smooth muscle, on the other cle of different organs. For example, the
hand, bears certain functional resemblances amounts of actin and myosin in smooth muscle
to cardiac muscle in that it has a myogenic of the uterus are under endocrine control.
autorhythmicity; the entire cell mass behaves Its cells hypertrophy during pregnancy, and
as though it were a single unit with impulses show a striking increase in the size of the
272 • MUSCULAR TISSUE

Figure 10-7. Cross section through human sartorius muscle, showing the connective tissue of the epimysium surrounding
the entire muscle, and the perimysium enclosing muscle fiber bundles of varying size. (Photograph by Muller, from
Heidenhain.)

Golgi apparatus and the extent of the gran¬ and so on. The individual muscle fibers, the
ular endoplasmic reticulum. Physiological fascicles, and the muscle as a whole are each
changes also occur during the normal estrus invested by connective tissue that forms a
cycle. Ribonucleic acid synthesis is one of the continuous stroma, but its different parts are
early responses of the uterus to estrogen designated by separate terms for convenience
stimulation, and the organelles concerned of description. The entire muscle is enclosed
with protein synthesis become more promi¬ by a connective tissue layer called the epimy¬
nent during estrus than at other times. Uter¬ sium (Fig. 10—7). Thin collagenous septa that
ine musculature in the terminal stages of extend inward from the epimysium, sur¬
pregnancy is also responsive to the hormone rounding each and all of the fascicles, collec¬
oxytocin, elaborated by the posterior lobe of tively make up the perimysium, and the ex¬
the hypophysis. Smooth muscle in other parts ceedingly delicate reticulum that invests the
of the body is relatively unresponsive to hor¬ individual muscle fibers constitutes the endo-
mones other than epinephrine. mysium. The connective tissue serves to bind
together the contractile units and groups of
units and to integrate their action; it also
allows a certain degree of freedom of motion
SKELETAL MUSCLE between them. Thus, although the muscle
fibers are very closely packed together, each
Histological Organization is somewhat independent of adjacent fibers,
and each fascicle can move independently of
The unit of histological organization of neighboring fascicles.
skeletal muscle is the fiber, a long cylindrical The blood vessels supplying skeletal muscle
multinucleate cell visible with the light micro¬ course in the connective tissue septa and
scope. Large numbers of parallel muscle fi¬ ramify to form a rich capillary bed around
bers are grouped into fascicles, which are the individual muscle fibers (Fig. 10-9). The
visible to the naked eye in fresh muscle. The capillaries are sufficiently tortuous to permit
fascicles are associated in various patterns to their accommodation to changes in length of
form the several types of muscles recognized the fibers, by straightening during elongation
by the anatomist—unipennate, bipennate, and contorting during contraction.
 MUSCULAR TISSUE • 273

and exemplified in the bulging biceps of the

boxer and the leg muscles of the ballerina.
Conversely, the fibers may become thinner
in muscle immobilized for long periods as in
the treatment of fracture, a change called
atrophy of disuse.

Cytology of the Muscle Fiber

The individual fibers can be separated by
teasing fresh muscle apart under a dissecting
microscope. In addition to the obvious trans¬
verse striations that give this type of muscle
its name, a more delicate longitudinal stria-
tion is also discernible within the muscle fiber.
The structural basis of this longitudinal stria-
tion becomes apparent when samples of mus¬
cle are treated with dilute nitric acid. In such
macerated specimens, the sarcolemma, the lim¬
iting membrane of the fiber, is destroyed;
the cytoplasmic matrix, called the sarcoplasm,
is extracted; and the contractile substance of
the muscle fiber separates into a large num¬
ber of thin, parallel, cross-striated myofibrils.
The fine longitudinal striation detectable
within the fresh muscle fiber is thus attrib¬
utable to the parallel arrangement of myriad
myofibrils within its sarcoplasm. The trans¬
verse striation comes about because each my¬
ofibril is made up of cylindrical segments or
bands of different refractility that alternate
regularly along its length. The corresponding
segments of the closely packed parallel myofi¬
brils are usually in register, so that the stria¬
Figure 10-8. Drawing of the relationships of skeletal
muscle fibers. (From Brodel, M. Bull. Johns Hopkins Hosp. tions appear to extend across the whole width
67:295, 1937.) of the fiber (Figs. 10-11, 10-12).
Each muscle fiber is invested by a delicate
membrane just visible with the compound
In muscles that do not taper at the ends, microscope. In teased fresh preparations,
such as the sartorius, the fibers apparently where the fiber has been torn or crushed, it
continue without interruption through the appears as a thin, transparent him (Fig.
entire length of the muscle. It is generally 10-8). This investment has traditionally been
believed, however, that in most muscles the called the sarcolemma. It has recently become
fibers are shorter than the muscle as a whole, apparent from electron microscopic studies
seldom extending from its origin to its inser¬ that this him, visible with the light micro¬
tion, but being connected at one end to scope, is not a single component but consists
connective tissue septa within the muscle and of the plasmalemma of the muscle fiber, its
at the other to the tendon. glycoprotein external coating, and a delicate
The thickness of the muscle fibers ranges network of associated reticular hbers. In cur¬
from 10 to 100 pm or more, depending on rent usage, the term sarcolemma is reserved
the species and the particular muscle exam¬ for the plasmalemma of the muscle hber, and
ined. Fibers within the same muscle may vary the other components of the traditional sar¬
considerably in their caliber. During the colemma are separately designated. The sar¬
growth of the organism, the diameter of the colemma differs in no essential respect from
fibers increases with age, and in the grown the limiting membrane of any other cell. It
individual it may undergo further increase should be realized that this membrane alone
in response to strenuous muscular activity—a is not resolved by the ordinary microscope
phenomenon referred to as hypertrophy of use under the usual conditions of observation,
274 • MUSCULAR TISSUE

Figure 10-9. A, Drawing of the blood supply of muscle bundles in the human rectus abdominis muscle. B, The capillary
network of the muscle fibers. C, The same at higher magnification. (From Brodel, M. Bull. Johns Hopkins Hosp. 61 \295,
1937.)

but with the added thickness of its associated other nuclei, of similar elongated form but
amorphous and fibrous investments, a limit¬ with a coarser chromatin pattern, are closely
ing layer is visible. associated with the surface of the muscle
The nuclei of the striated muscle fiber are fibers. These belong to elongated satellite cells,
numerous. No actual number can be speci¬ which are flattened against the muscle fiber
fied, for this depends on the length of the or occupy shallow depressions in its surface
muscle, but in a fiber several centimeters long and are enclosed within the same investing
there would be several hundred nuclei. They layer of glycoprotein and reticular fibers. The
are elongated in the direction of the fiber. cytoplasm of the satellite cell is scanty and its
Their position varies according to the type boundary with the muscle fiber usually can¬
of muscle and the animal species, but in the not be resolved with the light microscope.
great majority of skeletal muscles of mam¬ Skeletal muscle fibers have a limited capacity
mals the nuclei are located at the periphery for regeneration. After minor injury muscle
of the fiber, immediately beneath the sarco- fibers often regenerate. The satellite cells
lemma. This is especially apparent in trans¬ within the intact external lamina serve as
verse sections (Fig. 10—10). This characteris¬ stem cells that proliferate and differentiate
tic position is a helpful criterion for into myoblasts, which fuse to form myotubes
distinguishing skeletal from cardiac muscle, and ultimately new muscle fibers. If the in¬
in which the nuclei are centrally located. jury has resulted in disruption of the external
The nuclei of the muscle cells usually have lamina, regeneration does not occur; the area
one or two nucleoli and moderately abundant is invaded by fibroblasts and fibrous scar
chromatin distributed along the inner aspect tissue is formed.
of the nuclear envelope. A small number of The sarcoplasm of a muscle fiber corre-
 MUSCULAR TISSUE • 275

This network appeared to surround all of

the myofibrils and was called the sarcoplasmic
icticulum. It was demonstrated with difficulty
by the light microscope, and many observers
doubted its reality, but the presence of this
organelle has now been verified in electron
micrographs and will be described later in
this chapter.
Lipid droplets are found in varying num¬
bers in the muscles of some species. They
may be situated between the myofibrils or
among the clusters of mitochondria at the
poles of the nuclei and at the periphery of
the fiber. In appropriately stained prepara¬
tions, small amounts of glycogen can be dem¬
onstrated throughout the sarcoplasmic ma¬
trix. In addition to these microscopically
visible inclusions, the sarcoplasm of the living
muscle contains the oxygen-binding protein
myoglobin. In muscle at rest, oxygen probably
remains bound to myoglobin, but when de¬
mand for oxygen increases, it dissociates
from myoglobin and is available for oxida¬
tions. In man, myoglobin is of relatively little
significance in skeletal muscle, but in diving
mammals and in birds, it is especially abun¬
dant and probably of great physiological im¬
portance.
Figure 10-10. Photomicrograph of skeletal muscle fibers Most of the interior of the muscle fiber is
in cross section, illustrating their polygonal outline and the occupied by myofibrils 1 to 2 pm in diameter.
peripheral position of their nuclei.
In transverse sections they are resolved as
fine dots either uniformly distributed or
grouped in polygonal areas called the fields
sponds to the cytoplasm of other types and of Cohnheim. Whether this polygonal pattern
can be defined as the contents of the sarco- represents the true distribution of myofibrils
lemma exclusive of the nuclei. It consists, or is a consequence of shrinkage was long
therefore, of a typical cytoplasmic matrix and debated. The weight of evidence now favors
the usual cell organelles and inclusions as its interpretation as a shrinkage artifact, and
well as the myofibrils peculiar to muscle. no functional significance is now attached to
Although not visible in routine preparations, Cohnheim’s fields.
the Golgi apparatus can be demonstrated by In longitudinal sections of muscle the fea¬
special staining methods. As might be ex¬ ture of greatest interest is the cross striation
pected in a multinucleate syncytium, there of the myofibrils. The cylindrical segments
are multiple small Golgi bodies, which are of the myofibrils, which are markedly refrac-
located near one pole of each nucleus tile and dark in fresh muscle, stain intensely
throughout the muscle fiber. The mitochon¬ with iron-hematoxylin in histological sections,
dria (formerly called sarcosomes) are most while the less refractile, alternate segments
abundant near the poles of the nuclei and remain essentially unstained (Fig. 10-11).
immediately beneath the sarcolemma, but When muscle is examined with the polarizing
they also occur in the interior of the fiber, microscope, the contrast of the bands is re¬
where they are distributed in longitudinal versed. The dark-staining bands are now
rows between the myofibrils. birefringent or anisotropic (A bands) and
Several early cytologists examining prepa¬ therefore appear bright, whereas the light
rations of muscle that were impregnated with staining bands are isotropic (I bands) or only
heavy metals described a lacelike network of very weakly anisotropic and thus appear dark.
dark strands in the interhbrillar sarcoplasm. In the most commonly used terminology, the
276 * MUSCULAR TISSUE

Figure 10-11. Photomicrograph of three muscle fibers in longitudinal section. The transverse dark A bands and light I
bands are clearly shown. Preparation stained with iron-hematoxylin.

principal bands are named A band and I the A band. In its center is a narrow dark
band, according to their appearance in po¬ line, the M band or M line, located precisely
larized light. The relative lengths of the in the middle of the A band. Although all
bands vary, depending on whether the mus¬ these features of the cross-banded pattern of
cle is examined at resting length, during striated muscle can be seen with the light
contraction, or when passively stretched. The microscope, they can be demonstrated and
length of the A band remains constant in all interpreted more clearly in electron micro¬
phases of contraction, but the I band is most graphs, and will be discussed in greater detail
prominent in stretched muscle, is shorter at below in the section on the ultrastructure of
resting length, and is extremely short in con¬ the sarcoplasm.
traction. Both in stained preparations and in
living muscle viewed with phase contrast, a Cytological Heterogeneity of
dark transverse line, the Z line, bisects each I Skeletal Muscle Fibers
band. The repeating structural unit, to which
all the morphological events of the contractile When muscles are examined in the fresh
cycle are referred, is the sarcomere, which is condition by the naked eye, they differ some¬
defined as the segment between two succes¬ what in color. It has also been recognized
sive Z lines and therefore includes an A band since the late 1800s that the fibers making
and half of the two contiguous I bands. In up a single muscle are not uniform in their
histological sections of skeletal muscle, the A size or cytological characteristics. In red mus¬
bands, I bands, and Z lines are usually the cles, small dark fibers with a granular appear¬
only cross striations discernible, but in excep¬ ance predominate, whereas in white muscles,
tional preparations a paler zone, called the paler fibers of larger diameter predominate.
H band, may be seen traversing the center of Differences among the fibers were not con-
 MUSCULAR TISSUE 277

Figure 10-12. Electron micrograph of glycerin-extracted skeletal muscle. This treatment improves the contrast of the
myofibrils. Observe the uniform diameter of the,myofibrils and the peripheral location of the nucleus. Corresponding
bands of adjacent myofibrils are usually in register across the muscle fiber. Where they are out of register, as at upper
left, this is usually an artifact of specimen preparation.
278 • MUSCULAR TISSUE

Figure 10-13. Photomicrograph of skeletal muscle in

cross section stained for the enzyme succinic dehydro¬
genase. Three categories of fibers are distinguished: small Figure 10-14. Succinic dehydrogenase reaction in skel¬
red fibers rich in peripheral mitochondria; large white fibers etal muscle. A, Plantaris muscle of a control guinea pig.
with relatively few mitochondria; and fibers with interme¬ B, Same muscle of an animal after 8 weeks of running
diate characteristics. (Photomicrograph courtesy of G. on a treadmill. The fiber population of the exercised
Gautier.) muscle is more homogeneous—nearly all fibers are small
and rich in mitochondria. (Photomicrographs from Faulk¬
ner, J. A. In Podolsky, R. J., ed.: Contractility of Muscle
spicuous in routine histological preparations, Cells and Related Processes. Englewood Cliffs, NJ, Pren¬
but when histochemical methods for localiz¬ tice-Hall, 1944.)
ing enzymatic activity became available, the
distinctions between fiber types could be
clearly demonstrated and debned more ac¬ merous, large, and provided with many cris-
curately. Staining for the enzyme succinic tae. Such bbers, also called slow fibers, are
dehydrogenase clearly identibes the smaller easily stimulated but have a slow conduction
red bbers because of their great abundance rate (50—80 m/sec) and are innervated by
of mitochondria. Differences in myoglobin small axons. On the other hand, white bbers,
concentration and in myohbrillar adenosine also called fast fibers, are larger in diameter;
triphosphatase and phosphorylase activity their Z lines are relatively narrow; the pattern
can also be demonstrated. of sarcoplasmic reticulum is simpler and the
The multiplicity of bber types recognizable mitochondria are sparse, with subsarcolem-
by these methods has led to some terminol¬ mal and interhbrillar accumulations gener¬
ogical confusion. In the interests of simplicity ally absent. They have a rapid conduction
and continuity with tradition, we adopt here rate (70-110 m/sec); are less easily stimu¬
the terms red, intermediate, and white bbers. lated; are innervated by larger axons; and
The red bbers are small in diameter, rich in have a less rich blood supply. There are also
myoglobin (Figs. 10-13, 10-14, 10—15,) and differences in the neuromuscular junctions.
have a richer blood supply. In electron mi¬ The variations in color of different muscles
crographs, the Z lines are thicker; probles of are a rehection of the differing proportion
sarcoplasmic reticulum are more complex in of the three bber types. The proportions are
the region of the H band; and the mitochon¬ usually fairly constant for a given muscle,
dria located in the periphery of the bbers and it was formerly thought that the mechan¬
and in rows between the myobbrils are nu- ical and metabolic properties of muscle bbers
 MUSCULAR TISSUE • 279

Figure 10-15 Sections of the longissimus muscle of rabbit (A), pig (B), and ox (C), reacted for myoglobin. The species
differences in Lhe number of distribution of myoglobin-positive cells is obvious. (Photomicrograph from Cassens R G
J. Histochem. Cytochem. 78:364, 1970.) 1

were immutable. It is now known that, under other cell types, but in muscle it is largely
appropriate conditions, a fast muscle fiber devoid of associated ribosomes and exhibits
can be changed into a slow one and vice a highly specialized repeating pattern of local
versa. If the innervations of a postural and a differentiations that bear a constant relation¬
locomotor muscle are cross-transplanted, ship to particular bands of the striated myofi¬
there is a gradual change in the mechanical brils. The tubules of the reticulum overlying
and chemical properties of the muscles. the A bands have a prevailing longitudinal
Forced exercise training can also increase orientation but anastomose freely in the re¬
capillary number, mitochondrial density, and gion of the H band (Fig. 10—17). At regular
activity of oxidative enzymes (Fig. 10-14). intervals along the length of the myofibrils,
the longitudinal sarcotubules are confluent
The Ultrastructure of the Sarcoplasm with transversely oriented channels of larger
caliber called terminal cisternae. Pairs of par¬
The common organelles observed in the allel terminal cisternae run transversely
sarcoplasm do not depart significantly in fine across the myofibrils in close apposition to a
structure from those in other cell types. The slender intermediate element, the transverse
small Golgi complex found near many of the tubule, commonly called the T tubule. These
nuclei does not appear especially active. The three associated transverse structures consti¬
mitochondria are abundant at the poles of tute the so-called triads of skeletal muscle
the nuclei and beneath the sarcolemma. In (Figs. 10—16, 10-17). In amphibian muscle,
addition, a considerable number are lodged the triads are found encircling each I band
in narrow clefts between the myofibrils. In at the level of the Z line (Fig. 10-17). In
keeping with the high energy requirements mammalian muscle there are two triads to
for muscle contraction, the mitochondria each sarcomere, situated at the junctions of
have very numerous, closely spaced cristae. each A band with the adjacent I bands. The
Their intimate association with the contractile lumen of the slender F tubule does not open
elements brings the source of chemical en¬ into the adjacent cisternae and, strictly speak¬
ergy (ATP) close to the sites of its utilization ing, is not a part of the sarcoplasmic reticu¬
in the myofibrils. lum. Its limiting membrane is continuous
An important organelle that cannot prof¬ with the sarcolemma and its lumen commu¬
itably be studied with the light microscope is nicates with the extracellular space at the cell
the sarcoplasmic reticulum, a continuous system surface. It is therefore to be regarded as a
of membrane-limited sarcotubules that extend slender tubular invagination of the sarco¬
throughout the sarcoplasm and form a close- lemma penetrating deep into the interior of
meshed canalicular network around each my¬ the muscle fiber. To emphasize their separate
ofibril (Figs. 10—16, 10—17). This organelle identity and to distinguish the T tubules from
corresponds to the endoplasmic reticulum of elements of the sarcoplasmic reticulum, they
280 • MUSCULAR TISSUE

Figure 10-16, A, Micrograph of a longitudinal section of skeletal muscle, passing tangential to a myofibril. Observe the
longitudinal tubules of the sarcoplasmic reticulum and two transversely oriented triads at the level of the A-l junctions.
Glycoqen particles are present among the sarcotubules. B, Longitudinal section of muscle that has been immersed in
peroxidase. The dense-reaction product of the peroxidase is present in the lumen of the tubules, demonstrating
continuity of the lumen with the extracellular space. (Micrograph courtesy of D. Friend.)

are referred to collectively as the T system of tile material visible with the light microscope
the muscle fiber. (Fig. 10-18C,D), are found in electron micro¬
The longitudinal tubules and terminal graphs to be composed of smaller units, the
cisternae of the sarcoplasmic reticulum are myofilaments (Fig. 10— 18T). These are of two
now known to regulate the concentration of kinds, differing in dimensions and chemical
calcium ions in the microenvironment of the composition. The cross-banded pattern of
myofibrils. The limiting membrane of the striated muscle reflects the arrangement of
reticulum possesses an active calcium trans¬ these two sets of submicroscopic filaments.
port mechanism, and calcium is stored within The thicker myosin filaments, 15 nm in di¬
this organelle. Neurally induced depolariza¬ ameter and 1.5 \xm long, are parallel and
tion of the sarcolemma is conducted into the about 45 nm apart. The parallel arrays of
muscle fiber by the T tubules. An ATP- myosin filaments are the principal constituent
dependent mechanism at the junctions of the of the A band and determine its length (Fig.
T tubules with the terminal cisternae of the 10— 18T). The filaments are slightly thicker
triads results in release of calcium from the in the middle and taper toward both ends.
sarcoplasmic reticulum, triggering contrac¬ They are held in register by slender cross
tion of the myofibrils. When depolarization connections that are aligned at the midpoint
of the sarcolemma by nerve impulses ends, of the A band, giving rise to the transverse
calcium is actively transported back into the density recognized as the M line. In cross
sarcoplasmic reticulum. The lowering of the sections at the level of the H band, the
calcium concentration around the myofibrils filaments are disposed in an extremely reg¬
brings about cessation of their contraction. ular array (Fig. 10-18G). The thinner actin
The Substructure of the Myofibrils. The filaments, 5 nm in diameter, extend about 1
myofibrils, the smallest units of the contrac¬ |xm in either direction from the Z line and
 MUSCULAR TISSUE * 281

Myofibrils

Sarcolemma

Triad of the
reticulum

Z line Transverse tubule

Sarcoplasmic
reticulum

A band

Mitochondrion

I band
Transverse tubule

Terminal
cisternae

Sarcotubules
Figure 10-17. Schematic drawing of the distribution of the sarcoplasmic reticulum around myofibrils of amphibian
skeletal muscle. The longitudinal sarcotubules are confluent with transverse terminal cisternae. A slender transverse T
tubule extending inward from the sarcolemma is flanked by two terminal cisternae to form the “triads” of the reticulum.
In amphibian muscle (depicted here) the triads are at the Z lines. In mammalian muscle, there are two to each
sarcomere, located at the A-l junctions. (Modified from Peachey, L. J. Cell Biol. 25:209, 1965; from McNutt S. and D.
W. Fawcett. J. Cell Biol. 42:46, 1969.)
282 • MUSCULAR TISSUE

Figure 10-18. Diagram of the organization of skeletal muscle from the gross to the molecular level. F, G, H, and / show
the arrangement of filaments in cross section at the levels indicated. (Drawing by Sylvia Colard Keene.)
 MUSCULAR TISSUE • 283

1 ■ '■!

Figure 10-19. Micrograph of several juxtanuclear myofibrils labeled to indicate the various bands in the normal pattern
of cross striations in relaxed skeletal muscle.

thus constitute the I band. They are not myosin filament (Figs. 10-18/, 10-22). The
limited to this band, however, but extend depth to which the ends of the actin filaments
some distance into the adjacent A band, penetrate into the A band varies with the
where they occupy the interstices between degree of contraction (Fig. 10-20). In the
the hexagonally packed thick filaments. relaxed condition, the thin filaments that
Thus, in cross sections near the ends of the extend into the A band from opposite ends
A band, the punctate profiles of six thin actin do not meet. The distance between their ends
filaments are evenly spaced around each determines the width of the FI band, which

Figure 10-20. Left, a schematic

representation of the changing
appearance of the cross striations
in different phases of contraction.
Right, the differing degrees of in-
terdigitation of the thick and thin
filaments corresponding to the dif¬ REST LENGTH
ferent patterns of striation.

H
284 • MUSCULAR TISSUE

and this narrow interval is traversed by reg¬

ularly spaced cross bridges that extend radi¬
ally from each myosin filament toward the
I Band neighboring actin filaments (Figs. 10—18,
10-21, 10-24).
The details of the interrelation of filaments
of successive sarcomeres at the Z disc are still
a subject of debate, but certain points seem
to be adequately established. Each actin fila¬
ment approaching the Z line appears to be
continuous with four diverging thin strands
called Z filaments. Each of these runs
obliquely through the Z disc to one of the
actin filaments on the other side. The actm
filaments approaching the Z line from op¬
posite sides are offset, so that in longitudinal
sections the cross-connecting Z filaments pro¬
duce a characteristic zigzag pattern. In addi¬
tion to the Z filaments, there appears to be a
dense amorphous component simply re¬
ferred to as “Z disc material” or “Z disc
A Band matrix.” This component varies in abun¬
dance in different skeletal muscles and is
more easily extractable than the filaments. Its
association with the Z disc seems quite selec¬
tive for when the matrix material is added
back to extracted glycerinated muscle, it ac¬
cumulates around the Z filaments and re¬
stores the Z band density. The exact chemical
nature of the Z filaments is not yet clear. In
addition to the complex of actin filaments,
the Z band contains the protein a-actinin,
which contributes to its electron density and
probably plays a role in binding the actin
filaments together.
At the level of the Z bands, each myofibril
is surrounded by a honeycomb-like network
of desmin filaments and vimentin filaments. It
I Band is believed that the networks of 10-nm fila¬
ments extending across the muscle fiber serve
to link adjacent myofibrils together and thus
maintain the lateral register of the sarco¬
meres.
Figure 10-21, Electron micrograph of a sarcomere of
Although no further detail can be observed
rabbit psoas muscle. In the I band, only thin actin filaments
are present. In the A band (central portion of figure), the in electron micrographs of thin sections of
thin filaments interdigitate with a set of thicker myosin muscle, the analysis of the contractile mate¬
filaments. Cross bridges between the myosin and actin rial has been carried further by mechanical
filaments recur at regular intervals in the region of overlap.
disintegration of myofibrils under conditions
(Micrograph courtesy of H. Huxley.)
that permit the release of the individual my¬
ofilaments. These have been studied with the
is defined as the central region of the A band electron microscope after metal shadowing
that is not penetrated by the actin filaments. and with negative staining procedures.
In stretched myofibrils the H band is there¬ The isolated thick filaments are 1.5 |Jim
fore broad, whereas in the contracted state it long. They have a smooth central region, but
is very narrow or entirely absent (Fig. 10—20). toward either end they bear short lateral
In the region of their interdigitation at the projections corresponding to the cross
ends of the A band, the parallel thick and bridges seen between the thick and thin fila¬
thin filaments are only 10 to 20 nm apart, ments in intact myofibrils. When further dis-
 MUSCULAR TISSUE • 285

Figure 10-22. Micrograph of a cross section through the A band of insect flight muscle at high magnification, showing
the orderly arrangement of actin filaments around the larger myosin filaments. The general pattern is similar in vertebrate
muscle but usually does not exhibit such a highly ordered “crystalline” lattice. (Micrograph courtesy of H. Ris.)

mmimt m < > immmt

rmmmm

mi mmmMmim

•>»¥**»Sf0Mb>

"* 1 i WJj**#

•Sfrm
<WM' miS ^ -,.•.■<• 3waw-'jj»

u'WiP t^

nm***!?

pgipg

«*•

^ .'•***-*y

arses
iwwiifii»Mw<pi(iyf)^j

rnmmmi
ufyw ipPIWiMpMi
&H* .. ., ‘w*

>*• &U.
i «fc. «£
Figure 10-23. Micrograph of the major portion of a sarcomere of insect flight muscle. Cross bridges between the thick
and thin filaments are barely detectable, but are more apparent if an area of mammalian muscle comparable with that
in the square is examined in replica after rapid freezing, cleaving, and deep-etching (see Fig. 10-24). (High-voltage
micrograph courtesy of H. Ris.)
286 * MUSCULAR TISSUE

Figure 10-24. Micrograph of a region of skeletal muscle (comparable with that indicated in the square in Fig. 10-23)
prepared by rapid freezing in liquid"helium, deep-etching, and rotary shadowing. The cross bridges between the myosin
and actin filaments are evident. (Micrograph courtesy of N. Hirokawa and J. Heuser.)

sociation of the filaments is carried out, binding property, the S-l fragment is useful
myosin molecules are obtained. These are in detecting actin filaments by binding to
rodlike structures about 200 nm long and 2 them in morphologically distinctive arrow¬
to 3 nm in diameter, consisting of two heli¬ head configuration.
cally entwined polypeptides each terminating Under appropriate physicochemical con¬
in a globular head that diverges laterally from ditions, myosin molecules will self-assemble
the backbone of the molecule (Fig. 10—25A). in vitro to reconstitute myosin filaments. In
The heads of the molecule form the cross so doing they become arranged parallel with
bridges of the myosin filament and are the the rod portions oriented toward the center
site of ATP binding and myosin-ATPase ac¬ of the nascent filament and heads toward the
tivity. Upon enzymatic proteolysis the myosin ends (Fig. 10-255). The overlapping mole¬
molecule is cleaved into two fragments—a cules are staggered so that the projecting
straight fragment of 150,000 M.W. repre¬ heads recur at intervals of 14.3 nm, and each
senting the greater part of the length of the pair is rotated 120 degrees with respect to its
molecule and called light meromyosin (LMM), neighbors to give them a spiral course along
and a shorter fragment called heavy mero¬ the filament. The bare central segment, cor¬
myosin (HMM), which includes the heads and responding to the H band, is a region con¬
a short portion of the rodlike backbone of sisting only of the overlapping antiparallel
the molecule. Upon further proteolysis, rod portions of the molecules (Fig. 10—255).
heavy meromyosin yields two subfrag¬ Isolated thin filaments are about 1 pm in
ments—HMM-S2 (60,000 M.W.), a terminal length and are identified biochemically as
portion of rod, and HMM S-l, consisting of filamentous actin (F-actin). At very high mag¬
the two heads that extend at an angle from nifications they have a beaded appearance,
the rod (Fig. 10—25A). Because of its actin- and have been shown to consist of globular
 MUSCULAR TISSUE • 287

Ac 11 n
Act in filament

Figure 10-25. Drawings of the molecular organization of the myofilaments and their interaction. A, Configuration of an
isolated myosin molecule and the fragments obtained when the molecule is cleaved by controlled proteolysis. B,
Schematic representation of the antiparallel arrangement of myosin molecules in the thick filament. The filament is
greatly foreshortened here for illustrative convenience. C, The double helical configuration of the actin filament and its
associated tropomyosin and troponin. (Redrawn after Junqueira, L. C., and J. Carniero. Basic Histology. Los Altos, CA,
Lange Medical Publishers, 1983.) D and E, Current interpretation of the mechanism of translocation of the actin filament.
Calcium binding causes a change in configuration of the troponin-tropomyosin complex, exposing the myosin binding
site on actin. The myosin heads energized by ATP then change their angle, moving the actin filament. (Modified after
Ganong, W. F. In Review of Medical Physiology, 11th ed. Los Altos, CA, Lange Medical Publishers, 1983.)
288 • MUSCULAR TISSUE

subunits 5.6 nm in diameter polymerized to responsible for the cross striations but also
form two strands entwined in a helix with led to an entirely new concept of the mech¬
each gyre about 36 nm in length (Fig. anism of contraction. Basic to the new theory
10-25C). The filaments can be dissociated was the observation that the length of the A
into the globular monomeric subunits (G- band remains constant during contraction,
actin). When reassembled in vitro, the mon¬ while the lengths of the H band and the I
omers are consistently oriented to give the band both decrease. A possible explanation
filament a definite polarity. In the intact for these changes in the pattern of cross¬
myofibril the actin filaments on either side of banding became apparent when the electron
the Z line have opposite polarity. microscope revealed two interdigitating sets
Associated with the double helix of actin is of filaments. According to the sliding filament
a filament of tropomyosin that courses along hypothesis, when a muscle contracts, the thick
the groove between the two entwined strands and thin filaments maintain the same length
of actin (Fig. 10-25C). The tropomyosin mol¬ but slide past each other, so that the ends of
ecule is about 40 nm in length and consists the actin filaments extend farther into the A
of two polypeptide chains in alpha helical band, narrowing and ultimately obliterating
configuration. The molecules are arranged the FI band. As a consquence of the deeper
end to end to form the tropomyosin filament. penetration of the & band by the I filaments,
At regular intervals of about 40 nm along the Z disc is drawn closer to the ends of the
the actin filament, a molecular complex called adjacent A bands, and there is an overall
troponin is bound to each molecule of tropo¬ shortening of the myofibril (Fig. 10—20).
myosin (Fig. 10-25C). It consists of three Because the heads of the myosin molecules
subunits: one (TnT) that attaches it to tro¬ forming the cross bridges are the only visible
pomyosin; another (TnC) that has a binding structures by which a force could be devel¬
site for calcium; and a third (Tnl) that inhib¬ oped between the thick and thin filaments, it
its interaction of actin and myosin. When the is assumed that this is their function.
calcium concentration of the sarcoplasm is Contraction may involve displacement of
low, the tropomyosin filament is so situated as much as 300 nm in each half of the
that it blocks the myosin binding site on the sarcomere, while displacement of the distal
G-actin monomers. When the calcium con¬ ends of the bridges from the perpendicular
centration rises, calcium is bound to the TnC is no more than 10 nm. Therefore, during a
subunit of troponin, resulting in a configu¬ single contraction hundreds of bridge-form¬
rational change in the complex that changes ing and bridge-breaking cycles must take
the position of tropomyosin, exposing the place to produce the observed displacement.
underlying binding site, permitting interac¬ For a time some doubt was cast upon the
tion of the myosin heads with the G-actin involvement of the bridges in translocation
subunits of the thin filament, and triggering of the actin filaments by the observation in
the movement of the cross bridges that re¬ electron micrographs that the interhbrillar
sults in displacement of the actin filaments distance in some phases of contraction ap¬
toward the center of the A band (Fig. peared to be greater than the length of the
10-25 D,E). bridging heads of the myosin molecules. This
paradox was resolved by the discovery of
Sliding Filament Mechanism of flexible regions at the base of the heads and
Contraction at the junction of the heavy meromyosin
subunit with the linear backbone of the mol¬
Although classical cytologists described the ecule (Fig. 10-255). These two flexible cou¬
changes in the relative lengths of the bands plings permit the heads to attach to the actin
during muscle shortening, these observations filaments over a range of different interhla-
suggested no satisfactory explanation of the ment distances and still preserve the same
contractile mechanism. The most common orientation relative to the actin. What part of
speculation envisioned a process of shorten¬ the molecule is the site of development of
ing due to reversible folding and cross-link¬ the force that results in filament movement
ing of long molecules. In the past two dec¬ remains uncertain. But since it is unlikely
ades, however, detailed analysis of the that it could reside in the flexible region of
submicroscopic organization of muscle by the molecule, it is speculated that it is in the
electron microscopy and x-ray diffraction has head, and it is postulated that there is a force
not only revealed the structural relationships generating active change in the effective angle
 MUSCULAR TISSUE • 289

of the attachment of the heads to the actin were located only over the I band. There
filament. appeared, therefore, to be a structural com¬
In the sliding filament mechanism of mus¬ ponent at the center of each I band that is
cle contraction, the conversion of chemical responsible for inward conduction.
energy to mechanical work seems to involve The discovery of the sarcoplasmic reticu¬
the following train of events. In the resting lum and the finding of transversely oriented
muscle, ATP binds to the ATPase sites on triads at the level of each Z line led to the
the heads of the myosin molecules (Fig. suggestion that these might be the submi-
10—25D). However, actin is required as a croscopic structures involved in the inward
cofactor in the breakdown of ATP to release spread of activation. The demonstration that
energy, and interaction of myosin with actin the membranes of the T tubules are contin¬
is prevented in the resting muscle by the uous with the sarcolemma and that their
troponin-tropomyosin blockade of the bind¬ lumen is open to the extracellular space pro¬
ing sites on the actin filament. Release of vided the necessary final link in the evidence
calcium into the surrounding sarcoplasm in for the participation of the T tubules in
response to a nerve impulse is followed by excitation-contraction coupling. In muscle fi¬
binding of calcium ions to the TnC subunit bers of amphibians the T tubules are found
of troponin (Fig. 10-25T). This results in a at the level of the Z disc, while in the skeletal
change in configuration of the complex that muscle of mammals they are found at the
moves the tropomyosin deeper into the boundary between the A and I bands. Thus,
groove in the actin helix, exposing the myosin in mammals there are two T tubules to each
binding sites on the actin filament. Binding sarcomere.
activates the myosin ATPase with release of
energy from ATP. Active bending or shape
The Myoneural Junction
change induced in the myosin heads displaces
the actin filaments a short distance toward The specialized junctional region at the
the center of the A bands. This permits termination of a motor nerve on skeletal
alignment of other ATPase-prepared myosin muscle fibers is called the motor end plate. It
heads with another set of actin subunits for is recognized with the light microscope as a
binding in a new cycle of bridge-making and slightly elevated plaque on the muscle fiber,
breaking. Contraction continues until cal¬ marked by a local accumulation of nuclei
cium ions are taken up and sequestered in (Fig. 10-26A,B). The nuclei are of at least
the sarcoplasmic reticulum. Troponin-tro¬ two morphologically distinguishable types.
pomyosin complexes again cover the binding The so-called “arborization nuclei” belong to
sites on the actin filaments, restoring the the terminal sheath cells (Schwann cells) as¬
resting state (Fig. 10-25Z)). sociated with the motor nerve endings. These
cells are collectively referred to as the teloglia.
Coupling of Excitation to Contraction The second category of nuclei, usually larger
and less intensely stained, were called “sole
The attention of physiologists has long nuclei” in the classical literature. These are
been focused upon the problem of explaining simply the nuclei of the underlying muscle
how the myofibrils throughout the muscle fiber that congregate in the region of the
fiber are activated simultaneously and almost myoneural junction. With special methods of
instantaneously after arrival of a nerve im¬ impregnation, it can be shown that the axon
pulse at the sarcolemma. These events take of the motor nerve, after losing its myelin
place too rapidly to be explained by inward sheath, forms a terminal arborization over
diffusion of an activating substance liberated the clustered nuclei of the end plate. The
from an excitable surface membrane. In a terminal branches of the axons occupy re¬
new approach to this problem, a microelec¬ cesses in the surface of the muscle fiber called
trode was applied to the surface of an isolated synaptic troughs or primary synaptic clefts. When
muscle fiber and it was found that a local selectively stained, the surface of the under¬
reduction in membrane potential was not lying muscle fiber is found to be highly dif¬
equally effective at all points on the surface. ferentiated, forming what appear to be
It resulted in inward spread of an impulse, evenly spaced, ribbon-like lamellae attached
leading to contraction only if the tip of the to the sarcolemma by their edges and pro¬
microelectrode was over certain sensitive jecting from the myoneural interface into the
spots. In frog muscle, these sensitive points underlying sarcoplasm. This specialization of
290 • MUSCULAR TISSUE

Figure 10-26. Schematic representation of the motor end plate. A, End plate as seen in histological sections in the
long axis of the muscle fiber; B, as seen in surface view with the light microscope; C, as seen in an electron micrograph
of an area such as that in the rectangle on A. (Modified after R. Couteaux.)

the muscle fiber surface is called the subneural similar to that investing the rest of the surface
apparatus. of the muscle fiber. The axoplasm in the
Electron microscopy has greatly clarified nerve terminals contains mitochondria and a
the relationships at the myoneural junction. very large number of small vesicles (40-60
The teloglial cells cover the outer surface of nm), identical to the synaptic vesicles seen at
the axon terminal but never penetrate into axodendritic synapses in the nervous system
the synaptic clefts. Here the nerve and mus¬ (Fig. 10—29). These vesicles are the sites of
cle are directly exposed to one another. The storage of the neurotransmitter acetylcholine.
so-called “lamellae” of the subneural appa¬ It is estimated that each vesicle may contain
ratus are found instead to be narrow secondary 10,000 molecules of acetylcholine. In neural
synaptic clefts formed by infolding of the sar- transmission the contents of the synaptic ves¬
colemma lining the primary synaptic trough icles are released by exocytosis. This takes
(Fig, 10—26C). These relationships are ele¬ place at specialized sites in the presynaptic
gantly displayed in scanning electron micro¬ membrane, referred to as active zones (Fig.
graphs after enzymatic digestion of obscuring 10—30). When myoneural junctions are stud¬
connective tissue components. The terminal ied by the freeze-fracture method the active
arborization of the axon in the motor end zones are seen as linear specialization of the
plate is clearly shown (Fig. 10-27), and if the membrane running parallel to the ridges and
nerve ending is pulled away the primary and furrows of the subneural specialization of the
secondary synaptic clefts in the underlying muscle fiber. The synaptic vesicles cluster in
muscle fiber are revealed (Fig. 10—28). the axoplasm along these active zones (Fig.
In transmission micrographs the axo- 10—30£), and in preparations of endings fro¬
lemma and the sarcolemma are separated at zen with liquid helium within milliseconds of
all points by a glycoprotein boundary layer nerve stimulation, discharging synaptic vesi-
 MUSCULAR TISSUE • 291

Desaki, J., and Y. Uehara, J. Neurocytol. 70:101, 1981.)

:Suppl. 139, 1981.)

 *■*
Synaptic cleft
Mitochondria

<* r.vs^jssjR Axon

. t ** *,*wfc

Schwann
cell

Synaptic

Figure 10-29. Electron micrograph of the nerve ending at the myoneural junction, showing an accumulation of
mitochondria and large number of synaptic vesicles in the axoplasm. (Micrograph courtesy of T. Reese.)

Figure 10-30. Freeze-fracture preparation of the presynaptic membrane at a frog myoneural junction, showing the
linear active site. A, The nerve was electrically stimulated and the junction frozen within milliseconds; exocytosis of
several synaptic vesicles can be seen along the active site (at arrows). B, Similar preparation of an unstimulated nerve
ending. The fracture plane has broken into the axoplasm showing synaptic vesicles (at arrows) clustered near the active
site. (Micrograph courtesy of T. Reese.)
292
 MUSCULAR TISSUE • 293

cles can be seen along these active zones (Fig.

10-30A).
The membrane covering the ridges and
clefts of the underlying muscle cell contains
a high concentration of acetylcholine receptors.
In preparations rapidly frozen, deep-etched,
and rotary-shadowed these can be seen as
closely packed intramembrane particles (Fig.
10-31). When acetylcholine is released into
the synaptic cleft, it combines with the recep¬
tors and results in a transient increase in the
permeability of the membrane to sodium
ions. Influx of sodium depolarizes the mem¬
brane, generating an action potential that is
propagated over the sarcolemma and into
the T tubules, activating the intracellular re¬
lease of calcium, which triggers contraction.
The subneural sarcoplasm is unremarkable
except for the abundance of its mitochondria.
Histochemical studies demonstrate cholines¬
terase activity in the subneural appaiatus of
the motor end plate. The major part of this
activity is specifically acetylcholinesterase, lo¬
calized in basal lamina lining the secondary
clefts. The acetylcholine released at the mo¬
tor end plate is rapidly broken down by
acetylcholine esterase in the synaptic cleft,
thereby limiting the duration of the lesponse
to the neurotransmitter.
A reduction in the number of available
acetylcholine receptors in myoneural junc¬
tions is the basic defect in the human disease
myasthenia gravis, which is characterized by
weakness and fatigability of skeletal muscle.
There are normally 30 to 40 million receptors
per neuromuscular junction. In myasthenic
muscle there is a 70 to 90% reduction in their
number.

Neuromuscular Spindles
Skeletal muscle contains complex sensory
organs called muscle spindles. These fusiform,
encapsulated structures consist of several
modified striated muscle fibers and their as¬
sociated nerve endings enclosed in a common
sheath. The specialized muscle fibers in the Figure 10-31. Synaptic ridges and clefts of the subneural
interior of the organ, referred to as intrafusal portion of a motor end plate prepared by rapid freezing,
fibers, number from a few to as many as 20, deep-etching, and rotary shadowing. The etching exposes
large intramembrane particles in the postsynaptic mem¬
but there are usually about six (Figs. 10—32,
brane that are believed to be the acetylcholine receptors.
10-33). The fibers are 1 to 5 mm long and (Micrograph courtesy of N. Hirokawa and J. Heuser.)
attached at their ends to tendon or endomy-
sium. Two distinct types of fibers are present,
the nuclear bag and nuclear chain fibers. Nu¬ tural organization into three regions. The
clear bag fibers are subdivided into a cential central portion is usually devoid of obvious
or equatorial segment and two long tapering cross striations and contains an accumulation
polar segments. The equatorial segment can se of 40 to 50 spherical nuclei, which completely
further subdivided on the basis of its struc-
294 • MUSCULAR TISSUE

Figure 10-32. A, Photomicrograph of a muscle spindle

in the lumbrical muscle of a human hand. The equator of
the spindle is seen with its laminated capsule and large
periaxial space. There are nine intrafusal muscle fibers;
three of these are nuclear bag fibers. The other six small
muscle fibers, lying in a group, are nuclear chain fibers.
A blood vessel and several nerves are also seen. Trans¬
verse section. Holmes’ silver method. B, Muscle spindle
in human extrinsic eye muscle. Seven of the muscle fibers
are surrounded by a thin capsule and there is a small
nerve trunk attached. The muscle spindles are usually
smaller than those in the limb muscles and have no
nuclear bag fibers in man. Transverse section. Hematox¬
ylin and eosin. (Both photomicrographs courtesy of S.
Cooper.)

PERCAPSULAR FIBERS

INTRACAPSULA r
FIBERS

BRANCHES TO
’ EXTRAFUSAL
FIBERS

LAMELLAR AND
FIBROUS
LAYERS OF
CAPSULE
INSERTION OF
INTRA.CAPSULAR
FIBERS
/

TERMINATION OF
CAPSULE

PERCAPSULAR
FIBERS

Figure 10-33. Drawing of a muscle spindle and its innervation. The capsule has been cut open to show the periaxial
space and the sensory endings of the primary afferent nerves around the central portion of the intrafusal fibers. Near
the end of the spindle, fusimotor nerves penetrate, to terminate on the intrafusal fibers in typical motor end plates.
(Drawing courtesy of C. F. Bridgman.)
 MUSCULAR TISSUE • 295

fill and often slightly distend the fiber. This

region is referred to as the nuclear bag. Ex¬
tending from it toward either pole is a my-
otube region, in which oval nuclei are aligned
in a row in an axial core of sarcoplasm
surrounded by a peripheral layer of cross-
striated myofibrils. In the slender polar seg¬
ments the nuclei are scattered at irregular
intervals along the axis of the fiber. In nu¬
clear chain fibers, the nuclei form a single
longitudinal row in the central region. The
capsule closely invests the poles of the intra¬
fusal fiber but diverges from its surface in
the equatorial segment to enclose the periaxial
space, a fluid filled cavity up to 200 pm in
diameter that surrounds the nuclear bag and
myotube regions. It has been reported that
this space is continuous with the lymphatic
system, but this does not seem to have been
confirmed, and the space is usually regarded
as a closed cavity.
Special kinds of nerve endings are associ¬
ated with each of the three regions of the
intrafusal fiber. The sensory endings sup¬
plied by large myelinated afferent fibers are
confined to the equatorial segment. The pii-
mary sensory or annulospiral ending is associ¬
ated mainly with the nuclear bag or nucleai
chain region but may extend into the adja¬
Figure 10-34. Primary nerve ending of a muscle spindle
cent myotube region. The endings consist of in a cat’s plantaris muscle. Two branches of an afferent
a complex system of half rings and spiials nerve fiber supply the ending, consisting of two large
around the fibers (Figs. 10—33, 10—34). The spirals around the muscle fibers at their nuclear bag
secondary or flower-spray endings are primarily regions. Teased gold chloride preparation. (Courtesy of
S. Cooper.)
confined to the nuclear chain fibers and lo¬
calized on either side of the annulospiial
endings. Three types of motor terminals aie
associated with the spindles: fine fusimotor, or Sensory Nerve Endings in Tendons
'Y-efferents form both motor end plates neat
There appears to be more than one kind
the poles of the fibers and unspecialized trail
of nerve ending in tendons. In the simplest,
endings near the equatorial region. At the
unmyelinated nerve fibers ramify over the
very end of the nuclear bag fibers, collaterals
surface of the collagen bundles. These may
of the motor axons for the extrafusal, slow
possibly give rise to pain sensation on exces¬
muscle fibers form “en grappe terminals, so
sive stretch. Of greater interest are the en¬
named because they resemble a bunch of
capsulated tendon organs. These are believed
grapes. It must be noted, however, that the
to sense tensional stresses produced by mus¬
details of innervation vary greatly from spe¬
cle pull, and to act as sources of inhibition
cies to species. when muscle contraction becomes excessive.
The spindles scattered through skeletal
From reconstructions of tendon organs from
muscles appear to function like miniature
serial cross sections, it appears that they are
strain gauges, sensing the degree of tension
composed of specialized, encapsulated fasci¬
in the muscle. I he motor innervation of the
cles of dense collagen that are subdivisions
polar regions of the intrafusal fibeis main¬
or branches of the primary tendon of origin
tains the nuclear bag region under sufficient
or insertion of a muscle. Within this encap¬
tension for its stretch receptor endings to be
sulated region, branches of the entering sen¬
close to their threshold. A further stretch of
sory nerve are entwined among fine bundles
the equatorial region results in dischai ge of
of collagen coursing through delicate septa
the spindle afferent fiber, and the frequency
that subdivide the main fascicle of collagen
of its discharge is proportional to the tension
into smaller subunits. Toward the muscle end
exerted on the intrafusal fiber.
 296 • MUSCULAR TISSUE

of the organ, these finer bundles reorganize (3) The elongated nuclei of the cellular units
again into thicker bundles before they inter- are usually situated deep in the interior of
digitate with the ends of the extrafusal mus¬ the fiber instead of immediately beneath the
cle fibers. It is speculated that the spaces sarcolemma (Fig. 10-36). The principal phys¬
between the fine collagen bundles in the iological differences between cardiac and
relaxed state of the muscle spread open skeletal muscle are the spontaneous nature
slightly, reducing pressure on the nerve end¬ of the beat of cardiac muscle and its rhythm¬
ings lying between them. During muscular ical contraction, which ordinarily is not sub¬
contraction, these bundles would straighten ject to voluntary control.
and be drawn together, compressing the
nerve endings. The compression of the nerve The Cytology of Cardiac Muscle
terminals would generate electrochemical
events in sensory axons, resulting in trans¬ The thin sarcolemma of cardiac muscle is
mission of tension information to the central similar to that of skeletal muscle, but the
nervous system. sarcoplasm is relatively more abundant and
the mitochondria are much more numerous.
The longitudinal striation of the fibers is
quite obvious with tjie light microscope, ow¬
CARDIAC MUSCLE ing to the subdivision of the contractile ma¬
terial by rows of mitochondria in the interfi-
The heart consists of striated muscle fibers brillar sarcoplasm. The pattern of cross
that differ in several respects from those of striation of the contractile material and the
skeletal muscle. (1) The fibers are not syncy¬ designation of the A, I, M, H, and Z bands
tial but are made up of separate cellular units is identical to that of skeletal muscle (Fig.
joined end to end by special surface special¬ 10-37). The myofibrils diverge around the
izations, intercalated discs, that run trans¬ centrally placed nucleus to outline a fusiform
versely across the fiber (Fig. 10-35). (2) The axial region of sarcoplasm rich in mitochon¬
fibers are not simple cylindrical units but they dria. Near one pole of the nucleus is a small
bifurcate and connect with adjacent fibers to Golgi complex. In the conical regions of
form a complex three-dimensional network. sarcoplasm extending in either direction

Figure 10 35. Drawing of longitudinal section of human cardiac muscle, stained in thiazin red and toluidine blue to
show the intercalated discs. (From H. Heidenhain.)
Figure 10-36. A, Longitudinal section of human left ventricular muscle, illustrating the variable diameter and branching
of the fibers, as well as the central position of the nuclei. In routine hematoxylin and eosin preparations, intercalated
discs are not evident. B, Cross section of human cardiac muscle.

:iaure 10-37 Electron micrograph of a portion of a cardiac muscle cell in longitudinal section. The cross-banded
attern of the contractile material is similar to that of skeletal muscle. The numerous mitochondria occupy clefts or
jsiform spaces that appear in longitudinal section to subdivide the myofilaments in units comparable with the discre e
nyofibrils of skeletal muscle. They are, however, much more variable in their width.
297
 298 • MUSCULAR TISSUE

from the poles of the nucleus there are often cohesion of the successive cellular units of
a few droplets of lipid and, in older animals, the myocardium and for transmitting the
deposits of lipofuscin pigment. In aged hu¬ tension of myofibrils along the axis of the
mans this pigment may come to constitute as fiber from one cellular unit to the next.
much as 20 per cent of the dry weight of the
myocardium. In small animal species, lipid
The Submicroscopic Structure of the
droplets are plentiful and occur in the inter-
Sarcoplasm
fibrillar sarcoplasm throughout the fiber,
often located between the ends of the mito¬ In electron micrographs, cardiac muscle
chondria. The sarcoplasm of cardiac muscle bears a superficial resemblance to skeletal
is richer in glycogen than is that of skeletal muscle. Its contractile substance is composed
muscle. of two sets of myofilaments, thick and thin,
At fairly regular intervals along the length in the same interdigitating relationship. In
of the fibers, the intercalated discs appear as longitudinal section, the tubules of the sar¬
heavy transverse lines. These are relatively coplasmic reticulum and rows of mitochon¬
inconspicuous in hematoxylin and eosin dria appear to subdivide the contractile ma¬
stains but are clearly revealed in iron-hema¬ terial into myofibrils of variable width (Fig.
toxylin or phosphotungstic acid-hematoxylin 10—37). However, upon close examination of
preparations. The disc may extend uninter¬ transverse sections (Figs. 10-38, 10-39), it
ruptedly across the full width of the fiber, becomes evident that the myofilaments are
but more often it is divided into segments not organized in discrete myofibrils as they
that are offset longitudinally so as to give the are in skeletal muscle. In cross sections the
disc a steplike configuration. In the repeating circular profiles of the sarcotubules are
pattern of cross striations, the intercalated aligned in rows that partially demarcate po¬
discs invariably occur at the level of the I lygonal or irregular areas of myofilaments,
bands. They were formerly interpreted as but these are usually confluent with adjacent
local contraction bands or specializations for areas of myofilaments over some fraction of
intracellular conduction, but they are now their perimeter. Mitochondria often appear
known to be devices for maintaining firm completely surrounded by myofilaments (Fig.

tFh»U=,ri! ,T38' A'°w-P°wer e'ectran micrograph of portions of several cardiac muscle fibers in cross section illustration
to the captoes crp\pi!!a70m7sclethe '°Ca,i0n °' ^ "UCleUS' and ,he in,ima,e relation of the ™scle ^
 MUSCULAR TISSUE • 299

Figure 10-39. Electron micrograph of a small peripheral area of a cardiac muscle fiber in cross section. The myofilaments
are not associated in discrete myofibrils with clearly defined limits, but instead form a more or less continuous field
interrupted by mitochondria.

10—39). Thus, the contractile substance of but may occasionally be 7 or 8 pm long.

cardiac muscle forms a continuum that can Spherical lipid droplets are often located be¬
be thought of as a large cylindrical mass of tween the ends of the mitochondria. Glyco¬
parallel myofilaments incompletely subdi¬ gen occurs in the form of 30 to 40-nm dense
vided into irregular fascicles by deep inci¬ granules crowded into the interstices among
sures and by fusiform or lenticular clefts of the mitochondria. The bulk of the glycogen
sarcoplasm that are occupied by mitochon¬ is located in the interhbrillar sarcoplasm, but
dria and the other organelles essential to the particles may also be found aligned in rows
contractile mechanism. between the myofilaments (Fig. 10-41$).
The continuous nature of the contractile They are particularly numerous in the I band
mass is not peculiar to cardiac muscle but is and occur more sparsely in the H band. Both
also found in certain relatively slow, tonic the glycogen and the lipid may be used as
skeletal muscles, particularly in amphibia. energy sources for the contractile activity of
The German term Feldstruktur has been the myocardium.
adopted to describe this pattern of organi¬
zation of the myofilaments, and the term The T System and the Sarcoplasmic
Fibrillenstruktur is applied to the pattern of Reticulum
separate myofibrils that is typical of fast,
twitch muscles. The tubular invaginations of the sarco-
The large mitochondria of cardiac muscle lernrna that comprise the T system of cardiac
have very numerous, closely spaced cristae muscle are larger than the corresponding
that often exhibit a periodic angulation of intermediate elements of the triads in skeletal
their membranes, giving them a zigzag con¬ muscle. These tubules, representing inward
figuration. As a rule, the mitochondria are extensions of the extracellular space, are lo¬
about the length of one sarcomere (2.5 pm) cated at the level of the Z lines instead of at
300 * MUSCULAR TISSUE

T-tubule
Sarcoplasmic (sarcolemmal
reticulum Mitochondrion invagination)
Sarcolemma Transverse tubule

Contact of reticulum
with T-tubule

Contact of Mitochondrion T-tubule

reticulum with
T-tubules

Figure 10-40. Schematic representation of the disposition of the T system and sarcoplasmic reticulum in mammalian
cardiac muscle. The transverse tubules are much larger than those of skeletal muscle. The relatively simple sarcoplasmic
reticulum has no terminal cisternae and therefore there are no triads. Instead, small expansions to its tubules end in
close apposition to the sarcolemma, either at the surface of the fiber or at its inward extension in the T tubules. (From
Fawcett, D. W., and N. S. McNutt. J. Cell Biol. 42:1, 1969.)

the A-1 junction and penetrate to the center Similar contacts are made between small flat¬
of the muscle fiber. They are lined by a tened expansions of the reticulum and the
glycoprotein layer continuous with the exter¬ sarcolemma at the outer surface of the fiber.
nal lamina that coats the sarcolemma (Fig. The total surface area of the many small sites
10-42). Apparently no point in a cardiac of apposition of the reticulum to the sarco¬
muscle fiber is more than 2 to 3 pm from lemma of cardiac muscle is quite great, but
the extracellular space, either at the outer would seem to be considerably smaller than
surface of the fiber or in one of the transverse the area of contact between the terminal
tubules. In addition to playing a role in the cisternae and intermediate elements of the
coupling of excitation to contraction, these triads in skeletal muscle. It is noteworthy,
channels no doubt provide important addi¬ too, that the T tubules in cardiac muscle
tional surface for the exchange of metabolites occur only over the Z lines at the ends of the
between cardiac muscle and the extracellular sarcomeres, whereas in mammalian skeletal
space. muscle there are two triads located at the A-
The sarcoplasmic reticulum is not as highly I junctions of each sarcomere. The functional
developed as in skeletal muscle. It consists of significance of this difference in location of
a simple plexiform arrangement of tubular the T system is not fully understood.
elements occupying slender clefts within the
mass of myofilaments (Fig. 10-4IT). There
The Intercalated Disc
are no continuous transverse elements of the
reticulum comparable with the terminal cis¬ On the transverse portions of the interca¬
ternae of the triads in skeletal muscle. In¬ lated discs, the opposing ends of the cardiac
stead, small terminal expansions of the retic¬ muscle cells have a deeply sculptured surface.
ulum here and there are closely applied to A complex pattern of ridges and papillary
the membrane of the T tubes (Fig. 10-42). projections on each cell fit into corresponding
 MUSCULAR TISSUE • 301

Figure 10—41. Longitudinal sections of cardiac muscle. A, The section passes tangentially to an internal surface of the
mass of myofilaments and reveals the sarcoplasmic reticulum forming a loose network that continues across the level
of the Z lines without any terminal cisternae. B, Glycogen particles are seen around mitochondria and between filaments
in the I and H bands. The relaxed muscle in these two figures is stretched to different degree. Notice the constancy in
length of the I bands, indicated by brackets at right. (From Fawcett, D. W. and N. S. McNutt. J. Cell Biol. 42:1, 1969.)

grooves and pits in the other cell to form an disc, the opposing cell surfaces are specialized
elaborately interdigitated junction (Figs. as fasciae adherentes, with a mat of dense
10-43, 10-44). The entire junctional surface material occurring immediately subjacent to
of both cells is specialized in various ways for an otherwise unspecialized membrane. The
maintaining cell-to-cell cohesion, and one can thin filaments of the adjacent I bands termi¬
distinguish areas that are similar in their fine nate in this mat of dense material, which
structure to the desmosomes and the zonula evidently serves to bind the ends of myofila¬
adherens of epithelial junctions. However, in ments to the plasmalemma. At desmosomes
the mosaic of different types of surface spe¬ along the transverse portions of the disc, the
cialization that constitute the intercalated inner leaf of each of the opposing unit mem¬
disc, only the desmosomes are typical with branes is thickened and especially dense. A
respect to their shape. small condensation of sarcoplasm may be
The fine structural counterpart of the zon¬ associated with this dense plaque, but the
ula adherens actually is not beltlike, as is myofilaments usually diverge and do not ter¬
implied by the term zonula. Instead there are minate directly on desmosomes (Fig. 10-44).
multiple, moderately extensive but discontin¬ At irregular intervals along the disc, the
uous areas having irregular and variable out¬ opposing membranes approach to within 2
lines. A descriptive term that has been sug¬ nm of one another and run parallel for a
gested for these is fascia adherens. short distance. These sites of closer apposi¬
In longitudinal sections, the opposing cell tion were formerly interpreted as “tight junc¬
membranes at the intercalated disc can be tions” and were termed maculae occludentes.
identified as two parallel dense lines that Further study of these by the freeze-fracture
follow a sinuous course separated, for the method has revealed that these correspond
most part, by a 15 to 20-mm intercellular instead to the gap junctions of epithelia.
cleft. Over the greater part of the intercalated There is no fibrous layer or condensation of
 Figure 10-42. Longitudinal section of a small area of a
cardiac muscle fiber, illustrating a T tubule cut transversely
and a tubule of the reticulum in close apposition to it. The
T tubule is lined with a layer of protein-polysaccharide (at
arrows) like that coating the sarcolemma at the surface
of the fiber. The dense granules in the neighboring sar-
coplasma are glycogen.

Figure 10-43. Low-power electron micrograph of cardiac muscle in longitudinal section, showing a typical steplike
intercalated disc. The transverse portions are highly interdigitated and characterized by an abundance of dense material
at the insertions of the myofilaments into the end of the cell. The longitudinal portions of the cell boundary are smooth,
unspecialized, and difficult to see at this magnification.
302
 MUSCULAR TISSUE • 303

Figure 10-44. Electron micrograph of a transverse segment of an intercalated disc. The portion of the cell junction in
which the myofilaments terminate resembles the zonulae adherentes of epithelia, but is here called a fascia adherens.
Between sites of myofibril attachment are typical desmosomes. (From Fawcett, D. W., and N. S. McNutt. J. Cell Biol.
42:1, 1969.)

the adjacent sarcoplasm associated with these incomplete, but it is known that calcium ions
regions of close apposition. play an important role. If the beating, iso¬
In addition to the small gap junctions that lated heart of an experimental animal is per¬
occur here and there in the transverse por¬ fused for some time with a calcium-free me¬
tions of the intercalated discs, there are more dium, the heart will soon cease beating. If it
extensive areas of close membrane apposition is then fixed and examined in thin sections,
on the longitudinal portions of the steplike the individual cells of the muscle fibers are
cell-to-cell junctions, where overlapping found to have come apart at the intercalated
processes of successive cells are joined side discs. At high magnification it can be ascer¬
to side. Considerable emphasis is placed upon tained that the membranes, for the most part,
these gap junctions because they are areas of are intact but separation has taken place by
low electrical resistance that permit the rapid opening up of the intercellular space. How¬
spread of excitation from cell to cell through¬ ever, at the gap junctions where the mem¬
out the heart. Thus, they enable the myocar¬ branes are in intimate apposition, the two
dium to behave as though it were a syncy¬ elements are unable to separate in calcium-
tium. The other specializations of the free medium, and one or the other of the
transverse portions of the intercalated discs, cells is denuded of its membrane when the
where the surfaces do not have hexagonal ends of the cells are pulled apart in the
arrays of intramembranous particles and are agonal contractions of the muscle.
not in such close apposition, evidently have
a mechanical significance, being mainly con¬ Cytological Differences Between
cerned with maintaining cell-to-cell cohesion Atrial and Ventricular Cardiac Muscle
and transmitting the pull of one contractile
unit to the next along the axis of the muscle The fibers of atrial myocardium are basi¬
fibers. cally similar to those of the ventricle de¬
Our understanding of the nature of the scribed above, but they have a smaller aver¬
forces that bind cells together is still very age diameter and the T tubules are few or
304 • MUSCULAR TISSUE

absent. When found, they tend to be in the findings, the right atrium must now be con¬
larger fibers. It is possible that the smaller sidered an endocrine organ in addition to its
fiber diameter may make unnecessary a well- contractile and pacemaker functions, and
developed system of transverse tubules for may have a hitherto unsuspected role in the
inward conduction of the impulse to contract. regulation of blood pressure and fluid and
Despite their smaller diameter, conduction electrolyte balance. Its “myoendocrine” cells
of the action potential over the surface of clearly possess certain morphological and
atrial muscle fibers is said to be more rapid physiological properties in common with the
than over ventricular fibers. peptide-secreting cells of the gastrointestinal
Another noteworthy difference is the pres¬ and respiratory tracts.
ence of specific atrial granules in these cells. Cardiodilatin is a polypeptide of 126 amino
These are membrane-bounded spherical acids and has a M.W. of about 7500. Car¬
granules 0.3 to 0.4 pm in diameter ^with a dionatrin is a peptide of 28 amino acids
dense homogeneous interior (Fig. 10—45). having a sequence similar to a segment of
They are concentrated in the core of sarco¬ the cardiodilatin molecule. Other small pep¬
plasm extending in either direction from the tides have also been found in extracts of
poles of the nucleus, usually near the Golgi atrium. It is not yet clear whether the myoen¬
complex. They have the appearance of secre¬ docrine cells synthesize a large prohormone
tory granules but until recently their func¬ that is enzymatically cleaved to form multiple
tional significance was a mystery. Two poly¬ peptide hormones that are stored in the gran¬
peptide hormones have now been extracted ules, or whether the smaller peptides are
from atrial muscle: one, called cardionatrin, fragments produced by the isolation proce¬
has potent diuretic and natriuretic effects, dure.
and the other, cardiodilatin, acts upon vascular
smooth muscle, causing relaxation and vaso¬
Specialized Conducting Tissue
dilation. These polypeptides have been local¬
of the Heart
ized immunohistochemically in the granule-
containing cells, which are most abundant in In addition to those cells of the myocar¬
the right atrium but also occur in the left dium whose primary function is contraction,
atrium. In the light of these unexpected there is a specialized system made up of

Golgi
complex

Atrial Ij
granules

Figure 10-45. Electron micrograph of the juxtanuclear area of a cardiac muscle cell from cat atrium, showing dense
spherical atrial granules in the Golgi region. These are now known to contain peptide hormones involved in control of
sodium excretion and blood pressure. (From McNutt, N. S., and D. W. Fawcett. J. Cell Biol. 42:45, 1969.)
 MUSCULAR TISSUE • 305

modified muscle cells whose function is to establish the rate of beating of the rest of the
generate the stimulus for the heart beat and myocardium. In warm-blooded animals, the
to conduct the impulse to the various parts fibers of the sinoatrial node have the most
of the myocardium in such a way as to ensure rapid rhythm, and this node is therefore
the contraction of the atria and ventricles in referred to as the “pacemaker” of the heart.
the proper succession, so that the heart acts The evidence for this resides in the fact that
as an effective pump. This system consists of the electrical events associated with each beat
the sinoatrial node (node of Keith and Flack), begin at the sinoatrial node and travel from
the atrioventricular node (node of Tawara), and there over the atria. Warming or cooling the
the atrioventricular bundle (bundle of His). The node increases or decreases, respectively, the
sinoatrial node is located beneath the epicar- rate of the heart beat. Although the heart
dium at the junction of the superior vena will normally beat at a rate determined by
cava and the right atrium. The atrioventric¬ the inherent rhythm of its pacemaker, this
ular node is found beneath the endocardium rate can be modified by the autonomic ner¬
in the lower part of the interatrial septum vous system. Parasympathetic (vagal nerve)
between the attachment of the septal leaf of stimulation brings about a slowing of the
the tricuspid valve and the opening of the heart and sympathetic stimulation accelerates
coronary sinus. The atrioventricular bundle it.
originates from the anterior portion of the The fibers of the atrioventricular node are
node and enters the fibrous portion of the small, like those of the sinoatrial node. The
interventricular septum, where it soon di¬ fibers of the atrioventricular bundle are sim¬
vides into right and left bundles that are ilar at their origin, but more distally in the
ultimately distributed to the right and left right and left bundle branches they become
ventricles. Each of these bundles ramifies larger than ordinary cardiac muscle fibers
beneath the endocardium of its respective and take on a highly distinctive appearance.
chamber to form an extensive plexus, from These are the so-called Purkinje fibers (Figs.
which fine fibers penetrate the myocardium 10-46, 10—47). They have one or two nuclei
to come into intimate contact with the ordi¬ situated in a clear central mass of sarcoplasm
nary contractile fibers. that is rich in mitochondria and glycogen.
The specialized cells of the nodal tissue are The myofibrils are relatively sparse and dis¬
distinctly smaller than ordinary cardiac mus¬ placed to the periphery, and they are less
cle fibers and are arranged in a network consistent in their orientation than are those
embedded in an abundant and rather dense of ordinary cardiac muscle fibers.
connective tissue. In sections, the slender The Purkinje fibers of ungulates reach very
fusiform nodal cells coursing in various di¬ large size, and for this reason these have
rections among the collagen bundles may be been more extensively studied than those of
difficult to distinguish from the associated other mammals, but they do not seem to
fibroblasts, but careful examination reveals differ in any other important respect. Typical
their cross striations. In the mammal no con¬ intercalated discs are seldom seen in the
nection between the sinoatrial node and the conducting tissue. At their ends the Purkinje
atrioventricular node via specialized conduc¬ fibers are said to lose their specific cytological
tion tissue has yet been convincingly dem¬ features and to become continuous with the
onstrated. The nodal fibers appear to be ordinary muscle fibers of the myocardium.
continuous with ordinary atrial muscle fibers. In electron micrographs, the cytoplasmic
The node is richly innervated by both the matrix of the Purkinje cells is of relatively
sympathetic and parasympathetic divisions of low density and contains numerous randomly
the autonomic nervous system. In addition oriented mitochondria (Figs. 10—48, 10—49).
there are numerous nerve fibers closely as¬ The myofilaments do not form a continuous
sociated with the sinoatrial and atrioventric¬ contractile mass but are arranged in separate
ular nodes that are immunoreactive for the myofibrils that are relatively few in number.
peptides neurotensin and substance-P. The rate Although their prevailing orientation is par¬
and forcefulness of contraction of the heart allel to the long axis of the cell, they are very
are affected by these substances as well as by poorly ordered compared with those of or¬
the usual transmitters of the sympathetic and dinary cardiac muscle.
parasympathetic innervation. The cells have variable and unusual shapes,
In cardiac muscle, which beats rhythmically one often partially surrounding another or
without nervous or other external stimuli, sending a large process into a deep recess in
the cells with the most rapid inherent rhythm the adjoining cell (Fig. 10—49). As a conse-
306 • MUSCULAR TISSUE

Figure 10-46. Photomicrographs of the very large Purkinje fibers in the moderator band of the bovine heart. In A the
fibers are cut longitudinally; in B they are cut transversely. In both, it is evident that the myofibrils occupy only a small
part of the sarcoplasm. The large clear areas are rich in glycogen.

Figure 10-47. Photomicrograph of the specialized con¬

duction tissue of the human atrioventricular bundle. The
large Purkinje fibers seen in cross section at left of figure
can be compared with the smaller unspecialized heart
muscle cut longitudinally at right side of figure.
 MUSCULAR TISSUE 307

Figure 10-48. Electron micrograph of adjacent areas of two Purkinje fibers and an accompanying nerve in the
atrioventricular bundle of the cat heart. The mitochondria are abundant and pleomorphic, and the loosely organized
myofilaments occur only in scattered bundles.

Figure 10-49. Electron micrograph of the cell junctions in the atrioventricular bundle. The cells of the conduction tissue
are irregular in shape and have an extensive area of cell-to-cell apposition, on which aie numerous desmosomes and
nexuses. (Courtesy of D. Feldman.)
308 • MUSCULAR TISSUE

quence of their irregular shape, the cells are Slender axons merely pass near the surface
in extensive contact with one another. Al¬ of the cardiac muscle cells. That these are
though no typical intercalated discs are functional endings and not merely passing
found, numerous desmosomes are distrib¬ axons may be inferred from the fact that
uted at irregular intervals along the cell their axoplasm often contains large numbers
boundaries. There are also areas of close of small vesicles identical to those found at
membrane apposition corresponding to gap other nerve endings and at synapses in the
junctions of ordinary cardiac muscle. Sur¬ central nervous system.
prisingly, these do not appear to be as nu¬
merous or as extensive as in the unspecialized
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 —__

THE NER VO US TISSUE

Jay B. Angevine

The nervous system comprises the entire The sensory, integrative, and motor func¬
mass of nervous tissue in the body. The tions of nerve cells depend mainly upon
essential function of nervous tissue is com¬ irritability and conductivity. In addition,
munication, which depends upon special sig¬ however, some nerve cells possess secretory
naling properties of the nerve cells and their capabilities similar to those of the endocrine
long processes. These properties express two system, which carries out its integrative func¬
fundamental attributes of protoplasm: the tion by means of blood-borne hormones. The
capacity to react to various physical and secretory products of such nerve cells are
chemical agents (irritability) and the ability to released from axon terminals into a perivas¬
transmit the resulting excitation from one cular space, transported somehow into the
locality to another (conductivity). lumen of the vessel, and carried by the blood
Upon reception of a stimulus from the to the particular target organ. A neurosecre¬
external or internal environment, various tory system regulating the activity of the
forms of energy are transduced to electrical adenohypophysis has been thoroughly stud¬
energy by specialized cellular structures, the ied and in recent years great attention has
receptors. Patterns of electrical messages, or been devoted to functional interaction be¬
nerve impulses, are transmitted from recep¬ tween the nervous and endocrine systems.
tors to nervous centers, where they evoke in In the evolution of the nervous systems of
other nerve cells additional patterns of sig¬ higher organisms, it is believed that certain
nals that result in appropriate sensations or cells of primitive Metazoa developed to a high
responses. By these means, the organism degree the properties of irritability and con¬
reacts to the events in the world in which it ductivity, and as a result of their more effi¬
lives and coordinates the functions of its cient response and signaling gradually
organs. In addition, the nervous system pro¬ evolved into a rudimentary nervous system.
vides the structural and chemical basis of By further specialization, some nerve cells
conscious experience. It provides mecha¬ developed the capacity to react to special
nisms for behavior and the regulation of kinds of exogenous stimuli. These cells, with
behavior and for maintenance of the unity the corresponding accessory structures dis¬
of the personality. tributed throughout the body or near its
The central nervous system (CNS) consists of surface, gave rise to three systems of sensory
the brain and spinal cord and contains the receptors: the exteroceptive system, receiving
nerve cells or neurons and a variety of suppor¬ stimuli from the body surface; the interocep¬
tive cells, called collectively the neuroglia. tive system, receiving stimuli from the internal
Nerve impulses conveyed from all parts of organs; and the proprioceptive system, receiving
the body over the long processes of the nerve stimuli from the muscles, tendons, and joints.
cells, called axons, come together in the CNS. Other nerve cells became connected with
The peripheral nervous system comprises all the peripheral effector organs, principally the
nervous tissue outside of the brain and spinal muscles, forming neuromotor systems. Still
cord and functions to keep the other tissues other nerve cells, collected into a large central
of the body in communication with the CNS. mass, assumed the tasks of correlation and
The function of all parts of the organism are integration. These cells receive and modify
thus integrated by a central “clearing house” the impulses arising from the receptors and
that controls the activity of the individual as in turn appropriately influence the effector
a whole. organs.
311
 312 • THE NERVOUS TISSUE

The cells of the nervous system primarily nervous system. The neuron doctrine implies
involved in its special function are the neu¬ that the nervous system is entirely cellular;
rons. Each neuron has a cell body consisting that its cells are distinctive in structure and
of a nucleus and the surrounding cytoplasm, function; and that its cells are not in proto¬
which is called the perikaryon. Typically the plasmic continuity but are juxtaposed without
cytoplasm is drawn out into several short a significant amount of intervening extracel¬
radiating processes called dendrites and into a lular substance. Observations with the elec¬
single long process called the axon (Fig. tron microscope corroborate these basic as¬
11—1). The axon, which may attain great sumptions of the doctrine and show that the
length, often emits branches or axon collaterals nervous system is a highly specialized epithe¬
along its course and at its end may exhibit lium. The nervous system thus reflects its
additional fine ramifications. phylogenetic and ontogenetic origin from the
The size, shape, and other peculiarities of ectodermal epithelial layer of the body. Tike
the nerve cell body and the number and other epithelia, nervous tissue exhibits junc¬
mode of branching of its processes are all tional complexes, local specializations of the
subject to variation, which results in many surfaces of adjacent cells that serve to main¬
morphologically distinguishable kinds of tain the position of the nerve cells and to
nerve cells (Fig. 11—2). Functional speciali¬ stabilize those spatihl relations of their proc¬
zations correlate with the morphological di¬ esses that are essential to the signaling func¬
versity. The neurons are anatomically and tion of the nervous system.
functionally related by their processes to
other nerve cells, or to epithelial, muscular,
or glandular cells. At synapses, specialized sites
of contact between neurons, chemical or elec¬ THE NEURON
trical signals pass from cell to cell. Transmis¬
sion is usually in one direction but mixed The nerve cell, or neuron, is usually large
modes and reciprocal synapses do exist. The and complex in shape. The volume of cyto¬
countless neurons are morphologically and plasm in its processes is usually greater—
trophically independent but functionally in¬ often much greater—than in its perikaryon.
terrelated at synapses. This fundamental The nerve cell body in the CNS generally
generalization is the neuron doctrine, essen¬ has several processes. The outline of the
tially a restatement of the cell theory for the perikaryon is typically angular or polygonal,

Effector portion Conductile portion

Figure 1-I--I. Diagrammatic representation of the effector, conductile, and receptor portions of a typical large neuron.
he effector endings on skeletal muscle identify this as a somatic motor neuron. The effector endings of many neurons
may terminate on the receptor portions of a single neuron. The myelin sheath on the conductile portion of the neuron
ac s as insulation and serves to increase its conduction velocity. The discontinuity in the axon indicates that it is much
Tex t bookof °H i s to bg y* Sj ?t f? edS)°m etl m eS a motor neuron t0 a limb is 2 or 3 feet long. (Drawing after Bunge in Bailey’s
 THE NERVOUS TISSUE • 313

Figure 11-2. Drawing of some characteristic types of neurons whose axons (a) and dendrites remain within the central
nervous system, illustrating some of the remarkable diversity of cell form exhibited by neurons. A, Neuron of inferior
olivary nucleus. B, Granule cell of cerebellar cortex. C, Small cell of the reticular formation. D, Small gelatinosa cell of
the spinal trigeminal nucleus. E, Ovoid cell, nucleus of tractus solitarius. F, Large cell of reticular formation. G, Spindle-
shaped cell, substantia gelatinosa of spinal cord. H, Large cell of spinal trigeminal nucleus. /, Neuron, putamen of
lenticular nucleus. J, Double pyramidal cell, Ammon’s horn of hippocampal cortex. K, Cell from thalamic nucleus. L, Cell
from globus pallidus of lenticular nucleus. Golgi preparations, monkey brain. (Courtesy of Clement Fox, from Truex, R.
C., and M. B. Carpenter. Human Neuroanatomy. 6th ed. Baltimore, Williams and Wilkins, 1969.)
 314 • THE NERVOUS TISSUE

the surfaces between the processes being prominent and is located either near the
slightly concave (Figs. 11-3, 11-4). Motor nucleolus or at the periphery of the nucleus.
neurons in general and the pyramidal cells Although neurons usually contain only one
of the cerebral cortex (Fig. 11—5) are two of nucleus, binucleate cells are sometimes en¬
many examples of angular nerve cell bodies. countered in autonomic ganglia. In electron
Cell bodies in the dorsal root ganglia, on the micrographs, the nuclear envelope and its
other hand, are rounded, and only one proc¬ pores and the fine structure of the nucleolus
ess projects from the perikaryon (Fig. 11-6). and karyoplasm (Fig. 11-8) are not signifi¬
Regardless of shape, the neuron has a num¬ cantly different from the corresponding fea¬
ber of distinctive cytological characteristics. tures in other cells.

Nucleus Perikaryon
The nucleus is large, pale, spherical, or The cytoplasm of the nerve cell is crowded
slightly ovoid, and usually centrally placed with filamentous, membranous, and granular
within the perikaryon. In most cases there is organelles arranged more or less concentri¬
a single conspicuous nucleolus as well as very cally around the nucleus. These organelles
fine chromatin particles (Fig. 11-4). Because include the neuronal cytoskeleton, Nissl bod¬
of uniform dispersion of the chromatin, the ies, Golgi apparatus, mitochondria, cen-
nuclei of nerve cells, stained with basic dyes, trioles, and various inclusions.
appear empty and pale and are often de¬ Cytoskeleton. At the electron microscope,
scribed as “vesicular.” In smaller nerve cells, the chief components of the neuronal cyto¬
the concentration of chromatin may be skeleton are neurofilaments and microtu¬
greater and the vesicular character of the bules (Fig. 11—12). Neurofilaments are about
nucleus less obvious. In man, but not in all 10 nm in diameter and of indefinite length.
mammals, the sex chromatin of females is LJnlike other cytoplasmic filaments of com-

_JL tn hott ’ ISS?e cu.*ures of the nervous system, the three-dimensional configuration of the intact neuron can I
fn „ !° better a,,Van • han In sectl0ns- Shown here are multipolar neurons from the deep nuclei of the rat cerebellu
69455^"?966C)U UrG ^ neuroflbnls In the cel1 bodies- Holmes stain. (From Hild, W.: Zeitschr. f. Zellforsc
 THE NERVOUS TISSUE • 315

Figure 11-4. A, Two areas of section through the optic tectum of a leopard frog, showing blue-stained myelin sheaths
and the nerve cell bodies. The small dark nuclei are supporting cells. B, Section from pons of man, showing myelin
sheaths, nerve cell bodies, and glial cells. A, From a frozen section fixed in formalin; B, Paraffin section after postmortem
formalin fixation. Kluver and Barrera staining methods for cells and myelin sheaths. (Drawn by Esther Bohlman.)
 316 • THE NERVOUS TISSUE

Figure 11-5. Drawings of two principal cell types in cerebellar and cerebral cortex. Dendritic branches may provide a
very extensive area for attachment of synaptic terminals of many other cortical and subcortical neurons. Golgi
preparations, monkey brain. (Courtesy of Clement Fox, from Truex, R. C., and M. B. Carpenter. Human Neuroanatomy.
5th ed. Baltimore, Williams and Wilkins, 1966.)

parable dimensions, they appear at high mag¬ are different from the constituents of glial
nification as minute tubules with a dense wall filaments and intermediate filaments of other
3 nm thick and a clear center. They consist tissues. There is evidence that the 70,000-
of a triplet of proteins of molecular weights dalton protein forms the core of the neuro-
200,000, 150,000 and 70,000 daltons, which hlament, and the other two proteins are hel¬
ically wound around this core. Microtubules
(neurotubules) are long, straight protein
tubes with an outer diameter of about 25 nm
and a central, pale core of about 10 nm.
Their organization and protein composition
are the same as in non-neural cells. In the
perikaryon, both neurohlaments and micro¬
tubules are arranged in tracts that occupy
the spaces among Nissl bodies and Golgi
complexes and can be followed into the den¬
drites and axon.
Silver impregnation methods for the light
microscope stain a network of fine fibrils in
the perikaryon, which continue into the den¬
drites and axon, where they can be followed
to their finest ramifications (Fig. 11-7). These
neurofibrils arise from a deposition of silver
along bundles of neurohlaments. The neuro¬
Figure 11-6. Drawing of cells from the nodose ganglion
fibrils are much less stable than the neuro¬
of the vagus nerve. Like neurons in the dorsal root ganglia,
their cell bodies are rounded and only one process hlaments and would not be preserved in
projects from the perikaryon, which is surrounded by routine specimen preparation for light mi¬
satellite cells. (Redrawn from Ramon y Caial.) croscopy.
 THE NERVOUS TISSUE • 317

Figure 11-7. Drawings of a motor

neuron from the gray matter of the
ventral horn of the cat spinal cord
stained for Nissl substance (A) and
stained by the silver method for dem¬
onstrating neurofibrillae (B). The two
images are complementary, the net¬
work of neurofibrillae running be¬
tween the areas occupied by Nissl
bodies and continuing into the proc¬
esses. The Nissl bodies are largely
confined to the perikaryon but may
extend into the dentrites. They usu¬
ally are not found in the axon hillock.

Nissl Bodies. The Nissl bodies, or chrom- In form, size, and distribution, Nissl bodies
ophilic substance (Figs. 11-4, 11-7), stand vary considerably in different types of neu¬
out clearly in the cytoplasm of neurons rons. As a rule, they are coarser and more
stained with basic dyes and show important abundant in large neurons, especially motor
changes in some pathological conditions. neurons, and small and few in small neurons.
They are visible in living neurons with phase- Obvious exceptions are encountered, how¬
contrast microscopy, but are best demon¬ ever. The ganglion cells of the dorsal roots
strated by staining fixed cells with basic ani¬ of spinal nerves may attain large size yet
line dyes—toluidine blue, thionine, or cresyl typically display a uniform distribution of
violet. Thus stained, the bodies appear as very fine Nissl bodies. Under differing phys¬
deeply basophilic masses or blocks in the iological conditions, and in certain patholog¬
perikaryon. The study of Nissl substance in ical states, Nissl bodies change their appear¬
living cells with phase-contrast microscopy or ance.
by the freeze-drying method establishes the Golgi Apparatus. A Golgi complex is pres¬
fact that its clumped pattern in histological ent in all nerve cells, and when stained selec¬
sections accurately reflects its distribution in tively with osmium or silver for light micros¬
life. copy appears as a network of irregular, wavy
In electron micrographs, Nissl bodies are strands, coarser than' the neurofibrillar net¬
seen to consist of cisternae of rough-surfaced work. With the electron microscope, the
endoplasmic reticulum in ordered parallel Golgi network appears as clusters of closely
arrays (Figs. 11-8, 11-9). Ribosomes, ar¬ apposed, flattened cisternae arranged in
ranged in loops, rows, and spirals, are at¬ stacks and surrounded by myriad small vesi¬
tached to the outer surface of the mem¬ cles. The ends of the cisternae are frequently
branes, as in the basophilic bodies of other dilated. Areas of typical Golgi cisternae are
cell types. They also occur in clusters or interconnected by smooth-surfaced tubular
rosettes in the cytoplasm between cisternae. elements, often interpreted as agranular en¬
Nissl bodies, like the basophilic substance of doplasmic reticulum. These latter in turn are
pancreatic and hepatic cells, represent sites often continuous with tubules or cisternae of
of protein synthesis. the granular reticulum. The Golgi complex is
Nissl substance is abundant throughout the arranged in an arc or a complete circle
cytoplasm, including the dendrites. In the around the nucleus approximately halfway
latter it appears under the electron micro¬ between it and the surface membrane of the
scope as anastomosing slender tubules and perikaryon (Fig. 11—8). A cytochemical reac¬
short cisternae. Sites of dendritic branching tion for the enzyme thiamine pyrophospha¬
are frequently occupied by small Nissl bodies. tase yields a reaction product concentrated
They are usually absent from the most pe¬ in the cisternae on the maturing face of each
ripheral region of the perikaryon and from stack, and the resulting staining is coextensive
the area of the perikaryon in which the axon with the Golgi apparatus as classically delin¬
originates (the axon hillock), as well as from eated by silver or osmium impregnation
the axon itself (Fig. 11-7). methods.
 318 THE NERVOUS TISSUE

Figure 11-8. Electron micrograph of a portion of the perikaryon of a typical neuron, illustrating the principal organelles.
The Golgi complex of the neurons is highly developed and forms a continuous network in the perinuclear cytoplasm-
therefore, in a thin section such as this, it is transected at multiple sites. (Micrograph courtesy of Sanford Palay.)
 THE NERVOUS TISSUE • 319

Figure 11-9. Electron micrograph of a Nissl body at higher magnification. It consists of flattened cisternae of the
endoplasmic reticulum oriented parallel to each other. In addition to the ribosomes associated with the membranes,
there are many clusters of ribosomes in the cytoplasmic matrix between cisternae. (Micrograph courtesy of Sanford
Palay.)

Mitochondria. Rodlike or filamentous mi¬ occasionally encountered. Since neurons do

tochondria are scattered everywhere, inter¬ not proliferate, the role of this organelle in
mingling with Nissl bodies and neurohbrils. the adult nerve cell is unknown.
They are generally smaller than those of non- Inclusions. In addition to the organelles
nervous tissues, varying from 0.1 to 0.8 pm already described, there are inclusions in
in diameter, with a preponderance of slender nerve cells that are of more restricted occur¬
forms that generally measure close to the rence. Catecholamine-synthesizing neurons
smaller dimension. They can be demon¬ contain dense-core vesicles 80 to 120 nm in
strated in fresh nerve cells by supravital stain¬ diameter, filled with neurotransmitter and
ing. Their number varies from cell to cell enzymes involved in their synthesis. Neurose¬
and in different parts of the same cell; they cretory neurons of the hypothalamus are
are especially numerous in axon endings. characterized by granules 10 to 30 nm in
Their fine structure resembles that of mito¬ diameter, which contain the hormones vaso¬
chondria in other cells but displays two pe¬ pressin and oxytocin and their carrier pep¬
culiarities of unknown significance; the cris- tides neurophysins. These granules are trans¬
tae are not consistently transversely oriented ported via the axon to the neurohypophysis,
but often run parallel to the long axis of the where their hormonal contents are dis¬
mitochondrion, and the dense matrix gran¬ charged and diffuse into the bloodstream.
ules usually present in the inner mitochon¬ In recent years, isolated granule-containing
drial chamber are either absent or infre¬ cells of several kinds have been found widely
quent. dispersed in the CNS. The granules are im-
Centrioles. A pair of centrioles is charac¬ munoreactive for a number of peptides that
teristic of preneuronal multiplying cells dur¬ are also found in the gastrointestinal tract. The
ing embryonic development. In adult neu¬ physiological significance of these cells is now
rons of vertebrates, centrioles are only a subject of intensive investigation.
320 • THE NERVOUS TISSUE

Pigment granules are frequently encoun¬ the size of the perikaryon, but their pattern
tered in the neuronal perikaryon. The of branching is typical for each variety of
coarse, dark brown or black granules found neuron. The surface of many dendrites is
in neurons in the substantia nigra of the covered with innumerable minute projections
midbrain, the locus ceruleus near the fourth or spines, which serve as sites of synaptic
ventricle, the dorsal motor nucleus of the contact (Fig. 11—5).
vagus nerve, and the spinal and sympathetic In their initial portion, dendrites contain
ganglia are undoubtedly melanin. The phys¬ small Nissl bodies, free ribosomes, and mi¬
iological significance of melanin in these sites tochondria. In addition, they possess long,
is unknown. Of more general occurrence, straight, parallel microtubules and neurohia-
especially in man, are golden brown pigment ments (Figs. 11-11 and 11-12). With increas¬
granules termed lipofuscin. They are probably ing distance from the cell body, microtubules
a harmless by-product of normal lysosomal and neurofilaments become the dominant
activity that accumulates within the cyto¬ feature of the dendritic cytoplasm. The tu¬
plasm. Gradual increase in amount of lipo¬ bules of the endoplasmic reticulum decrease
fuscin with advancing age may displace the in number and the ribosomes become sparse,
nucleus and organelles far to one side of the whereas mitochondria remain more or less
neuron. Iron-containing granular deposits constant in number per unit length and may
are found in neurons of the substantia nigra, actually appear relatively increased in the
the globus pallidus, and elsewhere. Their finer dendritic ramifications.
number increases as the individual grows Through their synapses with axon termi¬
older. nals, the dendrites receive nerve impulses
Lipid, encountered as droplets in the cy¬ from other functionally related neurons. The
toplasm, may represent either normal meta¬ numbers of sources from which they are
bolic reserve material or a product of path¬ received may be very great. In a Purkinje cell
ological metabolism. Glycogen is found in of the cerebellar cortex (Figs. 11-10, 11-11),
embryonic neurons, in embryonic neuroglial the terminals upon the dendritic tree number
cells, and in embryonic cells of the ependyma in the hundreds of thousands. The dendrites
and choroid plexus, but is not present in a play a crucial role in the ability of the neuron
histochemically demonstrable quantity in to integrate information received from its
adult nervous tissue. many inputs. The arriving nerve impulses
Processes of Neurons. The cytoplasmic excite or inhibit electrical activity in localized
processes of nerve cells are their most re¬ regions of dendritic membrane and thus con¬
markable features. In almost all neurons tinuously shift the neuron toward or away
there are two kinds: the dendrites and the from its threshold for signaling a nerve im¬
axon. pulse of its own. Although the impulse car¬
The dendrites provide most of the recep¬ ried by the axon behaves in an “all-or-noth¬
tive surface of the neuron, although the cell ing” fashion, the integrative capacity of the
body, the initial segment of the axon, and dendrites depends on graded changes in elec¬
axon terminals also may receive afferent fi¬ trical potential. In certain instances dendrites
bers from other neurons. Dendrites may be may transmit, as well as receive, exerting
direct extensions of the perikaryon or remote influences upon adjacent dendrites through
arborizations, as in the peripheral branches specialized, reciprocal dendrodendritic syn¬
of a sensory ganglion cell, in which case a apses. They may also exhibit propagated all-
length of typical axon is interposed between or-nothing signaling within long dendritic
perikaryon and dendritic arborization. A shafts (see below). Such findings call for flex¬
neuron usually has several main dendrites; ibility in the functional characterization of
rarely there is only one. Where the dendrites dendrites, or for that matter, any part of the
emerge from the cell body they are thick, nerve cell, whose parts show great adaptabil¬
tapering gradually along their length toward ity to special requirements in particular situ¬
the ends. In most neurons the dendrites are ations.
relatively short and confined to the immedi¬ The axon differs considerably from the
ate vicinity of the cell body. Each dendrite dendrites. Whereas there are usually several
may divide into primary, secondary, tertiary, dendrites, there is only one axon to each
and higher orders of branches, of variable neuron or, in rare instances, no axon at all
shapes and sizes and distributed in diverse (e.g., the amacrine cells of the retina). This
patterns (Fig. 11—11). The number and cell process often arises from a small conical
length of the dendrites bear little relation to elevation on the perikaryon devoid of Nissl
 THE NERVOUS TISSUE 321

Figure 11-10. Electron micrograph of a small area of cerebellum. A small branch of the dendritic tree of a Purkinje cell
running vertically through the field contains several conspicuous mitochondria. Projecting laterally from the dendrite are
“spines” or “thorns” with bulbous tips and narrow stalks. Axons of granule cells form synapses with the Purkinje cel!
dendrite. (From Palay, S. L., and V. C. Palay. Cerebellar Cortex. Berlin, Springer-Verlag, 1974.)
322 • THE NERVOUS TISSUE

Figure 11-11. A, Photomicrograph of a Golgi preparation of a Purkinje cell, showing its highly branched dendritic tree.
B, C, Electron micrographs of small terminal dendrites of the Purkinje cell located in the molecular layer of the
cerebellum. D, Cross section of the primary dendrite of a Purkinje cell. Mitochondria, tubular elements of endoplasmic
reticulum, and punctate profiles of microtubules are found throughout the dendritic tree, but microtubules are more
numerous and more uniformly arranged in the primary dendrite. (Micrograph from Wuerker and Kirkpatrick. In Bourne,
G. H., and J. F. Danielli, eds.: International Review of Cytology. Vol. 33. New York, Academic Press, 1972.)
 THE NERVOUS TISSUE • 323

bodies, called the axon hillock. The axon may rons of this kind may combine to form a
arise, however, from the stem of a principal hbrous plexus of great complexity, envelop¬
dendrite. The portion of the axon interven¬ ing the perikarya of other nerve cells.
ing between axon hillock and the beginning The terminal arborization of the axon is
of the myelin sheath is called the initial seg¬ composed of primary, secondary, and higher
ment. In this region, the plasma membrane orders of branching, varying greatly in num¬
bears an undercoating of dense material and ber, shape, and distribution. Often its
the microtubules become bundled together branches assemble into networks that sur¬
into parallel fascicles. Within each fascicle, round the body of the postsynaptic neuron
each microtubule is connected to its neigh¬ in the form of a basket, or twist around the
bors by cross bridges resembling the rungs dendrites like a clinging vine. In simpler
of a ladder. The axon carries the response cases, the tips of one or two twigs make very
of the neuron in the form of a propagated local contact with the surface of a dendrite
action potential; the axon hillock and initial or the body of another neuron.
segment of the axon, from which this poten¬ The perikaryon, like the dendrites, offers
tial arises, are sometimes called the “trigger electrically excitable membrane upon which
zone. ’ The axon does not contain Nissl bod¬ excitatory or inhibitory influences from axon
ies and usually is thinner and much longer terminals of other neurons can be received
than the dendrites of the same neuron. The and integrated. The axon hillock, or subse¬
axoplasm contains longitudinally oriented tu¬ quent initial segment, may provide a critically
bules of the endoplasmic reticulum, long and placed receptive zone for inhibitory signals.
extremely slender mitochondria, microtu¬ The pseudounipolar neuron of the cranio¬
bules, and many neurohlaments. spinal ganglia transmits a nerve impulse di¬
The axons of many nerve cells have a rectly from the peripheral to the central
prominent sheath of material called myelin, branch of its long, single, T-shaped process
highly refractile in the fresh condition and (Fig. 11-6). Its perikaryon has no synaptic
appearing black in tissue fixed in osmium contacts from other neurons and is chiefly of
tetroxide. The myelin sheath of the axon is not trophic significance, even though its mem¬
part of the neuron but rather a part of an brane may reflect passage of the action po¬
ensheathing cell (see discussion later in this tential in the adjacent process.
chapter). Its presence or absence exerts an Through its ending, the axon transmits
important influence on the physiological nerve impulses to other neurons or to muscle
properties of the neuron. Because it is asso¬ fibers and gland cells. The response of the
ciated only with axons, it provides a criterion effector cells is always activity, but the re¬
for recognizing them. There are, however, sponse of neurons may be a varying degree
axons that are devoid of a myelin sheath of either excitation or inhibition. There are
(unmyelinated axons). In electron micrographs, many types of axon endings, and the same
unmyelinated axons and dendrites of large axon may terminate in several different ways
caliber can usually be distinguished by the and synapse with several different neurons.
fact that there is a much greater number of In general, dendrites display local changes
neurohlaments in the axon. The smaller in membrane potential in response to im¬
processes are more difficult to distinguish, pulses received. Certain neurons with very
because the neurohlaments upon which the long dendrites, however, exhibit propagated
identihcation largely depends are less nu¬ dendritic electrical potentials, similar to the
action potential characteristic of axons, which
merous in small axons.
Along its course, the axon may or may not appear to summate and convey to the peri¬
emit collaterals. Unlike the branches of den¬ karyon weak excitations from remote regions
drites, which diverge at an acute angle, ax¬ of the dendritic tree.
onal branches tend to depart at right angles. Neurons are unusual cells, because most of
In certain instances, a neuron may display an their synthesizing machinery is located in the
extensive system of axon collaterals, which cell body, but they possess long processes
whose maintenance depends on the activity
individually ramify into ever hner branches.
of the perikaryon. Material must therefore
In such cases, the total length of axon may
approach or even exceed that of the den¬ pass continuously from the cell body to the
drites and can extend the sphere of influence dendrites and axon or be returned from the
of the neuron to a greater number of othei periphery to the cell body. Fhe axon with its
terminal arborization largely depends on this
neurons. Axon collaterals from many neu¬
324 • THE NERVOUS TISSUE
\

Figure 11-12. Higher magnification of a dendrite from a ventral horn neuron of the spinal cord, showing microtubules
and clusters of neurofilaments. (Micrograph courtesy of Raymond Wuerker.)

traffic, whereas dendrites are more inde¬ the axonal cytoskeleton, and numerous pro¬
pendent, because they contain cytoplasmic teins of the cytomatrix including actin,
organelles, especially ribosomes, at least in clathrin, calmodulin, and various metabolic
their portion that is close to the ceil body. enzymes. For descriptive purposes the sub¬
Axonal transport is the process by which stances transported are now divided into two
materials are transferred to or returned from groups: slow component a (SCa), comprising
the axon and its terminal arborization; a the cytoskeletal proteins, and slow component
dendritic transport also takes place but has b (SCb), including the cytomatrix proteins. In
been studied in less detail. Two forms of radiolabeling experiments, both are localized
transport exist, an anterograde transport from throughout the cross section of the axons but
the perikaryon to the axon terminal and a the SCb proteins are found in highest con¬
retrograde transport in the opposite direction. centration in the subaxolemmal region.
Two components have been identified in the Many of the constituents carried by retro¬
anterograde transport, a fast and a slow one. grade transport are the same as those that
The velocity of the fast transport varies move in anterograde direction; in addition,
from 20 to 400 mm per day. The bulk of the retrograde transport conveys to the perikar¬
material transported is represented by mem¬ yon proteins and small molecules that have
brane-bounded organelles, such as tubules of been picked up by the axonal endings. Some
the smooth endoplasmic reticulum, vesicles of these proteins travel in tubules, multivesic-
of various kinds, and some of the mitochon¬ ular bodies, and vesicles that fuse with the
dria. In addition, materials of low molecular lysosomes when they reach the cell body.
weight are ferried by the fast transport: these Retrograde transport is also the pathway fol¬
include sugars, amino acids, nucleotides, and lowed by toxins, such as tetanus toxin, and
calcium. The slow anterograde transport has neurotropic viruses, such as those of herpes
a velocity of 0.2 to 4 mm per day and carries and rabies, to penetrate and invade the cen¬
microtubule and neurohlament proteins of tral nervous system. The velocity of retro-
 THE NERVOUS TISSUE • 325

grade transport varies in different axons and the basis of the shape, size, and position of
is in general slower, about half the rate of the cell body, as well as of the synaptic rela¬
the fast anterograde component. tionships. Neurons may have long axons that
The mechanisms of axonal transport are leave the gray matter, traverse the white
still poorly understood. The axonal cytoskel- matter, and terminate at some distance in
eton, especially microtubules, plays an im¬ another part of the gray matter. Alterna¬
portant role in the fast component. For the tively, axons may leave the CNS and end in
slow component, one hypothesis holds that the periphery. Such cells with long axons are
the entire cytoskeleton is slowly propelled termed Golgi Type I neurons; this type in¬
down along the length of the axon. cludes the neurons that contribute to for¬
One important function of axonal trans¬ mation of the peripheral nerves and those
port is the delivery to the axonal endings of whose axons form long fiber tracts of the
synaptic vesicles and enzymes involved in brain and spinal cord. In other neurons, the
transmitter metabolism. Synaptic vesicles can axon is relatively short and does not leave
travel along the axon as large dense-core the region of the gray matter where its cell
vesicles in the neurons that use biogenic body lies. These cells with short axons, which
amines as neurotransmitters; in other neu¬ are especially numerous in the cerebral and
rons, they probably derive from the tubules cerebellar cortices and in the retina, are Golgi
of the smooth endoplasmic reticulum. In Type II neurons.
addition to the enzymes necessary for their The shape of the perikaryon varies; it may
svnthesis, neurotransmitters themselves can be spherical, ovoid, pyriform, fusiform, or
also be transported along the axon, but their polyhedral. The absolute size of the cell body
quantity is small compared with the amounts also varies between extreme limits, from
synthesized in the endings. An exception to dwarf neurons of 4-pm diameter (smaller
this rule may be represented by neuropep¬ than an erythrocyte) to giants approaching
tides and neurosecretory granules produced 150 pm. The pyramidal cells of Betz in the
in the cell body. mammalian cerebral cortex and the paired
Mauthner neurons in the medulla oblongata
of certain fishes and amphibians are exam¬
DISTRIBUTION AND ples of exceptionally large neurons. Could
DIVERSITY OF NEURONS
they be isolated, they would be visible to the
The core of the central nervous system, naked eye.
the gray matter (Fig. 11-13), contains the cell True unipolar neurons are rare except in
bodies of the neurons, their dendrites, and early embryonic stages. In bipolar neurons, a
proximal portions of the axons. Clusters of process projects from each end of the fusi¬
nerve cell bodies in the gray matter are called form cell body. Typical bipolar neurons are
nuclei (not to be confused with cellular nuclei) found in the vestibular and cochlear ganglia,
and represent functional aggregates of neu¬ and in the olfactory nasal epithelium. In
rons. Surrounding the gray matter more or vertebrate embryos, all neurons of the cra¬
less concentrically is a zone, devoid of nerve niospinal ganglia are first bipolar, but during
cell bodies, which contains axons of neurons development the opposing processes shift
whose cell bodies are located either in the around the perikaryon and combine into a
gray matter or in ganglia outside the CNS. single process.These neurons are thus called
This zone is the white matter, so-called because pseudoumpolar. During embryonic stages, the
the axons here are invested by myelin, which perikaryon of such neurons is progressively
is glistening white in the fresh state. Bundles set apart from the region of fusion of the
of myelinated fibers in the white matter, two initial processes. The adult cell body is
which are cable-like functional groupings of globular or pear-shaped; a single process
nerve fibers, are called tracts. In the cerebral arises and divides like the letter T. One
hemispheres and cerebellum, additional gray branch is directed to the periphery and the
matter with nerve cell bodies arranged in other courses in a posterior nerve root to the
distinct layers forms the cortex surrounding CNS. The stem process may be relatively
the white matter (Figs. 11-14, 11-15). short, as illustrated in Figure 11-6, or may
A very great variety of neurons can be run a considerable distance before bifurcat¬
distinguished in the CNS (Fig. 11—2) on the ing, sometimes enveloping the cell body of
basis of the number, length, thickness, and origin in a complex tangle. Except in the
mode of branching of the processes, and on smallest examples, the initial single process
326 • THE NERVOUS TISSUE

Figure 11-13. Sections through human upper cervical spinal cord stained with thionine (A) to show cells, and with the
Weigert-Weil method (B) to show myelinated fibers. Note the external arrangement of the fibers (white matter) and the
central, cruciate area containing the cell bodies (gray matter). The ventral surface is below. A portion of the dorsal root
is seen in upper left. (Courtesy of P. Bailey.)
 THE NERVOUS TISSUE • 327

Granular layer
Outer molecular layer Purkinje cells Molecular layer

White matter White matter

Figure 11-14. Sections of human cerebellar folia stained with the Weigert-Weil method (A) for myelinated fibers and
with thionine (B) for cells. Note the central disposition of the white matter with its myelinated fibers, which stain black
with Weigert-Weil and pale with thionine; the outer molecular layer (pale gray), with scattered neurons and the large
Purkinje cells; and the intermediate, or granular, layer, composed of cells and fibers. (Courtesy of P. Bailey.)
328 • THE NERVOUS TISSUE

A B C
Figure 11-15. Sections from three areas of human cerebral cortex (gray matter), showing distribution of nerve bodies
(A) in the temporal eulaminate (associational) cortex, (B) in the precentral agranular cortex (motor area), and (C) in the
occipital koniocortex (striate visual cortex). Much stress has been laid on minute differences in lamination of the nerve
cells and fibers in these and other areas of the cortex, but there is now a tendency to minimize some of these
differences. (Courtesy of P. Bailey.)

and the peripheral and central branches are shaped neurons include the motor nerve cells
myelinated. The perikaryon of a pseudouni- of the ventral gray matter of the spinal cord
polar neuron is ensheathed by two cellular and of the motor nuclei of the brain stem.
capsules. The inner is made up of small, flat, Pyramidal neurons are characteristic ele¬
epithelium-like satellite cells that are continu¬ ments of the cerebral cortex.
ous with similar Schwann cells enveloping the Of remarkable shape are the graceful Pur-
peripheral process. The satellite cells have a kinje cells of the cerebellar cortex (Figs. 11-5,
relationship to the ganglion cells that is sim¬ 11-11A) One or two thick dendrites covered
ilar to that of neuroglial cells (oligodendro¬ with innumerable tiny spines arise from the
cytes) to neurons in the CNS. Satellite cells, upper end of the cell body. These branch
however, differ in structure and embryonic repeatedly to form a large dendritic arbori¬
origin from neuroglial cells. The outer cap¬ zation, oriented in one plane and shaped like
sule of pseudounipolar neurons is vascular a fan turned at right angles to the long axis
connective tissue, which extends along the of the surrounding cerebellar convolution.
cellular process and continues as the endo- The axon enters the white matter beneath
neurium of the nerve fiber. the cortex; hence, the Purkinje cell is a Golgi
In the majority of neurons, shape is deter¬ Type I neuron. Synaptic terminals upon the
mined by the number and arrangement of dendrites and body of the Purkinje cell num¬
the dendrites (Fig. 11-2). Stellate or star¬ ber hundreds of thousands per ceil and ex-
 THE NERVOUS TISSUE • 329

hibit, according to their source, specific places It has been estimated that well over half the
and modes of ending upon the postsynaptic cytoplasm of neurons is contained in the
surface. neuropil. The great variety of types of neu¬
Many more varieties of neurons are found rons and neuropils results in a striking de¬
in the cerebral and cerebellar cortices. In gree of regional heterogeneity in nervous
diminutive granule cells, a few short dendrites tissue.
radiate in all directions, while the axon and The number of nerve cells in the entire
its collaterals remain either in the immediate nervous system is astronomical, being esti¬
neighborhood of the cell or at least within mated at 14 billion in man. The tremendous
the cortical gray matter. Such neurons qualify increase in this number in the course of
as Golgi Type II. Neurons in the reticular evolution has involved chiefly the integrator
formation of the brain stem have large, var¬ cells or interneurons of the CNS. The number
iously shaped perikarya and extensive but of sensory neurons and associated receptors
poorly branched dendrites. Great attention has also increased, especially in the retina,
has been accorded these neurons in recent but to a much lesser extent. The number of
years because their morphology and synaptic motor neurons 'has remained relatively small
relationships suggest important integrative and in man probably does not exceed two
functions. The multitude of dendrites fre¬ million. The term final common pathway is
quently overlap in complex fashion and re¬ employed to designate the motor neurons by
ceive an input of axons and axon collaterals which nerve impulses from many central
derived from many sources. The typically sources pass to a muscle or gland in the
long axon may distribute impulses through periphery.
ascending and descending branches to a con¬
siderable portion of the length of the neu-
raxis and ramify into rich collateral plexuses
at different levels. At first glance, these
sprawling neurons convey an impression of
THE NERVE FIBER
disorder in the extreme, yet they are encoun¬
tered in the core of the brain stem, an area The nerve fiber is composed of an axon and
upon which the delicate and exquisite control certain sheaths of ectodermal origin. All pe¬
of homeostatic mechanisms depends. ripheral axons are enclosed by a sheath of
The few examples described give an in¬ Schwann cells, which invest the axon almost
complete picture of the wealth of different from its beginning to near its peripheral
kinds of neurons. Many more have been termination. The larger peripheral axons are
described by numerous investigators. Recent also enveloped in a myelin sheath, within the
studies, in which the electron microscope and sheath ofi Schwann. The smallest axons of pe¬
chrome-silver method of Golgi have played ripheral nerves lack a myelin sheath. It is
complementary roles, have further refined necessary, therefore, to designate axons as
the knowledge of neuronal types. It is appar¬ myelinated or unmyelinated. Fresh myelinated
ent that each ganglion, nucleus, or cortical fibers appear as homogeneous, glistening
area is composed of (1) a characteristic variety tubes. This refractile property of myelin ac¬
of neurons in differing proportions, each counts, as noted earlier, for the white color
type of cell designed to meet its special func¬ of fiber tracts of the brain and spinal cord
tional requirements, and (2) a complex and and of numerous peripheral nerves. In
highly ordered meshwork of dendritic, ax¬ stained preparations, the appearance of var¬
onal, and glial processes whose fine structure ious constituents of the nerve fiber differs
and relationships are adapted to provide a according to the technique applied. With
framework for a particular form of organized methylene blue vital staining the axon is
activity. The term used to designate this felt- stained blue, and with silver methods, brown
work of processes is neuropil. The details of or black, the myelin remaining unstained.
its dense entanglements cannot be resolved Unmyelinated fibers, often difficult to ob¬
in silver preparations and have only begun serve by routine histological methods, are
to be appreciated with the advent of the well demonstrated by these special tech¬
electron microscope. The neuropil is of great niques. Weigert’s method and osmium te-
importance in the communications function troxide darken the myelin, leaving the axon
of nervous tissue; it provides an enormous colorless or light gray (Fig. 11—16). Myelin
area for synaptic contact and functional in¬ sheaths are stained blue-green by the Kliiver-
teraction between the processes of nerve cells. Barrera method (Fig. 11—4).
330 • THE NERVOUS TISSUE

Torn area, showing

Node of Ran vie r Fibroblast Schwann’s sheath

Myelin Axon Nucleus of Lanterman n's

Schwann cell clefts
Figure 11-16. Two myelinated fibers of the sciatic nerve of a frog, treated with osmium tetroxide and picrocarmine and
teased. (After A. A. Maximow.)

The Sheath of Schwann The sheath of Schwann and the myelin

sheath are interrupted at regular intervals by
This sheath of flattened cells, sometimes nodes of Ranvier (Fig. 11—17), which are points
called the neurilemmal sheath, forms a thin of discontinuity between successive Schwann
sleeve around the myelin, which surrounds cells along the length of the axon. Here the
the axon. The Schwann cells, like the neu¬ axon is partially uncovered, being only in¬
rons, are of ectodermal origin and represent completely enclosed by a complex arrange¬
elements similar to neuroglial cells of the ment of Schwann cell processes. Myelinated
CNS, but they are adapted to the special axons thus have individual neurilemmal
conditions of the peripheral nervous system. sheaths, divided into segments. Each inter-
In embryonic life, Schwann cells accompany nodal segment of the sheath between two
outgrowing axons and migrate from branch consecutive nodes of Ranvier is composed of
to branch until they form complete neurilem¬ a Schwann cell with its myelin lamellae.
mal sheaths. In the adult their nuclei are
The internodal segments are shorter in the
flattened; a small Golgi apparatus and a few
terminal portion of the fiber. The length
mitochondria can be demonstrated in their
varies in different nerve fibers and in differ¬
attenuated cytoplasm. The myelin and the
ent species from about 200 to over 1000 pm;
Schwann sheaths appear distinct with the
the longer and thicker the fibers, the longer
light microscope and were formerly consid¬
the segments. If an axon gives off collateral
ered separate structures. The electron micro¬
branches, this takes place at a node of Ran¬
scope, however, shows that myelin is actually vier.
part of the Schwann cell, consisting of spirally
In fixed preparations of peripheral nerves,
wrapped layers of its surface membrane (dis¬
the myelin of each segment appears to be
cussed in more detail later). The outer mem¬
interrupted by oblique, cone-shaped discon¬
brane of the Schwann cell and the glycopro¬
tinuities, the incisures or clefts of Schmidt-Lan-
tein boundary layer on its outer aspect were
termann, several to each Schwann segment
resolved with the light microscope as a single
(Figs. 11-16, 11—17). These clefts, seen also
layer, traditionally called the neurilemma.*
in teased fresh or osmicated nerves, repre¬
sent areas of loosening or local separation of
*Originally the word neurilemma, as employed by Eu¬ the spirally wrapped myelin lamellae, which
ropean workers early in the twentieth century, meant are nevertheless continuous across the inci¬
the connective tissue tunic, continuous with the pia sures. The regions between the separated
mater, known today as the endoneurium. English-speak¬
lamellae consist of Schwann cell cytoplasm
ing authors, however, used the term to designate the
clear layer of Schwann membranes defined above in the continuous with that forming the outer sleeve
text. A related term, axolemma, originally referred to the of the nerve fiber on the one hand, and with
inner membrane of the Schwann cell but is now com¬ a thin, inconstant layer of cytoplasm next to
monly used to signify the plasmalemma of the axon. the axon on the other.
 THE NERVOUS TISSUE • 331

Incisure of Neurokeratin
Schmidt-Lantermann network
Node of Schwann cell
Ranvier nucleus

Figure 11-17. Diagrammatic representation of longitudinal sections and cross sections of a single myelinated nerve
fiber and its endoneurial sheath. Left half of drawing represents what would be seen after fixation with osmium tetroxide,
which preserves the lipid of myelin. Right half represents the appearance after ordinary methods of histological
preparation, which extract the myelin and leave behind an artifactitious network of residual protein described as
“neurokeratin.” (Redrawn from Ham, A. W. Histology, 5th ed. Philadelphia, J. B. Lippincott Co., 1965.)

The exact relationship of the Schwann be oriented with hydrocarbon chains extend¬
sheath to unmyelinated axons cannot be vis¬ ing radially and with polar groups aligned at
ualized with the light microscope, but in the aqueous interfaces, loosely bonded to the
electron micrographs it is evident that mul¬ proteins. In general electron microscopic
tiple axons, up to a dozen or more, may studies have supported this interpretation of
occupy deep recesses in the surface of the the molecular organization of myelin and
same Schwann cell (Fig. 11-18). The plas- have shown that the alternating layers of
malemma of the Schwann cell is closely ap¬ mixed lipids and proteins are in fact succes¬
plied to the axon and, as a rule, completely sive layers of the plasma membrane of the
surrounds it. At some point around the pe¬ Schwann cell wrapped spirally about the
riphery of each axon, however, the Schwann axon.
cell membrane turns back to form the mes- In electron micrographs at high magnifi¬
axon, a pair of parallel membranes marking cation, compact myelin presents as a series of
the line of edge-to-edge contact of the encir¬ light and dark lines in a repeating pattern of
cling sheath cell (Fig. 11—21). about 12 nm (Fig. 11-19). The dark line
Schwann cells are indispensable for the life bounding the repeating unit is called the
and function of the axons of peripheral nerve major dense line and is about 3 nm thick; it
fibers. In regeneration, the new axon grows represents the apposition of the inner (cyto¬
out of the proximal stump and follows the plasmic) surfaces of the unit membrane of
path formed by Schwann cells. the Schwann cell. Between major dense lines
is a less dense intraperiod line. This-has been
interpreted as representing the union of the
The Myelin Sheath
outer leaflets of the Schwann cell membrane.
Before the advent of biological electron However, in recent studies based on high-
microscopy, x-ray diffraction analysis sug¬ resolution micrographs of optimally fixed
gested that the myelin sheath was composed myelin sheaths in spinal roots, a narrow open
of concentrically wrapped layers of mixed cleft is resolved between the membranes (Fig.
lipids alternating with thin layers of neuro- 11-20). If this observation can be general¬
keratinogenic protein material. Within the ized, there exists between the outer leaves of
layers, the lipid molecules were thought to the opposing Schwann cell membranes an
332 • THE NERVOUS TISSUE

Figure 11-18. Electron micrograph of a small area of unmyelinated nerve from the rat mesentery, showing multiple
axons associated with the cross-sectional profile of each Schwann cell. Between these fascicles of unmyelinated axons
are unit fibrils of collagen of the endoneurium.

intraperiod gap or channel that is continuous movements could be initiated or controlled

through the myelin sheath from the periax¬ so as to result in formation of the precisely
onal to the endoneurial extracellular spaces. uniform laminated structure observed. Stud¬
Where the laminated myelin sheath is in¬ ies indicate that the myelin spiral does not
terrupted at each node of Ranvier, the axon develop because of any sort of corkscrew
is surrounded loosely by a collar of minute rotation of the axon during growth. If this
finger-like processes of the two adjoining were the mechanism, the direction of spiral
Schwann cells. A distinct gap, however, is for a particular axon would probably be the
found between all the membranes in this same in all its myelin segments, and such is
unmyelinated part of the node (Fig. 11-22). not the case. Apparently the Schwann cell
This gap probably is of significance in rela¬ alone actively produces the spiral, and
tion to current flow between axoplasm and no interaction occurs between individual
the exterior during propagation of the action Schwann cells so far as the direction of spiral
potential. is concerned. Formation of new membrane
In electron micrographs of myelinating pe¬ at the free edge of the original infolding of
ripheral nerves, successive stages can be the satellite cell membrane probably extends
found in the development of the sheath from the fold spirally around the axon without
a double-layered infolding of the Schwann significant change in the relative position of
cell membrane (Fig. 11-21). The mechanism the neuron and its satellite cells. Much re¬
of formation of the spiral, consisting of a few mains to be learned concerning the morpho¬
to 50 or more turns around the axon, is still genesis of myelin, but whatever the morpho¬
unsettled. It has been suggested that during genetic process, the result is that the axon
myelinization the spiral disposition of the becomes surrounded by a many-layered
myelin lamellae is established by rotation of sheath.
the sheath cells with respect to the axon. It In myelinated fibers of the central nervous
is difficult, however, to imagine how such system, certain neuroglial cells (oligoden-
 THE NERVOUS TISSUE 333

External
mesaxon

mmm wm

Schwann

Figure 11-19. Electron micrograph of a large myelinated nerve in a rat spinal root. The myelin sheath consists of a
multilayered spiral wrapping of Schwann cell membrane. Two apposed portions of Schwann cell membrane from the
internal mesaxon extend from the periaxonal space to the innermost layer of myelin. Similarly the two membranes
extending from the outermost turn of myelin to the endoneurial space are called the external mesaxon. (Micrograph
from Coggeshall, R. Anat. Rec. 794:201, 1979.)
334 • THE NERVOUS TISSUE

Periaxonal space
Axopl asm
Mesaxon

Schwann
cytoplasm
!&**><* . . :

Figure 11-20. At high magnification it can be seen that the periaxonal space continues into the cleft between the
opposing external leaflets of the membranes forming the internal mesaxon. This cleft in turn is continuous with a narrow
intraperiod gap in the innermost turn of the myelin sheath (see at arrows). (Micrograph from Coggeshall, R. Anat Rec
794:201, 1979.)

Mesaxon Myelin
Figure 11-21. Diagrams illustrating the development of nerve myelin. A, Earliest stage: axon enveloped by a relatively
large Schwann cell. B, Intermediate stage: unit membranes of mesaxon and to some extent of axon have come together,
line of contact representing future intraperiod line of myelin. C, Later stage: a few layers of compact myelin have formed
by contact of cytoplasmic surfaces of mesaxon loops to make major dense line of myelin. (Redrawn from Robertson J.
D. Prog. Biophys. 70:349, 1960.)
 THE NERVOUS TISSUE • 335

Basal lamina of Schwann cell

Major dense line of myelin sheath

Intraperiod line of myelin sheath

Axolemma-

Junctions between paranodal loops

Incisure
Axolemmal ridges of
Schmidt —
Bare axolemma of node Lantermann

Paranodal region
Mi crotubules
Subaxolemmal density
at the node

Schwann cell cytoplasm

Neurof ilamen Axoplasm

Figure 11-22. Diagrammatic representation of the myelin sheath, node of Ranvier, and incisures of Schmidt-Lantermann
in a peripheral nerve. (Courtesy of D. Kent Morest.)

droglia) play a role corresponding to that of tudinally arranged strands of collagenous fi¬
the Schwann cells in the peripheral nervous bers and fibroblasts pass into the spaces be¬
system. Nodes of Ranvier occur in the central tween the individual nerve fibers constituting
nervous system, but Schmidt-Lantermann the endoneurium. Where the nerve branches,
clefts have not been seen. the connective tissue sheaths become thinner.
In ontogenesis, myelin appears relatively The smaller branches lack epineurium, and
late, and the process of myelinization ends the perineurium cannot be distinguished
some time after birth. Different fiber systems from the endoneurium, being reduced to a
or tracts of the brain and spinal cord become thin, fibrillar layer covered with flat connec¬
myelinated at different times. tive tissue cells resembling endothelial cells.
Delicate reticular fibrils around each nerve
fiber form the tenuous endoneurium. Blood
PERIPHERAL NERVES vessels are embedded in the epineurium and
perineurium and more rarely in the thicker
In their course outside the central nervous layers of endoneurium.
system, nerve fibers of varying thickness It has become customary to classify nerve
(from 2 to up to 30 pm) are associated in fibers according to their diameter, because
fascicles and held together by connective tis¬ the speed conduction of the action potential
sue to form peripheral nerves. The outer varies with the diameter of the fiber. Diam¬
layer of the nerve, the epineurium, is made up eters cover a wide and continuous range from
of connective tissue cells and collagenous large myelinated to small unmyelinated fi¬
fibers, mainly arranged longitudinally (Fig. bers. In peripheral nerves the fibers fall into
11—23). Fat cells may also be found here. three distinct groups. The large fibers, group
Each of the smaller fascicles of a nerve is A, conduct at 15 to 100 meters per second
enclosed in concentric layers of connective and include motor and some sensory fibers.
tissue forming the perineurium. Fine, longi¬ Group B fibers conduct at 3 to 14 meters per
336 • THE NERVOUS TISSUE

Per ineurium
Epineurium

Adipose
Endoneurium tissue

Figure 11-23. Drawing of a histological cross section of a human ulnar nerve at very low magnification, illustrating the
perineural adipose tissue, epineurium, perineurium, and endoneurium. (From Bargmann, W. Histologie und mikrosko-
pische Anatomie des Menschen. Stuttgart, Georg Thieme, 1959.)

second and include mainly visceral sensory vous system. The dorsal root contains sensory
fibers. The C group, small unmyelinated fi¬ fibers from superficial and deep cutaneous
bers conducting at 0.5 to 2 meters per second, regions, sensory fibers from muscles and ten¬
carry autonomic and some sensory impulses. dons, and afferent fibers from viscera. More
Other systems of classification besides this than half of the dorsal root fibers are very
one are used in physiological studies. small axons; most of these distribute with the
Motor nerve fibers of the skeletal muscles cutaneous rami. The relative numbers of
are thick and heavily myelinated; those of myelinated and unmyelinated fibers vary
visceral smooth muscle are thin, lightly mye¬ widely in different spinal segments and in
linated, or without myelin. Tactile fibers are the same segment in different mammalian
medium-sized and moderately myelinated; species. In the mixed nerve trunks peripheral
pain and taste fibers are thinner, with less to the spinal ganglia, the fibers of motor and
myelin or none at all, and olfactory nerve sensory roots mingle, together with sympa¬
filaments are always unmyelinated. Such his¬ thetic fibers from the communicating rami.
tologically defined fiber aggregates constitute In mixed trunks stained with hematoxylin
distinct functional systems: somatic motor, vis¬ and eosin or by another routine method, the
ceral motor, tactile, gustatory, olfactory, and so lipid constituents of myelin are dissolved out,
forth. leaving a loosely arranged protein network
Clear segregation of functionally different called neurokeratin (Figs. 11 — 17, 11—24). A
nerve fibers is found in the spinal roots. In faintly stained axon can usually be seen in
the ventral root are motor fibers of several the center of this network. In such prepara¬
types: (1) coarse and heavily myelinated, in¬ tions, myelinated fibers of various sizes are
nervating ordinary skeletal muscle fibers; (2) readily identified by the clear zones of ex¬
small and myelinated, terminating on intra¬ tracted myelin surrounding the darkly
fusal muscle fibers; and (3) fine and lightly stained axons. In tissues that have been fixed
myelinated, belonging to the autonomic ner¬ with glutaraldehyde and osmium tetroxide,
 THE NERVOUS TISSUE • 337

Figure 11-24. Drawing of myelinated (A) and unmyelinated nerve (B) as they appear in cross section in routine
histological preparations. The lipid of the myelin sheaths in (A) has been extracted in specimen preparation.

the lipid of the myelin is preserved as a dark between the neuron and its related non-
rim around the axon (Fig. 11-25). nervous element. The physicochemical
In the brain and spinal cord, nerve fibers changes that mediate the transfer of the
also segregate into functional systems, the various stimuli from, or the nerve impulses
afferent (incoming) and efferent (outgoing) to, a peripheral non-nervous structure have
pathways (spinocerebellar, spinothalamic, been intensively studied. Three groups of
corticobulbar, corticospinal, and other fiber nerve terminations can be distinguished: (1)
tracts whose origins and terminations are endings in muscle, (2) endings in epithelium,
indicated by binomial nomenclature). Each and (3) endings in connective tissue.
has a special function.
Nerve Endings in Smooth and
Cardiac Muscle
NERVE ENDINGS
From complicated plexuses, thin unmyeli¬
Each peripheral nerve fiber, sensory, mo¬ nated nerve fibers depart and eventually con¬
tor, or secretory, sooner or later ends in some tact or approach the surface of the muscle
peripheral organ with one or several terminal cells. Visceral motor axons give rise to terminal
arborizations. Some nerve fibers ramify as branches that carry numerous boutons en pas¬
free endings among the non-nervous tissue sant filled with synaptic vesicles. These end¬
cells; others attach to tissue cells by means of ings remain at a variable distance from the
specialized terminations. The fibers ending surface of the smooth and cardiac muscle
in sensory receptors are dendrites; those with cells and do not form specialized junctions
motor or secretory endings are axons, their with their membrane. Visceral sensojy fibers
terminations being called axon endings. In ramify in the connective tissue between
general, the structure of the ending is smooth muscle bundles or contact the muscle
adapted to increase the surface of contact fibers themselves. In cardiac muscle the tissue
338 • THE NERVOUS TISSUE

Figure 11-25. A, Cross section of guinea pig sciatic nerve after glutaraldehyde and osmium fixation and embedding in
Epon-araldite. The myelin sheaths have been well preserved and appear as black rims around the axons. B, Longitudinal
section similarly prepared. Arrow indicates a node of Ranvier. (From Webster, H. In Hubbard, J., ed.: The Vertebrate
Peripheral Nervous System. New York, Plenum Press, 1974.)

is permeated by a multitude of thin fibers (Fig. 11-26). The terminal branches of the
passing between muscle trabeculae. They ap¬ nerve fiber ramify and occupy grooves or
pear to end near the surface of the muscle troughs in the surface of the muscle fiber.
fibers but form no specialized contacts with Within the expanded axon terminal are nu¬
them. merous mitochondria and synaptic vesicles, 40
to 60 nm in diameter. The neurofilaments
Motor Nerve Endings in and microtubules found within the axon usu¬
Striated Muscles ally do not continue into the terminal. The
apposed membranes of the axon and of the
The terminations of somatic motor nerves muscle fiber do not touch but are separated
have a more complex structure than those of everywhere by a glycoprotein layer continu¬
smooth and cardiac muscle. The motor end ous with the boundary layer investing the
plate has already been described in Chapter Schwann cell and the sarcolemma. This layer
10 and need only be reviewed here briefly. extends into the trough and the narrow gut¬
Each motor nerve fiber branches to supply ters formed by infolding of the sarcolemma
many muscle fibers. The motor neuron to¬ of the end plate. The gap between the sur¬
gether with the muscle fibers it innervates is faces of the axon and muscle fiber varies in
called a motor unit. The myelin sheath ends width up to 50 nm.
as a terminal branch of the nerve fiber nears
the muscle fiber. The enveloping sheath of
Sensory Nerve Endings in
Schwann cell cytoplasm continues beyond the
Striated Muscles
termination of the myelin and covers the
surfaces of the axonal branch. At the junction These are always present in considerable
of the nerve and muscle fiber there is a local numbers, some in the muscular tissue, others
accumulation of sarcoplasm rich in mitochon¬ on tendons or at muscle-tendon junctions.
dria and muscle nuclei, the motor end plate Some terminations are simple, others com-
 THE NERVOUS TISSUE • 339

Figure 11-26. Diagrammatic representation of a myoneural junction (motor end plate), illustrating a typical chemical
synapse in the peripheral nervous system. Synapses in the central nervous system have some features in common
with this, but they occur between neurons and have no basal lamina or postjunctional folds. (Courtesy of D. Kent
Morest.)

plex. The interstitial terminations are distrib¬ the nuclear bag. Another type of intrafusal
uted in the connective tissue; the epilemmal fiber displays a longitudinal array of nuclei,
terminations closely contact the muscle fibers. or nuclear chain in its nonstriated central
The interstitial terminations may be simple portion. The intrafusal muscle fibers are at¬
naked branches of the axons or encapsulated tached in parallel to the extrafusal fibers and
structures. The epilemmal endings likewise are stretched whenever the muscle is
may be simple: one or more tortuous axons, stretched. Each spindle is approached by two
after shedding their myelin sheath at approx¬ types of thick sensory nerve fibers. The larger
imately the midpoint of a muscle fiber, en¬ axons form annulospiral endings around the
velop the sarcolemma in continuous circular noncontractile segment of the intrafusal fi¬
and spiral twists. Their varicose twigs termi¬ bers. This primary receptor signals both the
nate with nodular swellings. More compli¬ rate and extent of muscle lengthening.
cated neuromuscular spindles, found only in Heightened activity of the primary receptors
higher vertebrates, are long (0.75 to 7 mm exerts an excitatory effect upon the motor
or more) narrow structures arranged parallel neurons to the same muscle by a direct reflex
to the bundles of ordinary muscle fibers, and connection in the CNS. The thinner axons
situated mainly near the junction of muscles have flower-spray endings, which predominantly
with tendons. Each spindle, enveloped by a innervate the nuclear chain fibers. This re¬
connective tissue capsule, consists of several ceptor signals the extent, more than the rate,
long striated muscle fibers, the intrafusal fi¬ of muscle lengthening.
bers, which are shorter and thinner than the Neuromuscular spindles are also supplied
other (extrafusal) muscle fibers (see also Chap¬ by thin motor nerves, gamma fibers, which
ter 10). Midway along each fiber, the stria- emanate from small gamma motor neurons in
tions are replaced by a collection of nuclei, the CNS. (The large motor neurons that send
340 • THE NERVOUS TISSUE

axons to the extrafusal muscle fibers are terminations in the epithelial layers of the
called alpha motor neurons.) The gamma fibers skin and mucous membranes are probably
terminate on the intrafusal muscle fibers with only sensory, and those in the epithelial
typical motor end plates. These fibers effect glands partly secretory, partly sensory. End¬
shortening of the striated, contractile por¬ ings in glands (lacrimal, salivary, kidneys, and
tions of the intrafusal fibers. These contrac¬ so on) are all unmyelinated sympathetic fibers
tions do not contribute significantly to the forming dense nets on the outer surface of
tension produced by the muscle but serve the basal lamina; branches penetrate the lam¬
instead to stretch the noncontractile region ina, often forming a second network on its
of the intrafusal fibers where the annulo- inner surface, and end between the gland
spiral endings are located. Such stretching cells.
causes the receptor to discharge more rap¬ Free sensory epithelial endings are abun¬
idly; hence, the function of the gamma fibers dant where sensitivity is highly developed: in
is to regulate the sensitivity of the neuromus¬ the epithelium of the cornea, in the mucous
cular spindle to stretch. membrane of the respiratory passages and
oral cavity, and in the skin. In the epidermis,
Sensory Nerve Endings in Tendons these branches do not penetrate farther than
the granular layer. Free nerve endings in
These are of several kinds and also may be hair follicles are important tactile organs.
either simple or encapsulated. In simple There are two sets of endings—one circularly
forms, the naked nerve fibers and their arranged in the middle layer of the dermal
branches spread over the surface of the ten¬ sheath, the other consisting of fibers running
don fibers in small, treelike figures of differ¬ parallel to the hair shaft and terminating in
ent types. Such endings in tendons and the outer root sheath.
fasciae probably carry pain sensation. Com¬
posite forms, the Golgi tendon organs, occur at
Nerve Endings in Connective Tissue
the junction of muscle fibers with intramus¬
cular connective tissue or with tendons. They These are numerous and of many forms,
are supplied by a single sensory fiber that particularly in the dermis, under the epithe¬
branches among the bundles of connective lium and mesothelium of the mucous and
tissue fibers and terminates with small sprays serous membranes, around the joints, in the
of endings. In contrast to neuromuscular endocardium, and elsewhere. They are either
spindles, however, which are placed in par¬ free or encapsulated endings, or are con¬
allel with the other muscle fibers, the Golgi nected with special tactile cells of epithelial
tendon organs are in series with the contrac¬ origin. More complex endings are found in
tile elements. These receptors respond to skin and hypodermis, mucous and serous
increase in tension; a heightened activity of membranes, endocardium, cornea, sclera,
the receptors exerts an inhibitory effect, and periosteum. Nonencapsuiated endings
through interneurons of the CNS, upon the are frequent in the papillary layer of the skin,
alpha motor neurons of the same muscle. connective tissue of mucous membranes
The physiological significance of the sen¬ (such as that of the urinary bladder), and
sory endings in muscles and tendons has been pericardium, endocardium, and periosteum;
clarified greatly in recent years, as has their the terminal branches form spherical or elon¬
morphology. These receptors participate in gated structures resembling glomeruli.
postural and phasic adjustments of skeletal
musculature. Intimate and complex connec¬
Encapsulated Terminal Sensory
tions in the CNS relate their activity to the
Endings
alpha and gamma motor neurons. This activ¬
ity, however, should not be described as In these, a special connective tissue capsule
“muscle sense” or “position sense”; awareness of varying thickness surrounds the actual
of the position of the body parts in space nerve endings. The capsule attains its great¬
appears to be mediated by receptors located est thickness in the corpuscles of Vater-Pacini
in joints. (Figs. 11-27, 11-28), found in the deeper
layers of the skin, under mucous membranes,
Nerve Endings in Epithelial Tissue in conjunctiva, cornea, heart, mesentery, and
pancreas, and in loose connective tissue.
Histologically, receptors and effectors can These structures are large (1 to 4 mm by 2
be distinguished only in rare instances. The mm) and white in color. Each is supplied with
 THE NERVOUS TISSUE 341

Lamellae

Inner bulb

Blood vessel between

superficial lamellae

Figure 11-27. Drawing of a cross section of a corpuscle of Vater-Pacini, from the dermis of the sole of the foot. (After
Schaffer.)

Figure 11-28. Electron micrograph of a cluster of three small encapsulated nerve endings. (Micrograph courtesy of E.
Weihe.)
342 • THE NERVOUS TISSUE

A B
Figure 11-29. A, Genital corpuscle from the glans penis Figure 11-30. Photomicrograph of palmar digital epider¬
of a 23-year-oid man. Silver preparation. (Courtesy of N. mis showing Meissner’s corpuscles in two neighboring
Cauna.) B, Lingual corpuscle from a filiform papilla on the dermal papillae. Hematoxylin and eosin.
tongue of a 21-year-old woman. Silver stain. (From
Cauna, N. Anat. Rec. 124:77, 1956.)

one or more thick myelinated fibers, which nomic nervous system. This system, as defined
lose myelin upon entering the corpuscle. Flat¬ long ago, unfortunately excludes the visceral
tened Schwann cells surround the nerve end¬ sensory neurons, or interoceptive system, which
ing, and these are in turn encircled by the form the afferent side of visceral arcs.
lamellae of the connective tissue capsule. Gen¬ The autonomic nervous svstem includes
/

ital corpuscles, in the skin of the external numerous small ganglia. The vertebral ga?iglia
genital organs and of the nipple, are similar form a chain, the sympathetic trunk, on either
(Fig. 11—29A). Meissner’s corpuscles (Figs. side of the spinal column; the sympathetic
11—30, 11—31, 11—32), occurring in connec¬ trunk is connected proximally with the ven¬
tive tissue of the skin of the palms, soles, and tral roots of the spinal nerves. Additional
tips of the fingers and toes, are elongated, ganglia lie at some distance from the central
pear-shaped, or elliptical formations with nervous system in certain nerve plexuses (col¬
rounded ends, located in the cutaneous pa¬ lateral or prevertebral ganglia) or in the walls
pillae, with the long axis vertical to the sur¬ of organs (terminal ganglia). The autonomic
face. Their size varies (40 to 100 pm by 30 ganglia contain motor nerve cell bodies,
to 60pm) Corpuscles of Golgi-Mazzoni and ter¬ which convey impulses originating in the
minal bulbs of Krause resemble corpuscles of brain and spinal cord to smooth muscle and
Vater-Pacini but are smaller and simpler in glands by way of the visceral or splanchnic
construction. nerves (Fig. 11—33). Fibers of some cell bod¬
ies in the sympathetic trunk join those in the
peripheral nerves and run to the sweat glands
and arrector pili muscles. Whatever the des¬
AUTONOMIC NERVOUS
tination, the autonomic nervous system me¬
SYSTEM diates activity by two motor neurons placed
in series; the first lies either in a nucleus of
Motor neurons of the central and periph¬ the brain stem or in a special territory of the
eral nervous systems concerned with the reg¬ spinal gray matter, and the second is located
ulation of visceral activities form the auto¬ in a ganglion outside of the CNS. In the
 THE NERVOUS TISSUE • 343

Epithelium

Thin myelinated i / Thick myelinated fiber

fiber-4

Figure 11-31 Figure 11-32

Figure 11-31. Meissner’s corpuscle of a dermal papilla of a human finger. Methylene blue stain. (Redrawn after Dogiel.)

Figure 11-32. Meissner’s corpuscle of an 11-year-old girl. Silver stain. (Courtesy of N. Cauna.)

peripheral nervous system, only one motor thoracic and upper lumbar spinal cord. The
neuron transmits activity to the effector or- axons pass into the ventral roots of spinal
gan. nerves and through the white communicating
The autonomic system consists of sympa¬ branches, to end either in a vertebral gan¬
thetic (thoracolumbar) and parasympathetic (cran¬ glion of the sympathetic trunk or in a pre-
iosacral) divisions: it influences the intrinsic vertebral ganglion. Most of these axons, the
activity of cardiac muscle and supplies nerve preganglionic fibers, with thin myelin sheaths,
fibers to smooth muscle in the viscera, sali¬ terminate in a sympathetic ganglion. Here
vary and sweat glands, blood vessels, and they synapse with secondary visceral motor
other structures. The peripheral nervous sys¬ neurons, whose axons—mostly unmyelinated
tem, in contrast, innervates striated skeletal postganglionic fibers—transmit activity to vis¬
muscle. Despite these distinctions, however, ceral muscles or glands. Some postganglionic
the traditional concept of an autonomic ner¬ fibers travel to internal viscera over sympa¬
vous system is justifiable only in terms of thetic nerves, such as the cardiac or splanch¬
convenience; its components and functions nic nerves; others extend from vertebral gan¬
are inextricably bound up with the rest of glia through gray communicating branches
the nervous system and do not possess auton¬ and spinal nerves to structures of the body
omy. wall and extremities; vasomotor fibers chiefly
The sympathetic trunks and their ganglia, to arteriolar muscles, pilomotor fibers to the
as well as the collateral ganglia, are the ave¬ small muscles of hair follicles, and sudomotor
nues of communication for the thoracolum¬ fibers to the sweat glands.
bar outflow between the CNS and the viscera. The craniosacral division of the autonomic
Each sympathetic trunk contains ganglia at system haS preganglionic neurons in the
the level of exit of most of the corresponding brain and spinal cord. Axons of the cranial
spinal nerves. The communicating branches component emerge in the oculomotor, facial,
(rami communicantes) pass between the trunk glossopharyngeal, and vagus nerves, to syn¬
and the spinal nerves. apse in terminal ganglia innervating the head
The cell bodies of the sympathetic neurons and trunk. From the second, third, and
lie in the intermediolateral gray column of the fourth sacral segments of the spinal cord,
344 • THE NERVOUS TISSUE

A. SOMATIC' B. VISCERAL

ventral roots to the spinal nerve and to one of the ganglia of the autonomic system. The pattern of afferent (blue) and
efferent (red) nerve fibers in both the somatic and visceral system may be directly compared. (From Copenhaver, W.
M., and R. Bunge, eds.: Bailey’s Textbook of Histology. 16th ed. Baltimore, Williams and Wilkins, 1971.)

axons leave via ventral roots and sacral nerves innumerable terminal boutons, as are the
to reach postganglionic neurons in terminal large nerve cell bodies of somatic motor neu¬
ganglia associated with pelvic viscera. rons. Instead, a relatively small number of
Postganglionic neurons may exercise a lo¬ axodendritic endings is found.
cal regulatory control over the viscera to Postganglionic neurons of the craniosacral
which they are related, secondarily subject to division lie, as a rule, close to the viscera
control by components of the CNS. innervated. The preganglionic fibers, accord¬
Distributed via both divisions of the auto¬ ingly, are relatively long—as in the vagus
nomic nervous system, the peripheral process nerve—and the postganglionic fibers short.
of visceral sensory neurons lead from the On the other hand, most synapses of the
viscera through the communicating rami, or thoracolumbar division are in the sympa¬
through cranial or sacral nerves, to sensory thetic trunk or collateral ganglia; therefore,
ganglia. Their cell bodies are morphologi¬ the postganglionic fibers are longer.
cally indistinguishable from those of the so¬ Sympathetic ganglion cells are generally
matic sensory neurons, with which they min¬ small and have diverse shapes. The cells are
gle in the craniospinal ganglia. usually multipolar, dendrites and axon some¬
times being distinguishable but in other cases
Autonomic Nerve Cells showing no obvious difference. Preganglionic
fibers often synapse with the dendrites of the
The cell bodies of preganglionic visceral ganglion cell in dense glomeruli. For a typical
efferent neurons are small, spindle-shaped example, see the description of the postgan¬
elements in the intermediolateral gray col¬ glionic neurons of the intestine (Chap. 26).
umn. Their perikarya are not studded with The cell body may be encapsulated by
 THE NERVOUS TISSUE • 345

satellite cells, which, like those of the cranio¬ limiting membrane of the CNS. Similar mem¬
spinal ganglia, are ectodermal elements sim¬ branes are formed around the blood vessels.
ilar to Schwann cells in the nerve sheaths. In The adult ependyma may represent the re¬
the outlying sympathetic ganglia these cap¬ maining epithelial cells of the embryonic ven¬
sules may be absent, but Schwann cells ac¬ tricular zone (see section on development) or
company the peripheral sympathetic fibers may be derived from a specialized subven-
everywhere. tricular zone that appears later in develop¬
ment and persists into adult life.

NEUROGLIA Neuroglial Cells

In any section of the central nervous sys¬
The number of nerve cells within the cen¬ tem prepared by ordinary histological meth¬
tral nervous system, although enormous, is ods, small nuclei are seen scattered among
exceeded, perhaps fivefold, by the number the nerve cells and their processes. The cy¬
of non-neural supportive cells, called the toplasm and long processes of these neuro¬
neuroglial cells or neuroglia. The term neu¬ glial elements are demonstrable by special
roglia is applied to the ependyma, which lines histological techniques.
the ventricles of the brain and spinal cord, Three types of neuroglia are distinguished:
and to neuroglial cells and their processes, astrocytes, oligodendrocytes, and microglia. The
which mingle with neurons in the CNS and hrst two, collectively macroglia, are ectoder¬
retina. The satellite or capsular cells of periph¬ mal in origin, as are the nerve cells. The
eral ganglia and the Schwann cells of periph¬ microglia are said to originate from meso¬
eral nerves may be considered peripheral dermal cells of the pia mater, which migrate
neuroglia. into the CNS along the blood vessels.
The astrocytes are of two varieties. The
Ependyma protoplasmic astrocyte has a larger nucleus than
oligodendrocytes and microglia, abundant
In early embryonic stages the wall of the granular cytoplasm, and numerous thick
neural tube is a simple epithelium. Certain processes (Fig. 11—344). Many processes at¬
thin, non-nervous parts of the brain retain tach to blood vessels and to the pia mater by
this structure throughout adult life, as, for means of expanded pedicles. In other cases,
example, the epithelial layer of the choroid the cell body lies directly on the wall of the
plexus. In most other parts, the wall is greatly blood vessel or on the inner surface of the
thickened by the accumulation of neurons pia mater. Some of the smaller cells of this
and neuroglial elements. The lining of the variety lie close to the bodies of the neurons
inner surface of the ventricular cavities al¬ and represent one type of satellite cell.
ways retains an epithelial character. This The fibrous astrocyte (Fig. 11—346) is distin¬
membrane, the adult ependyma, is composed guished by long, relatively thin, smooth, and
of the inner ends of epithelial cells, with their infrequently branched expansions. These
nuclei and some of their cytoplasm. cells also are often attached to blood vessels
The embryonic ependyma is ciliated; in by means of their processes. Embedded
some parts of the ventricular lining, the cilia within the cytoplasm are fibrillar structures.
may persist into adult life. In the mature Electron micrographs show that the neuroglial
brain, the broad bases of ependymal cells fibers of light microscopy are in fact aggre¬
taper to long, threadlike processes that gation of slender filaments present in pro¬
branch and are lost among other elements of fusion in the cytoplasm. Glial filaments are
the brain. In a few places where the wall is distinct from neurofilaments because they are
thin, as in the ventral fissure of the spinal more densely packed, possess a diameter of
cord, some ependymal cells span the entire only 8 nm, and have a different protein
distance between ventricular and external composition. Their main constituent is the
surfaces, as all do in the early embryonic glial fibrillar acidic protein, 51,000 daltons in
stages. In these cases, the ependymal cells molecular weight. Protoplasmic astrocytes are
form a dense internal limiting membrane at the found chiefly in gray matter and fibrous
ventricular end. At the external surface, un¬ astrocytes in white matter, insinuated be¬
der the pia mater, the ependymal threads and tween fascicles of nerve fibers. Mixed or
bars expand into pedicles that fuse into a plasmatofibrous astrocytes are occasionally en¬
thin, smooth, dense membrane, the external countered at the boundary between gray and
346 • THE nervous tissue

C D
Figure 11-34. Neuroglial cells of the central nervous system. A, Protoplasmic astrocyte; B, fibrous astrocyte; C,
microglia; D, oligodendroglia. (After del Rio-Hortega.)

white matter; processes that extend into the cance of this behavior in relation to their
gray have a protoplasmic character whereas normal function in the brain is not known.
those that pass into the white are fibrous. At the electron microscope astrocytes are pale
The oligodendrocytes (or oligodendroglia) cells with euchromatic nucleus and an incon¬
(Fig. 11 —34D) are akin to the astrocytes, spicuous complement of cytoplasmic organ¬
which they resemble in many respects. They elles. Oligodendrocytes, on the other hand,
are usually smaller and have smaller nuclei, have a heterochromatic nucleus, a dark cy¬
although there are many transitional forms. toplasm rich in ribosomes and cisternae of
As their name implies, their few and slender granular reticulum, a large Golgi apparatus,
processes have few branches. Oligodendro¬ and numerous mitochondria. Microtubules
cytes relate intimately to nerve fibers, along are prominent both in the perinuclear cyto¬
which they form rows or columns. Although plasm and in the cell processes.
it is difficult to demonstrate the connection In the microglia (Fig. 11-34C), the nucleus
of the oligodendrocyte with the myelin is small but deeply stained and surrounded
sheath in the adult, studies on the developing by scant cytoplasm. The few processes are
nervous system show that this cell forms short and, unlike the more or less straight
myelin in the CNS. Thus, it is the homologue extensions of the astrocytes, are twisted in
of the neurilemmal cell of Schwann. In gray various ways. Also, the processes and the cell
matter, oligodendrocytes adjoining nerve body are not smooth, but are covered with
cells are the principal type of satellite cell. In numerous tiny pointed twigs or spines. Mi¬
tissue cultures, oligodendrocytes exhibit croglia are scattered everywhere throughout
rhythmic pulsatile movements. The signifi¬ the brain and spinal cord.
 THE NERVOUS TISSUE • 347

In the mature CNS, and also it seems now forms, with active migration and phagocyto¬
during development, neuroglia provide an sis.
extremely complex framework for the neu¬
rons, a scaffolding for migratory young neu¬
rocytes and their cell processes. Like the
THE SYNAPSE AND THE
neurons, the supporting neuroglial cells do
not form a syncytium but retain their individ¬
RELATIONSHIPS OF
uality. In the chambers of a complex laby¬ NEURONS
rinth of glial cells, the nerve cells and their
processes are often individually encapsulated The nervous system consists of complex
and thus insulated from one another. An chains of neurons arranged to permit trans¬
interesting and probably significant fact re¬ mission of activity from one cell to the other.
vealed by electron microscopy is that wher¬ Each neuron receives excitatory and inhibi¬
ever nerve cell bodies and their processes are tory influences from other neurons or sen¬
not in synaptic contact with another neuron, sory receptors, integrates them, and (depend¬
they are generally enveloped by the cell bod¬ ing on their balance) generates and transmits
ies or processes of neuroglial cells. The dis¬ signals to other neurons or peripheral effec¬
tribution of these neuroglial processes thus tor cells. The site of signal transmission is the
appears neither to be random nor merely to synapse.
fulfill the requirements of mechanical sup¬ Two distinct modes of transmission are
port and nutrition of the neurons. Early in known: electrical and chemical. Electrical syn¬
this century, Ramon y Cajal proposed that apses are less common in the central nervous
neuroglial processes are always disposed to system of mammals; at the electron micro¬
prevent contact between processes of nerve scope they were identified as gap junctions,
cells at sites other than those appropriate to which permit movement of ions and thus
their specific signaling function. On the basis direct spread of current from one neuron to
of electron microscopic observations. This the adjoining one.
hypothesis has been revived. It has been At chemical synapses, the process of a neu¬
shown that each neuron has a characteristic ron, called presynaptic, secretes a neurotrans¬
pattern of neuroglial investment complemen¬ mitter substance that diffuses across the inter¬
tary to the specific pattern of its synaptic cellular space and binds to specific receptors
connections. Only at the synapses are the on the surface of a postsynaptic neuron,
neuroglial barriers interrupted, and only causing changes in ionic permeability of its
here is direct contact between the neurons membrane. Henceforth, chemical synapses
possible. Thus, by isolating and individualiz¬ will be simply referred to as synapses, because
ing the many diverse pathways that may they are by far the most common site of
converge upon a given neuron, the neuro¬ transmission between nerve cells.
glial cells may play an essential role in the Traditionally, the synapse was described as
communications function of the nervous sys¬ a place of contact between two neurons; it is
tem. usually established by the terminal arboriza¬
Neuroglial cells seem also to be an impor¬ tion of an axon on the dendrites (axodendritic),
tant mediator for the normal metabolism of the perikaryon (axosomatic), or the axon (axo¬
neurons, although little is known in this re¬ axonic) of one or more neurons. Dendroden-
spect. There is evidence that they remove dritic synapses and synapses between the ad¬
from the extracellular space the potassium jacent perikarya have also been found, but
that accumulates as a result of the neuronal they are uncommon. Thus, it appears that
activity. More is known about the activity of any part of a neuron can participate in the
neuroglia in states of injury or disease. When¬ formation of a synapse. Axons commonly
ever neurons are affected by a local or distant end by giving rise to terminal branches that
pathological process, the surrounding neu¬ carry tiny swellings along their course (boutons
roglial elements always react in some way. en passant) or at their end (boutons terminaux).
They are actively involved in degeneration These swellings or endings are applied to the
and regeneration of the nerve fibers, in vas¬ surface of the postsynaptic neuron. Some¬
cular disorders, and in various infectious dis¬ times, the terminal twigs may form “bou¬
eases. They are the chief source of tumors quets” or loose baskets adhering to the body
of the CNS. Under certain circumstances, or dendrites of other nerve cells. In rare
microglia may assume a great variety of instances, the axon gives rise to a single large
348 • THE NERVOUS TISSUE

Figure 11-35. The types and terminology of the synapses occurring on various parts of the neuron. (Redrawn after
Bunge, R. In Copenhaver, W. M., and R. Bunge, eds.: Bailey’s Textbook of Histology. 16th ed. Baltimore, Williams and
Wilkins, 1971.)

ending that occupies an extensive portion of run a parallel course, often gently convex
the surface of the postsynaptic cell; these toward the presynaptic cytoplasm, and they
endings are called end bulbs in the ventral are separated by an intercellular space or
cochlear nucleus, and calyces in the nucleus synaptic cleft 20 to 30 nm in width. The
of the trapezoid body. In the cerebellar and synaptic cleft is traversed by fine filaments
cerebral cortices, axon terminals contact the and is sometimes bisected by a tenuous
dendritic spines of the Purkinje and pyram¬ plaque of dense material. Synaptic vesicles, 40
idal cells, respectively. The number of syn¬ to 60 nm in diameter, cluster near the pre¬
apses on a neuron varies enormously. Some synaptic membrane; at this site, the mem¬
neurons, such as the granule cell of the brane carries dense material on its cyto¬
cerebellum, have only a few. A motor neuron plasmic surface. Finally, the cytoplasmic
may possess as many as 10,000 synapses. A aspect of the postsynaptic membrane is dec¬
Purkinje cell of the cerebellum may have orated by a layer of dense, fluffy material.
100,000 endings upon its dendrites alone. The cytoplasm of the presynaptic ending
The electron microscope demonstrates that contains randomly dispersed synaptic vesi¬
one or more specialized junctions occur at cles; a variable number of mitochondria, sit¬
the site of contact between presynaptic end¬ uated at some distance from the synaptic
ing and postsynaptic neuron; in addition, the junctions; and a few cisternae and tubules of
presynaptic ending contains a specific com¬ the agranular reticulum. Microtubules and
plement of cytoplasmic organelles (Figs. neurohlaments also are occasionally present.
11—36, 11—37). The synaptic junctions or Certain synaptic junctions have a cleft 30
complexes have the following general prop¬ nm in width: furthermore, they are charac¬
erties: the pre- and postsynaptic membranes terized by a prominent postsynaptic density
Figure 11-36. Tip of a dendritic spine, capped by a terminal bouton from the ventral horn of the spinal cord in a rat.
The typical features of synapses—mitochondria, clustered vesicles, and the cleft—are well shown. The terminal is
enclosed within a thin astrocytic process. (Courtesy of S. L. Palay.)

Figure 11-37. Electron micrograph of a typical synapse from the medial trapezoid nucleus of a cat, showing
neurofilaments and synaptic vesicles in the nerve ending, a typical synaptic cleft between the pre- and postsynaptic
membranes. (Micrograph courtesy M. Jean-Baptiste and D. Kent Morest.)
349
350 • THE NERVOUS TISSUE

so that the contact appears asymmetrical. that they may act as classical neurotrans¬
They are called by Gray Type I synapses and mitters has been obtained for the luteiniz¬
are consistently associated with round vesicles ing hormone-releasing hormone (LHRH),
in the presynaptic ending. In the cerebellar which, in sympathetic ganglia, acts on the
cortex, they mediate excitation; thus, it is postsynaptic neuron to produce a long-last¬
generally assumed that this type of synaptic ing depolarization. In addition to neurotrans¬
junction has excitatory function. In other mitters, the synaptic vesicles may also contain
synaptic junctions, called Type II by Gray, the enzymes involved in transmitter metabolism,
cleft is about 20 nm in width and the pre- such as dopamine-P-hydroxylase, which syn¬
and postsynaptic densities are nearly sym¬ thesizes norepinephrine.
metrical. Furthermore, they may be associ¬ In most synapses, the vesicles are spherical
ated with flattened synaptic vesicles in the in shape and have a clear content (agranular
presynaptic ending. In the cerebellar cortex, vesicles), but in some endings after aldehyde
they mediate inhibition; thus, it is often as¬ fixation and subsequent treatment with os¬
sumed that Type II synapses have inhibitory mium tetroxide, the vesicles appear flattened.
function wherever they occur. It must be Clearly, it is a preparation artifact, for in
noted, however, that types of synaptic junc¬ freeze-fractured specimens all vesicles are
tions exist whose structure is intermediate round. As mentioned above, in the cerebellar
between Gray’s Types I and II, so that it is cortex flattened vesicles are exclusively found
impossible to infer their physiological prop¬ at inhibitory synapses, whereas vesicles at
erties solely on the basis of their morpho¬ excitatory synapses appear spherical. This
logical appearance. correlation between structure and function
Synaptic vesicles are membrane-bounded cannot be generalized, because excitation and
organelles that contain neurotransmitter sub¬ inhibition are the result of the action of the
stances. Until recently, the monoamines (nor¬ neurotransmitter on the postsynaptic mem¬
epinephrine, dopamine, and serotonin), brane, and in different synapses the same
acetylcholine, and amino acids (glycine, gluta¬ neurotransmitter may have different post¬
mate, and y-aminobutyric acid or [GABA] ) synaptic effects.
were thought to be the only neurotransmit¬ A third variety of synaptic vesicles contain
ters involved in synaptic transmission. More a dense core after aldehyde and especially
recently, a large number of peptides appear after permanganate fixation; these granular
to act either as neurotransmitters or neuro¬ vesicles are typical of nerve endings that re¬
modulators; i.e., substances that modify the lease catecholamines as neurotransmitters.
action of classical neurotransmitters. Sub¬ The fact that soaking the tissue in biogenic
stance P, hypothalamic releasing hormones, en¬ amines or their analogues causes an increase
kephalin, vasoactive intestinal polypeptide (VIP), in the proportion of granular vesicles after
cholecystokinin (CCK), neurotensin are a few aldehyde fixation supports the idea that the
examples of this growing list of brain pep¬ dense core indeed contain catecholamines.
tides involved in synaptic transmission. It is The varieties of small synaptic vesicles just
interesting to note that some of these pep¬ described are often associated with a small
tides are also contained in secretory cells of population of large vesicles, 80 to 120 nm in
the gastrointestinal tract, where they act on diameter, which consistently contain a core
neighboring cells (paracrine secretion) and of dense material. In endings that use bio¬
may be released as hormones into the blood¬ genic amines as neurotransmitters, the large
stream by endocrine glands or liberated from dense-core vesicles probably contain cate¬
neurons into the blood of the hypophyseo- cholamines, but in endings that release dif¬
portal vessels to regulate the function of the ferent transmitters, their significance is un¬
pars distalis of the pituitary (neurohor¬ clear.
mones). In all these instances the basic func¬ At the site where synaptic vesicles cluster
tion of the molecule is intercellular commu¬ near the presynaptic membrane, its cyto¬
nication, and it is not surprising that the plasmic surface carries a set of dense projec¬
organism exploits the same peptide for signal tions arranged in a trigonal array. The vesi¬
transmission either locally at the synapse cles of the presynaptic cluster that directly
(neurotransmitters) or at a distance (hor¬ adjoin the presynaptic membrane nestle be¬
mones). Among the large list of peptides tween the projections and may actually con¬
synthesized by neurons, the best evidence tact the plasmalemma. In rare instances, one
 THE NERVOUS TISSUE • 351

of these vesicles is caught by the fixative fluid to its receptor increases the permeability of
in the act of fusing with the presynaptic the postsynaptic membrane to potassium or
membrane; the plasmalemma becomes con¬ chloride, or both; this drives the membrane
tinuous with the vesicular membrane, and potential away from threshold and thus coun¬
the lumen of the vesicle opens into the syn¬ teracts depolarization. After binding to the
aptic cleft. postsynaptic receptor, the transmitter is
Freeze-fracturing demonstrates two types either degraded by enzymes or taken up
of specializations in the interior of the pre¬ again by the presynaptic ending.
synaptic membrane: a population of ran¬ The structural counterparts of these phys¬
domly distributed large particles, and defor¬ iological events at the synapse have been
mations of the membrane that appear as actively investigated during the past decade.
dimples or craters on the inner leaflet or P- The specializations of the presynaptic mem¬
face and protrusions on the outer leaflet or brane and the associated cluster of synaptic
E-face. These deformations arise when the vesicles have been called the active zone, for it
fracture plane breaks through the neck of represents the site for transmitter release at
synaptic vesicles that are fusing with the pre¬ the surface of the presynaptic ending. The
synaptic membrane. synaptic vesicles contain the transmitter
In addition to a cytoplasmic layer of dense quanta, and upon calcium entry into the
material, the postsynaptic membrane may ending, they fuse with the plasma membrane
exhibit internal specializations that are re¬ at the synaptic junction, releasing their con¬
vealed by the freeze-fracturing technique. In tents by exocytosis into the synaptic cleft.
the cerebellum and olfactory bulb, excitatory Using the technique of instantaneously freez¬
synapses are characterized by a cluster of ing the presynaptic ending after the arrival
large intramembrane particles that remain of a nerve impulse, it has been shown that
consistently associated with the E-face, there is a good correlation between the num¬
whereas the postsynaptic membrane of inhib¬ ber of quanta released and that of vesicles
itory synapses contains the usual population undergoing exocytosis.
of randomly distributed particles, which are The large particles within the active zone
preferentially associated with the P-face. Syn¬ of the presynaptic membrane (Fig. 11-38)
apses with a cluster of particles on the P-face may represent the channels through which
of the postsynaptic membrane have also been calcium enters the presynaptic ending and
found; examples are the neuromuscular triggers vesicle fusion with the plasmalemma.
junction, the synapses in the superior cervical The dense material associated with the pre¬
ganglion, and those between photoreceptor synaptic membrane may trap synaptic vesicles
and horizontal cells in the retina, all excita¬ and concentrate them at the sites on the
tory. surface of the ending that face a cluster of
The physiological event that causes trans¬ postsynaptic receptors. It is interesting to
mitter secretion is depolarization of the mem¬ note that there is a correlation between the
brane of the presynaptic ending, produced configuration of this material that binds syn¬
by the arrival of a nerve impulse. As a con¬ aptic vesicles and the geometry of the syn¬
sequence of the depolarization, calcium en¬ apse. In the neuromuscular junction, where
ters the ending and triggers release of trans¬ acetylcholine receptors reside on the lips of
mitter. This is secreted in multimolecular the junctional folds of the muscle membrane,
packets or quanta of fixed size, and variations the dense material has the shape of a bar
in release depend on the number of quanta overlying the junctional folds. In the invagin-
secreted by the terminal. The transmitter ating synapses established by the endings of
diffuses across the synaptic cleft and com¬ the photoreceptor cells of the retina, the
bines with receptor molecules at the surface dense material has the shape of a thin plate
of the postsynaptic membrane. As a result of or ribbon, which bisects a wedge-shaped pro¬
the binding of the transmitter with its recep¬ jection of the presynaptic ending or synaptic
tor, ionic channels in the postsynaptic mem¬ ridge. Vesicles are thus positioned near the
brane open or close. In a typical excitatory slopes of the ridge, opposite the postsynaptic
synapse, the transmitter produces a simulta¬ specializations of two adjoining postsynaptic
neous increase in the permeability to sodium processes that belong to horizontal cells. In
and potassium, and this leads to depolariza¬ this fashion, two postsynaptic neurons may
tion of the postsynaptic membrane. At inhib¬ be activated simultaneously.
itory synapses, the binding of the transmitter The dense material contained within the
352 • THE NERVOUS TISSUE

\ \

s,- -■ .

WMi

Z i

Active zones

Figure 11-38. See legend on opposite page

 THE NERVOUS TISSUE • 353

synaptic cleft may provide direct mechanical surfaced cisternae (Fig. 11-39). From these
coupling between the pre- and postsynaptic endocytic organelles, new generations of syn¬
membranes and thus ensure cell-to-cell adhe¬ aptic vesicles are reformed and refilled with
sion. In fact, a number of experiments indi¬ transmitter, so that the constituents of their
cate that the synaptic junction is capable of limiting membrane are utilized many times
resisting stresses that elsewhere tear apart over again by the presynaptic ending. In
the pre- and postsynaptic surfaces. The ma¬ addition to this mechanism of local recycling
terial in the synaptic cleft may have additional of the vesicular membrane, new vesicles are
important functions in synaptic transmission: certainly contributed by the neuronal soma,
in the neuromuscular junction it contains the where they are probably formed in the Golgi
enzyme acetylcholinesterase, which hydro¬ apparatus and transported to the ending by
lyzes the transmitter acetylcholine and thus the axoplasmic flow. Since agranular vesicles
rapidly shuts off its effect on the postsynaptic are rarely seen in the axoplasm, they may
membrane. travel along the axon as tubules of the en¬
In the Torpedo electric organ, the postsyn¬ doplasmic reticulum, which bud new vesicles
aptic surface of the electroplax cell is covered upon their arrival to the ending.
with minute bumps that may correspond to The absence of cytoplasmic continuity be¬
the receptor molecules for the neurotrans¬ tween neurons forms the basic for the “neu¬
mitter. In the neuromuscular junction, using ron doctrine”—that each mature nerve cell
labeled a-bungarotoxin that irreversibly represents a cellular unit anatomically sepa¬
binds to acetylcholine receptors, these were rate from, and trophically independent of,
localized in the regions of the postsynaptic other neurons. The processes of a neuron
membrane that carry a coat of dense cyto¬ depend on the cell body and its nucleus.
plasmic material. Thus, there is no doubt When cut off from the cell body, the proc¬
that the specialized region of the postsynaptic esses die. New processes may, however, grow
membrane contains the receptor-ionophore out of the perikaryon. The body and nucleus
complexes responsible for the postsynaptic of the nerve cell are the trophic center of the
changes in ionic permeability. It is still un¬ entire neuron. If a nerve cell suffers irrepa¬
clear whether the intramembrane particles rable injury, other nerve cells are not neces¬
clustered within the postsynaptic membrane sarily affected. Nevertheless, certain groups
correspond to the ion channels that open or of neurons may atrophy, or degenerate, fol¬
close after transmitter binding to its recep¬ lowing interruption of afferent axons or
tors. death of other neurons to which they send
After fusion of the synaptic vesicles with their efferent output. This phenomenon,
the presynaptic membrane and release of transneuronal degeneration, has been consid¬
their transmitter contents into the synaptic ered exceptional for many years. It was es¬
cleft, the vesicular membrane is incorporated pecially evident in those neuronal subsys¬
into the plasmalemma. According to a widely tems, notably the visual pathway, in which
accepted hypothesis, the constituents of the one group of cells along the pathway receives
vesicular membrane diffuse laterally within almost all of its input from only one other
the plane of the plasma membrane and are type of cell. Recently, however, it has been
finally retrieved at the periphery of the active found that transneuronal effects are wide¬
zone through coated vesicles and smooth¬ spread, in both the central and peripheral

Figure 11-38. Freeze-fracture preparations of the synaptic membrane. A, Active zone in the presynaptic membrane of
a myoneural junction. The pits indicated by arrows are the cross-fractured necks of synaptic vesicles discharging their
transmitter. (Micrograph courtesy of Tom Reese.)
B, Synaptic junction from the inner plexiform layer of the macaque retina. The presynaptic active zone is characterized
by an aggregation of large intramembrane particles (arrowheads). A synaptic vesicle in exocytosis is indicated by arrow
with asterisk. (Micrograph courtesy of E. Raviola and G. Raviola.)
C, Specialization of the P-face of the presynaptic membrane in a synapse of the inner flexiforrn layer of the retina.
The fracture plane has passed through the cluster of synaptic vesicles and then cleaved the presynaptic membrane,
exposing its P-face at the active zone (arrowheads). The aggregation of large particles is in register with the underlying
cluster of synaptic vesicles. (Micrograph from Raviola, E., and G. Raviola. J. Comp. Neurol. 209:233, 1982.)
D, In one type of synapse in the central nervous system the postsynaptic membrane is characterized by an aggregation
of large particles that remain associated with the E-face (arrows). In this synapse from the inner plexiform layer of the
retina, two postsynaptic processes occur side by side, each with its aggregation of particles. (Micrograph from Raviola,
E., and G. Raviola. J. Comp. Neurol. 209:233, 1982.)
354 • THE NERVOUS TISSUE

Nerve

Figure 11-39. Hypothetical pathway for synaptic vesicle membrane recycling at the neuromuscular junction. Synaptic
vesicles coalesce with the plasma membrane in discharging their content of transmitter. Equal amounts of membrane
are retrieved by formation of clathrin-coated vesicles in regions of membrane adjacent to the Schwann sheath. The
coated vesicles coalesce to form cisternae where transmitter accumulates, and new synaptic vesicles are budded off to
return to the active zone of the nerve terminal. (After Heuser, J. E., and T. S. Reese. Reproduced from The Journal of
Cell Biology 57:315, 1973 by copyright permission of The Rockefeller University Press.)

nervous systems. Degeneration may be retro¬ the cerebral cortex, nuclei of the medulla
grade or anterograde, depending on the direc¬ oblongata, and elsewhere in the spinal cord.
tion, and may involve neurons more than Thus, the spinal motor cord cells serve as the
one synapse removed; hence, it may be pri¬ final common pathway by which activity from
mary, secondary, tertiary, or even quaternary. many sources is transmitted to effector or¬
Such findings raise important questions con¬ gans. In such arrangements, the response of
cerning the growth and maintenance of the the postsynaptic cell is determined by the net
nervous system, and also are crucial facts to effect of excitation and inhibition exerted
consider in interpreting patterns of neuronal upon it by the many afferent fibers. Thus, in
atrophy or death after natural or experimen¬ a spinal reflex arc the excitation of peripheral
tal damage to the nervous system. sensory elements is only one of many inputs
influencing the response of the motor neu¬
rons. The intricate connections between neu¬
Examples of Interrelationships of
rons and their enormous numbers (an esti¬
Neurons
mated 9 x 109 neurons in the cerebral cortex
Almost every neuron is connected with alone) indicate the complexity of neural
several or many other neurons. With the aid structure and function. Details of the orga¬
of the Golgi impregnation and other meth¬ nization of the central nervous system are
ods, many types of relationships between beyond the scope of this book and must be
neurons have been demonstrated, ranging sought in textbooks of neuroanatomy. It may
from complex tangles involving the processes be instructive, however, to refer back to the
of hundreds of cells, to a neuron synapsing photomicrographs of three different parts of
upon itself. As an example of the complex the nervous system. Figure 11—13, of the
type, there are attached to the body and spinal cord, illustrates the white matter, com¬
dendrites of large motor cells in the anterior promising masses of myelinated fibers that
gray columns of the spinal cord hundreds of surround the relatively small amount of gray
synaptic boutons of axons from neurons in matter containing the nerve cell bodies. Fig-
 THE NERVOUS TISSUE • 355

ures 11—14 and 11—15 show gray matter the connections between particular cells and
located outside the white in the cerebral cor¬ in the number, type, and location of synaptic
tex and cerebellum. White matter, made up terminals upon different parts of the same
of myelinated or unmyelinated axons, serves cell. Physiologically, particular neurons
to transmit patterns of nerve impulses from among the astronomical numbers making up
the body to the CNS or vice versa, and from the visual area of the cerebral cortex may
one part of the brain or cord to another part. respond to highly specific modes of visual
There is no evidence that any modification stimuli to the retina. Other principles of
of the nerve impulses occurs in nerves or organization of the nervous system emerge
tracts. from study of neuroanatomy. Among these
In gray matter, nerve cell bodies mingle are the concepts that the CNS is subdivided
with unmyelinated and myelinated fibers. A into a series of interdependent cellular en¬
microscopic preparation shows the bodies of sembles for analysis and control; that the
the cells arranged in some order, often in patterns of connections often permit the re¬
layers. The space between layers, and also ciprocal interactions necessary for modula¬
between individual cells, is packed with in¬ tion of both incoming and outgoing impulses;
numerable axons, dendrites, neuroglia, and and finally, that the cells of the CNS at all
blood vessels. The axons in these areas usu¬ levels from motor neuron to the cell of the
ally lack myelin sheaths, which accounts for cerebral cortex are designed for integrative
the gray appearance in fresh condition. In¬ action.
numerable reciprocal contacts between the
various types of neurons permit a variety of
mutual influences.
When stained with routine methods, the DEVELOPMENT OF THE
region between cell bodies has a punctate or
NEURONS AND OF THE
stippled appearance and corresponds to the
neuropil described above. In the cerebellar
NERVOUS TISSUE
and cerebral cortices and in the retina, cer¬
tain layers consist almost exclusively of naked The neurons of the nervous system de¬
neuronal and neuroglial processes. Huge velop from embryonic ectoderm. Also of ec¬
numbers of synaptic contacts take place in todermal origin are the neuroglial cells (ex¬
such synaptic fields. cept for the microglia), the Schwann cells of
The pattern of the cells and fibers (cytoar- peripheral nerves and corresponding satellite
chitecture) in gray matter varies from place to cells in peripheral ganglia, and certain ele¬
place. Every subcortical nucleus, peripheral ments of the meninges.
ganglion, and locality of the cerebral cortex In early embryonic stages, the future cen¬
has architectural features of its own. Thus, tral nervous system separates by folding from
the cortex in the precentral convolution of the primitive ectoderm to form the neural
the primate brain, the so-called motor area, tube. At the time the neural folds meet, some
differs noticeably from that of the postcentral cells leave the junctional region bilaterally to
convolution, where somatosensory function is form elongated cellular aggregations be¬
represented, and from all other parts of the tween the neural tube and the prospective
cerebral cortex. One important cortical re¬ epidermis. These bands, the neural crests,
gion having a characteristic cytoarchitecture soon become segmented and are the precur¬
is the visual area along the calcarine fissure sors of the sympathetic and part of the cran¬
of the occipital lobe. Another, in the sylvian iospinal ganglia; cells of the meninges; the
fossa, is the auditory area. However, careful adrenal medulla; part of the cranial mesen¬
attempts to correlate cytoarchitectonic and chyme; branchial cartilages; melanocytes;
functional findings have largely failed up to and odontoblasts. Schwann cells are also gen¬
the present time. erally regarded as derivatives of the neural
Anatomical and physiological studies pro¬ crests. Autoradiographic studies with triti-
vide compelling evidence that the nervous ated thymidine, however, show that some
system is not a random tangle of neurons Schwann cells originate in the neural tube.
and neuroglial cells and their processes. Neu¬ The early neural tube is a type of pseudo-
rons are sometimes considered redundant stratihed epithelium in which all cells border
but in many regions display remarkable on the lumen. Autoradiographic and electron
structural and functional individuality. Mor¬ microscopic studies demonstrate a single type
phologically the individuality is expressed in of columnar cell, now called a ventricular cell.
356 • THE NERVOUS TISSUE

During proliferation, the nuclei of ventricu¬ bodies of the somatic and visceral motor
lar cells undergo cyclical changes of position neurons remain within the brain or spinal
in the ventricular zone; nuclei of premitotic cord; their axons grow out as ventral roots
cells lie deep, progressively approaching the of the peripheral nerves and terminate in
lumen during prophase. Karyokinesis occurs muscles or autonomic ganglia.
only at the luminal surface, whereupon the The protoplasm of a growing axon shows
daughter nuclei move again to deeper posi¬ ameboid movements and insinuates itself be¬
tions. Even before the closure of the neural tween other tissue elements. At its advancing
tube, the nuclei of neuroblasts (immature neu¬ tip, a bulbous enlargement, or growth cone,
rons incapable of further division) derived thrusts slender, spinelike projections between
from ventricular cells migrate peripherally. obstructing cells and fibers. These features
Later, such cells will form an intermediate zone of neuronal development seen in fixed and
(future gray matter). A marginal zone (future stained material are confirmed in observa¬
white matter) is already present; it contains tions on living nerves in tissue culture and
the nucleus-free outer processes of the ven¬ by studies of growing nerves in the transpar¬
tricular cells. Neuroglial cells in general orig¬ ent tail of the living frog tadpole. Axons of
inate also from the ventricular zone but, neuroblasts grow into the intercellular spaces
unlike typical neuroblasts, continue to divide as slender, protoplasmic strands. In periph¬
after migration to the intermediate and mar¬ eral nerve fibers, all newly firmed axons are
ginal zones. In certain regions of the brain, at first devoid of Schwann and myelin
cells are produced that retain proliferative sheaths. The earliest myelin appears near the
ability after their origin in the ventricular Schwann cell nucleus, and from there it
zone; these give rise to additional neurons spreads proximally and distaliy.
and glial cells. One such secondary germinal The forces that direct the course of devel¬
matrix forms a subventricular zone immedi¬ opment of the complex nervous tissue of
ately beneath the ventricular cells; it is prom¬ vertebrates are largely unknown. The impor¬
inent in the lateral ventricles and persists into tance of an oriented microstructure as a
adult life. Its cells do not exhibit intermitotic guide, along whose channels developing ax¬
nuclear migrations. Another, similar matrix ons spread, has been confirmed. There has
is found in the development of the cerebel¬ been no confirmation, however, of the con¬
lum. A transient layer is formed on the ex¬ cept of “neurobiotaxis,” which assumes that
ternal surface of the embryonic cortex by differences in electric potential between den¬
cells that migrate from the ventricular zone drites and axons account for migration of
of the underlying brain stem. Proliferation nerve cell bodies in the direction of the
in this external layer produces neuroblasts source from which their stimuli come. No
that descend into the cerebellar cortex. Other clear-cut effects of electric currents have been
cerebellar neurons and most neurons of the seen on either the rate or direction of the
cerebral cortex arise directly from the ven¬ growth of nerves. Chemotactic influences
tricular zone and traverse intermediate and upon growing axons have been demonstrated
marginal zones to reach their destinations. clearly in the optic nerve. Regenerating optic
Autoradiographic studies demonstrate that nerve fibers from different parts of the gold¬
proliferation of neurons by the ventricular fish retina unerringly reach their former ter¬
zone is a rigorously timed and highly ordered minations in the optic lobes of the midbrain,
process, which is followed by active migration even after surgical cross-union of medial and
and frequent intermingling of neuroblasts as lateral optic tracts. There is evidence for
they proceed to their final positions. refined specificities of growing axons in other
The sensory neurons of the craniospinal regions of the CNS, but the role of chemo¬
nerves arise in the neural crests. The periph¬ tactic influences on the growth of peripheral
eral processes grow outward and become nerves remains controversial. There is also
axons of the sensory nerve fibers. The den¬ evidence to show that the peripheral organs
dritic region of these neurons is the receptive and tissues affect the development of CNS
zone at the periphery; here, the axon devel¬ in many ways after contact has been estab¬
ops a variable pattern of branches, which lished between the two regions. These effects
may or may not be encapsulated. The central are both quantitative and qualitative; they act
processes enter the CNS as the axons of to regulate the number and size of neurons
dorsal roots and establish connections with in specific regions of the neuraxis and to
interneurons or motor neurons. The cell influence the pattern of the connections. An-
 THE NERVOUS TISSUE • 357

other important finding has been the discov¬ derived from mesenchyme. The three layers
ery of specific nerve growth factors, identified enveloping the brain and cord are likewise
as proteins, which exert powerful effects composed chiefly of connective tissue. The
upon the generation, growth, and mainte¬ outermost, the dura mater or pachymeninx, is
nance of specific types of nerve cells. dense and firm (Fig. 11—41). The inner lay¬
Studies with the Golgi method show that ers, the innermost called the pia mater and
many neuroblasts retain for some time the the intermediate called the arachnoid (Fig.
primitive distal process they possessed as ven¬ 11—40), are composed of looser connective
tricular cells. These studies suggest that free tissue and constitute the leptomeninges.
ameboid motion of an entire cell may not
occur in neurogenesis. Instead, the nucleus
Dura Mater
may simply shift peripherally from the ven¬
tricular zone along this process until it The dura of the spinal cord and that of
reaches its definitive position, perhaps near the brain differ in their relationships to the
the external surface of the brain wall, as in surrounding bones. The inner surface of the
the prospective cerebral cortex. Retraction of vertebral canal is lined by its own periosteum;
the attachments of the internal and external a separate cylindrical dural membrane
processes of the neuroblast takes place from loosely encloses the cord. The wide epidural
the luminal surface and basal lamina of the space, between periosteum and dura, con¬
neural tube, respectively. Axon sprouting, tains loose connective and fatty tissue and
growth of dendrites, and other features of the epidural venous plexus. The dura is
neuronal differentiation appear to be inde¬ firmly connected to the spinal cord on each
pendent variables. Such events occur at dif¬ side by a series of denticulate ligaments. The
ferent times relative to the shift in position inner surface of the spinal dura is lined with
of the nucleus and in different sequences, squamous cells. Its collagenous bundles run
depending on the type of nerve cell and its for the most part longitudinally. Elastic fibers
region of the developing CNS. are less prominent than in the cerebral dura.
The dura mater of the brain in embryonic
development also has two layers, but in the
adult these are closely joined. Both consist of
MENINGES, VENTRICLES, connective tissue with elongated fibroblasts.
AND CHOROID PLEXUS The outer layer adheres to the skull loosely
except at the sutures and the base of the
In addition to the neurons and macroglia, skull, where it is more firmly adherent. It
both of which are of ectodermal origin, the functions as a periosteum, is richer in cells
brain and spinal cord contain blood vessels than the inner layer, and contains many

Arachnoid

Arachnoid
trabecula Cerebral
vein

Pia mater

Cortex Perivascular
Figure 11-40. Diagram of cerebral space
cerebri
pia-arachnoid, showing relations of
the subarachnoid space, perivas¬ Lining cells of
cular channels, and nerve cells. perivascular
(From Weed, L. H. Am. J. Anat. 31. space
1922.)

Capillary in
pericapillary
space
358 • THE NERVOUS TISSUE

Arachnoid trabecula Arachnoid villus Dura mater

Subdural space Superior sagittal sinus

Arachnoid
Endothelium
pia mater

Figure 11-41. Diagram of the organi¬

zation of the connective tissue sheaths
of the brain. Cerebrospinal fluid formed
in the choroid plexus circulates in the
subarachnoid space and is absorbed
by the venous sinuses through the
arachnoid villi, one of which is shown
here projecting into the sagittal sinus.
(From Weed, L. H. Am. J. Anat. 31,
1922.)

Subarachnoid space Falx cerebri Cortex cerebri

blood vessels; its thick collagenous fibers are their histological structure can best be de¬
arranged in bundles. The inner layer is thin¬ scribed together—in fact, these two mem¬
ner, with finer fibers forming an almost con¬ branes are often treated as a single entity,
tinuous sheet. Its fibers run from the frontal the pia-arachnoid.
region backward and upward, oriented op¬ The main elements of the arachnoid and
posite to those of the outer layer. The inner pia are interlacing collagenous bundles sur¬
surface of the dura is smooth and covered rounded by fine elastic networks. In the pia
with a layer of squamous cells. of the spinal cord an outer longitudinal and
an inner circular layer can be distinguished.
Arachnoid Among the cells are fibroblasts and macro¬
phages; the latter are especially numerous
The leptomeninges of the brain and spinal along the pial blood vessels. They correspond
cord are similar in structure. The arachnoid to macrophages of other parts of the body,
is a thin, netlike structure devoid of blood store vital dyes injected directly into the sub¬
vessels, resembling the transparent parts of arachnoid space, and in inflammation be¬
the omentum. Its outer surface is smooth, come large or transform into epithelioid cells.
but from its inner surface runs a multitude In man they often contain considerable
of thin, branching threads and ribbon-like amounts of a yellow pigment that sometimes
strands, attached to the underlying pia. Mic¬ reacts positively to tests for iron.
roscopically the tissue has a cobweb-like ap¬ Along the pial vessels lie scattered single
pearance (Figs. 11-40, 11-41). The arach¬ mast cells and small groups of lymphocytes.
noid bridges the sulci and fissures on the In certain pathological conditions the latter
surface of the brain and spinal cord, forming increase enormously in number and may
subarachnoid spaces of various extent. become plasma cells. In the pia, particularly
on the ventral surface of the medulla ob¬
Pia Mater longata, a varying number of melanocytes
can be found.
This layer is a thin connective tissue net The outer and the inner surfaces of the
closely adherent to the surface of the brain arachnoid, its trabeculae, and the outer sur¬
and spinal cord. It contains a large number face of the pia are covered with a layer of
of blood vessels, from which most of the squamous epithelial cells.
blood of the underlying nervous tissue is During development of the meninges two
supplied. Attached to the pia are the inner zones may be distinguished: an outer zone of
fibrous strands of the arachnoid; these two condensation of mesenchyme, which gives
membranes are so intimately related that rise to the periosteum, dura, and membra-
 THE NERVOUS TISSUE • 359

nous arachnoid; and an inner zone, which cisterna lies above the medulla oblongata and
becomes the pia. Between these two zones below the posterior border of the cerebellum
the mesenchyme remains loose and later (icisterna cerebellomedullaris, or cisterna magna).
forms spongy tissue, traversing the subarach¬ The fourth ventricle communicates with this
noid spaces. cisterna through three openings in the tela
choroidea: a medial foramen of Magendie, and
the two lateral foramina of Luschka.
Nerves of the Meninges
The dura and pia mater are richly supplied Ventricles
with nerves. All vessels of the pia and of the
choroid plexus are surrounded by extensive The central nervous system begins devel¬
nerve plexuses in the adventitia. Axons orig¬ opment as a neural tube with a wide cavity
inate in the carotid and vertebral plexuses throughout the length and remains a hollow
and in certain cranial nerves and belong to organ throughout life. The central canal or
the sympathetic system. Sensory, nonencap- ventricle of the spinal cord in the adult is
sulated nerve terminations, and even single minute, or it may be obliterated. It does not
nerve cells, are also present on the adventitia seem to perform any important function.
of the blood vessels. However, in the normal adult the ventricular
The cerebral dura contains, besides the cavities of the brain form a continuous chan¬
nerves of the vessels, numerous sensory nel for flow of cerebrospinal fluid. If part of
nerve endings in its connective tissue. The this channel is occluded by disease so as to
cerebral pia also contains extensive nervous prevent free circulation, intracerebral pres¬
plexuses, especially abundant in the tela cho- sure increases, with resulting hydrocephalus
roidea of the third ventricle. The fibers end or other serious consequences.
either in large pear-shaped or bulbous swell¬ The ventricular cavity is dilated in four
ings or in skeins and convolutions similar to regions: the two lateral ventricles in the cere¬
those of the corpuscles of Meissner. In the bral hemispheres, the third ventricle in the
spinal pia the vessels receive their nerves thalamic region, and the fourth ventricle in the
from the plexuses following the larger blood medulla oblongata and pons. Choroid plex¬
vessels to the cord. Afferent nerve endings uses develop in these four regions, and most
are also present, but these are unevenly dis¬ of the ventricular fluid is derived from the
tributed. blood vessels of these plexuses.
Both myelinated and unmyelinated nerve
fibers accompany the blood vessels into the Choroid Plexus
substance of the spinal cord and the brain,
ending on the muscle cells of the vessels. There are four places where the wall of
These come from similar nerves of the pial the brain retains its embryonic character as a
vessels, and the two nervous plexuses are thin, non-nervous epithelium. This part of
continuous. the brain wall is the lamina epithelialis. The
pia mater that covers it is extremely vascular
and is called the tela choroidea. Small arteries
Meningeal Spaces and capillaries of the pia form glomerular
Between the dura mater and the arach¬ tufts of vessels, covered by the lamina epithe¬
noid, the subdural space is a serous cavity lialis and protruding into the ventricle, con¬
containing a minimum of fluid; actually, it is stitute the choroid plexus.
scarcely more than a potential space. Between Choroid plexuses are found in the roof of
the outer sheet of arachnoid and pia is the the third and fourth ventricles and in part
subarachnoid space, traversed by cobwebby of the wall of the two lateral ventricles. In
connective tissue trabeculae, and containing each case, the tela choroidea is much folded
a large amount of fluid. Over the convolu¬ and invaginated into the ventricle, so that the
tions it is narrow, but in the sulci it is wide free surface exposed to the ventricular fluid
and deep. The subarachnoid space is espe¬ is large, with tortuous vessels and a rich
cially wide throughout the length of the capillary net (Fig. 11-42).
spinal cord. In the brain, it is greatly enlarged The epithelium early acquires a peculiar
in a few places (cisternae) where the arachnoid structure, different from that of the cells
is widely separated from the pia and the lining the ventricles elsewhere. In embryonic
trabeculae are rare or absent. The largest stages it contains glycogen and carries cilia.
360 • THE NERVOUS TISSUE

Epithelium

Connective
tissue

Blood vessel

Figure 11-42. Choroid plexus of the fourth ventricle of man. (After A. A. Maximow.)

In the adult its cells are cuboidal and ar¬

Cerebrospinal Fluid
ranged in a single regular layer. Each con¬
tains a large round nucleus and a varying The central nervous system is bathed in
number of rod-shaped mitochondria. cerebrospinal fluid as in a water bath. This
On the free surface the cells have a special¬ protects it from concussions and mechanical
ized border resembling a brush border. In injuries and is important for its metabolism.
electron micrographs, however, this border The subarachnoid spaces freely communi¬
is seen to consist of long microvilli that are cate; cerebrospinal fluid may pass through
irregularly oriented and often somewhat ex¬ them from one end to the other of the
panded at the tips (Fig. 11-43). Although neuraxis. The amount of the fluid is variable,
possibly these terminal expansions are arti¬ estimated at 80 to 100 ml, sometimes as much
facts of fixation, microvilli of other cell types as 150 ml. It is limpid and slightly viscous,
fixed similarly do not have this appearance. has a low specific gravity (1.004 to 1.008),
In animals repeatedly injected intrave¬ and contains traces of proteins, small quan¬
nously with vital dyes, such as trypan blue, tities of inorganic salt and dextrose, and few
the epithelium of the choroid plexus stores lymphocytes (about two or three, not more
large amounts of the dye. Also, in the peri¬ than ten, per milliliter). It resembles the
vascular connective tissue core of the plexus aqueous humor of the eye more closely than
many macrophages are found, which take up it does any other fluid of the body.
and store great amounts of dye. Cerebrospinal fluid is constantly renewed.
On the boundary between adjacent epithe¬ It circulates slowly through the brain ventri¬
lial cells in electron micrographs, a juxtalu- cles and through the meshes of the subarach¬
minal junctional complex appears to seal the noid space. If the space is opened to the
intercellular space. The capillaries beneath outside by injury, large amounts of fluid
the epithelium are unlike those found else¬ steadily drain off—200 ml or more a day.
where in the brain; they have thin walls and The sources of fluid are primarily the blood
fenestrations or pores closed by thin dia¬ vessels of the choroid plexus, the pia mater,
phragms. The junctions between endothelial and the brain substance. From the brain
cells also appear to be quite permeable. Fol¬ substance the flow is outward into the sub¬
lowing intravenous injection in mice, perox¬ arachnoid space; from the choroid plexus it
idase (a protein) can be shown to cross the is inward into the ventricles. Fluid may be
capillary walls and enter the stromal space. added to the ventricles in a few other places,
It moves between the epithelial cells but is notably the area postrema at the caudal mar¬
stopped near the lumen by the junctional gin of the fourth ventricle. The ependymal
complex. surfaces in general do not discharge fluid
 THE NERVOUS TISSUE • 361

Figure 11-43. Electron micrograph of choroid plexus at higher magnification than in Figure 11-42, illustrating the
unusual free border of bulbous or clavate microvilli.

into the ventricles. In contrast, absorption of sinuses permits passage of cerebrospinal fluid
fluid from the ventricles into nearby veins directly into venous blood, rather than into
takes place through the ventricular wall. The lymphatics. A small amount of fluid enters
plexuses are wholly secretory, not resorptive, lymphatics or venous vessels as just described,
and constitute the chief source of fluid. The but most of it passes directly into the large
major channel of ventricular fluid outward endocranial venous sinuses through local spe¬
into the subarachnoid space is through spe¬ cializations of the arachnoid called arachnoid
cially modified places in the membranous villi.
roof of the fourth ventricle.
Ventricular fluid normally flows from the Arachnoid Villi
lateral ventricles of the cerebral hemispheres,
where it derives chiefly from the lateral cho¬ The large endocranial venous sinuses are
roid plexuses, through the foramina of surrounded by thick walls of dura mater
Monro into the third ventricle. Here fluid is except in certain places, chiefly in the sagittal
added from the choroid plexus of the third sinus of the falx cerebri. Here the dura is
ventricle and the augmented flow passes perforated by numerous protrusions of the
through the aqueduct of Sylvius into the arachnoid, through each of which a finger of
fourth ventricle, where more fluid is added arachnoid epithelium, an arachnoid, villus (Fig.
from the choroid plexus there. Fluid then 11—41), evaginates into the lumen of the
passes into the cisterna magna and diffuses sinus. The cavity of the villus, containing a
in all directions through the subarachnoid small amount of loose arachnoid tissue, com¬
space. Some of the fluid may enter extracran¬ municates freely with the subarachnoid
ial lymphatics through perineurial spaces space, so that at this point fluid of the space
within the cranial nerve roots, part reaching is separated from blood in the sinus only by
the nasal cavity along sheaths of olfactory the thin epithelium of the arachnoid and the
nerve filaments. Around the spinal nerve vascular endothelium.
roots, an arrangement of dural veins and Arachnoid villi have been found in dogs,
362 • THE NERVOUS TISSUE

cats, monkeys, and humans. In man, with the tissue supplied. From this, it is clear that
advancing age, they enlarge and become pac¬ gray matter is metabolically much more ac¬
chionian corpuscles or granulations. tive; the amount of activity, however, varies
The villi provide the main pathway for from place to place, as reflected in the density
outflow of cerebrospinal fluid directly into of the capillary net.
the venous circulation. This flow is rapid. The linear extent of capillaries per unit
Dyes and other chemicals injected into the volume of brain substance has been meas¬
subarachnoid space can be detected in the ured in a number of representative parts of
bloodstream in 10 to 30 seconds. After only the CNS in various animals. In the rat, parts
30 minutes they can also be found in the of both white and gray matter differ in vas¬
lymphatics. cularity, but in all cases the gray is more
vascular than the white. Motor nuclei are less
vascular than sensory nuclei and groups of
Blood Vessels of the
interneurons.
Central Nervous System
There are no lymphatics in the CNS. Fluid
The arteries reach the spinal cord with the from the capillaries seeps through the tissue
ventral and dorsal nerve roots (anterior and but does not collect in lymphatic channels, as
posterior radicular arteries) and form a dense in most other parts of the body. The blood
arterial network in the spinal pia mater. Here vessels that penetrate from the pia are sur¬
several longitudinal arterial pathways can be rounded by perivascular spaces, which open
distinguished (spinal arterial tracts). The freely at the brain surface into the subarach¬
most important is the anterior spinal artery; noid space. Thus, cererospinal fluid derived
it emits many small branches (central arter¬ from blood drains from the brain outward
ies), which enter the ventral median fissure toward the meninges.
and penetrate to the right and left into the
medial part of the anterior gray columns,
thus supplying most of the gray matter with
Blood-Brain Barrier
blood. Numerous smaller branches of the pial When certain vital dyes are introduced into
arterial net (peripheral arteries) penetrate the circulating blood of adult animals, the
the white matter along its circumference. CNS remains colorless except for the choroid
Capillary nets in white matter are loose and plexuses and certain subependymal areas,
have meshes drawn out longitudinally; cap¬ where the dyes are found within extravascu-
illaries in gray matter are much more nu¬ lar cells. Thus, a blood-brain or hematoence¬
merous and dense. The course of the veins phalic barrier exists between the blood and the
does not correspond to that of the arteries. nervous system. The barrier is localized in
Numerous venous branches emerge from the the endothelium of the blood vessels. The
periphery of the cord and from the ventral zonulae occludentes between endothelial cells
median fissure and form a diffuse plexus in block movement of macromolecules along
the pia, which is especially prominent on the the intercellular clefts and there is little or
dorsal surface of the cord. From this plexus, no transport of solutes across the endothelial
blood is drained by veins accompanying the cells by plasmalemmal vesicles.
ventral and dorsal roots. In young animals given intravenous dye
The arterial supply of the brain derives injections, a distinct but small storage of dye
almost entirely from the carotids and the does occur in cells in parts of the brain stem.
large arteries at its base—the basilar artery The apparent impermeability of the brain to
and the circle of Willis. Most arteries from blood-borne macromolecules develops grad¬
these large vessels pass superiorly in the pia ually.
mater, from which small vessels dip into the
brain substance. These vessels are function¬
ally end arteries, since few anastomoses are
large enough to be effective in establishing
collateral circulation. RESPONSE OF THE
As in the spinal cord, the capillary net in NEURON TO INJURY
cerebral white matter is meager, with elon¬
gated meshes; in gray matter the net has a Two important responses of the neuron to
closer mesh. The density of capillaries is a injury are the progressive degeneration of an
crude indication of the metabolic needs of axon severed from its cell body and the
 THE NERVOUS TISSUE • 363

morphological changes that occur in the peri¬ Nerve fibers within the central nervous
karyon of a neuron after axonal transection. system also undergo wallerian degeneration.
These responses, basic to neuropathology, In young animals, degeneration is extremely
are included here because they exemplify the rapid, but in adult man it proceeds more
neuron doctrine and illustrate the trophic slowly. Loss of myelin in a heavily myelinated
influence of the nerve cell body over the tract, for example, as seen in Weigert stain,
axon and its other processes. may not be evident for two months. Special
When an axon is severed, trauma occurs staining methods for normal and abnormal
at both cut edges. Proximally, primary degen¬ lipids, however, show myelin catabolism more
eration extends only a short distance, depend¬ readily. In the CNS the microglia, equivalent
ing on the type of injury—one or two inter¬ to macrophages elsewhere, break down and
nodes in a clean cut, but as much as 2 or 3 remove the myelin and axonal fragments.
cm from a gunshot wound. Damage is soon Whether interfascicular oligodendrocytes un¬
repaired and new axonal sprouts appear. dertake similar functions to those of Schwann
Distally, however, the axon, its terminal ar¬ cells is not known. Wallerian degeneration is
borizations, and the myelin sheath completely widely exploited in neuroanatomical research
disintegrate. This secondary, or wallerian, de¬ to trace the course of connections following
generation in peripheral nerves usually pro¬ experimental ablations of nuclei and tracts.
ceeds simultaneously along the length of a Degeneration of terminal boutons, an essen¬
nerve fiber distal to the point of injury, but tial part of wallerian degeneration, offers a
a rapid centrifugal sweep of changes may more difficult but higher-resolution research
occur, The initial changes in axonal fine method, since it demonstrates the termina¬
structure consist in localized accumulations tion of degenerating axons and also serves
of mitochondria a few hours after injury; when brief survival precludes study of the
within 12 to 24 hours, neurofilaments and slower reactions of axon or myelin.
mitochondria vesiculate and disintegrate, and The fleeting and restricted traumatic de¬
later dense granules aggregate. At about two generation of the proximal stump of a pe¬
days, the axon becomes beaded and shortly ripheral nerve is usually soon followed by
begins to fragment and dissolve. Concomi¬ axonal regeneration, in the form of fine
tantly, myelin degeneration begins, breaking sprouts with protoplasmic growth cones or fil-
down into fatty droplets of varying size and opodia at their advancing tips. Regenerating
concentric lamellar structures, often enclos¬ fibers resemble embryonic ones and grow
ing an axonal fragment. Macrophages, de¬ along the outer edge of a band of Biingner.
rived somehow from the endoneurial or There they become progressively enfolded
Schwann cells, progressively remove the de¬ by Schwann cells in a fashion similar to the
bris. In accord with the neuron doctrine, the multiple envelopment found in unmyeli¬
process of degeneration does not usually nated nerves. The rate of growth of the fibers
spread across the synapse. Nevertheless, is rapid—3 to 4 mm a day in mammals—but
many exceptions are known and additional the distance that must be traversed to the
ones have been suspected. The Schwann cy¬ terminations may be 1 m or more.
toplasm remains and, after axonal and mye¬ Functional recovery depends on reestab¬
lin degeneration, forms a tube surrounded lishment of appropriate sensory and motor
by endoneurium and filled with fluid and connections at the periphery. The abundance
scattered fragments. The wall of the tube of regenerative sprouts from the original
thickens, reducing the original bore until it neuron and the capacity of bands of Biingner
may be obliterated, and the tube shrinks to to accommodate hundreds of fibers favor
perhaps less than half of its original diame¬ successful reinnervation. Although surgical
ter; meanwhile, its constituent Schwann cell alignment of individual axons is impossible,
nuclei multiply. Ultimately these changes and regenerating sprouts can negotiate hia¬
produce a solid, nucleated band of Biingner. tuses of considerable distance, prompt ap¬
As myelin disappears, a flexible, ribbon-like proximation of the two ends of a cut periph¬
peripheral nerve condenses into a dull gray, eral nerve is nevertheless of cardinal
hardened, and rounded cord. The bands of importance. In addition to the abundance of
Biingner may remain lor many months, sprouts and their accommodation by the
awaiting reinnervation. If regeneration of bands of Biingner, peripheral deletion of
axons does not occur, the bands gradually maladaptive terminations and central remod¬
become reduced by encroaching endoneurial eling of reflex arcs offer additional possibili¬
connective tissue. ties for functional recovery.
364 • THE NERVOUS TISSUE

Following the regeneration of a peripheral superabundance, beginning as a basophilic

nerve, the axon, myelin, and Schwann cells rim around the nucleus. Return to normal
slowly return to their normal size and con¬ appearance takes several months. Very large
dition. During these months, large amounts amounts of axonal cytoplasm may have to be
of axoplasm are resynthesized—possibly 250 regenerated, entailing an enormous meta¬
times that of the parent cell body. bolic effort on the part of the cell body and
Retrograde changes in the cell body of synthesis of immense amounts of cytoplasm
origin of a severed axon were first described relative to the size of the perikaryon. Sensory
by Nissl in 1892. Chief among these is retro¬ ganglion cells react in a manner similar to
grade chromatolysis, the apparent disappear¬ motor cells, but show less obvious changes
ance of the Nissl substance, obvious and well after lesions of the peripheral nerves and still
studied in motor cells, but occurring in vary¬ slighter, perhaps undetectable, changes after
ing extent and rate in other types of neurons. section of the dorsal roots. The finely divided
With light microscopy, breakup and dissolu¬ Nissl substance in the normal dorsal root
tion of Nissl material is first seen near the ganglion cell and the great length of axon it
axon hillock and in a rim around the nucleus, possesses probably tend to minimize the vis¬
subsequently spreading to other parts of the ible response. In many central neurons, such
cell. In addition, the perikaryon swells, bal¬ as Betz cells of the cerebral cortex, retrograde
looning the normally concave boundaries be¬ chromatolysis is seldom observed. Unless de¬
tween dendrites, and the nucleus shifts from struction of the axon and all its branches
its usually central position to a peripheral takes place, intact sustaining collaterals are
one away from the axon hillock. Fine struc¬ thought to remain functional and to provide
tural investigations have shown dispersion or enough axoplasm to offset overt retrograde
disappearance of ribosomes and of the cister¬ cell change.
nal membranes of the granular endoplasmic
reticulum, with appearance of numerous
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 p
12
BL 00D AND L YMPH
VASCULAR SYSTEMS

Multicellular animals require a mechanism this portion of the circulation that carries out
for distribution of oxygen, nutritive mate¬ the primary function of the vascular system.
rials, and hormones to the tissues, and to
collect from them carbon dioxide and other
products of metabolism for transmission to ARTERIES
the excretory organs. In vertebrates, this
function is carried out by the blood vascular
Blood is carried from the heart to the
system. This consists of a muscular pump, the
capillary networks in the tissues and organs
heart, and two systems of blood vessels. One
by arteries. These constitute an extensive
of these, the pulmonary circulation, carries
system of vessels beginning with the aorta and
blood to and from the lungs; the other, the
pulmonary artery, which emerge from the left
systemic circulation (peripheral circulation), dis¬
and right sides of the heart, respectively. As
tributes to all the other tissues and organs of
they course away from the heart, these vessels
the body. In both, the blood pumped from
branch repeatedly and thus give rise to large
the heart passes successively through arteries
numbers of arteries of progressively dimin¬
of diminishing size to networks of minute ishing caliber.
capillaries and back to the heart via veins of
The basic organization of the wall of all
increasing caliber.
arteries is similar in that three concentric
The heart delivers about 80 ml of blood layers can be distinguished: (1) an inner
into the pulmonary artery and aorta at each layer, the tunica intima, consisting of an endo¬
beat or about 6 liters a minute. The initial
thelial tube whose cells generally have their
velocity of flow is about 33 cm/min, but as long axis oriented longitudinally; (2) an in¬
the total cross-sectional area of the vascular termediate layer, the tunica media, composed
system is increased by progressive dichoto¬ predominantly of smooth muscle cells dis¬
mous branching of the arteries, the rate of posed circumferentially; and (3) an outer
flow gradually decreases. A further expan¬ coat, the tunica adventitia, made up of fibro¬
sion of the cross-sectional area of the system blasts and fibrous elements that are oriented,
occurs quite abruptly at the level of the cap¬ for the most part, longitudinally (Fig. 12-1).
illaries, resulting in a decrease in rate of flow This layer gradually merges with the loose
to about 0.3 cm/sec. The extensive capillary connective tissue that accompanies all blood
networks of the body have a total surface vessels. The boundary between the tunica
area of 700 m2 for exchange of metabolites. intima and tunica media is marked by the
It is only in capillaries and small venules that internal elastic lamina (elastic interna), which is
the vessel walls are thin enough to permit especially well developed in arteries of me¬
passage of substances to and from the sur¬ dium caliber. Between the tunica media and
rounding tissues. The other vessels are con¬ the tunica adventitia, a thinner external elastic
cerned with distribution of blood to the cap¬ lamina (elastica externa) is also present in many
illaries. At any given moment, only about 5 arteries.
per cent of the total blood volume is in the
capillaries and 95 per cent is on its way to or Elastic Arteries
from them. The structure and biochemical
properties of the capillary wall are subjects There is a continuous gradation in size and
of great physiological importance, for it is in character of the vessel wall from the largest

367
368 • BLOOD AND LYMPH VASCULAR SYSTEMS

A B
Figure 12-1. Drawings of the wall of a small artery in cross section, showing the concentric arrangement of tunica
intima, media, and adventitia. A, Stained with hematoxylin and eosin; B, stained with orcein to reveal the elastic tissue
component.

arteries down to the capillaries, but it is gitudinally. Adjacent endothelial cells are at¬
customary to designate several categories of tached by simple occluding junctions and
arteries on the basis of their size, the predom¬ occasional communicating (gap) junctions.
inant structural component of their tunica The cells possess all of the common organ¬
media, and their principal function in the elles in limited numbers, usually located in
arterial system. The large elastic or conducting the thicker region of the cytoplasm around
arteries, such as the aorta, innominate, subcla¬ the flattened nucleus. The endothelium is a
vian, common carotid, and common iliac, very slowly renewing population of cells that
have walls containing many fenestrated layers are rarely found in division. Their adluminal
of elastin in the tunica media (Fig. 12-2). and abluminal plasmalemmae have numer¬
Their walls may be distinctly yellow in the ous associated vesicles that are believed to be
fresh state because of the abundance of their involved in transendothelial transport of
elastic elements. These major conducting ves¬ water, electrolytes, and certain macromole¬
sels near the heart are distended during cules. Short blunt processes occasionally ex¬
contraction of the heart (systole), and the tend from the base of endothelial cells
subsequent elastic recoil of their walls during through fenestrae in the elastica interna and
diastole serves as a subsidiary pump main¬ establish communicating junctions with
taining continuous flow in the system despite smooth muscle cells of the media.
the intermittency of heartbeat. Rodlike cytoplasmic inclusions are ob¬
The tunica intima of these arteries consists served in electron micrographs of arterial
of the endothelium, a thin squamous epithe¬ endothelium. These were first reported 20
lium, separated from the elastica interna by years ago and have since been called Weibel-
loose connective tissue containing a few fibro¬ Palade bodies after the investigators who first
blasts, occasional smooth muscle cells, and drew attention to them. Their chemical na¬
sparse collagenous and elastic fibers. The ture and functional significance remained
endothelium provides a smooth lining layer obscure until recently. It was then found that
for the vessel and a partially selective diffu¬ their electron density diminished in response
sion barrier between the blood and the other to epinephrine administration and it was pos¬
tunics of the vessel wall. Its cells are polygonal tulated that they might contain a procoagu¬
in outline, 10 to 15 pm wide and 25 to 50 lant factor. It has now been shown that a
pm long, with their long axis oriented lon¬ Factor VUI-related antigen (von Willebrand
 BLOOD AND LYMPH VASCULAR SYSTEMS • 369

Subendothelial layer

Intima Long itud i na I ly

striated layer

Figure 12-2. Longitudinal section

through the posterior wall of the human Fenestrated
descending aorta. Elastic tissue is Media
elastic
black; the other elements are not
membrane
shown clearly. Elastic fiber stain. (After
Kolliker and von Ebner.)

Vei n

Elastica externa

Adventitia

Vei n

factor) can be localized immunocytochemi- ration of the elastic component of arterial

cally in the Weibel-Palade bodies. This coag¬ walls is difficult to visualize from study of
ulating factor is believed to be synthesized by histological sections, but it is possible to ex¬
arterial endothelial cells and secreted into the tract all other components with hot formic
blood plasma. acid and examine the elastin with scanning
The elastica interna is a less prominent electron microscopy (Fig. 12-3). In large elas¬
feature of elastic arteries than of muscular tic arteries, the internal elastic lamina is not
arteries. It is merely the first of many laminae as continuous a sheet as in other conducting
of elastin found in the media and has rather and muscular arteries, but may have numer¬
large fenestrae traversed by a network of ous large fenestrations traversed by an irreg¬
finer elastic fibers. The media is up to 500 ular network of elastin strands (Fig. 12-5A).
pm thick in the larger vessels and composed The tunica adventitia of elastic arteries is
of 40 to 70 fenestrated laminae of elastin 5 relatively thin, and consists of fibroblasts and
to 15 pim apart and interconnected by radi¬ collagen bundles of predominant longitudi¬
ally oriented finer strands of elastin. The nal orientation and a loose network of thin
spaces between elastic laminae are occupied elastic fibers. The walls of the larger elastic
by long branching smooth muscle cells, oc¬ arteries are too thick to be nourished by
casional fibroblasts, with associated collagen diffusion from the vessel lumen. Such vessels
fibers and other extracellular matrix compo¬ have a microvasculature of their own—small
nents. The elastica externa is relatively thin vessels call vasa vasorum that run in the ad¬
and inconspicuous, being simply the most ventitia and may penetrate some distance into
peripheral of the multiple elastic laminae of the media. Their penetration is deeper in
the media. The three-dimensional configu¬ large veins than in arteries.
370 • BLOOD AND LYMPH VASCULAR SYSTEMS

Figure 12-3. Scanning electron micrograph of the three-dimensional architecture of the elastin in a cross section of the
wall of rat aorta extracted with hot formic acid to remove all other tissue components. The elastic tissue consists of
multiple concentric sheets or laminae interconnected by radially oriented strands and fenestrated septa. In the intact
vessel, smooth muscle cells occupy the spaces demarcated by these elastic elements. (Micrograph from Wasano, K.
J. Electron Microsc. 32:33, 1983.)

junctions with the innermost smooth muscle

Muscular or Distributing Arteries
cells of the tunica media. The fenestrations
Elastic arteries gradually give way to mus¬ in the elastica interna are believed to be
cular or distributing arteries such as the bra¬ essential for nutrition of the avascular media
chial, femoral, radial, popliteal, and their by permitting diffusion of metabolites from
branches. Their tunica media consists mainly the lumen. The communicating junctions of
of smooth muscle that can actively change the cell processes that traverse some of the
the diameter of the vessel to adjust the vol¬ fenestrae serve to maintain metabolic cou¬
ume of blood delivered to meet the needs of pling of the endothelium to the smooth mus¬
the region supplied. This category includes cle of the media.
the majority of the vessels in the arterial The thickness of the media varies from 40
system and spans a wide range of sizes down layers of helically or circumferentially ori¬
to half a millimeter in diameter. ented smooth muscle cells in large arteries to
The intima is somewhat thinner than that three or four layers in small arteries. The
of the elastic arteries and lacks smooth muscle individual cells are surrounded and sepa¬
cells but is otherwise similar in its organiza¬ rated from one another by a moderately thick
tion. Beneath the intima is a well-developed external lamina comparable with the basal
elastica interna, which often has an undulat¬ lamina of epithelia (Fig. 12-7). Short proc¬
ing contour in the contracted vessels nor¬ esses extend from cell to cell through small
mally encountered in histological sections discontinuities in this layer to form commu¬
(Figs. 12—4, 12—6). The endothelium closely nicating junctions. These low-resistance junc¬
conforms to all the irregularities in the un¬ tions are probably essential for coordination
derlying elastica and sends processes through of contraction throughout the media. The
its fenestrations to establish myoendothelial external lamina investing the smooth muscle
 BLOOD AND LYMPH VASCULAR SYSTEMS • 371

Figure 12-4. Photomicrograph of a muscular artery from the rat, showing the elastica interna, the media, the elastica
externa, and a thick adventitia. Areas comparable with those enclosed in the rectangles are illustrated in electron
micrographs in Figures 12-6 and 12-8. Aldehyde fuchsin stain for elastin.

cells stains deeply with the periodic tochondria and numerous synaptic vesicles.
acid—Schiff reaction. Embedded within this The nerves ordinarily do not penetrate into
abundant interstitial material are small bun¬ the media but appear to terminate at the
dles of collagen fibrils corresponding to the elastica externa (Fig. 12-8). The neural stim¬
network of delicate reticular fibers seen sur¬ ulation of the muscle cells evidently results
rounding individual muscle fibers in silver- from diffusion of the neurotransmitter
stained preparations viewed with the light through fenestrations in this layer. The re¬
microscope. Loose networks of thin elastic sulting depolarization of the peripheral mus¬
fibers also course circumferentially in the cle cells is propagated throughout the media
media and can be recognized as dark wavy via low-resistance cell-to-cell contacts (gap
lines among the smooth muscle cells in prep¬ junctions or nexuses) between the muscle
arations stained with aldehyde-fuchsin or re- cells.
sorcin-fuchsin (Fig. 12—4). In electron micro¬ The tunica adventitia of muscular arteries
graphs they appear as unstained elongated may be thicker than the media (Figs. 12—4,
profiles of irregular outline. 12—9) and consists of fibroblasts, elastic fibers,
The elastica externa often appears in his¬ and bundles of collagen, oriented, for the
tological sections to be a continuous lamina most part, longitudinally. These continue
at the boundary between the media and the into the surrounding connective tissue with¬
adventitia (Fig. 12—4), but in electron micro¬ out a clearly defined boundary. The loose
graphs it is an interrupted layer of irregular consistency of the tunica adventitia and pre¬
sheets of elastin considerably thinner than dominant longitudinal orientation of its com¬
the elastica interna. Closely applied to the ponents permits the continual changes in the
elastica on its outer aspect are numerous diameter of the vessel and limits the amount
small fascicles of unmyelinated nerve axons, of retraction that takes place when an artery
some containing local accumulations of mi¬ is cut.
Figure 12-5. A, Scanning micrograph showing a surface view of the aortic internal elastic lamina of the rat. It is
characterized by large fenestrations traversed by an irregular meshwork of elastin strands. B, In contrast, a similar view
of the internal elastic lamina of the femoral artery shows small round fenestrations. (Micrographs from Wasano K.
J. Electron Microsc. 32:33, 1983.)

Lumen

Endothelium

Elastica
inferno

Smooth
muscle

Figure 12-6. Transmission electron micrograph of a section through the wall of a small muscular artery (for orientation
see upper box in Fig. 12-4). The elastica interna is scalloped owing to agonal contraction of the vessel wall. It is
traversed at intervals by small fenestrations through which processes of endothelial cells extend to contact smooth
muscle cells of the media.
372
Figure 12-7. Electron micrograph of a portion of the wall of a small artery in longitudinal section. The elastica interna
does not stain and therefore appears as a clear area between the endothelium and the smooth muscle of the media.

Figure 12-8. Electron micrograph of the junctional zone between the media and adventitia of a small muscular artery
(see lower box in Fig. 12-4). The smooth muscle layer is limited on its outer aspect by a discontinuous elastica externa.
Closely applied to this are numerous small nerves, some of whose axons contain many synaptic vesicles.
373
374 • BLOOD AND LYMPH VASCULAR SYSTEMS

Innominate Artery

Thoracic Aorta

Arch of Aorta

Anterior Cerebral Artery Radial Artery Femoral Artery

Figure 12-9. Photomicrographs of the walls of elastic and muscular arteries of a macaque, illustrating variations in
relative thickness and the differing amounts and distribution of elastic tissue, which has been selectively stained with
resorcin fuchsin. (From Cowdry, E. V. Textbook of Histology. Philadelphia, Lea and Febiger, 1950.)

Transitional and Specialized Arteries separate or interrupt the elastic laminae in

many places. The visceral arteries that arise
In the gradual transition of one type of from the abdominal aorta are also of mixed
artery to another, it is sometimes difficult to type. For a varying distance, in the transi¬
classify the intermediate region. Some arter¬ tional region, the tunica media may consist
ies of intermediate caliber (e.g., popliteal, of two different layers—an internal muscular
tibial) have walls that resemble larger arteries, layer and an external, composed of typical
while some large arteries (e.g., external iliac) elastic laminae.
have walls not unlike those of medium-sized The thickness of the media of arteries is
arteries. The transitional regions between adapted to the internal pressure and external
elastic and muscular arteries are often des¬ forces to which they are subjected. The cor¬
ignated arteries of mixed type. Examples are the onary arteries of the heart are subjected to
external carotid, axillary, and common iliac high pressure and have a wall that is thicker
arteries. Their walls contain islands of than other muscular arteries of comparable
smooth muscle fibers in the tunica media that size. Similarly, in the arteries of the lower
 BLOOD AND LYMPH VASCULAR SYSTEMS • 375

limbs, the tunica media is thicker than in of circumferential fibers. In many places the
corresponding arteries in the upper limbs. longitudinal muscle raises longitudinal ridges
Blood pressure in the pulmonary circula¬ protruding into the lumen. The extra-ab¬
tion is considerably lower than in the systemic dominal portion of the umbilical artery is
circulation. Accordingly, the blood vessels in provided with numerous rounded swellings
the lungs are relatively thin walled and dis¬ or varicosities. In these regions the wall be¬
tensible. A unique feature of the vessels of comes thin and consists almost exclusively of
the lung is the presence of cardiac muscle the circularly oriented muscle.
extending from the heart a short distance
into the wall of the pulmonary artery.
Arterioles
Within the cranial cavity, where vessels are
protected from external pressures and ten¬ The small arteries and arterioles form an
sion, the dural and cerebral arteries have important segment of the circulation, for
relatively thin walls. The elastica interna is they constitute the principal component of
well developed but the media is thin and the peripheral resistance to flow that regu¬
devoid of elastic fibers (Fig. 12-9). lates the blood pressure. Arterioles range in
Where vessels are subject to bending or diameter from 300 (am down to less than 50
stretching, as in the popliteal and axillary (am. The tunica intima consists of a continu¬
arteries, longitudinal bundles of smooth mus¬ ous endothelium, a thin basal lamina, and a
cle fibers may be found in the intima. very thin subendothelial layer consisting of a
The umbilical artery, carrying blood from few reticular and elastic fibers. The elastica
the fetus to the placenta, has an atypical interna is thin and fenestrated and is absent
structure. Its intima consists only of endothe¬ in the terminal arterioles. The tunica media
lium, and an internal elastic layer is lacking. of larger arterioles may consist of two layers
The tunica media contains a few elastic fibers of smooth muscle cells, but in most arterioles
and two thick muscular layers that are quite there is a single layer of helically arranged
distinct. The inner layer is composed of lon¬ smooth muscle cells (Figs. 12-10, 12-11,
gitudinally oriented fibers; the outer layer, 12—12). In the short transitional region be-

Figure 12-10. Scanning micrograph of a branching arteriole showing the circumferential arrangement of the single
layer of smooth muscle cells. (Micrograph from Uehara, Y., and K. Suyama. J. Electron Microsc. 27:157, 1978.)
Figure 12-11. Electron micrograph of a typical small arteriole with one or two layers of smooth muscle cells around the
endothelium.

Figure 12-12. Arteriole from the corticomedullary boundary of the mouse thymus, 5 minutes after intravenous injection
of peroxidase. The endothelium is freely permeable to this protein. The dense reaction product is seen in the lumen,
between endothelial cells and filling the fenestrations in the elastica interna and the extracellular space around the
single layer of smooth muscle cells. (Micrograph from Raviola, E., and M. J. Karnovsky. J. Exp. Med. 136:466, 1972.)
376
 BLOOD AND LYMPH VASCULAR SYSTEMS • 377

tween arterioles and capillaries, sometimes skin by blood at body temperature is in¬
called metarterioles, there are single muscle creased manyfold, resulting in a greater loss
cells spaced at intervals. Their contraction is of heat. Consistent with a role in temperature
believed to give this region a sphincter-like regulation is the finding of significant differ¬
function, intermittently opening and closing. ences in distribution and numbers of arterio¬
The adventitia of arterioles consists of venous anastomoses in various animal species
loose connective tissue with occasional asso¬ related to their mode of thermal regulation.
ciated fibroblasts, macrophages, and small For example, in sheep in which the insulating
unmyelinated nerves. fleece limits evaporative heat loss from much
of the body surface, the ears, nose, and lower
legs, which are devoid of wool, are of great
Arteriovenous Anastomoses
importance in regulation of body tempera¬
In many parts of the body the terminal ture. The density of arteriovenous anasto¬
ramifications of arteries are connected with moses per square centimeter of skin (72) over
veins, not only by capillaries, but also by the lower leg is nearly five times that over
direct arteriovenous anastomoses of larger the trunk (15). Similarly, in fur seals, which
caliber. They usually arise as side branches are provided with an internal insulating layer
from terminal arterioles and run directly to of subcutaneous blubber and thick fur for
small venules. Their walls are muscular, re¬ external insulation, the density of arteriove¬
markably thick for the caliber of the vessel, nous anastomoses is seven to ten times
and richly supplied with vasomotor nerves. greater in the bare flippers, which can be
Observation of living vessels has shown that used to dissipate heat when the seal is out of
they contract markedly on stimulation of the its cold aquatic environment.
sympathetic nerves. When the arteriovenous Although most arterial vessels are inner¬
anastomosis is contracted, blood passes along vated by sympathetic nerves, both adrenergic
the arteriole into the capillary network; when and cholinergic nerves are found in arterio¬
it relaxes, blood can bypass the capillaries venous anastomoses, and the presence of
and go directly into a thin-walled venule. nonadrenergic, noncholinergic nerve end¬
The arteriovenous anastomoses are therefore ings has been reported. The contractions of
considered important structures for regulat¬ arteriovenous anastomoses are described as
ing the supply of blood to many tissues. rapid, forceful, and relatively independent
In addition to these simple direct shunts, of the constrictions of neighboring arteries.
more highly organized connections have There is evidence that they are under control
been described as part of a small organ called of thermoregulatory centers in the central
the glomus. These are found in the nailbed, nervous system, whereas peripheral arteries
the pads of the fingers and toes, and the are more responsive to locally generated
ears. The afferent arteriole enters the con¬ stimuli.
nective tissue capsule of the glomus, loses its The glomus type of arteriovenous anasto¬
internal elastic lamina, and develops a heavy moses that possess modified smooth muscle
epithelioid muscle coat and narrow lumen. cells with an epithelioid appearance, multiple
The arteriovenous anastomoses within the convoluted channels, and an exceptionally
glomus may be branched and convoluted, rich innervation are considered by many in¬
and they are richly innervated by sympathetic vestigators to be far more complex than
nerves. The anastomoses empty into a short, would be required for simple shunting of
thin-walled vein with a wide lumen, which blood, and are suspected of having an addi¬
drains into a periglomic vein and then into tional function yet to be discovered.
the ordinary veins of the skin.
Arteriovenous anastomoses are a device for Sensory Organs of Arteries
shunting arterial blood directly into the ve¬
nous system. By opening or closing, they can Sensory nerves are associated with arteries
vary the amount of flow through the associ¬ throughout the vascular system, but at certain
ated capillary bed. Their physiological signif¬ sites there are specialized neural organs
icance in many organs is still poorly under¬ whose function is to monitor the pressure
stood, but there is considerable evidence that and composition of the blood. These are of
those in the integument on the extremities great importance in regulating respiration,
of some species have an important role in heartbeat, and the vasomotor activity con¬
thermoregulation. When arteriovenous an¬ trolling blood pressure.
astomoses open, the rate of perfusion of the The carotid bodies are inconspicuous flat-
378 • BLOOD AND LYMPH VASCULAR SYSTEMS

tened bodies about 3 mm wide and 5 mm that glomus cells also secrete a hormone
long, associated with the vessel walls at the cannot be ruled out. On the basis of their
bifurcation of the common carotid into inter¬ ultrastructure and cytochemical properties,
nal and external carotid arteries. They are some authors consider them paracrine cells
richly innervated by the carotid sinus branch probably secreting a polypeptide, but as yet
of the glossopharyngeal nerve and by a no peptide hormone has been identified in
plexus of sympathetic components from the the carotid body.
vagus and glossopharyngeal nerves. The ca¬ Other chemoreceptor organs, the aortic
rotid bodies are highly vascular structures bodies, are situated near the arch of the aorta
with a large blood flow in relation to their between the angle of the subclavian and the
small volume of parenchyma. They consist common carotid on the right, and medial to
of irregular clumps of pale-staining epithe¬ the origin of the subclavian artery on the
lioid cells in intimate relation to sinusoidal left. Their structure and function appear to
capillaries lined with fenestrated endothe¬ be identical to those of the carotid bodies.
lium. Two types of cells can be distinguished At the bifurcation of the common carotid,
with the light microscope on the basis of the vessel exhibits a slight dilatation called
characteristic differences in their nuclei. the carotid sinus. This local specialization usu¬
They are more clearly distinguishable in elec¬ ally involves the proximal part of the internal
tron micrographs. carotid. The tunica media is thinner than
The Type I cell (glomus cell) contains numer¬ elsewhere, while the adventitia is thicker and
ous dense-cored vesicles and occurs in clus¬ contains a large number of sensory nerve
ters that are surrounded by Type II cells, endings derived from the glossopharyngeal
which contain no dense-cored vesicles and nerve. These nerve endings are stimulated
are commonly regarded as sheath cells. by stretching. The carotid sinus therefore
These may yet prove to have a more signifi¬ serves as a baroreceptor reacting to changes in
cant function than is implied by that term. blood pressure and initiating reflexes that
The Type I cells were originally considered bring about appropriate modifications of sys¬
to be a homogeneous population but sub- temic pressure. A few baroreceptors are also
types have now been identified in several found in the wall of the aorta and other large
mammalian species. Type A cells usually have arteries in the thorax and neck, but these are
a smooth globular contour with a few proc¬ not visually identifiable.
esses, while Type B is irregular in outline
with several long thin processes. The dense-
cored vesicles of A glomus cells are twice as PHYSIOLOGICAL IMPLICATIONS OF
abundant as those of B cells and about 30 THE STRUCTURE OF ARTERIES
per cent larger.
Many nerves ramify throughout the ca¬ The flow of blood in the arteries results
rotid body and have endings rich in synaptic from intermittent contraction of the heart
vesicles that terminate mainly on type A glo¬ and is therefore pulsatile. If the wall of the
mus cells. There is continuing controversy as arteries were inflexible, the flow of blood in
to whether these nerves are afferent (con¬ the capillaries would be intermittent. How¬
ducting to the brain) or efferent (conducting ever, because the large arteries near the heart
to the carotid sinus). The bulk of recent have distensible elastic walls, only part of the
evidence indicates that over 90 per cent of force of cardiac contraction is dissipated in
them are afferent and have reciprocal syn¬ advancing the column of blood in the vessels;
apses with the glomus cells. The precise role the rest of the energy goes to expanding the
of the glomus cell in chemoreceptor function walls of the large elastic arteries. The poten¬
is not clear. The traditional interpretation tial energy accumulated in the stretching of
considered them to be chemoreceptors stim¬ the walls of these vessels during contraction
ulated by hypoxia, hypercapnia, or elevated of the heart (systole) is dissipated in the elastic
hydrogen ion concentration in the blood. recoil of the vessel wall during the period
They were assumed to pass information on when the heart is inactive (diastole). This
the associated afferent nerves. An alternative release of tension in the arterial wall serves
interpretation postulates that the afferent as an auxiliary pump, forcing the blood on¬
nerve endings are the chemoreceptors, and ward during diastole when no forward pres¬
that the glomus cells are dopaminergic inter-, sure is exerted by the heart. Thus, although
neurons that modulate the sensitivity of che- the flow of blood is pulsatile throughout
moreceptive nerve endings. The possibility much of the arterial system, the elasticity of
 BLOOD AND LYMPH VASCULAR SYSTEMS • 379

the walls of the large vessels ensures contin¬ tion. If the vessels are then rapidly fixed in
uous flow through the capillaries. this condition and sections are subsequently
The musculature in the media of the dis¬ cut through the open and the adjacent con¬
tributing arteries is normally in a state of stricted segments of the same vessel, one can
partial contraction, referred to as tone, but clearly observe the great decrease in caliber
the degree of contraction can be modulated of the lumen and the striking change in the
in response to changes in pressure in the character of the wall that accompanies vaso¬
system and variations in activity of various motor activity (Figs. 12—13, 12—14). Such
tissues and organs. The vasoconstriction and contractions and relaxations of the muscular
vasodilation of arteries is regulated in part by walls of arteries obviously influence the dis¬
the autonomic nervous system. In the major¬ tribution of blood to the various organs. They
ity of arteries seen in routine histological also change the peripheral resistance to flow
sections, the smooth muscle of the tunica and therefore affect the blood pressure. Con¬
media has undergone some degree of con¬ traction of arteries may be the result of stim¬
traction, either after death or in response to ulation by sympathetic nerves, or of direct
the stimulus of immersion in a chemical fix¬ action of local products of injury. This latter
ative. One therefore gets a somewhat erro¬ effect is important in limiting hemorrhage
neous impression of the thickness of the from torn vessels in sites of tissue injury.
arterial wall in relation to the size of the There are other local factors acting at the
lumen. The remarkable capacity of the arte¬ level of small arteries and arterioles that tend
ries to change their caliber is observed best in to regulate blood flow. If blood flow is inter¬
the living anesthetized animal. If a micro¬ rupted briefly, the lack of oxygen and accu¬
droplet of norepinephrine is placed on an mulation of carbon dioxide and lactic acid
artery, the underlying portion of the vessel tend to cause relaxation of smooth muscle
wall will undergo marked local vasoconstric¬ and vasodilation, so that when circulation is

Figure 12-13. Low-power micrographs of two cross sections less than 1 mm apart in the same frog arteriole. A
microdroplet applied to the living vessel caused local vasoconstriction in the area indicated by the brackets (inset). The
vessel was then fixed in situ and sectioned. The two sections offer a dramatic demonstration of the structural correlates
of vasoconstriction and vasodilation. (From Phelps, P. C., and J. H. Luft. Am. J. Anat. 125:399, 1969.)
380 • BLOOD AND LYMPH VASCULAR SYSTEMS

Figure 12-14. A, Micrograph of a typical small artery fixed with its lumen open. B, Micrograph of an artery of comparable
size that has undergone agonal vasoconstriction. The cell bodies of the endothelial cells have increased in height, and
they now almost completely occlude the lumen. Such extreme vasoconstriction probably occurs only rarely in normal
physiological conditions, but it is evident that at sites of injury this could effectively reduce hemorrhage. (Micrograph
courtesy of R. Bolender.)

restored the rate of flow may be two to six endothelium has proved to be quite erro¬
times greater than it was before. This impor¬ neous. Endothelial cells have now been
tant local autoregulation or reactive hyper¬ shown to be metabolically quite active and
emia correcting metabolic deficits does not capable of synthesizing and releasing a vari¬
depend on the nervous system. ety of substances into the blood. They are
The maintenance of normal blood pres¬ believed to secrete the A and B blood-group
sure depends in large measure on the pe¬ antigens and certain of the clotting factors of
ripheral resistance to flow in the system, the blood plasma. They also synthesize pros¬
which in turn is a function of the smooth tacyclin, a potent inhibitor of platelet aggre¬
muscle tone in the walls of arterioles and gation. The endothelium thus contributes
small arteries. Halving the diameter of arte¬ both to maintenance of the fluidity of the
rioles can increase the resistance as much as blood in the intact circulation and to the
16-fold, and a constriction to one fourth their limitation of blood loss in the event of injury
original diameter may increase resistance as to the vessels. Enzymes in the plasmalemma
much as 250 times. This is the basis for the of endothelial cells inactivate norepineph¬
powerful effect of catecholamines and vaso¬ rine, serotonin, and bradykinin and trans¬
active peptides, such as angiotensin, on blood form angiotensin I to the potent vasoactive
pressure. substance angiotensin II.
The entire vascular system is lined by en¬
dothelium. The function of this cellular layer
was long believed to be limited to providing CHANGES IN ARTERIES WITH AGE
the vessels with a smooth internal surface
that would facilitate flow and prevent activa¬ The structure of the wall of large arteries
tion of the intrinsic coagulation mechanism undergoes a gradual process of further dif¬
of the blood. It was not thought to have ferentiation from birth to age 25. In elastic
significant metabolic or biosynthetic func¬ arteries there is a progressive thickening and
tions. This limited view of the role of the development of increasing numbers of elastic
 BLOOD AND LYMPH VASCULAR SYSTEMS • 381

laminae, while in muscular arteries the thick¬ proteoglycan results in a local thickening of
ness of the media increases with little or no the intima. With progression of the disease,
addition of elastin. From middle age onward, local extravasation of blood, necrosis, and
there is a relative increase in the collagen calcification result in what are described as
and proteoglycans, and the walls of the larger complicated lesions. These become sites of ero¬
arteries become less pliant. Perhaps the most sion of the endothelium, and aggregation of
significant age-related changes are in the in- blood platelets to form a mural thrombus
tima. At birth it consists of endothelium and (clot). In advanced atherosclerosis, there may
the internal elastic lamina, with little inter¬ be necrosis and calcification of the tunica
vening connective tissue and only occasional media of the muscular arteries in addition to
smooth muscle cells. With increasing age, the lesions in the intima.
extracellular matrix components slowly ac¬ The principal processes involved in arteri¬
cumulate and smooth muscle cells become osclerosis thus appear to be proliferation of
more numerous. smooth muscle cells; their production of ex¬
Arteries are constantly exposed to mechan¬ cess collagen, elastin, and proteoglycan; and
ical stresses associated with oscillations of the intracellular and extracellular accumula¬
pressure and pulsatile flow in their lumen, tion of lipid. Research on the pathogenesis
and seem to be more subject to wear and of the disease is now concentrating on the
tear than other organs. The late stages of respective roles of the endothelial barrier;
development of their walls are not sharply the various plasma lipoprotein fractions; and
differentiated from the early regressive the mitogenic agents released by aggregated
changes associated with aging and the onset blood platelets.
of atherosclerosis (“hardening of the arteries”).
Some authors view these early changes as
physiological; others consider them patholog¬
ical. The larger arteries—particularly the
CAPILLARIES
aorta, iliac, femoral, coronary, and cerebral
arteries—are especially prone to atheroscle¬ The branches of terminal arterioles pass
rosis. This disease is the principal cause of through a short transitional region in which
myocardial infarction (heart attack) and cerebral scattered smooth muscle cells persist around
thrombosis (stroke) and is the leading cause of the endothelial tube. The vessels then con¬
death in America and Europe. It is charac¬ tinue as true capillaries that branch and an¬
terized by a patchy, irregular thickening of astomose with little or no change of diameter
the intima with intracellular and extracellular forming extensive capillary networks (Fig.
deposits of lipid. The pathogenesis of the 12-15).
disease is still poorly understood. By the age The capillary wall consists of a layer of
of 10 years, small focal accumulations of extremely attenuated endothelial cells, a basal
intimal smooth muscle cells containing and lamina, and a sparse network of reticular
surrounded by deposits of cholesterol-rich fibers. Occasional undifferentiated mesen¬
lipid form yellow “fatty streaks” that are chymal cells may be associated with the cap¬
visible to the naked eye on the luminal aspect illary wall. In certain sites the pericapillary
of the aorta. These lesions that are asymp¬ cells are more highly differentiated and have
tomatic increase to occupy 30 per cent or long branching processes that extend circum¬
more of the aortic intimal surface by the age ferentially around the capillary (Figs. 12—16,
of 25. Whether these fatty streaks are invar¬ 12—17). It has been speculated that these may
iably precursors of the more advanced lesions be contractile. It is customary to refer to all
of atherosclerosis is still controversial. More such cells as pericytes without attributing to
patently pathological are lesions called fibrous them a special role in regulating the caliber
plaques in older individuals. These are white of the vessel (Fig. 12-18). The endothelial
and elevated so that they project slightly into cells themselves may contract after mechani¬
the lumen. There is substantial agreement cal stimulation and it is therefore unnecessary
that these arise by proliferation of intimal to ascribe variations in size of the lumen to
smooth muscle cells and by migration of such special cells in the capillary wall.
cells from the media through fenestrations The caliber of the capillaries in different
in the internal elastic lamina. Lipid accumu¬ parts of the body varies within relatively
lates in and around these cells, and their narrow limits and averages about 8 to 12 pm,
deposition of excess collagen, elastin, and which permits unimpeded passage of the
382 • BLOOD AND LYMPH VASCULAR SYSTEMS

Figure 12-15. Normal human retinal blood vessels. These have been isolated by tryptic digestion of the neural and
receptor elements, leaving behind only the vessels. At left is an arteriole, at right a venule, and between them is a
network of capillaries of very uniform caliber. (Courtesy of T. Kuwabara.)

Pericytes Figure 12-16. Photomicrograph of an intact capillary in

a whole mount of rat mesentery. The nuclei of the flattened
endothelial cells lining the capillary can be distinguished
from those of the pericytes, which bulge outward. May-
Grunwald-Giemsa stain.
 BLOOD AND LYMPH VASCULAR SYSTEMS • 383

Figure 12-17. Photomicrograph of a capillary in skeletal

muscle cut longitudinally. The nuclei of a pericyte, an
endothelial cell, and a fibroblast can be distinguished.
Several erythrocytes in the lumen give an indication of its
dimensions. (Courtesy of J. Venable.)

Figure 12-18. Scanning micrographs of small blood vessels showing pericytes with processes encircling the vessel
wall. A, Pericyte of a capillary with primary processes directed longitudinally and secondary processes running
circumferentially. B, Arterial capillary with numerous associated pericytes. C, Terminal arteriole with pericyte and circular
smooth muscle cells. (Micrographs from Uehara, Y„ and K. Suyama. J. Electron Microsc. 27:157, 1978.)
384 • BLOOD AND LYMPH VASCULAR SYSTEMS

cellular elements of the blood. In organs that by circular fenestrations or pores 80 to 100
are in a state of minimal functional activity, nm in diameter closed by a very thin pore
many of the capillaries are narrowed so that diaphragm with a punctate central thickening
little blood circulates through them. Nor¬ (Fig. 12—246). When seen in surface view in
mally only about 25 per cent of the total scanning micrographs or freeze-fracture
capillary bed in the body is patent, but with preparations, the pores are very uniformly
increased activity, these open and flow is distributed with a center-to-center spacing of
restored to meet local needs. about 130 nm. In these fenestrated capillaries
In cross sections of small capillaries, a sin¬ (visceral capillaries), the areas exhibiting pores
gle endothelial cell may extend all around make up only a fraction of the vessel wall,
the lumen (Fig. 12-19). In larger capillaries the remainder resembling the endothelium
the wall may be made up of portions of two of muscle-type capillaries. In this mosaic of
or three cells. The nucleus of endothelial fenestrated and unfenestrated areas, the rel¬
cells is greatly flattened and thus appears ative proportions vary in capillaries of differ¬
elliptical in section. The thicker nuclear re¬ ent organs. Among fenestrated capillaries,
gion of the cell bulges into the lumen. The those of the renal glomerulus appear to be
attenuated peripheral portion of the cell is exceptional in that the pores are not closed
extremely thin, with the adluminal and ablu- by pore diaphragms*, and their basal lamina
minal membranes separated by a layer of is as much as three times thicker than that of
cytoplasm only 0.2 to 0.4 pm thick. A small other capillaries. Fluid traverses the wall of
Golgi complex and a few mitochondria are glomerular capillaries as much as 100 times
found in the juxtanuclear region, with mean¬ more rapidly than in muscle capillaries.
dering tubular elements of the endoplasmic
reticulum extending into the thin peripheral
cytoplasm. Tysosomes are rare but multives- STRUCTURAL BASIS OF
icular bodies are not uncommon. A conspic¬ TRANSENDOTHELIAL EXCHANGE
uous feature of endothelial cells is the pres¬
ence of a large population of plasmalemmal An important concern of physiologists has
vesicles about 70 nm in diameter with narrow been the mechanism of exchange across the
necks, present on both cell surfaces and capillary wall. The observed rates of passage
opening onto the lumen and to the extravas- of water-soluble molecules can be accounted
cular space (Fig. 12-20). for by postulating two fluid-filled systems of
The luminal surface of the cells is usually pores traversing the endothelium: one of
smooth contoured, but edges of adjacent cells “small pores” about 9 nm in diameter and of
often overlap and a thin marginal ridge or relatively high frequency, and the other of
flap may project a short distance into the “large pores” up to 70 nm in diameter and
lumen (Fig. 12-21). Desmosomes and a zon¬ of lower frequency. Pores as such are not
ula adherens are usually absent but there is seen in electron micrographs of muscle cap¬
a shallow occluding junction (Fig. 12-22), illaries and the structural equivalent of the
which in freeze-fracture preparations ex¬ postulated two sets of pores has been a sub¬
hibits one to three parallel intramembranous ject of lively debate. To clarify this issue,
strands on the E-face. electron-dense molecules of known dimen¬
At the resolution afforded by the light sions greater than 10 nm are introduced into
microscope, capillaries in different tissues the circulation. In muscle capillaries, these
and organs appear quite similar, but with the molecules are rapidly taken up by open ves¬
electron microscope at least two morpholog¬ icles on the adluminal surface of the contin¬
ically distinct types can be distinguished on uous endothelium, ferried across the cyto¬
the basis of differences in the endothelium plasm, and discharged into the extravascular
(Fig. 12—23). In muscle, nervous tissue, and space by fusion of the vesicles with the ablu-
connective tissues of the body, the endothe¬ minal plasmalemma (Fig. 12—26T).
lium forms a thin uninterrupted layer The uptake of materials in small vesicles is
around the entire circumference of the cap¬ a form of endocytosis common to many cell
illary. These are designated continuous capil¬ types and is generally referred to as micropi-
laries (muscle-type capillaries). In pancreas, in¬ nocytosis. In this process, the vesicles generally
testinal tract, and endocrine glands the fuse with lysosomes or are incorporated in
endothelium varies in thickness, and some multivesicular bodies within the cytoplasm.
extremely attenuated regions are interrupted The use of vesicles to ferry fluid and solutes
 BLOOD AND LYMPH VASCULAR SYSTEMS 385

Perivascular

Basal
X / lamina
Endothelial
nucleus
t ■' '

Capillary lumen

Endothelial
cell junction

Collagen

Figure 12-19. Electron micrograph of a typical capillary from guinea pig pancreas. The entire circumference is made
up of a single endothelial cell. There is a thin basal lamina and a few associated collagen fibrils. No pericyte is present
in this cross section. (Micrograph from Bolender, R. J. Cell Biol. 61:269, 1974.)
 Capillary lumen

Nucleus

Plasmaiemmal
vesicles

; 81 rm- ^
V I* *

f #. • >»
Muscle
fiber , .
^ r& %
..
v * i r* «
i * ->

:
. - * ' 4 fc * * v
; . >jor * ' ■ - ..
J? * %Ti , ** * -
"" \
*
Figure 12-20. Electron micrograph of capillary endothelium, illustrating the small vesicular inpocketings of the luminal
and basal surfaces that are characteristic of capillaries in muscle. (From Fawcett, D. W. J. Histochem. Cytochem. 73:75,
1965.) Inset shows two such vesicles at high magnification. (From Bruns, R., and G. E. Palade. J. Cell. Biol. 37:244,
1968.)

Basal
lamina

Endothelial junction

: V. ■, 'u# JP 4#%*:

!« v Hf
MwScle fiber
4- V,

, VS *■ 4 V •: > ,
<•' * - > * » * x*' •• 4
S. 88 , x * * US

Figure 12-21. Capillary from cardiac muscle, illustrating the interdigitating cell junction and a marginal fold.
386
 BLOOD AND LYMPH VASCULAR SYSTEMS • 387

Figure 12-22. Micrograph of the junction of two endothelial cells in a muscle-type capillary. At the arrows the opposing
membranes are joined to form an occluding junction. (Micrograph courtesy of E. Weihe.)

Figure 12-23. Schematic representation of the two most common types of capillaries. A, The continuous or muscle
type with an uninterrupted endothelium. B, The fenestrated type, in which the endothelium varies in thickness and the
thinnest areas have small pores closed by an exceedingly thin membranous diaphragm. (After Fawcett, D. W. In
Orbison, J. L., and D. Smith, eds.: Peripheral Blood Vessels. Baltimore, Williams and Wilkins, 1962.)
 iiS

Figure 12-24. Micrographs of segments of endothelium from the two types of capillary. A, Endothelium of the muscle
capillary endothelium has vesicular invaginations of both adluminal and abluminal plasma membranes (at arrows). B,
Endothelium of fenestrated capillary from the lamina propria of the colon is extremely thin and has pores closed by thin
diaphragms (at arrows). (Micrographs courtesy of E. Weihe.)

Figure 12-25. Replica of a freeze-fractured preparation of a fenestrated capillary. This extensive surface view of the
cleaved membrane of an endothelial cell shows fenestrated areas separated by nonfenestrated areas. Notice the
uniform size and spacing of the pores. (Micrograph courtesy of S. McNutt.)
388
 BLOOD AND LYMPH VASCULAR SYSTEMS • 389

VESICULATION OF THE PLASMALEMMA

FORMATION OF CHANNELS AND FENESTRAE

Figure' 12-26. Schematic representation of alternative models for transport of water-soluble molecules across the
endothelium. A, Continuous formation of plasmalemmal vesicles followed by detachment, transit, and fusion with the
membrane on the other front of the cell. B, Transport mediated by a separate cytoplasmic population of vesicles,
possibly of Golgi origin, which undergo transient fusion and fission first at one surface and then at the other without
mixing of their membrane with the plasmalemma. This would not require massive movement of membrane from one
surface to the other. C, Transcellular passage involving fusion of vesicles to form channels or formation of fenestrae in
thin areas of endothelium. (Redrawn after Simionescu, N., and M. Simeonescu. In Ussing, H. H., N. B. Bindslev, and
O. Sten-Knudsen, eds.: Water Transport Across Epithelia. Copenhagen, Munksgaard, 1981.)

across the cell is largely confined to endo¬ uous vesiculation of the adluminal plasma¬
thelial cells and is an expression of their lemma, detachment, movement across the
specialization for transport. The term transcy- cytoplasm, and fusion with the opposite
tosis has been suggested to distinguish this membrane. To account for the measured
process from pinocytosis. In addition to the albumin clearance, this would involve a con¬
translocation of vesicles from one surface of tinuous massive translocation of membrane
the endothelium to the other, serial sections from the luminal to the abluminal surface of
reveal that transient transendothelial chan¬ the endothelium. Some recent evidence
nels may be formed by fusion of several strongly suggests that the transport vesicles
vesicles, or in extremely thin areas a single are not all newly formed at the expense of
vesicle may open at both surfaces of the cell the luminal plasmalemma, but may be a sep¬
(Fig. 12-26C). arate and relatively stable population, arising
There is now quite general agreement that possibly from the Golgi complex, and simply
in continuous capillaries, the transcellular shuttling back and forth, undergoing alter¬
traffic of vesicles and the channels occasion¬ nate fusion and fission without intermixing
ally formed by vesicle fusion together corre¬ of membrane constituents at the plasma¬
spond to the “large-pore” system postulated lemma (Fig. 12-2621). This model would be
by physiologists. The process of transcytosis consistent with the images observed in elec¬
was originally envisioned as involving contin¬ tron micrographs and would not involve
390 • BLOOD AND LYMPH VASCULAR SYSTEMS

translocation of large amounts of membrane there was little evidence of their movement
from the luminal to the abluminal surface of into the cytoplasm or of their discharge on
the endothelium. the abluminal side of endothelium. Thus, the
There is still disagreement as to the struc¬ blood-brain barrier is attributable to a paucity
tural equivalent of the postulated small pores of transport vesicles in cerebral capillaries
in muscle capillaries, permitting passage of arid tighter occluding junctions than in mus¬
molecules smaller than 9 nm, Some investi¬ cle-type capillaries elsewhere in the body. A
gators localize them in the intercellular junc¬ blood-ocular barrier and a blood-thymus barrier
tions. In the case of fenestrated capillaries, have been shown to depend on similar prop¬
there is general agreement that the fenestrae erties of the capillary endothelium in these
are the equivalent of the “large pores,” while organs. In the thymus, segments of the mi¬
the “small pores” may be in the intercellular crovasculature only a few millimeters apart
clefts or in the diaphragms of the fenestrae. have different permeability properties. The
The permeability properties of the pore dia¬ endothelium prevents access of circulating
phragms are still largely unexplored. macromolecules to the lymphoid cells of the
Factors other than molecular size influence cortex, while the capillaries of the medulla
transendothelial transport-—the chemical na¬ are freely permeable to electron-opaque
ture of the molecules and their net charge, probes.
as well as the charge in the pathways in¬
volved. In general the surface of the endo¬
thelium is negatively charged. When tracers SINUSOIDS
of opposite charge, such as cationized ferri¬
tin, are perfused they bind randomly on the Sinusoids are endothelium-lined vascular
plasmalemma, and especially tenaciously to channels of relatively large caliber and irreg¬
diaphragms of fenestrae, but not to the plas- ular cross-sectional outline. Unlike capillaries
malemmal vesicles and channels involved in that are cylindrical, sinusoids conform in
transcytosis, which appear to be neutral. shape to the interstices among the epithelial
Thus, the endothelial surface presents to the sheets and cords of the organ they supply.
blood a mosaic of microdomains of differing Their unusual configuration is a consequence
charge and can probably sort macromole¬ of their mode of development. Capillaries
cules according to their differing charge. develop as dichotomously branching cellular
Cationic tracers are taken up mainly by ad¬ cords that secondarily acquire a lumen, and
sorptive endocytosis in coated vesicles, they grow by addition of vasoformative cells
whereas anionic or neutral tracers are shut¬ at their ends. Sinusoids, on the other hand,
tled across the endothelium in smooth vesi¬ develop during organogenesis by ingrowth
cles. of the epithelial parenchyma into large, thin-
There are marked differences in the walled embryonic blood vessels. This ac¬
permeability of capillaries of the same type counts for their close conformity to the ge¬
in different organs. For example, intrave¬ ometry of the organ. The most typical ex¬
nously injected dyes that readily escape from ample of sinusoids is to be found in the
capillaries in most tissues are retained in the microvasculature of the liver, but sinusoids
cerebral capillaries, and only a few special are also found in spleen, bone marrow, ad¬
areas of the brain are stained. This observa¬ renal cortex, adenohypophysis, and certain
tion, first made 70 years ago, gave rise to the other endocrine glands. The sinusoidal en¬
concept of a blood-brain permeability barrier. Its dothelium in these organs is more active in
structural basis was thought to reside in spe¬ endocytosis and has a better-developed lyso¬
cial relationships of the foot processes of somal system than capillary endothelium else¬
perivascular astrocytes to each other and to where. For this reason these vessels were
the capillary walls. When this was reinvesti¬ traditionally grouped together as elements of
gated by electron microscopy, the capillaries the so-called reticuloendothelial system.
of the brain were found to have a continuous The endothelium of sinusoids in some
endothelium like that of muscle capillaries, lymphoid organs is extremely attenuated but
but when peroxidase was administered intra- continuous. In endocrine glands, it is a mo¬
vascularly, the tight junctions did not permit saic of fenestrated and unfenestrated areas
escape of the molecules by an intercellular as in visceral capillaries. The endothelium of
path. Occasional vesicles containing peroxi¬ the hepatic sinusoids is unique in having
dase were found at the luminal surface but large fenestrations of varying size and shape.
 BLOOD AND LYMPH VASCULAR SYSTEMS • 391

Here the blood plasma has direct access to media, and tunica adventitia. However, the
the liver cells with no interposed permeability boundaries of these layers are frequently
barrier. indistinct, and in certain veins these coats,
particularly the tunica media, cannot be dis¬
tinguished. The muscular and elastic tissue
is not nearly as well developed in the veins
VEINS as in the arteries, whereas the connective
tissue component is much more prominent.
The blood is carried from the capillary
networks toward the heart by the veins. In Venules and Veins of Small Caliber
progressing toward the heart, they gradually
increase in caliber, while their walls become When several capillaries unite, they form
thicker. The veins usually accompany their a venule, a cylindrical vessel 15 to 20 \im in
corresponding arteries (Fig. 12-27). The diameter, consisting of a layer of endothe¬
veins are more numerous than the arteries lium surrounded by thin, longitudinally ori¬
and their caliber is larger, so that the venous ented reticular fibers with occasional fibro¬
system has a much greater capacity than the blasts (Fig. 12—28). Although the caliber of
arterial system. The walls of the veins are the vessel is larger, the structure of its wall is
always thinner, more supple, and less elastic not very different from that of a capillary
than those of the arteries. Thus, in sections, (Fig. 12-29). It appears to be more permea¬
the veins are usually collapsed, and their ble to intravenously injected dyes. Not all of
lumen is irregular and slitlike unless a special the exchange between the blood and the
effort has been made to fix them in disten¬ tissues takes place in the capillaries. The
tion. venules appear to have a significant role in
It is customary to distinguish three types: this, and they are particularly important in
veins of small, medium, and large caliber. the changes associated with inflammation.
This subdivision is often unsatisfactory, for Early observations on the properties of
the caliber and structure of the wall cannot venules have now been amplified by electron
always be correlated. Veins in the same cat¬ microscopic studies. When particulate mark¬
egory show much greater variations than do ers are injected intravascularly, the particles
the arteries, and the same vein may show are usually found not outside of the walls of
great differences in structure in different the capillaries but along somewhat larger
parts. vessels interpreted as venules. This same cat¬
Most authors distinguish three layers in egory of vessels is also most susceptible to
the walls of the veins: tunica intima, tunica histamine, serotonin, and other substances

Endothelium

Muscl

Figure 12-27. Cross section through a small artery and its accompanying vein from the submucosa of a human
intestine. (After A. A. Maximow.)
392 BLOOD AND LYMPH VASCULAR SYSTEMS

Figure 12-28. Photomicrograph of a thin-spread mesentery showing a nerve, venule, and capillaries. May-Grunwald-
Giemsa stain.

Figure 12-29. Electron micrograph of a portion of the wall of a small venule from the myocardium. The thin continuous
endothelium is essentially the same as that of a capillary. The nuclear region of the endothelial cell bulges into the
lumen.
 BLOOD AND LYMPH VASCULAR SYSTEMS • 393

known to increase vascular permeability (Fig. Veins of Medium Caliber

12—30). If one of these substances is injected
locally into an animal that has previously The veins of medium caliber (2 to 9 mm)
been injected intravenously with a particulate include the cutaneous and deeper veins of
marker, it induces the appearance of small the extremities distal to the brachial and the
intercellular gaps in the endothelium. The popliteal, and the veins of the viscera and
particles, held back by the basal lamina, ac¬ head, with the exception of the main trunks.
cumulate in the gaps, thus marking the ves¬ In the tunica intima of these veins, the endo¬
sels and permitting identification of the sites thelial cells in surface view are polygons with
of increased permeability. Although leaks can highly irregular outlines. Sometimes the tun¬
occasionally be found in capillaries, the vast ica intima also contains an inconspicuous con¬
majority are in small venules. There seems nective tissue layer with a few cells and thin
to be a gradient of permeability from the elastic fibers. Externally, it is sometimes
bounded by a network of elastic fibers. Be¬
arterial to the venous side, which reaches a
maximum in the venules and then diminishes cause the tunica intima is often poorly devel¬
oped, some authors consider the inner and
abruptly in vessels of larger size.
When the caliber of venules has increased middle coats as forming one layer.
The tunica media is much thinner than in
to about 50 pm, partially differentiated
the arteries and consists mainly of circular
smooth muscle cells appear between the en¬
smooth muscle fibers separated by many lon¬
dothelium and the surrounding connective
gitudinal collagenous fibers and a few fibro¬
tissue. These cells are at first some distance
blasts. The tunica adventitia is usually much
apart (Fig. 12-31). Farther along, in small
thicker than the media and consists of loose
veins, they become arranged closer and closer
connective tissue with thick longitudinal col¬
together. In small veins with a diameter of
lagenous bundles and elastic networks. In the
200 pm, these elements form a more or less
layers adjacent to the media, it often contains
continuous layer and have a typical long
a number of longitudinal smooth muscle
spindle shape.
bundles.
In still larger veins, thin networks of elastic
fibers appear. Their tunica intima consists
only of endothelium, and one or several Veins of Large Caliber
layers of smooth muscle cells form the media.
The tunica adventitia consists of fibroblasts The tunica intima has the same structure
and thin elastic and collagenous fibers. Most as in the medium-sized veins. In some of the
of these fibrous elements run longitudinally, larger trunks its connective tissue layer is of
but some penetrate among the muscle cells considerable thickness (45 to 68 pm). The
of the tunica media. tunica media, in general, is poorly developed

ABC
Figure 12-30. Photomicrographs illustrating the greater permeability of venules induced by serotonin. A, Cremaster of
the normal rat injected with carbon to demonstrate the entire vascular system. B, Vascular labeling resulting from
leakage of opague particles from the vessels from local injection of serotonin. The black vessels are venules, the
permeability of the many small capillaries visible in A has not been enhanced by this treatment. C, Higher magnification
of a venule after 7 days, showing intracellular mass of particulate matter in the vascular wall. (From Majno, G., G. E.
Palade, and G. I. Schoefl. J. Biophys. Biochem. Cytol. 11:607, 1961.)
394 • BLOOD AND LYMPH VASCULAR SYSTEMS

Figure 12-31. Scanning micrograph of a large venule joined by a smaller tributary at upper right. The smooth muscle
cells are more irregular in shape, and are spaced farther apart than arterioles. Elements of the perivascular nerve net
can be seen branching over the vessel. The fibrous and amorphous connective tissue components have been digested
enzymatically in this preparation to reveal the underlying structures. (Micrograph from Uehara, Y., J. Desaki, and T.
Fujiwara. Biomed. Res. 2(Suppl.):139, 1981.)

and is sometimes absent. Its structure is the In a considerable portion of the inferior
same as in the veins of medium caliber. The vena cava, the tunica media is absent and the
tunica adventitia makes up the greater part well-developed longitudinal muscle bundles
of the venous wall and is usually several times of the tunica adventitia are directly adjacent
as thick as the tunica media. It consists of to the intima. In the pulmonary veins, the
connective tissue containing thick elastic fi¬ media is well developed, with circular smooth
bers and longitudinally oriented collagenous muscle. It is like an artery in this respect.
fibers. The tunica adventitia contains promi¬ Smooth muscle is particularly prominent in
nent longitudinal bundles of smooth muscle all the layers of the veins in the pregnant
and elastic networks. This is the structure of uterus.
the inferior vena cava and of the portal, Other veins are entirely devoid of smooth
splenic, superior mesenteric, external iliac, muscle tissue and consequently lack a tunica
renal, and azygos veins. media. In this group belong the veins of the
maternal part of the placenta, the spinal pia
mater, the retina, as well as the sinuses of the
Special Types of Veins
dura mater, the majority of the cerebral
In the iliac, femoral, popliteal, saphenous, veins, and the veins of the nailbed and the
cephalic, basilar, and umbilical veins, there trabeculae of the spleen. The last two are
are longitudinal smooth muscle fibers in the simply channels lined by endothelium, with
subendothelial connective tissue layer of the a fibrous connective tissue covering.
tunica intima. In certain veins the longitudi¬ The adventitia of the vena cava and partic¬
nal orientation of cellular elements is also ularly of the pulmonary vein is provided for
noticed in the innermost layers of the tunica a considerable distance with a layer of cardiac
media. muscle fibers arranged in a ring, with a few
 BLOOD AND LYMPH VASCULAR SYSTEMS • 395

longitudinal fibers where these vessels enter endothelium. The endothelial cells lining the
the heart. In the rat, the pulmonary veins up surface toward the lumen of the vessel are
to their radicles contain a considerable elongated in the axis of the vessel; those that
amount of cardiac muscle in the tunica me¬ line the surface of the valve facing the sinus
dia. are transversely elongated.

Valves of the Veins Portal Systems of Vessels

Many veins of medium caliber, especially As a general rule, a capillary network con¬
those of the extremities, are provided with nects the terminal ramifications of the arterial
valves that prevent the blood from flowing system with those of the venous system, and
away from the heart. These are semilunar the transition from arterioles to capillaries to
pockets on the internal surface of the wall, venules is gradual. In a number of regions,
with their free edges pointing in the direction however, modifications of this vascular plan
of the blood flow (Fig. 12-32). In man the are adapted to the special functional require¬
valves are usually arranged in pairs, one ments of the particular organ.
opposite the other. Between the valve and In one physiologically important modifi¬
the wall of the vein is the sinus of the valve, cation of the general plan, the flow from one
where the wail of the blood vessel is usually capillary bed passes through a larger vessel
thin and somewhat distended. or vessels having histological characteristics
The valve itself is a thin connective tissue of veins to a second capillary network before
membrane. On the side toward the lumen of the blood returns to the heart via the systemic
the vessel, it contains a network of elastic venous circulation. Such a set of vessels in¬
fibers continuous with those of the tunica terposed between two capillary beds is called
intima of the vein. In the thinner region of a portal system. For example, the capillary
the wall, comprising the sinus of a valve, the networks of the intestines and certain other
intimal and medial tunics contain only lon¬ abdominal viscera drain via the portal vein to
gitudinal smooth muscle fibers. These do not the liver. There the portal vein ramifies into
enter into the substance of the valve in man. a network of sinusoids throughout the organ.
Both surfaces of the valve are covered by Blood passes from these through a system of

Leaflet of valve

th its
ra ne

membrane

Figure 12-32. From a cross section of the femoral vein of man. The section passes through the origin of a valve. E,
Elastic fiber network in the intima on the inner surface of the valve leaflet; L, longitudinal muscles of the base of the
valve. Acid orecin stain. (After Schaffer.)
396 • BLOOD AND LYMPH VASCULAR SYSTEM.

converging vessels of gradually increasing The wall of the heart, in both the atria and
caliber to the hepatic vein and thence back the ventricles, consists of three main layers:
to the heart via the inferior vena cava. This an internal, the endocardium; an intermediate,
arrangement permits nutrient material ab¬ the myocardium; and an external, the epicar-
sorbed in the intestines to be exposed to, and dium. The internal layer is directly exposed
processed by, the liver cells before being to the blood; the myocardium is the contrac¬
distributed throughout the body by the gen¬ tile layer, and the epicardium is the visceral
eral circulation. layer of the pericardium, a serous membrane
The capillaries in the median eminence of that forms the pericardial sac, the serous
the hypothalamus are continuous with a cavity in which the heart lies.
plexus of small veins, the hypophyseoportal sys¬ The endocardium is generally regarded as
tem, which courses along the hypophyseal homologous to the tunica intima of the blood
stalk and then divides into the sinusoids of vessels, the myocardium to the tunica media,
the anterior lobe of the hypophysis. This and the epicardium to the tunica adventitia.
arrangement permits releasing factors, which
are liberated at the ends of neurosecretory Endocardium
axons in the hypothalamus, to be carried
downstream to activate endocrine secretory The endocardium is lined with ordinary
cells in the hypophysis (see Chapter 19). endothelium, which is continuous with that
An artery may ramify into a set of capillar¬ of the blood vessels entering and leaving the
ies, which are then collected into a larger heart. This endothelium consists of polygonal
arterial vessel. An example of this is found squamous cells. Directly under the endothe¬
in the kidney, in which an afferent arteriole lium in most places is a thin sub endothelial
suddently breaks up into a mass of contorted layer that contains fibroblasts and collagenous
capillaries comprising the glomerulus. These fibers and a few elastic fibers. External to this
capillaries do not empty into veins but coa¬ loose layer is a thick layer of denser connec¬
lesce to form the efferent arteriole, which tive tissue, which comprises the main mass of
goes on to ramify into another set of capil¬ the endocardium and contains great numbers
laries around the kidney tubules. In this case of elastic elements (Fig. 12—33). Bundles of
the efferent arteriole is a portal vessel (see smooth muscle fibers are found in varying
Chapter 30). numbers in this layer, particularly on the
interventricular septum.
A subendocardial layer, absent from the pap¬
illary muscles and from the chordae tendi-
THE HEART neae, consists of loose connective tissue that
binds the endocardium to the myocardium
The heart is a thick, muscular, rhythmically and is directly continuous with the interstitial
contracting portion of the vascular system. It tissue of the latter. It contains blood vessels,
lies in the pericardial cavity within the me¬ nerves, and branches of the conduction sys¬
diastinum. It is about 12 cm long, 9 cm wide, tem of the heart. In the spaces between the
and 6 cm in its anteroposterior diameter, and muscular bundles of the atria, the connective
consists of four chambers: a right and left tissue of the endocardium continues into that
atrium and a right and left ventricle. The of the epicardium, and the elastic networks
superior and inferior venae cavae bring the of the two layers intermingle.
venous blood from the body to the right
atrium, from which it passes to the right Myocardium
ventricle. From here the blood is forced
through the lungs, where it is aerated, and it The histology and fine structure of the
is then brought to the left atrium. From there cardiac muscle has been described in Chapter
it passes to the left ventricle and is distributed 10. In the embryos of the higher vertebrates,
throughout the body by the aorta and its the myocardial fibers form a spongy network.
branches. The orifices between the atria and In the adult stage, however, they are bound
the ventricles are closed by the tricuspid valve by connective tissue into a compact mass.
on the right and the mitral valve on the left This condensation of the myocardium pro¬
side. The openings to the pulmonary artery gresses from the epicardium toward the en¬
and the aorta, from the right and left ventri¬ docardium. Many embryonic muscle fascicles
cles, respectively, are closed by the aortic and remain in a more or less isolated condition
pulmonary semilunar valves. on the internal surface of the walls of the
 BLOOD AND LYMPH VASCULAR SYSTEMS • 397

Endothelium

Connective
tissue

Cardiac
muscle

Figure 12-33. Section of the endocardium of the ventricle of man.

ventricular cavities. These muscle fiber bun¬ another during contraction and relaxation of
dles are covered with endocardium and are the heart. When they become adherent and
called trabeculae carneae. the potential space between them is obliter¬
Elastic elements are scarce in the myocar¬ ated by disease (pericarditis), they may im¬
dium of the ventricles of adult mammals, pose considerable restraint upon the action
except in the tunica adventitia of the larger of the heart.
blood vessels in the walls of these chambers.
In the myocardium of the atria, however, Cardiac Skeleton
there are networks of elastic fibers, which
run everywhere between the muscle fibers The central supporting structure of the
and are directly connected with similar net¬ heart, to which most of the muscle fibers are
works in the endocardium and epicardium. attached and with which the valves are con¬
They are also continuous with the elastic nected, is called rather inappropriately the
networks in the walls of the large veins. A cardiac skeleton. It has a complicated form and
large part of the interstitial connective tissue consists, for the most part, of dense connec¬
of the cardiac muscle consists of extensive tive tissue. Its main parts are the septum
networks of reticular fibrils. membranaceum, the trigona fibrosa, and the an¬
nuli fibrosi encircling the atrioventricular and
arterial foramina.
Epicardium
In man the fibrous rings around the atri¬
The epicardium is covered on its free sur¬ oventricular foramina contain some fat and
face by a single layer of mesothelial cells. elastic fibers but are mainly dense connective
Beneath the mesothelium is a thin layer of tissue. The structure of the septum mem¬
connective tissue with networks of elastic fi¬ branaceum suggests that of an aponeurosis,
bers, blood vessels, and many nerves. In the with its more regular orientation of collagen¬
loose connective tissue along the coronary ous bundles in layers. The connective tissue
blood vessels, considerable amounts of adi¬ of the trigona fibrosa contains islands of
pose tissue are found. cartilage-like tissue (chondroid) consisting of
The parietal layer of the pericardium is a globular cells resembling chondrocytes. The
serous membrane of the usual type—a flat interstitial substance stains deeply with basic
layer of connective tissue that contains elastic aniline dyes and hematoxylin, and is pene¬
and collagenous fibers, fibroblasts, fixed mac¬ trated by collagenous fibers. In aged persons
rophages, and a covering layer of mesothelial the tissue of the cardiac skeleton may become
cells. The smooth, moist, apposed surfaces of calcified in places and sometimes even ossi¬
the epicardium and the parietal pericardium fied. In bovine species, bone is normally
permit these layers to glide freely over one found in the trigona fibrosa.
398 • BLOOD AND LYMPH VASCULAR SYSTEMS

Cardiac Valves bers. In the vicinity of the annulus hbrosus,

the subendocardial layer is quite loose, and
Each atrioventricular valve consists of a sup¬ the musculature of the atrium penetrates far
ple sheet of connective tissue, which begins into it. On the ventricular side, the endocar¬
at the annulus hbrosus and is reinforced dial layer has a similar structure but is much
internally by thin ligamentous strands. It is thinner. In many places the chordae tendi-
covered on its atrial and ventricular surfaces neae, which extend from the edge of the
by a layer of endocardium. At the free edge valve to the papillary muscles, enter this layer
of the value these three layers blend (Fig. and mingle with the deeper-lying connective
12-34). tissue.
The ground plate of connective tissue con¬ The aortic and pulmonic valves have the
sists largely of dense chondroid tissue, with same general structure as the atrioventricular
small spindle-shaped or rounded cell$ and a valves. In the middle of the valves are plates
basophilic interstitial substance. The endo¬ of chondroid tissue with collagenous and thin
cardial layer is thicker on the atrial side. Here elastic fibers. At the root of the valve these
the subendothelial layer has a small amount all continue into the annulus hbrosus. At the
of chondroid tissue and rests upon a connec¬ middle of the free edge they form a thick¬
tive tissue layer, which contains many elastic ening called the nodulus Arantii.
fiber networks and some smooth muscle fi- The aortic and pulmonary valves are nor¬
mally avascular structures. The mitral and
tricuspid valve leaves may be penetrated by
small vessels, but only to a distance of a few
millimeters from their origin.

Myocardium

IMPULSE CONDUCTING SYSTEM

In the adult mammalian heart, the motor

impulse arises in the part of the heart that
develops from the embryonic sinus venosus,
Elastic fibers
astic fibers
an area in which the superior vena cava
Endocardium of enters the right atrium. There is a specialized
ventricular side mechanism by which the contraction spreads
to the atria and then to the ventricles.
At the boundary between the right atrium
Dense Myocardium and the superior vena cava, in the region of
connective tissue the sulcus terminalis, is the sinoatrial node, 1
cm in length and 3 to 5 mm in width. Al¬
Endocardium of though not sharply outlined, it can be seen
atrial side with the naked eye. It consists of a dense
Elastic fibe
network of interwoven Purkinje fibers. An
impulse beginning at the sinoatrial node,
which is the pacemaker of the heart, activates
ocarciium the atrial musculature and is conducted to
the atrioventricular node. A continuous tract of
Elastic fibers atypical muscle fibers extends down the in¬
terventricular septum from this node to both
ventricles, sending branches to the papillary
muscles and other portions of the myocar¬
dium. This system thus serves to initiate and
Myocardium
transmit the contractile impulse. The micro¬
scopic and submicroscopic structure of this
specialized conduction tissue has been de¬
scribed in Chapter 10. The conduction sys¬
Figure 12-34. Cross section through the mitral valve of
man. Atrial surface on right, ventricular on left. In upper tem, even up to the terminal ramifications in
left is the attachment of the aortic valve; on left, below, is the ventricles, is enclosed within a connective
the passage of chordae tendineae into the valve. Low tissue layer that separates it from the working
magnification. (After Sato.) musculature of the myocardium.
 BLOOD AND LYMPH VASCULAR SYSTEMS • 399

The atrioventricular node is a flat, white both of its nodes are abundantly supplied
structure about 6 mm long and 2 to 3 mm with blood from special branches of the cor¬
wide; it is located in the posterior lower part onary arteries.
of the interatrial septum below the posterior 1 hree groups of lymphatic vessels are de¬
ieaf of the aortic valve. The node consists of scribed in the heart: (1) large lymphatic ves¬
Purkinje fibers, which form a tangled dense sels, which lie in the grooves of the heart
network whose meshes are filled with con¬ together with the blood vessels—these drain
nective tissue. These fibers pass into (or be¬ to the lymphatic nodes beneath the arch of
tween) the usual myocardial fibers, so that the aorta and at the bifurcation of the tra¬
the boundary of the node is indistinct. To¬ chea; (2) lymphatic vessels of the epicardial
ward the ventricles the substance of the node connective tissue; and (3) lymphatic vessels
converges abruptly into a band about 1 cm of the myocardium and the endocardium.
long, the atrioventricular bundle. It is located In the subepicardial connective tissue, net¬
in the dense connective tissue of the tri- works of lymphatic capillaries may easily be
gonum fibrosum dextrum and continues into demonstrated and larger vessels provided
the septum membranaceum, where it divides with valves. Lymphatic capillaries have also
into two branches. been described in the atrioventricular and
The first branch, a cylindrical bundle 1 to semilunar valves, but their presence here in
2 mm thick, runs downward along the pe¬ the absence of blood vessels raises some
riphery of the membranous septum and is doubts about the validity of these observa¬
located in part directly under the endocar¬ tions.
dium of the right ventricle. It proceeds along The lymphatic network in the endocar¬
the interventricular septum and splits into dium can be confused with the finer ram¬
many branches, which spread along the en¬ ifications of the sinoventricular conduction
tire internal surface of the right ventricle and system, for both structures may be demon¬
into the papillary muscles. strated by the same injection method. How¬
The left branch is a wide, flat band that ever, the conducting system forms much
comes forward under the endocardium of wider meshes, and its cross connections are
the left ventricle in the upper portion of the thicker than those of the lymphatic network.
interventricular septum, under the anterior The myocardium is penetrated by an abun¬
edge of the posterior cusps of the aortic valve. dant lymphatic network, which is everywhere
It divides into two main branches at the connected with the subendocardial plexus
border between the upper and middle thirds and also communicates with the pericardial
of the septum; then it divides, as in the right lymphatic network.
ventricle, into numerous anastomosing thin The numerous nerves of the heart are in
threads, which are lost to view in the myo¬ part ramifications of the vagus nerve and in
cardium. part sympathetic nerves. Some nerve endings
in the heart apparently are of effector type,
Blood Vessels, Lymphatics, and while other endings are of receptor or sen¬
Nerves of the Heart sory character.

The blood supply to the heart is carried by

the coronary arteries, usually two in number,
HISTOGENESIS OF THE
which arise from the aorta in the sinuses BLOOD VESSELS
behind two of the valve cusps. They are
distributed to the capillaries of the myocar¬ In mammals the first vessels are laid down
dium. The blood from the capillaries is col¬ in the area vasculosa on the surface of the
lected by the cardiac veins, most of which yolk sac, where they develop from the mes¬
empty by way of the coronary sinus into the enchymal cells. In the embryo proper, the
right atrium. A few small cardiac veins empty blood vessels and the heart appear later; at
directly into the right atrium. first they contain no blood cells. In the spaces
In the coronary arteries of the human between the germ layers, groups of mesen¬
heart, the tunica media, which is limited on chymal cells flatten around spaces filled with
both sides by the usual internal and external fluid, which are thus surrounded by a thin
elastic membranes, is divided by a thick fe¬ endothelial wall. In this way, the primordia
nestrated membrane into an inner and an of the heart and the main blood vessels, such
outer layer. as the aorta and the cardinal and umbilical
The conduction system and, particularly, veins, are laid down. These originally inde-
400 • BLOOD AND LYMPH VASCULAR SYSTEMS

pendent primordia then rapidly unite with blood. The mesenchymal cells outside the
one another and with the vessels of the area endothelium become smooth muscle cells.
vasculosa, after which the blood circulation Soon more layers of smooth muscle join the
is established. The endothelial cells in these first layer; these arise in part by the addition
first stages are merely mesenchymal cells of new mesenchymal cells and in part by
adapted to the new function of bounding the division of smooth muscle cells.
blood vessel lumen. The factors that cause the larger arteries
After the closed blood vascular system has and veins to develop in more or less constant
developed and the circulation has begun, new patterns in particular places and in definite
blood vessels always arise by “budding” from directions are not completely understood. It
preexisting blood vessels. The new formation is probable that in the earliest embryonic
of blood vessels by budding may be studied stages the formation of the major vessels is
in sections of young embryos or in the living genetically determined, while in the later
condition in the margin of the tail in larval stages the pattern and growth of the blood
amphibians, in the mesentery of newborn vessels are determined by local hemodynamic
mammals, or in the thin layer of inflamed factors.
tissue that grows between two coverslips in¬
troduced under the skin of an animal. A
method has been devised for the continued
observation of such chambers in the living
LYMPHATIC VESSELS
rabbit for weeks and even months.
In the process of budding, a protrusion In most tissues and organs, with the excep¬
appears on the wall of the capillary and is tion of the central nervous system, cartilage,
directed into the surrounding tissues. From bone and bone marrow, thymus, teeth, and
the beginning it often appears to be a simple, placenta, the blood capillaries are paralleled
hollow expansion of the endothelial wall but by a plexus of lymph capillaries. The blood
in other cases it is at first a solid accumulation vessels form a closed “circulatory system”
of endothelial cells. This vascular bud or with a central pump (the heart), an outflow
sprout enlarges, elongates, and assumes tract (arteries), and a return system (veins).
many shapes. Most frequently it appears as a The lymph vascular system, on the other
solid strand of cells. It later becomes hollow hand, is a “drainage system” whose smallest
and thus represents a lateral extension of the vessels, the lymphatic capillaries, end blindly
blood vessel into which blood cells penetrate. and conduct a clear fluid, called lymph, from
An endothelial bud may encounter another the extracellular spaces in the periphery back
bud and fuse end on, or it may come into through successively larger lymphatic vessels
lateral contact with another bud or another that ultimately converge upon lymphatic ducts
capillary. A lumen then forms among the (thoracic duct and right lymphatic duct),
fused endothelial cells and unites the two which are confluent with the great veins at
capillaries. In this way a new mesh is formed the base of the neck. Along the course of the
in the capillary network, and blood begins to lymphatic vessels are encapsulated accumu¬
circulate through it. Later, new buds may lations of lymphoid tissue comprising small
arise from the newly formed vessels. The organs called lymph nodes (see Chapter 15).
developing vascular buds are often accom¬ The lymph stream entering the node via
panied by undifferentiated mesenchymal afferent lymphatic vessels breaks up within it,
cells and fibroblasts, deployed parallel to the percolating through a labyrinthine system of
long axis of the buds. minute channels lined with endothelium and
Arteries and veins of all types are always phagocytic cells. Lymph then emerges from
formed first as capillaries. The primary endo¬ the node through efferent lymphatic vessels. The
thelial tube then expands and thickens as lymph, exposed to an enormous number of
new elements are added to the outside of the phagocytes, is cleared of foreign matter as it
wall. These elements originate from the sur¬ filters through the lymph nodes. Many lym¬
rounding mesenchyme in the embryo and phocytes that enter the lymph nodes from
play an important part in the formation of the blood are added to the efferent lymph
new arteries and veins from capillaries, as and are thus carried back to the bloodstream.
well as in the formation of large vessels from Lymph is essentially an ultrafiltrate of the
smaller ones during the development of blood plasma formed by continual seepage
pathways for “collateral circulation” of the of fluid constituents of the blood across the
 BLOOD AND LYMPH VASCULAR SYSTEMS • 401

capillary walls into the surrounding intersti¬ Lymphatic Capillaries

tial spaces. It contains water, electrolytes, and
variable amounts of protein (2 to 5 per cent) The thin-walled, endothelium-lined lym¬
depending upon the site and conditions of phatic capillaries differ from blood capillaries
its formation. The walls of blood capillaries in several respects. They branch and anasto¬
are normally freely permeable to water and mose freely, and although they are generally
small molecules. They are less permeable to cylindrical, they are far more variable in
plasma proteins, which therefore tend to shape and caliber than are their counterparts
maintain a significant colloid osmotic pres¬ in the blood vascular system (Figs. 12—35,
sure in the blood. At the arterial end of blood 12-36). Except in the perinuclear region, the
capillaries (where the hydrostatic pressure endothelium usually is extremely thin. In
exceeds the colloid osmotic pressure of the electron micrographs, microvilli projecting
blood), water, solutes, and some plasma pro¬ into the lumen are not uncommon. The thin
teins move across the wall into the tissue. At edges of adjacent endothelial cells often over¬
the venous end, the hydrostatic pressure is lap for some distance. There is a distinct
lower and the colloid osmotic pressure tends intercellular cleft throughout most of this
to draw water, electrolytes, and products of region of overlap, but one or two punctate
tissue catabolism back into the blood. How¬ areas of closer apposition and adherence are
ever, some of the fluid and much of the usually seen along the boundary. There are
plasma protein that have left the blood do no pericytes associated with lymphatic capil¬
not return directly but are drained off in the laries, and a continuous basal lamina is usu¬
lymph and returned to the blood via the ally lacking (Fig. 12—37). Extracellular bun¬
lymph vascular system. A delicate balance is dles of filaments (5 to 10 nm) have been
thus maintained, which keeps the volume of described, terminating on the abluminal
extracellular fluid reasonably constant and plasma membrane of the endothelium and
conserves the small amount of plasma protein extending outward into the ground substance
that continually escapes through the walls of and between the collagen bundles of the
the blood capillaries. surrounding connective tissue. These have
It follows from this mechanism of lymph been called lymphatic anchoring filaments, and
formation that the flow of lymph will be it is suggested that they have a mechanical
increased by (1) an increase in blood capillary role in maintaining the patency of the vessels.
permeability, (2) an increase in hydrostatic The filaments appear to insert in the outer
pressure, or (3) a decrease in colloid osmotic leaflet of the membrane or to terminate in
pressure of the blood plasma. Any of these patches of amorphous material on the outer
will increase the transudation of fluid. If fluid surface of the plasmalemma. These patches
accumulates in the tissues in excess of the bear a superficial resemblance to the material
capacity of lymphatic vessels to drain it away, composing the continuous basal lamina of
the resulting swelling of the tissues is referred endothelium in blood capillaries. The chem¬
to as edema. Thus, increased resistance to ical nature of the anchoring filaments has
venous return in congestive heart failure may not been established, but they very closely
raise pressure in the blood capillaries, result¬ resemble the filaments associated with elastic
ing in edema of the ankles. Similarly, obstruc¬ fibers and may possibly be identical to them.
tion to the lymphatic vessels of an extremity The terminal elements of the lymphatic
owing to parasitic disease or radical surgery system are quite variable in their form from
may result in persistent edema. one organ to another. In the skin and mucous
The principal function of the lymph vas¬ membranes, a plexus of tubular lymphatic
cular system is to return to the blood the capillaries generally occurs parallel to the
fluid and plasma protein that escape from network of blood capillaries but tends to be
the circulation; to return to the blood the more deeply situated. In the lamina propria
lymphocytes of the recirculating pool; and to of the intestine, a single vessel or a simple
add to the blood immune globulins (antibod¬ network of lymphatic capillaries extends
ies) that are formed in lymph nodes. We are from the submucous plexus into the core of
concerned here only with the structure of each villus and ends blindly near its tip. In
the vessels; the organization of the associated the lining of the oviduct, the terminal lym¬
lymphoid tissue will be discussed in Chapter phatics are not tubular but are narrow, flat¬
15. tened sinusoids extending for considerable
402 • BLOOD AND LYMPH VASCULAR SYSTEMS

m 39
til: ^ s '"\*L1

Silifti
lit

«iiiili
■ a,

«5r
Blood
Lymphatic capillary
■
m * l
O*'"** ■
MM
.
m* <7 »
\

■ .

Figure"!2-35. Photomicrograph of guinea pig skin, illustrating a typical lymphatic capillary in the dermis. (Photograph
courtesy of L. V. Leak.)

Lymphatic

Figure 12-36. A, Photomicrograph of a lymphatic in the interstitial tissue of a ram testis. B, Lymphatic in interstitium of
a bull testis. Both these preparations have been fixed by vascular perfusion. The blood capillaries are therefore empty,
whereas the lymphatics have a light gray content, representing precipitated protein of the lymph. (From Fawcett, D. W.,
W. B. Neaves, and M. N. Flores. Biol. Reprod. 9:500, 1973.)
 BLOOD AND LYMPH VASCULAR SYSTEMS • 403

Figure 12-37. Electron micrograph of a subcutaneous lymphatic capillary from guinea pig in cross section. Notice the
irregular outline, the thin wall, and the slight variations in thickness of the endothelium. The absence of a basal lamina
cannot be verified at this magnification. (Micrograph courtesy of L. V. Leak.)

distances along the axis of each fold of mu¬ often indistinct, so that the division is some¬
cous membrane. In the testis of some ro¬ what artificial. The intima consists of endo¬
dents, the lymphatics form labyrinthine peri¬ thelium and a thin layer of interlacing lon¬
tubular sinusoids that have no consistent gitudinal elastic fibers. The media is a layer
geometry, but conform to the shapes of the or two of smooth muscle cells, while the
intertubular spaces that they occupy and to adventitia is composed of elastic fibers and
the contours of the blood vessels and peri¬ collagenous bundles continuous with those of
vascular clusters of Leydig cells that they the surrounding connective tissue.
surround. In man and other large species, Valves are a conspicuous feature of lym¬
the testicular lymphatics are not sinusoidal phatics. They occur in pairs with the two
but occur as one or two tubular vessels in members on opposite sides of the vessel and
each interstitial space. These are usually their free edges pointing in the direction of
much larger than the blood capillaries (Fig. lymph flow. As in veins, the valves are folds
12-36). of the tunica intima and therefore consist of
back-to-back layers of endothelium sup¬
Larger Lymphatic Vessels ported near their base by a thin intervening
layer of connective tissue. They occur at
These vessels are easily distinguished from much closer intervals along the length of the
blood vessels by the large size of their lumens lymphatic vessel than do the valves of veins.
in relation to the thickness of their walls. The Immediately distal to a valve, the lymphatic
wall is somewhat thicker than that of lym¬ is often slightly expanded. The periodic fu¬
phatic capillaries and is invested by thin col¬ siform expansions at the sites of valves give
lagenous bundles, elastic fibers, and occa¬ the lymphatic vessels a highly characteristic
sional smooth muscle cells. In lymphatics with appearance.
a diameter greater than 0.2 mm, three layers The walls of lymphatic vessels are inner¬
of wall are recognizable corresponding to vated and there is cinematographic evidence
intima, media, and adventitia of blood ves¬ that, in some small mammals, rhythmic con¬
sels. The boundaries between the layers are traction of smooth muscle cells in the vessel
404 • BLOOD AND LYMPH VASCULAR SYSTEMS

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THYMUS BARRIERS
Biol. 42:794, 1969.
deCastro, F.: Sur la structure et l’innervation du sinus Brightman, M. W.: Morphology of blood-brain inter¬
carotidien de Fhomme et des mammiferes. Trav. faces. Exp. Eye Res. 25(Suppl) 1, 1977.
Lab. Recherches. Biol. Univ. Madrid 25:331, 1927. Brightman, M. W., I. Klatzo, Y. Olsson, and T. Reese:
Duncan, D., and R. Yates: Ultrastructure of the carotid The blood-brain barrier to proteins under normal
body of the cat as revealed by various fixatives and and pathological conditions. J. Neurol. Sci. 79:215,
the use of reserpine. Anat. Rec. 757:667, 1967. 1970.
McDonald, C. M., and R. A. Mitchell: The innervation Crerr, H. F.: Physiolosry of the charoid plexus. Physiol.
of glomus cells, ganglion cells and blood vessels in Rev. 57:273, 1971.'
the rat carotid body: a quantitative structural anal¬ Dobbing, J.: The blood-brain barrier. Physiol. Rev.
ysis. J. Neurocytol. 4:177, 1975. 47:130, 1961.
Ross, L. L.: Electron microscopic observations of the Pappenheimer, J. R.: Passage of molecules through
carotid body of the cat. J. Biophys. Biochem. Cytol. capillary walls. Physiol. Rev. 55:387, 1953.
6:253, 1959. Rapoport, S. I.: Blood-brain barrier in physiology and
Yates, R. D., 1-Li Chen, and D. Duncan: Effects of sinus medicine. New York, Raven Press, 1976.
nerve stimulation on carotid body glomus cells. J. Raviola, E., and M. J. Karnovsky: Evidence for a blood-
Cell Biol. 46:544, 1970. thvmus barrier using electron opaque tracers. J.
Exp. Med. 756:466, 1972.
ARTERIOVENOUS ANASTOMOSES Raviola, G.: Effects of paracentesis on the blood-aqueous
Bryden, M. M., and G. S. Molyneux: Arteriovenous barrier: an electron microscopy study on Macaca
anastomoses in the skin of seals. II. The California mulatta using horseradish peroxidase as a tracer.
sea lion, Zalophus californianus, and the northern fur Invest. Ophthalmol. 75:826, 1974.
seal, Callorhinus ursinus. Anat. Rec. 797:253, 1978. Reese, T. G., and M. J. Karnovsky: Fine structural
Cauna, N.: The fine structure of the arteriovenous localization of a blood-brain barrier for exogenous
anastomosis and its nerve supply in the human nasal peroxidase. J. Cell Biol. 54:207, 1967.
respiratory mucosa. Anat. Rec. 168:9, 1970. Simionescu, N., and M. Simionescu: The hydrophilic
Hales, J. R., A. A. Fawcett, J. W. Bennett, and A. D. pathway of capillary endothelium, a dynamic sys¬
Needham: Thermal control of blood flow through tem. In Transport Across Epithelia. Copenhagen,
capillaries and arteriovenous anastomoses in skin of Munksgaard, 1980.
sheep. Pflugers Arch. 378:55, 1978. Wissig, S. L., and M. C. Williams: Permeability of muscle
Molyneux, G. S., and M. M. Bryden: Comparative as¬ capillaries to microperoxidase. J. Cell Biol. 76:341,
pects of arteriovenous anastomoses. In Harrison, R. 1978.
J., ed.: Progress in Anatomy. Vol. 1. Cambridge
VENULES AND VEINS
University Press, 1981.
Majno, G., G. E. Palade, and G. Schoeffl: Studies on
CAPILLARIES inflammation. II. The site of action of histamine
Bennett, H. S., J. H. Luft, and J. C. Hampton: Mor¬ and serotonin along the vascular tree. A topograph¬
phological classification of vertebrate capillaries. ical study. J. Biophys. Biochem. Cytol. 77:607, 1961.
Am. J. Physiol. 796:381, 1959. Movat, H. Z., and N. V. P. Pernando: The fine structure
Brightman, M. W.: Morphology of the blood-brain in¬ of the terminal vascular bed. IV. Venules and their
terfaces. Eye Res. (Suppl), 1977. perivascular cells. Exp. Mol. Pathol. 5:98, 1964.
Bruno, R. R., and G. E. Palade: Studies on blood Rhodin, J. A. G.: Ultrastructure of mammalian venous
capillaries. I. General organization of muscle capil¬ capillaries, venules and small collecting venules. J.
laries. J. Cell Biol. 57:244, 1968. Ultrastruct. Res. 25:452, 1968.
Florey, H.: Exchange of substances between the blood Simionescu, N., M. Simionescu, and G. E. Palade: Open
and tissues. Nature 792:908, 1961. junctions in the endothelium of post-capillary ven¬
Jennings, M. A., U. T. Marchesi, and H. Florey: The ules of the diaphragm. J. Cell Biol. 79:27, 1978.
transport of particles across the walls of small blood
LYMPHATIC VESSELS
vessels. Proc. R. Soc. Lond. [Biol.] 756:14, 1962.
Karnovsky, M. G.: Morphology of capillaries with special Drinker, C. K., and J. M. Yoffey: Lymphatics, Lymph
reference to muscle capillaries. In Crone, C., and and Lymphoid Tissue. Cambridge, MA, Harvard
N. A. Lassen, eds.: Capillary Permeability. New University Press, 1941.
York, Academic Press, 1970. Leak, L. V.: Normal anatomy of the lymphatic vascular
Maul, G. G.: Structure and formation of pores in fenes¬ system. In Meessen, H., ed.: Handbuch der Allge-
trated capillaries. J. Ultrastruct. Res. 56:768, 1971. meine Pathologie. Berlin, Springer-Verlag, 1972.
Revel, J. P., and E. Raviola: Evidence for a blood-thymus Leak, L. V.: Electron microscopic observations on lym¬
barrier usine electron opaque tracers. T. Exp. Med. phatic capillaries and structural components of the
756:466, 1972. connective tissue-lymph interface. Microvasc. Res.
Palade, G. E.: Transport in quanta across the endothe¬ 2:361, 1970.
lium of blood capillaries. Anat. Rec. 756:254, 1960. Rouviere, H.: Anatomie des Lymphatiques de l’Homme.
Palade, G. E., M. Simionescu, and N. Simionescu: Struc¬ Paris, Masson, 1932.
 13
THE IMMUNE SYSTEM
Elio Raviola

Organisms of different species and the on the surfaces of other cells that have chem¬
various individuals belonging to the same ical patterns different from the normal con¬
species, with the exception of genetically stituents of the individual they inhabit, and
identical twins, possess a unique chemical are able to set up against them a specific
identity, because the macromolecular constit¬ defensive reaction, the immune response. Anti¬
uents of their cells and body fluids have a gen is the term used to define any material
different composition. The immune system that bears a surface configuration (antigenic
protects the individual from exogenous ma¬ determinant) capable of eliciting an immune
cromolecules, either introduced as such into response upon entry into the internal envi¬
the body or deployed at the surface of invad¬ ronment of the body. The clonal selection
ing viruses, microorganisms, or cells; fur¬ theory holds that during ontogeny of the im¬
thermore, it exerts a surveillance function on mune system, and possibly throughout life,
the appearance in the body of endogenous, lymphocytes arise continuously, each geneti¬
abnormal constituents. The immune system cally programmed to respond to a single
encompasses the lymphoid organs (thymus, antigen. Thus, the versatility of the immune
lymph nodes, spleen, and tonsils); all the system in disposing of myriad endogenous
aggregates of lymphoid tissue occurring in or exogenous materials is innate and resides
nonlymphoid organs; the lymphocytes of the in a population of lymphocytes, all of which
blood and lymph; and the whole population are identical from a morphological point of
of lymphocytes and plasma cells dispersed view, but each endowed with the capacity to
throughout the connective and epithelial tis¬ react to a different antigen. This multiplicity
sues of the body. The various components of of antigen specificities is generated during
the immune system are kept in communica¬ lymphocyte development and results from
tion by a continuous traffic of lymphocytes, recombination of coding sequences dispersed
like a mobile army continuously patrolling on the lymphocyte’s DNA. Upon first meet¬
the body. The cells of the immune system ing with the appropriate antigen (primary
can be thought of as having “academies,” the response), the lymphocyte is stimulated and
thymus and the bursa analogue in which they undergoes a series of morphological and bio¬
are trained; their “battlefields” are the pe¬ chemical changes (transformation) that result
ripheral lymphoid organs and the connective in proliferation and differentiation. Proliferation
tissues of the body; their “lines of communi¬ leads to amplification of the population of
cation” are the blood and lymph. They dis¬ relevant cells: this is called clonal expansion.
pose of the “invaders” either by attacking Differentiation results in the appearance of
them directly or through highly selective both effector cells and memory cells. Effector
weapons, the antibodies, and they are assisted cells (activated lymphocytes and plasma cells) are
by “mercenary troops” of macrophages. instrumental in causing antigen disposal.
The science of immunology has developed Memory lymphocytes revert to the inactive
a rather complex and specialized terminol¬ state, but are then capable of setting up an
ogy; therefore, we must present at the outset immune response with greater efficiency
some concepts and definitions that will be upon encountering their specific antigen
needed to understand the organization and again (secondary response).
functioning of the immune system. As lymphocytes displaying variable degrees
Lymphocytes have the ability to recognize of affinity for a given antigen may preexist
macromolecules on viruses, on bacteria, or in the individual, this antigen will represent

406
 THE IMMUNE SYSTEM • 407

a better stimulus for those cells that have aphylaxis. This latter is the result of release
greater avidity for its surface determinants. of pharmacologically active substances by tis¬
Antigen, therefore, exerts a selective pres¬ sue cells that have bound antigen-antibody
sure upon lymphocyte proliferation, thus complexes, the most typical example being
simulating on a microscopic scale “natural the release of histamine by mast cells.
selection’' by which the environment favors Antibodies are proteins found in the glob¬
the multiplication of genetically fit individu¬ ulin fraction of the plasma. They are also
als. called immunoglobulins and are subdivided
Contact with antigen, however, is not nec¬ into several classes. The immunoglobulins G
essarily followed by lymphocyte stimulation (IgG) represent the bulk of the immunoglob¬
and consequent immunization. In special cir¬ ulins of the normal human blood, and their
cumstances, the body may become tolerant to structure is known in great detail. The IgG
the foreign material. The dose of the antigen, molecule has a weight of about 150,000 dal-
its degree of foreignness, and the mode of tons. It consists of about 1400 amino acid
its presentation (in solution, for example, residues and contains 2 to 3% carbohydrate.
rather than at the surface of another cell) are It comprises four polypeptide chains, paired
among the factors that may result in toler¬ in such a way that the molecule has identical
ance rather than stimulation. The impor¬ halves, each consisting of one long or heavy
tance of this phenomenon is considerable, chain (H chain) and one short or light chain
because self-tolerance rather than innate un¬ (L chain). The four chains are held together
responsiveness is the basis for lymphocytes’ by disulfide bonds and noncovalent interac¬
inability to attack constituents of the body of tions. In the free IgG molecule, the subunits
which they are a part. are folded into a roughly cylindrical unit,
Antigen disposal is based on two different about 3.5 X 20 nm in dimensions, but the
mechanisms: (1) in the humoral immunologi¬ molecule displays a considerable degree of
cal responses, such as those evoked by most plasticity, and upon combination with antigen
bacterial antigens, lymphocytes and plasma it may acquire a characteristic Y or T shape.
cells synthesize and release proteins called Papain splits the antibody molecule into three
antibodies, which specifically combine with the parts: one, the Fc fragment containing two
antigen; (2) in the so-called cellular or cell- half H chains, is crystallizable, and does not
mediated immunological responses, such as combine with the antigen. It enables the
those triggered by transplantation of foreign antibody molecule to bind to complement
tissues, application of a chemical sensitizing and to the surface of cells that carry appro¬
agent to the skin (delayed hypersensitivity), priate membrane receptors. Each of the
many viral diseases, and infection with facul¬ other two parts, the Fab fragments, contains
tative intracellular bacteria (such as Mycobac¬ a combining site for antigen and consists of
terium tuberculosis), clones of lymphocytes the whole L chain and the remaining portion
arise that either release small molecules with of the H chain; both chains contribute to the
a variety of pharmacological actions on mac¬ combining site for the antigen. Therefore,
rophages, granulocytes, or other lymphocytes the IgG molecule as a whole is bivalent (i.e.,
(lymphokines) or attack directly the foreign it is capable of combining with two entities),
cells (cytolytic lymphocytes). and this property permits formation of poly¬
Upon binding to the antigen, antibodies meric aggregates of antigen and antibody,
may neutralize its harmful effects (if the which facilitate the attachment of comple¬
antigen is a toxin); inhibit its entry into the ment and the uptake of the complex by
cells of the body (if the antigen is a virus); phagocytes. The IgG antibodies have high
enhance the uptake and destruction of bac¬ affinity for the antigenic determinant; thus,
teria by phagocytic cells such as neutrophils they are very effective in neutralizing bacte¬
or macrophages (a process called opsoniza¬ rial toxins or in protecting the body against
tion); or induce lysis of bacteria or cells by virus infections; they exchange readily be¬
activating complement, a system of proteins tween blood and extravascular space. How¬
that are normally present in the plasma and ever, they are produced late during the pri¬
are necessary for lysis of foreign cells in mary immune response and are rather
immunological defenses. Antigen-antibody ineffective in activating complement.
reactions may also have a detrimental effect The immunoglobulins M (IgM) are large,
upon the host by causing either a severe complex molecules with a weight of about
inflammatory response (Arthus reaction) or an¬ 900,000 daltons and a 10 per cent content of
408 • THE IMMUNE SYSTEM

carbohydrate. They consist of 20 polypeptide sent the activity of two morphologically

chains, joined by disulfide bonds. They rep¬ similar, but functionally distinct, classes of
resent a pentamer of subunits, each resem¬ lymphocytes, commonly referred to as thy¬
bling IgG in its general organization, but they mus-dependent or T lymphocytes and bursa-de¬
contain an additional polypeptide chain, pendent or B lymphocytes. T lymphocytes have
called J or joining chain. The IgM molecules been preconditioned under the influence of
are not as specific as IgG for the antigenic the thymus to respond to antigen by differ¬
determinant and therefore are not very ef¬ entiating into lymphokine-secreting and cy¬
fective in neutralizing the harmful effects of tolytic lymphocytes; B lymphocytes, on the
toxic antigens. However, even a single IgM other hand, have acquired the ability to re¬
molecule is capable of activating comple¬ spond to antigen by differentiation into an¬
ment; thus, IgM readily induces lysis of for¬ tibody-secreting lymphocytes and plasma
eign cells. Furthermore, it is much more cells. In birds, the site of production or pre¬
effective in antigen agglutination and opson¬ conditioning of B lymphocytes is the bursa of
ization. Fabricius, an appendix-like diverticulum of
The immunoglobulins A (IgA), when pres¬ the cloaca.
ent in the plasma, are 170,000 daltons in Both T and B lymphocytes manifest im¬
molecular weight and, like IgG, consist of munological specificity; that is, both are ge¬
four polypeptide chains, but they may be netically programmed to respond to a specific
linked to each other in a polymeric form. antigen and both carry “memory/’ The func¬
IgA is also contained in a variety of secretory tions of T and B lymphocytes are not inde¬
products, such as colostrum, saliva, tears, and pendent of each other, however, but are
nasal and tracheobronchial mucus, and it is intimately interrelated. Only certain antigens
released into the intestinal lumen. The secre¬ directly stimulate B cells and lead to antibody
tory form of IgA has a molecular weight of secretion without participation of T cells.
390,000 daltons because two antibody mole¬ With most antigens that elicit a humoral
cules become united to each other by a J response, T lymphocytes cooperate with B
chain, and the resulting complex is bound to cells, helping them to become stimulated and
a glycoprotein with a high carbohydrate con¬ regulating their differentiation (“helper” T
tent, called the secretory piece, which is pro¬ lymphocytes). Furthermore, another class of T
duced by the epithelial cells. Secretory IgA is lymphocytes (“suppressor” T lymphocytes) spe¬
very resistant to proteolytic enzymes; there¬ cifically suppresses antibody responses by ex¬
fore, it has been speculated that the secretory erting an inhibitory feedback control on
piece confers stability on the immunoglobulin helper T lymphocytes or acting on antibody-
molecule and prevents its catabolism in the secreting B lymphocytes. There is finally a
intestinal lumen. IgA probably plays a pro¬ third category of lymphocytes, which have
tective role at the surface of mucous mem¬ different functional properties from both T
branes. and B lymphocytes; they have been denomi¬
Two additional types of immunoglobulins nated null lymphocytes. Their precise signifi¬
have been described: the IgE, which induce cance, however, is still under discussion. Al¬
release of histamine by mast cells and are though lymphocytes are the only cells that
responsible for certain allergic reactions, and confer specificity on the immunological re¬
the IgD, glycoproteins with a molecular sponse, they are assisted by macrophages in
weight of 170,000 to 200,000 daltons, whose the process of antigen recognition. Phago¬
concentration in the serum varies greatly in cytic cells are also involved in antigen dis¬
different individuals. The biological function posal, and eosinophils seem to destroy the
of this serum IgD is poorly understood. How¬ complexes of antibody with soluble antigens.
ever, IgD, together with monomeric IgM, is Lymphocytes originate from stem cell pre¬
present as an integral component of the cursors which, in the embryo, arise from the
plasma membrane of one class of lympho¬ mesenchyme intervening between the endo-
cytes (B lymphocytes), where they function derm of the yolk sac and the splanchnopleu-
as antigen receptors. In contrast, the number ric mesoderm. During postnatal life, the
of lymphocytes bearing surface IgG is very source of the stem cells becomes the bone
small. marrow. The differentiation of the stem cells
Although a cellular immune response and into T lymphocytes takes place in the thymus,
secretion of antibody may both occur upon a central or primary lymphoid organ, made up
introduction of a single antigen, they repre¬ of a special variety of lymphoid tissue. In birds,
 THE IMMUNE SYSTEM • 409

the differentiation of stem cells into B lym¬ their functional performance, life span, de¬
phocytes occurs in the bursa of Fabricius; in gree of differentiation, and sensitivity to ion¬
mammals, where the analogue of the bursa izing radiations and hormones. Lymphocytes
has not been identified, differentiation of B circulate with blood and lymph and infiltrate
lymphocytes takes place in the embryonal connective tissues and epithelia; they are
liver and in the bone marrow. In both the present in the bone marrow and compose
thymus and the avian bursa (or its mammal¬ the bulk of the thymus, lymph nodes, white
ian equivalent), the stem cell precursors pulp of the spleen, and lymphoid masses
undergo antigen-independent proliferation and associated with the digestive, respiratory, and
differentiation into lymphocytes that are ge¬ urinary passages.
netically programmed to set up a specific When suspended in a fluid medium,
type of immune response upon meeting their nonmotile lymphocytes are round. When
appropriate antigen. They subsequently pop¬ crowded together in tissues, they mutually
ulate the blood and lymph, are disseminated deform one another into polyhedral shapes.
throughout the connective tissues of the Motile lymphocytes display a slow, ameboid
body, and infiltrate most epithelia. Together progression and conform to the shape of the
with macrophages and with plasma cells, interstices through which they are advancing.
which arise from differentiation of B lym¬ When moving on a solid, flat substrate, they
phocytes, they become associated with retic¬ acquire a characteristic hand-mirror shape,
ular cells and reticular fibers, thus giving rise with the nucleus ahead, followed by a tail of
to a second variety of lymphoid tissue, which cytoplasm in which most organelles are con¬
represents the bulk of the peripheral or sec¬ centrated.
ondary lymphoid organs such as the lymph The size of the lymphocytes varies in dif¬
nodes, spleen, and tonsils. In the secondary ferent organs and in various functional situ¬
lymphoid organs, T and B lymphocytes ations. Lymphocytes circulating in the blood
undergo antigen-dependent proliferation and have a diameter of 4 to 8 pm but upon
differentiation into effector and “memory” flattening, as when they are smeared and
cells. dried on a slide, this increases to 7 to 10 pm
(Fig. 13—1A). In lymphoid organs and tissues
not involved in an acute immunological re¬
sponse, lymphocytes range between 4 and 15
CELLS OF THE IMMUNE pm in diameter, the larger forms being
SYSTEM rather uncommon. It is customary to subdi¬
vide them into small (4 to 7 pm), medium¬
sized (7 to 11 pm), and large lymphocytes
CYTOLOGY OF THE CELLS OF THE (11 to 15 pm) on the basis of cell size, nuclear
IMMUNE SYSTEM morphology , and intensity of the cytoplasmic
basophilia (Fig. 13-2); such a subdivision is
useful for descriptive purposes, but is rather
Lymphocytes
arbitrary, because lymphocyte diameter and
This cell type has already been considered organization vary in a continuous fashion.
as a component of the blood (Chapter 4) and According to this classification, blood lym¬
connective tissue (Chapter 5), but the fore¬ phocytes are represented exclusively by small
going descriptions need review and amplifi¬ and medium-sized cells; lymphocytes of the
cation in the context of the immune system. lymph include a variable proportion of large
The term “lymphocyte” actually refers to a cells, whereas lymphoid organs and tissues
family of cells characterized by lack of specific contain the whole spectrum of cell dimen¬
granules; a round, centrally located nucleus; sions. As a consequence of stimulation by
and a cytoplasm displaying various degrees antigen, mononuclear cells arise that are up
of basophilia, due to the presence of free to 30 pm in diameter (Figs. 13-1C, F). These
ribosomes (Figs. 13—1 to 13—4). As previously were variously termed blast cells, immunoblasts,
stated, although they are morphologically large pyroninophilic cells, hemocytoblasts, and
similar, lymphocytes are physiologically het¬ lymphoblasts. There is now good evidence that
erogeneous; not only does this cell type in¬ these large elements result from transfor¬
clude two major classes of cells, the T and B mation of small lymphocytes and that they
lymphocytes, but also within these two classes, may in turn generate small lymphocytes.
individual lymphocytes have the ability to Therefore, these cells will be referred to as
recognize different antigens and may vary in lymphoblasts.
410 • THE IMMUNE SYSTEM

Figure 13-1. A, Smear preparation of small lymphocytes from the thoracic duct lymph of the rat stained with the May-
Grunwald and Giemsa mixtures. B, Autoradiograph of rat thoracic duct lymphocytes obtained 2 weeks after 3H-thymidine
administration. Only long-iived small lymphocytes are labeled. C and F, When the lymphocytes from the rat thoracic
duct lymph are cultured 3 days on a monolayer of mouse embryo cells, the contact with the foreign cells stimulates a
proportion of the small lymphocytes to transform into lymphoblasts and large lymphocytes. Transformation is followed
by proliferation, as shown by the dividing cell in F. D and E, Autoradiograph of the transformed lymphocytes that arose
from the labeled long-lived small lymphocytes of the rat after 2 days’ culture on monolayer of mouse embryo cells. Note
that the grain count is essentially the same as that of the labeled small lymphocytes in S; this shows that transformation
by antigen precedes cell division. (Courtesy of N. B. Everett.)

Small lymphocytes have a dense nucleus Medium-sized lymphocytes have a nucleus

surrounded by a thin rim of cytoplasm (Fig. with a larger nucleolus and more abundant
13—3). The nucleus is central, round, or euchromatin; the cytoplasm displays more
slightly indented and very rich in randomly basophilia, due to a greater abundance of
dispersed, heterochromatic masses; the nu¬ free ribosomes. In large lymphocytes and
cleolus is small and scarcely identifiable in lymphoblasts, the nucleus is largely euchro-
smear preparations. The cytoplasm is slightly matic and contains one or two prominent
basophilic and contains a variable number of nucleoli (Fig. 13—4). The cytoplasm is abun¬
azurophilic granules when stained with the dant and intensely basophilic, owing to the
Giemsa mixture. The electron microscope presence of large numbers of free polyribo¬
shows a diplosome located at the nuclear somes. Cisternae of the granular endoplasmic
indentation, surrounded by a small Golgi reticulum are, however, scarce. The Golgi
apparatus and by a few mitochondria. Free apparatus is moderately enlarged and mito¬
ribosomes in moderate numbers are scattered chondria and lysosomes are slightly increased
as single units throughout the cytoplasm; in number.
cisternae of the granular endoplasmic retic¬
ulum are found only exceptionally. Small
Plasma Cells (Plasmacytes)
numbers of lysosomes, which represent the
ultrastructural counterpart of the azurophilic The term “plasma cell” includes a range of
granules, complete the list of cytoplasmic immature and mature elements, character¬
organelles. An occasional small lipid droplet ized by the presence of considerable but
may also be observed. varying numbers of cisternae of the granular
 Mitosis in Dividing
medium-sized Reticular large
lymphocyte cells lymphocytes

Medium -sized
lymphocyte

Reticular
cells Large
lymphocyte

mall
lymphocytes

Medium-sized
lymphocytes

Figure 13-2. Various types of lymphocytes in a section of a human lymph node. Hematoxylin-eosin-azure II. (After A.
A. Maximow.)

Figure 13-3. A typical lymphocyte from blood. The nucleus has a coarse pattern of heterochromatin and has an
indentation that is not obvious with the light microscope. A pair of centrioles and a small Golgi complex are located in
or near the concavity of the nucleus.
412 • THE IMMUNE SYSTEM

Figure 13-4. Micrograpns illustrating the transformation of T lymphocytes to lymphoblasts. A, An unstimulated

lymphocyte. B, From the same culture, a lymphoblast 36 hours after transformation induced by treatment with the lectin
concanavalin A. These micrographs at the same magnification show the great increase in cell volume and development
of a euchromatic nucleus with a prominent nucleolus. Transformation induced by phytohemagglutinin or concanavalin
A is similar to that which occurs in response to antigenic stimulation.
 THE IMMUNE SYSTEM • 413

endoplasmic reticulum. Their function is the have a centrally located nucleus, but which
synthesis and release (secretion) of antibody. in electron micrographs display the abundant
They represent the late stages of differentia¬ granular endoplasmic reticulum typical of
tion of the B lymphocytes. Plasma cells are the plasma cell line (Fig. 13-5).
found in the medullary cords of resting Plasma cells are 6 to 20 jam in diameter
lymph nodes, in the marginal zone and cords and have a rounded, elongate, or polyhedral
of the resting spleen, and scattered through¬ form, depending on their location. Seen with
out the connective tissues of the body. They the light microscope, mature elements are
are especially numerous in the lamina pro¬ small; they possess an eccentric, rounded
pria of the intestinal mucosa, where most of nucleus with a small nucleolus and radially
them have been shown by immunofluores- arranged coarse heterochromatic masses ad¬
cent methods to produce immunoglobulins jacent to the nuclear envelope, resulting in a
A. During the acute phase of a humoral cartwheel configuration. The cytoplasm is
immune response, large numbers of imma¬ strongly basophilic, except for a conspicuous
ture plasma cells appear in the deep portion juxtanuclear, pale area, which contains the
of the cortex of lymph nodes and at the diplosome and surrounding Golgi apparatus.
boundary between white and red pulp of the It is evident in electron micrographs that
spleen. Mature plasma cells are sessile ele¬ the cytoplasmic basophilia of plasma cells is
ments and apparently never enter blood and due to their highly developed granular en¬
lymph. After an antigenic challenge, how¬ doplasmic reticulum, often distended with
ever, immature forms appear in the lymph; flocculent material (Fig. 13-6). Experiments
furthermore, a limited number of elements involving immunolabeling with ferritin or
appear in the blood that at the light micro¬ horseradish peroxidase have shown that the
scope level resemble lymphocytes in size and content of the cisternae of the granular retic-

Figure 13-5. An immature plasma cell from peripheral blood. The cell has developed an extensive endoplasmic
reticulum. In its further maturation the pattern of nuclear chromatin would become coarser, the cytoplasm would increase
in volume, and the reticulum would become ordered into closely spaced, parallel arrays of cisternae. For an illustration
of a mature plasma cell, see Figure 5-20.
414 • THE IMMUNE SYSTEM

Figure 13-6. A, Electron micrograph of a plasma cell from the rabbit spleen. The eccentric, rounded nucleus contains
masses of heterochromatin adjacent to the nuclear envelope. The cytoplasm displays a highly developed granular
endoplasmic reticulum. B, Plasma cell from the spleen of a rabbit, which was injected with the enzyme horseradish
peroxidase as an antigen. The antibody-containing spleen cells were subsequently treated with the antigen and stained
with the histochemical method for demonstration of peroxidase activity. Dense reaction product is seen in the lumen of
the cisternae of the granular endoplasmic reticulum, indicating the presence of anti-horseradish peroxidase antibody.
(Courtesy of E. H. Leduc and S. Avrameas.)

ulum consists largely of antibody. The Golgi antibody. The intermediate stages in the
apparatus of mature plasma cells is large; the course of this differentiation are often re¬
mitochondria are few and unremarkable in ferred to as proplasmacytes.
their internal structure. In a small percentage
of plasma cells, one or more cisternae of the Macrophages
granular reticulum are greatly distended with
a mass of dense material. These inclusions The structure of macrophages was dis¬
(Russell bodies) are readily seen with the light cussed in Chapter 5 and will not be repeated
microscope and consist of incomplete im¬ here.
munoglobulin molecules. It has been sug¬
gested that Russell bodies are indicative of
an aberrant synthesis or faulty intracellular HISTOPHYSIOLOGY
transport of antibody, but this speculation
lacks conclusive evidence. Surface Properties of T and B
Very immature precursors of the plasma Lymphocytes
cell line (plasmablasts) are difficult to distin¬
guish from lymphoblasts or large lympho¬ The terms “T” and “B lymphocyte” de¬
cytes. The nucleus is rich in euchromatin and scribe two functionally distinct types of lym¬
provided with a large nucleolus; the cyto¬ phocytes that circulate with blood and lymph
plasm contains many free polyribosomes as and inhabit the peripheral lymphoid tissues.
well as narrow cisternae of the granular en¬ The lymphocytes of the thymus have differ¬
doplasmic reticulum. The transition from ent properties from those of T lymphocytes,
plasmablasts to plasmacyte involves progres¬ but they represent the precursors of the T
sive condensation of the chromatin, reduc¬ cells. Although B lymphocytes cannot be dis¬
tion in size and complexity of the nucleolus, tinguished from T lymphocytes by light or
disappearance of the free polyribosomes, en¬ transmission electron microscopy, they have
largement of the Golgi apparatus, and ap¬ distinct surface properties demonstrable by
pearance of a highly organized granular en¬ indirect methods. If antibody is produced
doplasmic reticulum. The cisternae of the against immunoglobulins by injecting the an¬
reticulum may form parallel, concentric ar¬ tibody of one species into an animal of a
rays or become distended with accumulated different species, the anti-immunoglobulin
 THE IMMUNE SYSTEM • 415

Figure 13-7. A, B, and C, B lymphocytes from the mouse

spleen stained with anti-immunoglubulin antibody conju¬
gated to fluorescein isothyocyanate and photographed
with the fluorescence microscope. In A, the lymphocyte
was reacted with the labeled antibody at 4° C. The anti¬
immunoglobulin is dispersed over the entire cell surface,
although some patching of the marker is already in
progress. B, and C, Upon warming at 37° C, the fluores¬
cent antibody becomes concentrated over one pole of the
cell (capping). For comparison, D illustrates the intense
staining of the antibody in the cytoplasm of splenic plasma
cells treated with fluorescent anti-immunoglobulin. (Cour¬
tesy of E. R. Unanue.)

antibody produced by the recipient can be

isolated, purified, and conjugated to a visible
marker. For example, the anti-immunoglob-
ulin antibody can be conjugated with a flu¬
orescent dye and then, after interaction with
the lymphocyte surface, it can be localized
with the fluorescence microscope (Fig. 13—
7A, B, C). Alternatively, the antibody can be
labeled with radioiodine and its localization
studied with light or electron microscopic
autoradiography (Fig. 13—8); finally, in a
third method, the antibody can be conjugated
to the electron-opaque particulate ferritin or
hemocyanin (Figs. 13—9, 13—10) or to the
enzyme horseradish peroxidase, and visual¬
ized with the electron microscope either di¬
rectly or after appropriate histochemical re¬
action. With these techniques, it has been Figure 13-8. Electron microscope autoradiography of
shown that B lymphocytes incubated at 0° C mouse spleen B lymphocytes treated with 125l-labeled
with labeled anti-immunoglobulin bind the rabbit anti-immunoglobulin antibody. A, At 4° C the marker
is randomly distributed over the entire cell membrane. B,
antibody over their entire surface (Figs. 13—
When the lymphocytes are incubated at 37° C, the label
7A, 13—8). This property is attributable to becomes concentrated at one pole of the cell, forming a
the presence of immunoglobulins, predomi¬ cap. C, Finally, the label is interiorized by endocytosis.
nantly monomeric IgM and IgD, bound to (From Unanue, E. R., et al. J. Exp. Med. 136:885, 1972.)
416 • THE IMMUNE SYSTEM

Figure 13-9. B lymphocyte of the mouse spleen treated with rabbit anti-immunoglobulin antibody conjugated with
hemocyanin, a respiratory pigment present in the hemolymph of many invertebrates. The hemocyanin molecule has the
shape of a short cylinder and is readily recognized with the electron microscope. This lymphocyte was reacted with the
labeled anti-immunoglobulin at 4° C and then warmed to 37° C for 10 min. The label is concentrated in a cap over the
cell pole in which the Golgi apparatus is contained. (From Karnovsky, M. J., et al. J. Exp. Med. 736:907, 1972.)

the cell membrane. Compared with the se¬ moiety of the antibody heavy chain. Some T
creted, pentameric IgM, the heavy chain of lymphocytes have surface receptors for the
the surface IgM has an added hydrophobic Fc region of IgM and others for the Fc region
sequence, which anchors the molecule to the of IgG; a few have receptors for the C3b
plasmalemma. There is evidence that this component of complement. In the human, a
antibody at the cell surface represents the large proportion of T lymphocytes, and pos¬
receptor that combines with antigen. The sibly all of them, bind sheep erythrocytes,
plasma membrane of B cells is also able to and to a lesser extent pig erythrocytes, form¬
bind antibody by means of the Fc fragment ing characteristic clusters or rosettes. The
of their molecule and the C3b component of significance of this phenomenon, which lacks
the complement system. immunological specificity, is poorly under¬
B cells have lower electrophoretic mobility stood. Nevertheless, spontaneous rosette for¬
and lower density than T cells, and they mation provides a reliable clinical test for
adhere preferentially to nylon wool at 37°C evaluation of the size of the T cell population
in the presence of serum. These properties in human patients.
have been exploited in attempts to separate When T lymphocytes are transferred from
the B- from the T-cell components of a mixed a donor mouse into a recipient of the same
lymphocyte population. species but with a slightly different genetic
T lymphocytes do not possess immuno¬ constitution, they elicit production of anti¬
globulins as integral membrane proteins. bodies that combine with T but not with B
Their antigen receptor is a polypeptide chain lymphocytes. Thus, murine T cells possess a
consisting of a constant and a variable region, surface antigenic determinant called Thy-1
similar or identical to the antigen-combining or theta, which is lacking on B cells. Anti-
 THE IMMUNE SYSTEM • 417

Hemocyanin
molecules

Figure 13-10. Same specimen as in Figure 13-9. Hemocyanin-labeled anti-immunoglobulin antibody is attached to the
membrane immunoglobulins of the B lymphocyte. The cell has begun to interiorize the label in endocytotic vacuoles
(From Karnovsky, M. J., et al. J. Exp. Med. 136:907, 1972.)

Thy-1 antibodies, when injected into mice diation, and to cortisone; their distinctive
belonging to the strain whose T lymphocytes distribution in the organs of the immune
carry the Thy-1 antigen, lead to specific com¬ system; and their different pattern of recir¬
plement-mediated destruction of T cells. It culation are discussed on later pages.
is thus possible to study the distribution of T
lymphocytes in the immune system and the
Response of T Lymphocytes to
functional impairment caused by their selec¬
Antigen
tive elimination. Furthermore, anti-Thy-1 an¬
tibody conjugated to a visible marker, such Both T and B lymphocytes manifest im¬
as fluorochrome or ferritin, or labeled with munological specificity; i.e., both are geneti¬
radioiodine, binds to the surface of T lym¬ cally programmed to respond to an antigen
phocytes and permits their morphological that is specific for each individual cell. This
identification. Other antigens expressed on commitment is expressed by the presence of
the surface of T lymphocytes belong to the receptors on the lymphocyte plasma mem¬
Ly (lymphocyte) system. Of the peripheral brane, which combine with the antigenic de¬
murine T lymphocytes, about 50 per cent terminants. This process of specific binding
bear Ly 1,2 antigens, 30 to 40 per cent the is called antigen recognition. Antigen binding
Ly 1 antigen, and 10 per cent the Ly 2 by T lymphocytes can be studied in labora¬
antigen. Helper and lymphokine-secreting T tory rodents by mixing lymphocyte suspen¬
lymphocytes express the Ly 4-1, —2 pheno¬ sions with foreign erythrocytes, usually those
type, suppressor and cytolytic T lymphocytes of sheep. This technique cannot be applied
express the Ly - l, + 2 phenotype, and both to human T cells because most of them bind
differentiate from Ly 4- 1,4-2 lymphocytes. sheep erythrocytes nonspecihcally. Antigen¬
The different reactivity of T and B lym¬ binding T cells of laboratory rodents can also
phocytes to various mitogens, to x-ray irra¬ be studied by combined autoradiography and
418 • THE IMMUNE SYSTEM

immunofluorescence, using radioiodine-la- vors the view that antigen, upon combining
beled antigen and fluorochrome-conjugated with its receptor at the cell surface, somehow
anti-Thy-1 antibody. It has been shown by triggers the transformation of the small lym¬
these experimental strategies that antigen¬ phocyte into a proliferating lymphoblast
binding T lymphocytes in animals not previ¬ (Figs. 13-1, 13-4). The size of the cell in¬
ously exposed to the antigen are very few in creases, its nucleus becomes euchromatic, the
number and belong to the category of the nucleolus enlarges, a great number of poly¬
small or medium-sized lymphocytes. They ribosomes appear in the cytoplasm, and the
increase in number following immunization, Golgi apparatus becomes more prominent.
probably because the challenge with the an¬ Probably related to antigen recognition are
tigen induces amplification of the small frac¬ the behavioral phenomena called peripolesis
tion of the lymphocyte population that car¬ and emperipolesis—i.e., the tendency of lym¬
ries surface receptors specific for that phocytes in cell culture to move about, in¬
particular antigen. The number of receptor denting and even penetrating other cells.
sites for antigen on the membrane of T Antigens that elicit a humoral response
lymphocytes is very small, probably a few stimulate a class of T lymphocytes character¬
hundred, in contrast to the several thousands ized by the Ly +1,-2 phenotype and these,
on B lymphocytes. called helper T lymphocytes, at some stage of
To become stimulated, T lymphocytes their transformation into lymphoblasts, inter¬
must interact with histocompatibility mole¬ act with B lymphocytes and trigger their
cules, a class of membrane glycoproteins, differentiation into antibody-secreting cells.
which are present on the surface of eukar¬ The mechanism underlying this cooperation
yotic cells and are coded by a set of genes is not fully understood; it involves intimate
known as major histocompatibility complex. His¬ surface association and complex interactions
tocompatibility molecules belong to two among antigen, macrophages, T cells, and B
types, one expressed on the surface of all cells. At the onset, the helper T cell is acti¬
cells and the other exclusively represented vated by the antigen presented by a macro¬
on the cells of the immune system. Those phage in association with an autologous his¬
expressed on the surface of all cells differ tocompatibility molecule. The activated
from individual to individual and their ge¬ helper T cell releases factors that stimulate
netic diversity provides biological uniqueness the resting B cell, in turn bearing antigen
for every subject in a normal population. bound to the plasma membrane through its
They represent the specific antigen recog¬ IgM and IgD receptors. The B cell trans¬
nized by T lymphocytes in graft rejection and forms into a lymphoblast that has receptors
activate T cells directly, without participation for soluble growth factors. During a subse¬
of macrophages as a third partner cell. The quent phase, which is antigen independent,
other type of histocompatibility molecules, the interacting T lymphocyte and macro¬
named la in the mouse, are expressed on the phage produce growth factors that stimulate
surface of macrophages, B lymphocytes, and the B lymphoblast to proliferate and mature
some activated T lymphocytes. When the into antibody-secreting effector cells. Prolif¬
antigen is a foreign substance, different from eration of the helper T lymphocytes leads to
the histocompatibility molecules recognized amplification of the response and to the dif¬
in graft rejection, it can stimulate T lympho¬ ferentiation of memory T cells, which revert
cytes only when deployed at the surface of a to the state of small lymphocytes.
cell in association with an autologous la mol¬ Antigens that elicit a delayed type of hy¬
ecule. This requirement is met by macro¬ persensitivity reaction, such as chemical skin-
phages, which present the T lymphocyte with sensitizing agents or products of facultative
the antigen bound to their plasma membrane intracellular bacteria, are presented by mac¬
and their own surface histocompatibility mol¬ rophages to another class of T lymphocytes,
ecules. These observations explain why eu¬ which also express the Ly +1,-2 phenotype,
karyotic cells can stimulate T cells directly, and stimulate them to transform into lym¬
whereas soluble, particulate, or bacterial an¬ phoblasts. As a result of this transformation,
tigen requires the participation of macro¬ T cells begin to produce lymphocyte media¬
phages for effective immunization. tors or lymphokines. Lymphokines are mol¬
The sequence of events following antigen ecules of 8000 to 80,000 daltons in molecular
binding by T lymphocytes is not fully under¬ weight, which display a great variety of phar¬
stood. However, circumstantial evidence fa¬ macological activities; they are not immuno-
 THE IMMUNE SYSTEM • 419

globulins and their biochemical characteri¬ their development and those that suppress
zation is still incomplete. The best known all antibody responses. The mechanism of
lymphokine identified to date is the macro¬ action of suppressor T cells has not yet been
phage inhibitory factor (MIF), a glycoprotein fully elucidated: they elaborate mediators,
20,000 to 40,000 daltons in molecular weight, 10,000 to 50,000 daltons in molecular weight,
which immobilizes macrophages and may which have the ability to bind to the antigen
possibly lead to accumulation of these cells at that stimulated their production. In humoral
the site of the antigen. A macrophage-activating immunity, they exert an inhibitory feedback
factor enhances the bactericidal capacity of on helper T cells or act on antibody-secreting
macrophages. A chemotactic factor recruits B cells and plasma cells, decreasing their
phagocytes to the site of the antigen, and immunoglobulin secretion. Suppressor T
lymphotoxin causes cell lysis. cells also participate in cellular immune re¬
Antigens such as tissue grafts and cells sponses, such as delayed hypersensitivity, and
bearing tumor antigens or infected with vi¬ responses to tumor antigens.
ruses stimulate directly (i.e., without partici¬
pation of macrophages) a third class of T
Response of B Lymphocytes to
lymphocytes, which express the Ly -l, + 2
Antigen
phenotype. In this instance, lymphoblast pro¬
liferation and differentiation lead to the ap¬ Antigen receptors on the surface of B
pearance of cytolytic lymphocytes, which lymphocytes are clearly immunoglobulins,
have the ability to cause the lysis of the for it can be shown that specific binding of
foreign or altered cells after making contact radioactive antigen to the cell membrane is
with their surface. The mechanism of this blocked by anti-immunoglobulin antibody.
direct cytotoxicity is poorly understood; it is The number of immunoglobulin receptors
specific for the cells that caused immuniza¬ varies between 50,000 and 150,000 per cell;
tion, for nearby cellular elements are not thus, the number of sites is much higher than
affected and it requires close cell-to-cell con¬ on T lymphocytes. Despite the abundance of
tact. Three phases have been identified in binding sites at the cell surface, direct stim¬
the interaction between cytolytic T lympho¬ ulation of B lymphocytes is possible only by
cytes and target cells; the first is adhesion: it polymeric antigens—i.e., by molecules that
is temperature dependent and requires the display identical groupings in a linear, repet¬
presence of Mg^ + , but not Ca+ + . Adhesion itive sequence such as the pneumococcal
is prevented by drugs that affect the cytoskel- polysaccharide. Furthermore, the response
eton, inhibitors of ATP production, and local only occurs within a narrow range of antigen
anesthetics. The second phase has been dose and results predominantly, if not solely,
termed the lethal hit: during this stage the in production of IgM antibodies. With most
cytolytic T lymphocyte causes irreversible antigens that elicit a humoral response, B-
damage to the membrane of the target cell. cell stimulation also requires the participation
It is temperature dependent and requires of helper T lymphocytes and macrophages.
Ca"~ but not protein synthesis. The third The influence of helper T lymphocytes is
phase, called killer cell independent lysis, is char¬ also necessary for efficient production of IgG
acterized by the osmotic swelling of the target antibodies and for differentiation of memory
cell as a result of the membrane lesions cells within the B-cell population.
caused by the lethal hit. The activity of helper As a result of the cooperation between
T cells has been extended in the recent past stimulated T and B lymphocytes, antibody is
to other immune responses in addition to secreted and plasma cells appear. That
antibody production. Thus, the development plasma cells synthesize and release antibody
of Ly — 1, + 2 cytolytic T cells is enhanced by has been firmly established by comparative
the influence of specific Ly +1,-2 T cells, immunological and histological studies, by
which therefore regulate most immune re¬ immunofluorescent (Fig. 13—7) and autora¬
sponses positively. diographic techniques, and by antibody assay
Antigen also stimulates a separate T cell in microdroplets containing individual
population with a suppressor function that plasma cells. However, B lymphocytes are
belongs to the Ly — l, + 2 phenotype. Two also capable of synthesizing and releasing
types of suppressor T cells have been iden¬ antibody. Evidence for this was obtained
tified: those that specifically suppress the im¬ from antibody assay in microdroplets con¬
mune response to the antigen that stimulated taining single cells and from electron micro-
420 • THE IMMUNE SYSTEM

scopic identification of single cells that had forms having cytological characteristics inter¬
demonstrated their capacity to produce he¬ mediate between lymphoblasts and immature
molytic antibody by forming a plaque of lysis plasma cells are found consistently. Further¬
in a layer of erythrocytes dispersed in agar more, all experiments in which lymphocyte
(Figs. 13-11, 13-12). Antibody-producing populations have been transferred from im¬
lymphocytes are typical lymphoblasts or large munized animals into syngeneic* unprimed
cells that in addition to polyribosomes contain recipients, or from unprimed donors into
a small amount of cisternae of the granular allogeneic recipients, have led to the appear¬
endoplasmic reticulum. These cells have been ance of plasma cells in the recipient.
regarded as transitional forms between lym¬ It is thus widely accepted that the small
phocytes and immature plasma cells. lymphocytes of the B type, stimulated by
Two schools of thought have developed on antigen under the influence of T lympho¬
the interrelationships between lymphocytes cytes, undergo transformation into lympho¬
and plasma cells, one regarding the lympho¬ blasts. During this process, antibody having
cyte as the ancestor of the plasma cell, the identical specificity to that deployed on the
other considering plasma cells as a separate cell surface is synthesized in increasing
cell line arising from independent, still un¬ amounts and is released as a secretory prod¬
identified, stem cell precursors. A great deal uct instead of being inserted into the plasma
of circumstantial evidence, however, favors
the view that lymphocytes represent the pre¬
*Syngeneic in the field of transplantation immunity
cursors of the plasma cells. Especially persua¬
refers to individuals of the same species that are genet¬
sive are the fact that lymphocytes can also ically identical, such as monozygotic twins or inbred
secrete antibody and the observation that laboratory animals. Allogeneic refers to individuals of the
during the immune response transitional same species that are not genetically identical.

Figure 13-11. Hemolytic plaque-forming cell. A diluted suspension of lymphoid cells from an animal immunized with
foreign erythrocytes was plated in agar along with the erythrocytes that served as antigen. The lymphoid cell at the
center has synthetized and released hemolytic antibody into the surrounding agar, and the antibody has combined with
the erythrocytes embedded in it. Complement has subsequently been added and the erythrocytes carrying the antibody
have lysed, leaving a clear halo or plaque around the antibody-secreting cell. (Courtesy of A. A. Nordin and N. K.
Jerne.)
 THE IMMUNE SYSTEM • 421

cells with central nucleus, scanty cytoplasm,

but abundant granular endoplasmic reticu¬
lum, and that these cells enter the blood and
propagate the response throughout the body
(Fig. 13-5).
During the primary response to antigen,
the first antibody to appear in the blood
belongs to the IgM type; later, much larger
amounts of the more efficient IgG are pro¬
duced. The switch from IgM to IgG produc¬
tion is dependent on the regulatory influence
of T lymphocytes. Both types of immuno¬
globulins can be produced by either lympho¬
cytes or plasma cells.
Antibody synthesis and release involve an
intracellular pathway that has been found to
be typical of all other protein-secreting cells
studied to date. The heavy and light chains
are transcribed from separate messengers on
polyribosomes bound to the membranes of
the granular endoplasmic reticulum. They
Figure 13-12. Electron micrograph of an antibody-se¬ are subsequently transferred into the lumen
creting cell from the rabbit popliteal lymph node identified of the cisternae, either free or combined with
by the hemolytic-antibody plaque technique illustrated in each other. In lymphocytes, the first detect¬
Figure 13-11. The cell has the morphology of a large
able antibody appears in the perinuclear cis-
lymphocyte with euchromatic nucleus, prominent nucleo¬
lus, and abundant cytoplasm lacking an organized gran¬ terna and later, as differentiation to plasma
ular endoplasmic reticulum. (From Harris, T. N., et al. J. cells proceeds, antibody is produced and
Exp. Med. 723:161, 1966.) stored throughout the granular endoplasmic
reticulum. In mature plasma cells, antibody
is no longer detected in the space within the
membrane. Antibody-secreting lymphoblasts nuclear envelope and it disappears from
are actively proliferating cells; their progeny some of the cisternae of the granular endo¬
include (1) memory B cells, which revert to plasmic reticulum. Some carbohydrate com¬
the state of small lymphocytes; and (2) tran¬ ponents of the antibody molecules (e.g., N-
sitional elements, which display an enlarged acetylglucosamine) are incorporated into the
Golgi apparatus and increasing amounts of nascent H chain. The polypeptide backbone,
granular endoplasmic reticulum. These tran¬ carrying part of the carbohydrate moiety, is
sitional cells, in turn, undergo further differ¬ subsequently transported to the Golgi appa¬
entiation into plasma cells, passing through ratus, where additional saccharides (e.g., ga¬
the stages in which they are identified as lactose) are added. From the Golgi, antibody
plasmablasts and proplasmacytes. Differen¬ is carried to the cell surface by some un¬
tiation into plasma cells is accompanied by known mechanism and is released from the
decrease in amount of surface membrane- cell.
bounded immunoglobulins, loss of prolifer¬ The amount of antibody produced by lym¬
ative capacity, and loss of motility. Experi¬ phocytes may be smaller than that produced
ments involving labeling with 3H-thymidine by plasma cells, but it is released at an early
indicate that the process of differentiation of stage of the response and at the very site of
lymphocytes into plasma cells takes about one antigen stimulation, where it can be much
day. The life span of plasma cells is a few more effective than the circulating antibody,
weeks. Lymphoblasts and immature plasma which is confined largely to the intravascular
cells also have the capacity to enter the effer¬ space. Plasma cells may synthesize much
ent lymph of the lymph nodes draining the larger amounts of antibody, but they store a
site of antigen injection and to colonize ad¬ large proportion of this immunoglobulin in
ditional lymph nodes along the same path of the distended cisternae of their granular re¬
lymphatic drainage. There are also indica¬ ticulum and may release it only upon cell
tions that they may transform into smaller death and disintegration.
422 * THE immune system

The Role of Macrophages in Immune Macrophages certainly act in general by re¬

Responses moving and digesting excess of antigen. If
antigen interacts indiscriminately with lym¬
The only immune responses in which lym¬ phocytes in a location unfavorable for cell
phocytes are directly stimulated by antigen cooperation, tolerance is induced instead of
are those triggered by eukaryotic cells carry¬ immunity. Thus, only a small fraction of the
ing histocompatibility molecules on their sur¬ antigen escapes destruction by macrophages
face, and the special case of certain polymeric and triggers immunization. Macrophages,
antigens that stimulate B cells directly, with¬ however, seem to play a more intimate and
out participation of helper T cells. In all important role in lymphocyte stimulation by
other instances, effective immunization re¬ antigen. According to a widely accepted hy¬
quires an additional cell that carries the an¬ pothesis, in addition to ingesting and destroy¬
tigen on its surface and presents it to the T ing antigen, macrophages retain a small
lymphocyte in association with autologous amount of antigen bound to their surface
histocompatibility molecules. However, the and present it in a concentrated form to
specificity of the immune response is deter¬ lymphocytes (a phenomenon referred to as
mined by the lymphocyte, since other cells antigen presentation). In addition, to be im¬
are unable to distinguish foreign determi¬ munogenic antigen must be presented by
nants from the normal body constituents. A macrophages in association with their surface
subject of controversy over the past few years histocompatibility molecules (la molecules in
concerns the question whether macrophages the mouse); only macrophages that possess
belonging to the mononuclear phagocyte li¬ la molecules are capable of cooperating with
neage represent the partner cell that collab¬ T lymphocytes. The function of the histo¬
orates with the lymphocyte during the induc¬ compatibility molecules is to mediate in some
tive phase of the immune response. Since this way the recognition of antigen by T cells,
view is widely accepted by immunologists, it either by ensuring cell-to-cell contact or by
will be discussed first. binding the antigen at the macrophage sur¬
The need for macrophage participation in face. This function implies close topograph¬
the initiation of immune responses has ical relationships between macrophages and
emerged from a wide variety of experiments, lymphoid cells, and this intimate association
all showing that antigens which are avidly has been demonstrated with the electron mi¬
taken up by macrophages are good immu¬ croscope in the cell clusters that produce
nogens, whereas those which are not phago- antibody against foreign red blood cells in
cytized are poor immunogens. Blockade of vitro.
the macrophages of the body by administra¬ As a result of the interaction among anti¬
tion of inert particulate suspensions signifi¬ gen, macrophage, and T lymphocyte, it has
cantly depresses the immune response. If a been suggested the macrophages secrete
donor animal is injected with an antigen that growth and differentiation factors that affect
is feebly immunogenic, and its macrophages the adjacent lymphocytes.
are then transferred to a syngeneic recipient, These views on the identification of mac¬
this latter responds with a vigorous immune rophages as partner cells for the lymphocytes
response, possibly because the small amount in immune induction are vigorously chal¬
of antigen bound to the surface of the mac¬ lenged by some cell biologists, who regard
rophages is a much stronger immunogen the evidence presented by immunologists as
than the antigen in its original dispersed inconclusive. According to these investiga¬
form. The strongest evidence for macro¬ tors, radioresistance and adherence to glass
phage participation in lymphocyte stimula¬ surfaces are not sufficient criteria for identi¬
tion comes from experiments in which the fication of the partner cell as a macrophage.
response to antigen has been reproduced in They also argue that macrophages com¬
vitro; in these systems it has been clearly pletely degrade the ingested material, that
shown that in addition to T and B lympho¬ the membrane of their lysosomes is imperme¬
cytes, a third partner cell is required. This able to macromolecules, and that they are
cell has many properties of macrophages; it unable to release undigested material by ex-
is radioresistant, adheres to glass surfaces, ocytosis. Thus, they deem it unlikely that in
and does not synthesize antibodies. macrophages a small fraction of the antigen
The precise mechanism of macrophage escapes destruction and is transferred to the
function in the induction of an immune re¬ cell surface for presentation to the lympho¬
sponse by lymphocytes is poorly understood. cyte.
 THE IMMUNE SYSTEM • 423

Candidates for the antigen-presenting cells The enhancement of phagocytosis by an¬

in immune induction are the lyynphoid dendritic tibody and complement is not an exclusive
cells, identified in cell suspensions of lymph¬ property of macrophages; granulocytes also
oid organs. These cells are provided with have an enhanced capacity for engulfing bac¬
numerous irregular processes, which may ap¬ teria when the bacteria are complexed with
pear as spiny branches, blunt pseudopods, or antibody and complement.
thin veils. The cytoplasm has many mito¬ The effect on macrophages of the lym-
chondria, but few ribosomes, lysosomes, or phokines released by T cells in the course of
secretory inclusions. The nucleus is irregular delayed hypersensitivity reactions has been
in shape, lined with a rim of heterochroma¬ discussed previously (p. 418).
tin, and provided with distinct nucleoli. Den¬
dritic cells adhere to plastic or glass during
the first few hours in culture, but show little Nonspecific Stimulation of
or no phagocytic activity. They derive from Lymphocytes
the bone marrow, possess histocompatibility
(la) molecules at their surface, and seem A number of agents besides antigen, com¬
capable of cooperating with lymphocytes in monly referred to as mitogens, stimulate lym¬
the induction phase of certain immune re¬ phocytes, inducing their transformation into
sponses. Possibly related to lymphoid den¬ actively proliferating lymphoblasts. The ma¬
dritic cells are other cell types, all character¬ jor difference between antigen and mitogens
ized by irregular shape, inconspicuous cell resides in the fact that the former reacts with
organelles, and lack of phagocytic activity. individual lymphocytes that on a genetic basis
These include the follicular dendritic cells, have developed membrane receptors specific
identified in germinal centers, which retain for its surface determinants, whereas mito¬
antigen-antibody complexes on their surface; gens are effective on much larger lymphocyte
the interdigitating cells, identified in the thy¬ populations. Mitogens include a variety of
mus-dependent areas of lymph nodes and substances extracted from plants or seeds,
spleen; the “veiled” cells, identified in the constituents of bacteria or products of their
afferent lymph of lymph nodes in rodents metabolism, and antilymphocyte sera, pro¬
and man; and the Langerhans cells of the duced by immunizing animals with exoge¬
epidermis. At present, it is unclear whether nous lymphocytes. All mitogens whose mech¬
all these dendritic cells represent a homoge¬ anism of action has been studied in detail
neous population with novel biological prop¬ have the capacity to bind chemical groupings
erties and important functions in immune on the plasma membrane of lymphocytes;
induction, or a subpopulation of macro¬ many of them also combine with the surface
phages in a special state of differentiation. of other cells, but only lymphocytes respond
Macrophages are certainly involved in the with transformation and mitosis. As agents
terminal events of antigen disposal. Their also exist that bind to the surface of lympho¬
role includes removal of foreign cells or bac¬ cytes without stimulating them, mitogens can
teria whose viability has been impaired by be regarded as belonging to a larger class of
lymphocytes or lytic antibody in presence of substances whose common property is an
complement. Furthermore, phagocytosis of affinity for chemical groupings on the sur¬
foreign material, cells, and bacteria is pow¬ faces of cells, and which are therefore gener-
erfully enhanced when the antigenic deter¬ ically named ligands. The effects of mitogenic
minants are opsonized, i.e., complexed with ligands on lymphocytes has great theoretical
antibody or with antibody and complement. importance because their mechanism of ac¬
Opsonization depends on the presence of tion, although' nonspecific, appears to be
receptors on the macrophage membrane that identical to that of antigens. Therefore, they
bind the Fc part of the antibody molecule or have become a useful laboratory tool in stud¬
the C3b component of the complement sys¬ ies aimed at an understanding of lymphocyte
tem. Only certain classes of immunoglobulins stimulation. Especially well studied are the
bind directly to the macrophage surface (IgG chemical and biological properties of extracts
in humans, IgG and IgM in mice), and the of certain plants or seeds, which have long
interaction is especially strong when they are been known to cause agglutination of eryth¬
combined with antigen. IgM, which does not rocytes and leukocytes. These substances are
bind to the membrane of human macro¬ called lectins. Agglutination depends on the
phages directly, becomes cytophilic in the fact that lectins are multivalent ligands and
presence of complement. thus form bridges between neighboring cells,
424 * THE IMMUNE SYSTEM

causing formation of clumps. Plant lectins lymphocytes are poorly understood, but
are proteins or glycoproteins that have strong there is evidence that in seven- to ten-day-
chemical affinity for the oligosaccharide res¬ old cultures of PWM-stimulated lymphocytes,
idues on the plasma membrane of mammal¬ plasma cells develop.
ian cells. They can be conjugated to an ultra- It is evident that binding of molecules to
structural marker such as ferritin or the plasma membrane is not effective per se
hemocyanin, and in this way the distribution in causing lymphocyte stimulation, but satu¬
of the lectin-binding oligosaccharides at the ration of specific chemical groupings at the
cell surface can be visualized with the electron cell surface is probably required. Once a
microscope. mitogenic ligand has bound to the plasma
Ligands do not stimulate all lymphocytes membrane, it somehow triggers lymphocyte
indiscriminately. Phytohemagglutinin (PHA), transformation. Lymphocytes are unique
a lectin extracted from the red kidney bean among mammalian cells in that they respond
(Phaseolus vulgaris), and concanavalin A (Con with cell division and differentiation to a
A), extracted from the jackbean (Canavalia variety of exogenous substances, including
ensiformis), stimulate T lymphocytes. Poke- the antigen for which they carry specific
weed mitogen (PWM), extracted from the receptors.
root of Phytolacca americana, activates both T Studies using ligands conjugated to a visi¬
and B cells, whereas a lipopolysaccharide ble marker have cast some light upon the
extracted from the bacterium Escherichia coli cellular events that immediately follow the
(LPS) is specific for B cells. ligand’s combination with the lymphocyte
T lymphocyte stimulation by PHA has been membrane (Figs. 13—7 to 13—10). When anti¬
studied in great detail and provides a satis¬ immunoglobulin antibody conjugated to a
factory means of imitating the morphological fluorescent dye (or to ferritin or hemocyanin)
and biochemical phenomena that follow com¬ is reacted at 4° C with a B lymphocyte bearing
bination of antigen with T cells. The earliest the immunoglobulin, the label appears uni¬
event after addition of PHA to a culture of formly dispersed over the entire cell surface.
small lymphocytes (15 min) is enhanced en- This demonstrates that antibody molecules
docytotic activity, as evidenced by increased on B cells are uniformly distributed through¬
uptake of neutral red, and increased synthe¬ out the plasma membrane. With the passage
sis of RNA (15 to 30 min). Soon afterward, of time, the marker becomes aggregated,
the nucleolus begins to enlarge (four hours). resulting in formation of interconnected
At 24 to 36 hours, the nucleus becomes more patches. This phenomenon, called patching,
euchromatic, the nucleolonema is clearly vis¬ depends on the fact that surface immuno¬
ible in the nucleolus, and the cell volume globulins move randomly in the fluid domain
increases. The cell begins DNA synthesis, of the plasma membrane and that when they
thus entering the S phase of the cell cycle, approach sufficiently close to one another,
which lasts six to ten hours. Then, it enters they are cross-linked by the multivalent anti¬
the G2 phase, characterized by active RNA immunoglobulin antibody. However, if the
and protein synthesis. There is a striking cell suspension is warmed to 37° C, the
concomitant increase in cytoplasmic polyri¬ marker molecules become aggregated to
bosomes, whereas the granular endoplasmic form a continuous cap localized in the region
reticulum remains relatively sparse. The of the cell surface overlying the Golgi appa¬
Golgi apparatus enlarges and the lysosomes ratus, a behavior called capping. That portion
are moderately increased in number. At the of the plasma membrane bearing the marker
end of G2, which lasts two to four hours, the subsequently becomes interiorizecl by endo-
lymphocyte has undergone a fourfold in¬ cytosis, or the label is shed from the cell
crease in volume and enters mitosis. DNA surface. This process leaves the B lymphocyte
synthesis in cultures of PHA-stimulated lym¬ denuded of its surface immunoglobulins and
phocytes reaches a peak at 72 to 120 hours, antigen receptors for several hours.
then slowly declines over a period of five to Capping is an energy-dependent process
seven days. Beginning at about 24 hours after by which the cell eliminates the ligand bound
PHA exposure, the T lymphocyte develops to its surface; its relationship to lymphocyte
the capacity to impair nonspecifically the vi¬ stimulation is still poorly understood. Proba¬
ability of other cells, such as fibroblasts. This bly, capping occurs upon binding of antigen
property is mediated by production and re¬ to the surface of the B lymphocyte, but stim¬
lease into the culture medium of various ulation also requires the cooperative interac¬
lymphokines. The effects of lectins on B tion with helper T cells.
 THE IMMUNE SYSTEM • 425

Other Functional Properties of tory tracts, they occur together with plasma
Lymphocytes cells and macrophages as densely packed
masses in loose connective tissue. Further¬
Lymphocyte heterogeneity is not limited to
more, the thymus, lymph nodes, white pulp
the three classes of T, B, and null cells.
of the spleen, and tonsils consist mainly of
Within each class, lymphocytes may differ
lymphocytes. The terms “lymphoid” and
considerably in other functional properties, “lymphatic tissue” have been widely em¬
such as immunocompetence, life span, and
ployed to define the common features of all
sensitivity to ionizing radiation or to adrenal
aggregates of lymphocytes and lymphocyte-
steroids. In T lymphocytes, which have been rich organs. Although in the past these terms
more thoroughly studied, these functional were often used with quite different conno¬
differences are related to the degree of cell tations, modern advances in immunology
differentiation. Most lymphocytes of the thy¬
have rendered the traditional distinctions ob¬
mus, which represent the precursors of the solete. On the other hand, our knowledge of
T lymphocyte, are not immunocompetent— the cell-to-cell interactions at a tissue or organ
i.e., they lack the capacity to respond to level is still largely incomplete, and a solid
antigen or to lectins such as PHA by trans¬
basis for a rational classification of the lym¬
formation and proliferation. They are also
phocyte collections of the body is lacking.
readily destroyed by x-ray irradiation or The term “lymphoid tissue” or “lymphoid
administration of cortisone; furthermore, organ” will be used here to define regions of
some of the thymic lymphocytes have a very the body in which lymphocytes, with or with¬
short life span (see Chapter 14). Upon leav¬
out associated plasma cells, represent the
ing the thymus, T lymphocytes become im¬ chief cellular constituent. It must be empha¬
munocompetent and more resistant to irra¬ sized, however, that this definition is purely
diation and cortisone. Their life span is descriptive and includes cell aggregates that
unknown, but upon interaction with antigen may have very different functions.
they can give rise to memory cells, which One morphologically and functionally dis¬
have the morphology of small lymphocytes tinct type of lymphoid tissue is that found in
and a life span in man of up to several years. the thymus and in the medulla of the nodules
The changes in functional properties of B of the avian bursa of Fabricius. It consists of
lymphocytes during their development are lymphocytes and a few macrophages, con¬
still poorly understood. There is evidence, tained in the meshes of a tridimensional
however, that their bone marrow precursors network of stellate cells joined by desmo-
are resistant to corticosteroids, whereas pe¬ somes. These stellate stromal elements are
ripheral B lymphocytes are sensitive to both called reticular cells simply because they form
x-ray irradiation and adrenal steroids. Mem¬ a network. Unlike the mesenchymal stroma
ory cells of the B type also have a long life of most other organs, these cells arise from
span. an epithelial outgrowth of the endoderm,
An important property of lymphocytes is which becomes secondarily invaded by lym¬
their motility, which enables them to cross phopoietic stem cells. Reticular fibers are
the walls of the postcapillary venules in order scarce in this variety of lymphoid tissue.
to enter or leave the bloodstream. They also A much more common type of lymphoid
move about in the parenchyma of lymph tissue makes up the bulk of the lymph nodes,
nodes and can leave the nodes by migrating the white pulp of the spleen, and the tonsils
into the lymph. They penetrate epithelia and and forms more or less discrete masses scat¬
freely wander through the connective tissues tered in the connective tissues of the body.
of the body. Differentiation into plasma cells From a descriptive point of view, “diffuse”
is accompanied by a loss of motility. and “nodular” subvarieties of this second
type of lymphoid tissue can be distinguished
(Fig. 13-13).
LYMPHOID TISSUE
Diffuse Lymphoid Tissue
Lymphocytes occur as individual cells in
blood, in lymph, and throughout the connec¬ Lymphoid tissue of this description is typ¬
tive and epithelial tissues of the body. How¬ ically found in the internodular, deep corti¬
ever, in most organs, but especially in the cal, and medullary regions of lymph nodes,
lamina propria of the digestive and respira¬ in the periarterial lymphoid sheaths of the
426 • THE IMMUNE SYSTEM

Diffuse Germinal center Diffuse

lymphoid tissue lymphoid tissue
0

Figure 13-13. Diffuse lymphoid tissue and a germinal center in the outer cortex of a mesenteric lymph node of a dog.
Hematoxylin-eosin.

spleen, and in the internodular regions of

the tonsils and Peyer’s patches. It consists of
a spongelike stroma with lymphocytes in the
meshes. The stroma, in turn, is made up of
reticular fibers and reticular cells of mesen¬
chymal origin (Fig. 13-14). Reticular fibers,
best shown by the silver impregnation meth¬
ods, are intimately associated with the retic¬
ular cells and often occupy deep recesses or
grooves in their surface. In ordinary histo¬
logical preparations the reticular cells appear
as stellate or elongate elements with oval,
euchromatic nucleus and scanty acidophilic
cytoplasm. In vitally stained animals, some of
the reticular cells take up colloidal dyes avidly
while others do not. By this functional crite¬
rion, some of them can actually be regarded
as macrophages. With the electron micro¬
scope, reticular cells display cisternae of gran¬
ular endoplasmic reticulum in varying
amounts and a moderately well-developed
Golgi apparatus, whereas the other cell or¬
ganelles are inconspicuous. The cell periph¬
Figure 13-14. Section of a lymph node after the lympho¬
ery is often devoid of organelles and inclu¬
cytes have been removed, showing the network of retic¬
sions and contains bundles of filaments. ular cells and their intimate relations with the reticular
Thus, some of the reticular cells of the dif- fibers. Mallory-azan stain. (Redrawn after Heidenhain.)
 THE IMMUNE SYSTEM • 427

fuse lymphoid Ussue are tissue macrophages, tally and should be discarded in favor of
while others are not very different from the “germinal center.”
fibroblasts of the connective tissues elsewhere Germinal centers are a highly organized,
in the body. Experiments involving labeling widely distributed component of the lymph¬
with ^H-thymidine have shown that in lymph oid tissue absent only in the normal thymus
nodes, reticular cells have a very slow turn¬ (Figs. 13—15, 13—16). In their fully developed
over rate. Moreover, during the regeneration form, the germinal centers appear as a spher¬
of the lymph node following irradiation, no ical mass with a dark, or densely populated,
transformation of labeled reticular cells to pole and a pale, or less densely populated,
the free elements of the lymphoid paren¬ pole. The germinal center is surrounded by
chyma is seen. Thus, contrary to the tradi¬ a capsule of elongated cells, which is in turn
tional teaching, it seems unlikely that reticu¬ partially invested by a crescentic cap of small
lar cells can give rise to other cell types. There lymphocytes. Germinal centers display a
seems to be no evidence supporting the time- clear-cut morphological polarity, inasmuch as
honored view that reticular cells are primitive the lymphocyte cap is especially thick over
or undifferentiated elements, capable of giv¬ the light region and becomes gradually thin¬
ing rise to lymphocytes or other connective ner toward the darker pole. Furthermore,
tissue elements. It now appears that reticular they show a consistent orientation with re¬
cells, as seen with the light microscope, rep¬ spect to the neighboring structures. In the
resent either fibroblasts, concerned with the lymph nodes, the light region and lympho¬
synthesis and maintenance of the reticular cyte cap of the germinal centers are directed
fibers, or occasional tissue phagocytes, be¬ toward the marginal sinus; in the spleen they
longing to the mononuclear phagocyte sys¬ are directed toward the red pulp. In the
tem. The free cells of the diffuse lymphoid digestive and respiratory passages they are
tissue are lymphocytes of various sizes, mac¬ oriented toward the nearest epithelial sur¬
rophages, and a variable number of plasma face. When the plane of a histological section
cells. passes through a germinal center in a direc¬
tion perpendicular to its axis of symmetry,
the polarity just described is not seen, for the
Lymphoid Nodules
cap of small lymphocytes appears as a circular
Lymphoid nodules or follicles are compact, rim of uniform width surrounding the ger¬
circumscribed collections of cells within the minal center. For this reason, the cap has
diffuse lymphoid tissue. They are typically often been described as a mantle or corona.
found in the cortex of lymph nodes, at the In the dark region of the germinal center
periphery of the white pulp of the spleen, (Fig. 13—15), the intense staining results from
and in the lamina propria of the digestive the nuclei and basophilic cytoplasm of nu¬
and respiratory passages. They are very nu¬ merous closely packed elements of the
merous in the tonsils, Peyer’s patches, and lymphoid cell line—namely, lymphoblasts,
appendix. There is much disagreement in large and medium-sized lymphocytes, and
the literature as to the nomenclature for cells in transition to the plasma cell line. All
lymphoid nodules, since the terms “primary” these cells are actively proliferating and they
and “secondary nodule” and “germinal cen¬ contain antibody within the perinuclear space
ter” have been used to define different enti¬ and occasional cisternae of the granular en¬
ties. Primary nodule is most commonly em¬ doplasmic reticulum. In the dark zone, mac¬
ployed to designate a rounded collection of rophages are also found consistently, loaded
tightly packed small lymphocytes, whereas with debris of phagocytized lymphocytes.
secondary nodule (also called germinal center) The free cells are contained in the meshes of
describes ovoid structures consisting of a a cellular framework composed of stellate
spherical cluster of larger, pale-staining cells elements joined by desmosomes. These cells
invested by a cap of small lymphocytes. The display little cytoplasmic specialization; they
precise significance of primary nodules is are stained by silver methods and were called
unknown; moreover, there is no evidence dendritic cells because of their numerous
that secondary nodules, with their cap of radiating processes. The transition between
small lymphocytes, arise by transformation the light and the dark region at the equator
of preexisting primary nodules, as the terms of the germinal center is a gradual one; the
would imply. Thus, the term “secondary nod¬ large basophilic cells of the lymphoid line
ule” does not seem to be justified experimen¬ progressively give way to small lymphocytes,
428 • THE immune system

Light
region

Dark
region

Figure 13-15. Germinal center in the outer cortex of an inguinal node of a dog. Hematoxylin-eosin.

mitotic figures disappear, and macrophages In very large germinal centers, lymphocyte
decrease in number. The dendritic cells ac¬ phagocytosis is intense; thus, they acquire a
quire abundant eosinophilic cytoplasm and characteristic appearance with the light mi¬
myriad peripheral interdigitating processes. croscope, because the macrophages loaded
The capsule of the germinal center consists with residual bodies are seen as light areas
of a few layers of flattened reticular cells on a background of tightly packed nuclei
joined by desmosomes. This investment is (starry sky). The life span of germinal centers
disorganized over the light pole because of and the precise sequence of events leading
the presence of many highly deformed small to their disappearance are unknown.
lymphocytes, allegedly fixed in the course of Very little is understood about the function
their migration toward or from the overlying of germinal centers. They are the site of
small lymphocyte cap. Mature plasma cells active production of lymphocytes, but a pro¬
are scarce in germinal centers except in those portion of the newly formed cells die locally
of the tonsils. Reticular fibers are sparse and are disposed of by macrophages; the fate
within the center, but they form a concentric of the survivors is unknown. On the one
envelope around its periphery. hand, autoradiographic studies on the tonsil
Germinal centers are thought to pass after 3H-thymidine injection seem to indicate
through a sequence of developmental that lymphocytes arise in the germinal center,
changes and ultimately to involute and dis¬ move outward entering the small lymphocyte
appear. They seem to arise from small nests cap, and finally migrate into the overlying
of large lymphocytes or lymphoblasts, which epithelium. On the other hand, in the ger¬
progressively gain in size and complexity to minal centers of lymph nodes and spleen, no
form aggregations, up to 1 mm in diameter. such centrifugal cell movement is observed
 THE IMMUNE SYSTEM • 429

Medium-sized

Medium-sized lymphocyte

lymphocytes Debris in mitosis Large lymphocytes

Macrophage

Small
lymphocytes
Dividing
large
lymphocyte

Large
lymphocyte

Small
Medium lymphocytes
lymphocytes
Dividing
medium-sized
lymphocyte

Macrophage Large Medium-

lymphocytes sized
lymphocytes

Figure 13-16. Portion of a germinal center of a human lymph node. Hematoxylin-eosin-azure II. (After A. A. Maximow.)

and the small lymphocytes of the cap seem thymectomized at birth and in patients with
to belong to the long-lived variety. congenital thymic aplasia. In birds, their ap¬
The functional significance of the dendritic pearance is prevented by bursectomy, and
cells is not clear. They do not seem to be they are absent in humans with congenital
capable of phagocytosis, for which reason the agammaglobulinemia. Intravenously injected
term “dendritic macrophages” originally ap¬ B lymphocytes localize both in germinal cen¬
plied to these cells is no longer tenable. How¬ ters and in their small lymphocyte cap. Thus,
ever, they have been shown to trap antigen they are probably involved in some stage of
in the presence of antibody and to retain the the development or functional differentia¬
antigen-antibody complex for long periods tion of B lymphocytes.
of time. It has been suggested that the nu¬ The appearance of germinal centers is
merous peripheral processes of these cells closely correlated with the evolution of hu¬
bind the complex to their surface membrane, moral immunological responses. They are
but inert particles such as carbon, titanium formed de novo during the primary response
oxide, and saccharated iron oxide are also to antigen and increase explosively in num¬
retained. The possibility that dendritic cells ber during the secondary response, whereas
may participate in immune induction has they are very few in animals reared in a
been discussed previously. germ-free environment. Furthermore, their
Germinal centers also develop in rodents lymphoid cells synthesize antibody of the IgG
430 • THE IMMUNE SYSTEM

variety, although they do not differentiate found anywhere in the body, with the single
into plasma cells or do so only to a very exception of the central nervous system. A
limited extent. Each germinal center seems special situation is found in the bone marrow,
to produce monospecific antibody; thus, the which in sensu stricto does not belong to the
suggestion has been advanced that the whole immune system, but which represents the
lymphoid population of an individual ger¬ only source for the stem cell precursors of
minal center may represent a clone of cells, lymphocytes in late fetal and postnatal life.
all committed in the response to a single The various regions of the immune system
antigen. Despite the good correlation be¬ have precise functional interrelationships
tween the appearance of germinal centers and are interconnected by an orderly traffic
and the humoral response to antigen, ger¬ of lymphocytes, which exploit blood and
minal centers are not essential for antibody lymph as circulatory pathways. To under¬
production nor for plasma cell formation. In stand these relationships, one must first re¬
the human fetus, there is antibody secretion capitulate the developmental history of the
before germinal centers develop; in the lymphocyte; stem cell precursors arising
course of the primary response to antigen, from the yolk sac in the embryo and from
antibody appears in the blood and plasma the bone marrow in the adult migrate
cells develop before antibody-producing ger¬ through the bloodstream into the thymus and
minal centers can be demonstrated. It has the unknown mammalian analogue of the
been claimed, however, that during the sec¬ avian bursa of Fabricius. Under the influence
ondary response, germinal center formation of thymus and bursa equivalent, these stem
precedes the rise of circulating antibody. On cell precursors undergo antigen-independent
the basis of this observation and experiments proliferation and differentiate into immu¬
on antibody secretion in vitro, transfer of nocompetent T and B lymphocytes, respec¬
immune responses, and effects of drugs or tively. These reenter the bloodstream and
x-ray irradiation, it has been speculated that populate the lymph nodes, the spleen, and
germinal centers develop following repeated the connective tissues of the body. Upon
contact with antigens eliciting antibody secre¬ meeting their appropriate antigen, T and B
tion and that they may therefore be involved lymphocytes are stimulated to transform,
in the long-term memory of the IgG re¬ proliferate, and differentiate, thus giving rise
sponse. At the present state of our knowl¬ to activated T lymphocytes and antibody-
edge, however, it must be admitted that the secreting B lymphocytes and plasma cells. As
precise function of the germinal center, a a result of antigen stimulation, lymphocytes
conspicuous and ubiquitous component of also arise that propagate the response
the lymphoid tissue, is still unknown. throughout the immune system or carry
memory of the primary response, and these
are capable of mounting an enhanced reac¬
tion upon successive exposures to the same
HISTOPHYSIOLOGICAL antigen.
OVERVIEW OF THE An efficient immunological surveillance of
the body is only possible if lymphocytes, each
IMMUNE SYSTEM
endowed with the property to respond to a
single antigen, are able to move freely
LYMPHOCYTE CIRCULATION throughout the body, thus increasing the
chance for them to encounter their appro¬
The immune system consists of (1) specific priate antigen. That this is the case has been
lymphoid organs, (2) masses of lymphoid proved by a series of elegant studies that
tissue embedded in other organs, (3) isolated have disclosed the existence of a continuous
lymphoid cells inhitrating the epithelial and traffic of lymphocytes between the various
connective tissues of the body, and (4) lym¬ lymphoid organs via the blood and lymph.
phocytes circulating with blood and lymph. There are two main patterns of migration of
Among the organs of the immune system, lymphocytes through the body—slow and fast
the thymus and lymph nodes are composed (Fig. 13-17). The movement of stem cells
exclusively of lymphoid tissue, whereas the from bone marrow to thymus and bursa and
spleen possesses an additional component, the subsequent seeding of lymphocytes to the
the red pulp, which is primarily concerned peripheral lymphoid organs are measured in
with nonimmune functions. Aggregates of weeks, during which cells undergo sequential
lymphoid tissue and lymphoid cells can be steps of differentiation. Superimposed upon
 THE IMMUNE SYSTEM • 431

Figure 13-17. Diagram of lymphocyte circulation and differentiation. Stem cell precursors arising from the yolk sac in
the embryo or the bone marrow in the adult enter the blood and migrate into the thymus and the unknown mammalian
analogue of the avian bursa. Here they proliferate and differentiate into T and B lymphocytes. These enter the blood
and seed the peripheral lymphoid organs (lymph nodes, spleen, gut-associated lymphoid tissue), where they undergo
antigen-dependent proliferation and differentiation. SC, stem cell; T, thymus-dependent lymphocyte; B, bursa-dependent
lymphocyte; A, antigen; M, memory lymphocyte; E, effector lymphocyte, which propagates the immune response
throughout the body; ma, macrophage.

this slow traffic is a second type of migratory The plasma cell precursors localize in great
phenomenon, by which long-lived small lym¬ numbers in the lamina propria of the mucosa
phocytes rapidly move from blood to periph¬ of the gut, where they undergo differentia¬
eral lymphoid organs and tissues and back tion into mature plasma cells; a proportion
into the blood. This latter process, called of them may be involved in synthesis and
recirculation, does not involve lymphocyte release of immunoglobulins A.
proliferation and is measured in hours. Fi¬ Recirculation was demonstrated by exper¬
nally, there are hints of a third pattern of imental drainage of lymphocytes from a
cell migration, which may be especially prom¬ chronic fistula of the thoracic duct. This
inent in the course of an acute immune lymphatic channel collects most of the lymph
response; effector lymphocytes and plasma of the body and returns it to the bloodstream;
cell precursors are seeded by lymph and the thoracic duct lymph contains variable
blood throughout the immune system and numbers of cells in different mammals (2 to
connective tissues of the body, thus bringing 30 X 103/mnT in man) predominantly rep¬
about a propagation of the immune response. resented by small lymphocytes (90 to 95 per
432 • THE IMMUNE SYSTEM

cent) (Fig. 13—IA). The remaining cells are seem to recirculate at a slower rate than T
large lymphocytes that do not recirculate and cells and are mobilized with difficulty by
possibly represent plasma cell precursors that prolonged drainage of the thoracic duct
are released into the blood and later selec¬ lymph, possibly because they are inherently
tively localize in the mucosa of the gut. The sluggish or because they are somewhat seg¬
output of small lymphocytes from the tho¬ regated from the main recirculatory pathway.
racic duct is sufficient to replace all blood Drainage of the thoracic duct lymph causes
lymphocytes several times daily; thus, since a selective lymphocyte depletion in specific
the number of blood lymphocytes remains territories of the lymphoid organs and tis¬
constant, they must continuously leave the sues: at the beginning of the experiment,
blood at the same rate as they enter it lymphocytes disappear from the deep cortex
through the thoracic duct. of the lymph nodes, the central region of the
Prolonged drainage of the thoracic duct periarterial lymphoid sheaths of the spleen,
lymph causes pronounced lymphopenia and and the internodular regions of Peyer’s
extreme depletion of the lymphocyte popu¬ patches. These territories are the same that
lation of the spleen, lymph nodes, and gut- appear devoid of lymphocytes in neonatally
associated lymphoid tissue. The bone marrow thymectomized rodents and which were
is not affected; the thymus responds with a therefore designated thymus dependent. This
decrease in weight, but this effect seems to observation can be explained by the fact that
be nonspecific. T lymphocytes represent the main compo¬
If thoracic duct lymphocytes are recovered, nent of the recirculating pool and that they
labeled radioactively in vitro, and injected are rapidly mobilized from the peripheral
intravenously into a syngeneic recipient, it lymphoid organs. More prolonged drainage
can be shown that they rapidly leave the of the thoracic duct lymph leads to lympho¬
bloodstream and localize in the peripheral cyte depletion of the thymus-independent ter¬
lymphoid organs. They do not enter either ritories of the peripheral lymphoid organs:
the thymus or the bone marrow. The idea namely, the superficial cortex and medullary
therefore emerged that a pool of small lym¬ cords of lymph nodes and the peripheral
phocytes continuously migrate from the regions of the white pulp of the spleen. This
blood into the peripheral lymphoid organs finding may reflect the late mobilization of
and tissues, but leave them again to reenter the recirculating B lymphocytes. The recir¬
the blood, either directly or through the culating T and B lymphocytes, labelled in
lymphatic system. This being true, it should vitro and injected into a syngeneic recipient,
be possible to deplete the recirculating pool localize respectively in the thymus-dependent
by destroying lymphocytes at any point along and thymus-independent (or bursa-depend¬
their migratory pathway. This has been ent) territories of the peripheral lymphoid
shown to be the case through local irradiation organs. The mechanism by which recirculat¬
of the hilus of the spleen. ing small lymphocytes leave and subsequently
Recirculation is very rapid, as the average reenter the bloodstream in the spleen or
transit time of the small lymphocytes through leave the blood and enter the lymph in lymph
the blood is 0.6 hours; transit time through nodes will be discussed in the chapters de¬
the spleen is five to six hours, and through voted to these organs (Chapters 15 and 16).
the lymph nodes 15 to 20 hours. Recirculat¬ In normal subjects, despite the continuous
ing lymphocytes represent a substantial frac¬ exchange of lymphocytes between the various
tion of the body’s small lymphocyte popula¬ districts of the immune system, a steady state
tion (about half of it in rats); most, if not all, is reached that ensures a consistent propor¬
of these cells are long-lived (Fig. 13—If?). tion of T and B lymphocytes in blood, lymph,
Since the blood contains a much higher pro¬ and major lymphoid organs. Thus, in the
portion of short-lived small lymphocytes (30 mouse, 65 to 85 per cent of the small lym¬
to 50 per cent in rats) than the thoracic duct phocytes of the lymph nodes and thoracic
lymph, only part of the blood lymphocytes duct lymph and 30 to 50 per cent of those in
must belong to the recirculating pool. The the spleen belong to the T variety. In the
origin and fate of these short-lived blood human blood, 69 to 82 per cent of the lym¬
lymphocytes are poorly understood. The vast phocyte population are T and the remaining
majority of the recirculating lymphocytes be¬ 20 to 30 per cent are B lymphocytes. The
longs to T variety (85 per cent in mice), the significance of the recirculation of the small
remaining being B lymphocytes. These latter lymphocytes is still open to investigation, but
 THE IMMUNE SYSTEM • 433

it has been speculated that they represent, at Berenbaum, M. C.: The autoradiographic localization
least in part, memory cells “patrolling” the of intracellular antibody. Immunology 2:71, 1959.
Bjorneboe, M., and H. Gormsen: Experimental studies
immune system, ready quickly and efficiently
on the role of plasma cells as antibody producers.
to set up a secondary response upon meeting Acta Pathol. Microbiol. Scand. 29:649, 1943.
their appropriate antigen. Bussard, A. E., and J. F. Binet: Electron micrography
of antibody-producing cells. Nature 295:675, 1965.
Cantor, H., and E. Boyse: Lymphocytes as models for
the study of mammalian cellular differentiation.
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Good, R. A., and D. W. Fisher, eds.: Immunobiology. and macrophages. Adv. Immunol. 9:163, 1968.
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 14
THYMUS
Elio Raviola

The thymus is a median organ situated in HISTOLOGICAL

the superior mediastinum anterior to the ORGANIZATION
great vessels as they emerge from the heart.
It extends from the pericardial sac, caudally,
to the root of the neck, cranially. It consists Each thymic lobe is invested by a thin
of two lobes, arising in the embryo as separate capsule of loose connective tissue and is sub¬
primordia on each side of the midline, but divided by primary connective tissue septa
later becoming closely joined by connective that carry blood vessels into a number of
tissue. The thymus attains its greatest relative parenchymal lobules that appear polyhedral
weight at the end of fetal life, but its absolute in shape and are 0.5 to 2 mm in diameter
weight continues to increase, reaching 30 to (Fig. 14—1). The thymic lobules are not, how¬
40 g at about the time of puberty. It then ever, completely independent of one an¬
begins to undergo an involution that pro¬ other. By serial sectioning, one can demon¬
gresses rapidly until in the adult the organ strate continuity from lobule to lobule via
becomes largely replaced by adipose cells. narrow parenchymal bridges. Thus, each
The thymus is the only primary lymphoid lobe of the thymus actually consists of a
organ thus far identified in mammals. It is convoluted parenchymal strand with irregu¬
the first organ to become lymphoid during lar expansions corresponding to the lobules.
embryonic life, being seeded by blood-borne The principal cellular constituents of the
stem cells from the yolk sac, which then thymus are lymphocytes (thymocytes), retic¬
differentiate into lymphocytes within the spe¬ ular cells, and a smaller number of macro¬
cial environment of the thymus. Thymic lym¬ phages. At the periphery of the lobule, lym¬
phocytes undergo intensive, antigen-inde¬ phocytes are numerous and densely packed,
pendent proliferation. For reasons that are whereas at the center of the lobule, lympho¬
not understood, a portion of them degener¬ cytes are fewer in number and reticular cells
ate within the organ, while the remainder have more abundant acidophilic cytoplasm.
enter the bloodstream, populate the periph¬ Thus, each lobule comprises a darkly stained,
eral lymphoid organs, and ultimately differ¬ peripheral region, the cortex, and a lighter
entiate into thymus-dependent or T lympho¬ staining central portion, the viedulla (Figs.
cytes. These are capable of performing a 14—1,14—2). Secondary connective tissue
variety of immunological functions that col¬ septa, carrying blood vessels, extend inward
lectively constitute the cellular immune re¬ from the surface of the cortex and reach as
sponses, and they also cooperate with B lym¬ far as the corticomedullary boundary.
phocytes in humoral responses. Germinal The thymic parenchyma consists of a tri¬
centers are lacking in the thymus and there dimensional network of stellate reticular cells
is no antibody production. Although the ma¬ bounding irregular compartments filled with
jority of the thymic lymphocytes have not yet lymphocytes that are closely aggregated and
acquired immunological competence, the re¬ in direct contact with each other, without
moval of the organ before the immune sys¬ intervening connective tissue. This compact
tem has completed development causes a cellular mass is comparable to an epithelium
specific irretrievable impairment of the or to the brain in its paucity of intercellular
body’s immunological defenses. substance, but small blood vessels do thread

436
 THYMUS • 437

£■, •' ’ ' .. VjfeCk Thymic lobules

Medulla

* :;ft;v<^onneCtive
ATS ^

"Masa'aM s,
corpuscle

Figure 14-1. Section through the thymus of a guinea pig. The thymic lobes consist of polyhedral lobules separated
from each other by connective tissue septa. Each lobule comprises a densely staining peripheral region or cortex and
a lighter staining central portion, the medulla. The dark areas in the medulla are Hassall’s bodies or corpuscles.
Toluidine blue. (Courtesy of G. B. Schneider and S. Clark, Jr.)

their way through it bringing a minimal The Cortex

amount of connective tissue in their adven¬
titia. The stellate reticular cells in the cortex
The reticular cells of the thymus, like those have a scanty acidophilic cytoplasm and a
of the lymph nodes and spleen, are stellate large, oval nucleus, 7 to 11 jam in diameter,
in shape, but their embryonal origin is en- which is smooth-contoured, is lightly staining,
dodermal instead of mesenchymal. They are and contains one or two small nucleoli. In
in places associated with connective tissue electron micrographs (Figs. 14-3,14-4), the
fibers of the reticular type. They occasionally processes of the reticular cells are seen to be
display more obvious epithelial features in joined by small desmosomes. Their cytoplasm
the medulla of the lobule, where they may contains bundles of intermediate filaments,
bound cysts or be organized into concentric some of which seem to insert on the desmo¬
arrays of squamous epithelial cells, compris¬ somes. Their cytoplasmic organelles are un¬
ing the Hassall’s bodies or thymic corpuscles. The remarkable: sparse mitochondria, a few ri¬
thymic reticulum is often referred to as cy- bosomes, either free or attached to rare
toreticulum, to emphasize the cellular nature cisternae of the granular endoplasmic retic¬
of the parenchymal framework of the thymic ulum, and a small Golgi apparatus. Mem¬
lobules. Thymic lymphocytes are morpho¬ brane-bounded vacuoles that contain a trans¬
logically indistinguishable from the lympho¬ parent matrix and a variable amount of
cytes of the blood, lymph, and peripheral debris are also found. Although these are
lymphoid organs. scarce in the fetal thymus and in the super¬
The thymic lobule is a highly dynamic ficial cortex, they increase in number with
structure. Lymphocytes are continuously age, especially in the deep cortical regions of
produced in the cortex; some of them die the lobule. They are probably lysosomes, but
and are destroyed by macrophages, others their significance is obscure, inasmuch as re¬
migrate toward the medulla and enter the ticular cells are incapable of phagocytosis.
bloodstream through the walls of the post¬ At the periphery of the cortex (Fig. 14-3)
capillary venules. and around the blood vessels, attenuated
438 • THYMUS
 THYMUS • 439

Figure 14-3. Electron micrograph of the periphery of a rat thymic lobule. Large and small lymphocytes are seen in the
superficial portion of the cortex. These are separated from the connective tissue of the interlobular septum by a
continuous layer of attenuated processes of the reticular cells (arrows). (Courtesy of E. Raviola.)

reticular cell processes form a continuous ribosomes, whereas cisternae of the granular
limiting sheath that separates the thymic par¬ endoplasmic reticulum are exceedingly rare.
enchyma from the interlobular and adventi¬ A diplosome surrounded by a small Golgi
tial connective tissue. A boundary layer of apparatus is located near a slight indentation
amorphous material analogous to the basal of the nuclear envelope. Mitochondria are
lamina of epithelia intervenes between this few and tend to be grouped near the Golgi
limiting cellular sheath and the connective apparatus. Multivesicular bodies, small dense
tissue. granules, and lipid droplets are seen only
The vast majority of the cell population of exceptionally. The small lymphocytes (Fig.
the cortex is made up of lymphocytes. These 14-4) have a round, darkly staining nucleus,
include large, medium-sized, and small 4 to 5 fxm in diameter, with a small nucleolus.
forms. The largest lymphocytes have a round The rim of cytoplasm is very thin and con¬
or oval nucleus, 9 pirn in diameter, rich in tains a few free ribosomes, mostly dispersed
euchromatin, and containing one or two as single units. The cytocentrum and associ¬
prominent nucleoli. The cytoplasm is rela¬ ated minute Golgi apparatus slightly indent
tively abundant and strongly basophilic. In the nucleus. Mitochondria and granular en¬
electron micrographs (Fig. 14—3), the most doplasmic reticulum are encountered even
prominent feature of the cytoplasm of large less often than in large lymphocytes. Occa¬
lymphocytes is the abundance of free poly¬ sional multivesicular bodies, a rare lipid

Figure 14-2. Sections through the thymus of monkey. A, On left is the cortex of a lobule with densely packed
lymphocytes. On right, and in B, is the medulla; the lymphocytes are fewer in number and the reticular cells have more
abundant acidophilic cytoplasm. The pink, homogenous areas are Hassall’s bodies. Hematoxylin-eosin.
440 • THYMUS

Figure 14-4. Electron micrograph from the deep cortex of a rat thymic lobule, showing among the crowded smail
lymphocytes two reticular cell processes joined by a desmosome (at arrow). (Courtesy of E. Raviola.)

droplet, and small dense granules complete consistent component of the cell population
the list of cytoplasmic organelles. Large and of the cortex. They are scattered throughout
small lymphocytes are at the extremes of a the cortex, and in most mammals except the
continuous spectrum of cells displaying inter¬ mouse, they increase in number at the
mediate gradations of nuclear and cyto¬ boundary region between the cortex and the
plasmic organization. Consistent features of medulla. With the light microscope, they are
cortical lymphocytes are their smooth surface distinguished from reticular cells with some
contour and their polyhedral shape, due to difficulty, but with the electron microscope,
mutual deformation. they can be easily recognized by their lack of
Large lymphocytes make up only a small desmosomes and the presence within the
proportion of the lymphoid population of cytoplasm of phagocytized lymphocytes or
the lobule and tend to be concentrated at the the remnants of their digestion. The cells
periphery of the cortex; progressively smaller that have been described by light microscop-
forms are found in increasing number to¬ ists as containing PAS-positive inclusions are
ward the center of the lobule, and the deep actually macrophages loaded with residual
cortex consists chiefly of tightly packed small bodies.
lymphocytes. Both dividing and degenerat¬ A few plasma cells are present within the
ing lymphocytes are commonly found in the parenchyma and the interstitial connective
cortex. Mitoses are more frequent at the tissue of the involuting thymus. They occur
periphery of the lobule, whereas degenerat¬ at the extreme periphery of the cortex and
ing cells with pyknotic nuclei are most abun¬ along the blood vessels; their significance is
dant in the deep cortical areas. not known. Mast cells may also be found, but
The macrophages represent a minor but they are mainly extralobular.
 THYMUS • 441

The Medulla small variety. They also differ from cortical

small lymphocytes in their irregular shape
In the medulla, the reticular cells are ex¬ and have a somewhat greater amount of
tremely pleomorphic. In some areas they cytoplasm containing relatively few ribo¬
maintain a stellate shape and contain numer¬ somes.
ous bundles of intermediate filaments (Fig. Macrophages are only rarely found in the
14-5); in other areas, they are much larger medulla of the thymus. Granulocytes, espe¬
and have a pale cytoplasm and myriad cyto¬ cially eosinophils, may be found in small
plasmic processes. Some of them contain numbers. Plasma cells are absent from the
granules of unknown nature; others are filled medulla.
with vacuoles. Some are rounded; others are The significance of the pleomorphism of
flattened and wrapped around one another, reticular cells in the medulla is not under¬
giving rise to the structures known as thymic stood. It may well represent an abnormal,
corpuscles or Hassall’s bodies (Fig. 14-6). local response of the thymic reticulum to the
These may reach 100 jam or more in diam¬ loss of surface relationships with the lympho¬
eter, and consist of a concentric array of cytes.
squamous cells joined by many desmosomes In the thymus of nonmammalian verte¬
and containing keratohyalin granules and brates, especially reptiles and birds (and also
conspicuous bundles of cytoplasmic fila¬ rarely in mammals), the medulla of the
ments. The cells in the central part of a thymic lobules displays an extraordinary con¬
Hassalfs corpuscle may degenerate or be¬ geries of seemingly extraneous components,
come calcified. such as striated muscle cells (Hammar’s
Lymphocytes are much less abundant than myoid cells); cysts lined by epithelial cells
in the cortex and are predominantly of the provided with a brush border or with cilia;

Figure 14-5. Electron micrograph from the medulla of rat thymus, showing portions of several reticular cells joined by
desmosomes and containing conspicuous bundles of tonofilaments. (Courtesy of E. Raviola.)
442 • THYMUS

Hassall's body
Figure 14-6. Hassall’s body in the medulla of the thymus of an 8-year-old boy. It consists of a concentric array of
squamous reticular cells. Eosin-azure.

mucus-secreting cells; and reticular cells with still within the cortical parenchyma, the cap¬
large vacuoles lined by microvilli. It is not illaries form a network of branching and
clear whether these unusual constituents anastomosing arcades and turn back toward
have functional significance or are simply the interior of the lobule. In their recurrent
embryonic rests or errors of differentiation. course through the cortex, the capillaries join
Germinal centers may appear in the med¬ to form larger vessels, which can still be
ullary region of the lobules as a consequence classified as capillaries on the basis of their
of certain diseases for which an autoimmune fine structure. These vessels are confluent
pathogenesis has been postulated. with postcapillary venules at the corticomed¬
ullary boundary and in the medulla. As an
exception to this basic pattern, capillaries may
Vessels and Nerves
leave the periphery of the cortex and join
The arteries supplying the thymus arise superficial veins coursing within the interlob¬
from the internal thoracic arteries and their ular connective tissue. The postcapillary ven¬
mediastinal and pericardiacophrenic branches. ules of the corticomedullary boundary and
They ramify in the interlobular connective medulla leave the thymic parenchyma via the
tissue, and their ultimate subdivisions follow secondary connective tissue septa and join to
the secondary connective tissue septa, which form interlobular veins. The majority of
extend inward from the surface of the lob¬ these are ultimately drained by a single
ules; thus, they penetrate the lobule at the thymic vein, a tributary of the left brachioce¬
corticomedullary boundary, without coursing phalic vein.
through the cortex. The arterioles, following Because of the peculiar arrangement of
the boundary between cortex and medulla, the parenchymal blood vessels, the various
give off capillaries that ascend into the cortex, segments of the vascular tree appear to be
joined to each other by collateral anasto¬ spatially segregated within the lobules, the
moses. At the periphery of the cortex, but cortex being exclusively supplied by capillar-
 THYMUS • 443

ies, and the corticomedullary boundary and technique and antisera to surface antigens on
the medulla also containing arterioles and thymic lymphocytes have clearly shown that
venules. There is very little movement of the blood-borne stem cells originating from
macromolecules from blood to thymic par¬ the yolk sac in the embryo migrate into the
enchyma across the capillary walls in the thymic primordium and there differentiate
cortex (Fig. 14—7), whereas the large medul¬ into lymphocytes. These subsequently in¬
lary vessels are highly permeable to sub¬ crease in number, blood vessels penetrate the
stances in the plasma. Thus, only the lymph¬ rudiment, and the parenchyma is gradually
oid population of the cortex is protected converted into a meshwork of stellate cells of
from the influence of circulating macro¬ endodermal origin attached by desmosomes
molecules. This is the structural basis for the and bounding a labyrinthine system of spaces
so-called blood-thymus barrier to antigens. occupied by proliferating lymphocytes. The
Great numbers of lymphocytes enter the medulla arises relatively late in the deep
bloodstream by traversing the walls of the region of the lobules, by disappearance of
postcapillary venules of the corticomedullary many lymphocytes and an enlargement of
junction and those of the medulla. The en¬ the reticular cells.
dothelium of these venules is low, in contrast In man, the thymus is the first organ of
to the cuboidal endothelium of the postcap¬ the immune system in which lymphocytes
illary venules of lymph nodes. It must be appear, and it continues to be the most active
noted, however, that in lymph nodes lym¬ lymphocytopoietic tissue of the body
phocytes migrate in the opposite direction, throughout embryonal life. The rate of
namely from blood to parenchyma. growth of the thymus, when expressed in
Lymphatics are found in the connective relation to the body weight, levels off at the
tissue septa, but they seem to be lacking beginning of the third fetal trimester; a grad¬
within the lobular parenchyma; their lymph ual decline follows, which is already percep¬
is drained by the sternal, tracheobronchial, tible at birth, and continues thereafter. In
and anterior mediastinal nodes. laboratory rodents, on the other hand, the
The thymus receives branches from the thymus continues to grow until the second
vagus and sympathetic nerves. Sympathetic week of postnatal life.
fibers are distributed to the blood vessels, but
the manner of termination of the vagal fibers
is unknown.
NORMAL, ACCIDENTAL, AND
EXPERIMENTAL INVOLUTION
Histogenesis
In man, the thymus arises from an out¬ The thymus undergoes a slow physiological
growth of the endodermal lining of the third process of involution with age; lymphocyte
branchial pouch on each side of the midline; production declines, the cortex becomes thin¬
the fourth branchial pouch often gives rise ner, and the parenchyma shrinks and be¬
to some thymic tissue. The primordium has comes replaced by adipose tissue, which is
a cleftlike lumen continuous with that of the thought to arise from precursors in the in¬
embryonic pharynx and a wall composed of terlobular connective tissue. It is generally
several layers of columnar epithelium. The assumed that the onset of this normal process
lumen disappears as the endodermal bud of age involution is coincidental with puberty,
proliferates, giving rise to solid epithelial out¬ but if relative reduction of the cortical par¬
growths that invade the surrounding mes¬ enchyma is taken as an index of declining
enchyme. Round, basophilic cells have been functional activity, age involution in humans
described in the mesenchyme surrounding actually begins in early childhood. In adults,
the thymic rudiment at very early stages of the thymus is transformed into a mass of
development. adipose tissue, containing scattered islands of
The two separate primordia, after consid¬ parenchyma consisting mainly of enlarged
erable elongation caudally and medially, reticular cells. The parenchyma, however,
meet in the midline in embryos of about does not disappear completely, even in old
eight weeks and acquire a common mesen¬ age. Moreover, experiments involving thy¬
chymal investment. At about the same time, mectomy in adult rodents that have been
lymphocytes appear within the epithelium. deprived of their lymphoid population show
Studies using both the chromosomal marker that the thymus maintains its functional com-
444 • THYMUS

Figure 14-7. Cortical capillary in the mouse thymus. When horseradish peroxidase (molecular weight, 40,000 daitons)
is injected as a tracer into the bloodstream, its progression along the intercellular clefts of the capillary endothelium is
blocked by an impermeable tight junction (arrow). Very little tracer is transported across the endothelium by micropino-
cytotic vesicles, and this is readily sequestrated by perivascular macrophages (arrowheads). RB, residual bodies. Inset.
In addition, a much smaller molecule, such as cytochrome c (molecular weight, 12,000 daitons), is arrested by the
interendothelial junctions (arrow). (From Raviola, E., and M. J. Karnovsky. J. Exp. Med. 136:466, 1972.)
 THYMUS • 445

petence throughout life and can reacquire Much of the current knowledge on the
full lymphocytopoietic capacity. The same functions of the thymus began with experi¬
may be true in man, although it has not been ments in rodents involving thymectomy at a
demonstrated. critical stage of the development of the im¬
The process of gradual age involution can mune system. The removal of the thymus in
be complicated or accelerated by acute invo¬ adult animals has but little effect on periph¬
lutional changes constituting the so-called ac¬ eral lymphocyte populations or immune re¬
cidental involution, which occurs in response sponsiveness, but in newborn rodents, thy¬
to a wide variety of stimuli, such as disease, mectomy causes lymphocytopenia, marked
severe stress, dietary deficiencies, ionizing decrease in the population of long-lived re¬
radiation, injection of colloidal substances, circulating lymphocytes, impairment of cel¬
bacterial endotoxin, adrenocorticotrophic lular immune reactions, and severe depres¬
hormone, and adrenal and gonadal steroids. sion of those antibody responses requiring
Under these conditions, the thymus rapidly the cooperative participation of the thymus-
diminishes in size, primarily because of mas¬ dependent lymphocyte. The inner cortex of
sive death of cortical small lymphocytes and the lymph nodes and the periarterial lymph¬
their destruction by macrophages. Medullary oid sheaths in the spleen fail to develop, but
lymphocytes are less sensitive, so that the plasmacytopoiesis and germinal center for¬
usual pattern of the lobule, with a densely mation are not affected.
staining cortex and a lighter medulla, may In neonatal rodents, the thymus has com¬
be reversed. Acute involution in experimen¬ pleted development, but the peripheral pop¬
tal animals is followed by intensive regener¬ ulation of thymus-dependent lymphocytes
ation, so that the thymus rapidly returns to has not yet been established. The appearance
its former size. of this population is therefore prevented by
removal of the thymus. Once the periphery
has become populated with T lymphocytes,
thymectomy is no longer followed by a dra¬
HISTOPHYSIQLOGY matic deficiency of cellular immune re¬
sponses, unless the peripheral stock of thy¬
The thymus is essential to the development mus-dependent lymphocytes is destroyed by
of the class of lymphocytes that is responsible sublethal, total body irradiation. A notable
for cellular or cell-mediated immunological exception to this rule is represented by sup¬
responses, i.e., homograft rejection, delayed pressor T cells, which are short-lived and con¬
cutaneous reactions to protein antigens (de¬ tinuously generated in the adult thymus.
layed hypersensitivity), immune attack of a Adult thymectomy therefore abolishes T-cell
nonresponsive host (graft-versus-host reac¬ suppressor functions, whereas helper, lym-
tion), and immune response to fungi, facul¬ phokine-producing, and cytolytic T lympho¬
tative intracellular bacteria, and certain vi¬ cytes can only be depleted by neonatal thy¬
ruses. Thymus-dependent lymphocytes do mectomy. In other mammals, including man,
not release conventional antibody, but act as the immune system is fully mature at birth
helper cells in humoral responses. All these and less dependent on the integrity of the
functions are carried out by lymphocytes that thymus.
are circulating in the blood and lymph or In neonatally thymectomized rodents, the
residing in the spleen, lymph nodes, and immunological defect can be corrected by
connective tissues of the body. The vast ma¬ grafting a thymus or injecting lymph node
jority of the lymphocytes located in the thy¬ or spleen cells. Injection of thymic cell sus¬
mus are functionally inert. There is evidence pensions, on the other hand, is relatively
that thymic lymphocytes become immuno¬ ineffective, probably because very few ma¬
competent when they move into the blood or ture lymphocytes are present in the thymus.
peripheral lymphoid organs, but the mecha¬ Experiments in animals that were thymecto¬
nism triggering this further differentiation is mized, irradiated, “reconstituted” with bone
unknown. Thus, the thymic lymphocyte or marrow cells bearing a chromosomal marker,
thymocyte must be regarded as a type of cell and grafted with an unmarked thymus have
functionally distinct from the peripheral thy¬ shown that the original lymphocyte popula¬
mus-dependent (T) lymphocyte, but it rep¬ tion of the grafted thymus is replaced by a
resents the immature precursor of the latter. new population of cells bearing the bone
446 • THYMUS

marrow karyotype; this provides further evi¬ lymphocytes do migrate from the cortex to
dence that the lymphocyte population of the the medulla of the lobule and there enter the
thymus arises from immigration and differ¬ bloodstream through the walls of the post¬
entiation of blood-borne stem cell precursors, capillary venules.
arising in this instance from the bone mar¬ The lymphocytes that leave the thymus
row. “home in'’ to the so-called thymus-dependent
Two possible explanations exist for the regions of the peripheral lymphoid organs,
results of the reconstitution experiments. (1) namely, the inner cortex of lymph nodes, the
The thymus may provide stem cell precursors periarterial lymphoid sheaths of the spleen,
immigrating from the embryonic yolk sac or and the internodular areas of the tonsils,
postnatal marrow with a uniquely favorable appendix, and Peyer’s patches. All these re¬
environment for differentiation into periph¬ gions represent the portal of entry of blood-
eral, thymus-dependent lymphocytes^ The borne lymphocytes into the peripheral sta¬
thymus would therefore seed the immune tions of the immune system and the sites in
system with lymphocytes capable of express¬ which the thymus-dependent lymphocytes
ing the thymus-dependent immune func¬ are preferentially concentrated.
tions. (2) Alternatively, the thymus may pro¬ Although the vast majority of thymocytes
duce one or more factors that act at the are not immunocompetent, a small pro¬
periphery on stem cell precursors, triggering portion of them seem to be capable of
their differentiation into thymus-dependent transformation upon interaction with phyto¬
lymphocytes. Whereas little doubt exists that hemagglutinin or allogeneic cells. These lym¬
the thymus seeds the peripheral lymphoid phocytes are less sensitive to x-ray irradiation
organs with lymphocytes, no definitive evi¬ and to the cytolytic effect of adrenal steroids,
dence is available so far that thymic factors and they have therefore been identified with
are released into the circulation and act on a the lymphocytes located in the medulla of
peripheral target. the lobule. These observations seem to sug¬
Once stem cells have migrated into the gest that the small lymphocytes of the cortex
thymus, they differentiate into thymic lym¬ of the lobule are functionally inert and ac¬
phocytes, possibly under local inductive influ¬ quire immunological competence as they mi¬
ences, and begin a sustained proliferation. At grate to the medulla.
the peak of its activity, which corresponds to Indirect evidence supporting this hypoth¬
the perinatal period in laboratory rodents, esis comes from a growing body of animal
the thymus has the highest rate of lympho¬ experiments with antisera to surface antigens
cyte production in the whole immune system. on the thymic or thymus-derived lympho¬
Larger lymphocytes proliferate in the super¬ cytes. In the mouse, thymus lymphocytes
ficial cortex and give rise to generations of possess membrane constituents, such as the
smaller cells that accumulate in the deep Thy-1, TL, GIX, and Ly antigens, which can be
cortical areas of the lobule. This lymphocyte detected by immunological methods; their
proliferation does not depend on antigen significance is poorly understood, but they
stimulation, in contrast with the situation in can be usefully employed as cell markers for
peripheral lymphoid organs. However, the investigating cell interactions, differentiation,
mechanism controlling the rate of lympho¬ and migration. Stem cells lack these surface
cyte production is unknown. antigens; as they move into the thymus and
The intrathymic life span of the newly differentiate into thymic lymphocytes or thy¬
formed small lymphocytes has been shown to mocytes, they acquire the Thy-1, TL, GIX,
be very short (two to three days). It follows and Ly 1,2 surface antigens. Upon migration
that lymphocytes either die within the organ out of the thymus and differentiation into
or migrate out of the thymus. Lymphocyte peripheral T cells, the Thy-1 antigen de¬
death does occur in the cortex, and these creases in amount on the lymphocyte surface,
cells are disposed of by macrophages. The TL and GIX disappear altogether, and Ly
extent of this cell death and its physiological 1,2 persists on about 50 per cent of the
significance are not understood. However, peripheral T lymphocytes. Ly + l, + 2 lym¬
evidence coming from autoradiographic and phocytes undergo further differentiation at
chromosome marker labeling, from experi¬ the periphery into Ly +1,-2 cells, which
ments involving thymectomy, and from lym¬ can either express helper function or pro¬
phocyte counts in arterial and thymic venous duce lymphokines, and into Ly — l, + 2 cells,
blood shows that a proportion of the thymic which can express either cytolytic or sup-
 THYMUS • 447

pressor functions. From these studies the Cantor, H., and E. Boyse: Lymphocytes as models for
hypothesis has emerged that the differentia¬ the study of mammalian cellular differentiation.
tion of thymic lymphocytes into peripheral, Immunol. Rev. 55:105, 1977.
Cantor, H., M. A. Mandel, and R. Asofsky: Studies of
thymus-dependent lymphocytes consists of
thoracic duct lymphocytes of mice. II. A quantitative
an orderly succession of maturational steps: comparison of the capacity of thoracic duct lympho¬
first, undifferentiated stem cell precursors of cytes and other lymphoid cells to induce graft-
yolk sac or bone marrow origin enter the versus-host reactions. J. Immunol. 704:409, 1970.
Claman, H. N., and F. H. Brunstetter: The response of
thymus and differentiate into thymic lym¬
cultured human thymus cells to phytohemagglu¬
phocytes. These proliferate in the cortex, tinin. J. Immunol. 700:1127, 1968.
then move to the medulla and acquire im- Claman, H. N., and F. H. Brunstetter: Effects of anti¬
munocompetence either at the very moment lymphocyte serum and phytohemagglutinin upon
of leaving the organ or after they have en¬ cultures of human thymus and peripheral blood
lymphoid cells. I. Morphologic and biochemical
tered the bloodstream through the wall of
studies of thymus and blood lymphoid cells. Lab.
the postcapillary venules. Invest. 18:757, 1968.
It was reported that immunological re¬ Clark, S. L., Jr.: The thymus in mice of strain 129/J
sponsiveness can be partially restored in studied with the electron microscope. Am. J. Anat.
772:1, 1963.
thvmectomized animals by grafts of thymus
Clark, S. L., Jr.: Incorporation of sulfate by the mouse
enclosed in chambers that are permeable to thymus: its relation to secretion by medullary epi¬
molecules but not to cells. In addition, active thelial cells and to thymic lymphopoiesis. J. Exp.
factors were isolated, which simulate the Med. 128:927, 1968.
Cohen, M. W., G. J. Thorbecke, G. M. Hochwald, and
function of the thymus both in vivo and in
E. B. Jacobson: Induction of graft-versus-host re¬
vitro, and were variously called thymosin, thy¬ action in newborn mice by injection of newborn or
mopoietin, thymic humoral factor, and thymic adult homologous thymus cells. Proc. Soc. Exp. Biol.
serum factor (FTS). Of these, FTS was char¬ Med. 774:242, 1963.
acterized biochemically as a nonapeptide 847 Colley, D. G., A. Malakian, and B. H. Waksman: Cellular
differentiation in the thymus. II. Thymus-specific
daltons in molecular weight and localized in
antigens in rat thymus and peripheral lymphoid
the thymic reticular cells by immunocyto- cells. J. Immunol. 704:585, 1970.
chemistry. It has been claimed that these Colley, D. G., A. Y. Shih Wu, and B. H. Waksman:
factors are actually hormones and that the Cellular differentiation in the thymus. III. Surface
properties of rat thymus and lymph node cells
thymus exerts its functions on the immune
separated on density gradients. I. Exp. Med.
system by releasing them into the blood¬ 752:1107, 1970.
stream as an endocrine gland. The evidence Dardenne, M., and J. F. Bach: Studies on thymus prod¬
supporting this view is controversial, how¬ ucts. I. Modification of rosette-forming cells by
ever, and it seems more likely that the various thymic extracts. Determination of the target RFC
subpopulation. Immunology 25:343, 1973.
thymic factors act within the organ as media¬
Davies, A. J. S., E. Leuchars, V. Wallis, R. Marchant,
tors of short-range cellular interactions. and E. V. Elliott: The failure of thvmus-derived
cells to produce antibody. Transplantation 5:222,
1967.
Defendi, V., and D. Metcalf, eds.: The Thymus. The
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in the thymus of the mouse: qualitative and quan¬ Wallis: The thymus and circulating lymphocytes of
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Immunogenetics and Immune Regulation. Milano, cytopoietic factor (thymosin). Proc. Natl. Acad. Sci.
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Blomgren, H., and B. Andersson: Evidence for a small Good, R. A., and A. E. Cabrielsen, eds.: The Thymus
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Haelst, U. van: Light and electron microscopic study of Owen, J. J. T., and M. A. Ritter: Tissue interaction in
the normal and pathological thymus of the rat. 1. the development of thymus lymphocytes. J. Exp.
The normal thymus. Z. Zellforsch. 77:534, 1967. Med. 729:431, 1969.
Hoshino, T., M. Takeda, K. Abe, and T. Ito: Early Parrott, D. M. V., M. A. B. de Sousa, and J. East:
development of thymic lymphocytes in mice, studied Thymus-dependent areas in the lymphoid organs
by light and electron microscopy. Anat. Rec. 764:47, of neonatally thymectomized mice. J. Exp. Med.
1969. 725:191, 1966.
Ishidate, M., and D. Metcalf: The pattern of lympho¬ Raff, M. C.: Evidence for subpopulation of mature
poiesis in the mouse thymus after cortisone admin¬ lymphocytes within mouse thymus. Nature New
istration or adrenalectomy. Aust. J. Exp. Biol. Med. Biol. 229:182, 1971.
Sci. 47:637, 1963. Raviola, E., and M. J. Karnovsky: Evidence for a blood-
Izard, J.: Ultrastructure of the thymic reticulum in thymus barrier using electron-opaque tracers. J.
guinea pig. Cytological aspects of the problem of Exp. Med. 756:466, 1972.
the thymic secretion. Anat. Rec. 755:117, 1966. Raviola, E., and G. Raviola: Striated muscle cells in the
Kennedy, J. C., L. Siminovitch, j. E. Till, and E. A. thymus of reptiles and birds: an electron micro¬
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rocytes. Proc. Soc. Exp. Biol. Med. 726:868, 1965. cytes from the thymus: a review and study of the
Leckband, E., and E. A. Boyse: Immunocompetent cells problem. Rev. Can. Biol. 56:51, 1971.
among mouse thymocytes: a minor population. Sci¬ Sanel, F. T.: Ultrastructure of differentiating cells dur¬
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Lundin, P. M., and U. Schelin: Ultrastructure of the rat Schwarz, M. R.: Transformation of rat small lympho¬
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cortex of guinea pig thymus. Z. Zellforsch. 92:159, Warner, N. L.: The immunological role of different
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Miller, H. C., S. K. Schmiege, and A. Rule: Production logical competence of thymic cell suspensions. Aust.
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with thymic factor. J. Immunol. 777:1005, 1973. Weber, W. T.: Difference between medullary and cor¬
Miller, J. F. A. P., and P. Dukor: Die Biologie des tical thymic lymphocytes of the pig in their response
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Moore, M. A. S., and J. J. T. Owen: Experimental studies H. Waksmann: Antigenic markers on cells leaving
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Murray, R. G., A. Murray, and A. Pizzo: The fine Winkelstein, A., and C. G. Craddock: Comparative re¬
structure of the thymocytes of young rats. Anat. sponse of normal human thymus and lymph node
Rec. 757:17, 1965. cells to phytohemagglutinin in culture. Blood
Order, S. E., and B. H. Waksman: Cellular differentia¬ 29:594, 1967.
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Med. 752:1216, 1970.
 L YMPH NODES
Elio Raviola

Lymph nodes are small organs occurring structural differences according to their lo¬
in series along the course of lymphatic ves¬ cation in the body. Extreme examples are,
sels. Their parenchyma consists of a highly on the one hand, the small, poorly developed
organized accumulation of lymphoid tissue, popliteal node of unstimulated laboratory
which recognizes antigenic materials in the rodents and, on the other hand, the highly
lymph that percolates through the node, and organized human mesenteric nodes, which
builds up against them a specific immune are continuously bombarded by a variety of
reaction. Lymph nodes are also very rich in antigens of intestinal origin. If, however, mi¬
macrophages, which clear the lymph of un¬ croorganisms or a high dose of any foreign
desirable cells, invading microorganisms, and macromolecule is injected subcutaneously, or
other particulate matter. if a foreign tissue is transplanted into the
Large numbers of lymph nodes occur, usu¬ body, the lymph nodes draining the site of
ally in groups, scattered throughout the pre- injection or grafting undergo profound
vertebral region, along the large blood vessels structural changes typical of an acute primary
of the thoracic and abdominal cavities, be¬ or secondary immune response.
tween the leaves of the mesentery, and in the
loose connective tissue of the neck, axilla,
and groin. They are commonly flattened and
ovoid or reniform in shape, varying from 1
HISTOLOGICAL
to 25 mm in diameter, with a slight indenta¬ ORGANIZATION
tion, the hilus, where blood vessels enter or
leave the organ. In most mammals, as afferent The lymph node consists basically of a
lymphatic vessels approach the node they give parenchymal mass of lymphoid tissue, trav¬
rise to a number of branches that enter the ersed by specialized lymph vessels or sinuses
node at multiple sites over its convex surface (Figs. 15—1, 15—2). Its collagenous framework
(Fig. 15-1). A smaller number of efferent consists of a capsule, which invests the whole
lymphatic vessels leave the node at the hilus. organ, but which is greatly thickened at the
The afferent vessels are provided with valves hilus. From the capsule a variable number of
that open toward the node, whereas the branching connective tissue trabeculae extend
valves of the efferent lymphatics point away into the substance of the node. The lymphoid
from the hilus. This arrangement of valves parenchyma between the trabeculae is sup¬
ensures unidirectional lymph flow through ported by a tridimensional network of retic¬
the node. ular fibers with associated reticular cells. The
Lymph nodes vary somewhat in their struc¬ meshes of this network are filled with lym¬
ture from species to species, but their varia¬ phocytes, plasma cells, and macrophages.
tion in appearance within species depends The lymph sinuses are irregular channels
primarily upon their state of activity. In a transformed into a labyrinth of intercom¬
healthy subject, each lymph node reflects in municating chambers by a loose network of
its organization both the background activity tissue strands that traverse their lumen.
of the immune system as a whole and the Under low magnification, the sectioned
local response of the node to small amounts lymph node is seen to have an outer, densely
of antigens reaching it from the body terri¬ staining cortex and an inner, paler medulla.
tory drained by its afferent lymphatics. Rest¬ The difference in appearance of these two
ing lymph nodes therefore show pronounced regions is due mainly to differences in num-
449
450 • LYMPH NODES

afferent
germinal center /lymphatic vessel

postcapillary
venules

outer
cortex intermediate
sinus

marginal
sinus
inner''
cortex

trabecula
medullary cord

medullary sinus

efferent
lymphatic
vessel
artery
Figure 15-1. Diagram of lymph node. Blood supply is depicted on right side.

Medullary cord Medullary sinus

Trabecula

Capsule

Germinal
center

Subcapsular
sinus

Inner or deep
cortex

T rabecula Subcapsular sinus

Germinal center

Figure 15-2. Section through small jugular lymph node of man. (Redrawn and slightly modified from Sobotta.)
 LYMPH NODES • 451

ber, diameter, and arrangement of the lymph Nodes deep in the body, as in the abdominal
sinuses, 4 he relative amounts of cortical and cavity, are distinguished by the relatively
medullary substance and their distribution poor development of their trabeculae com¬
vary within wide limits. The nodes of the pared with that of the more peripheral
abdominal cavity are especially rich in med¬ nodes. In some cases, a hilus may be absent;
ullary substance. The cortical substance in in others, it is highly developed, with its
some nodes may surround the medulla com¬ connective tissue penetrating far into the
pletely, but in others the medullary substance node and partitioning it extensively.
may border directly on the capsule for long
distances. In some cases the medulla and
Lymph Sinuses
cortex may accumulate at opposite poles of
the node. In the pig, the “cortical” substance The afferent lymphatic vessels approach
is collected in the central portion of the node, the convex surface of the node, pierce its
while “medullary” tissue is located at the capsule obliquely, and open into the marginal
periphery. In the ox, the cortex is extensively or sub capsular sinus (Figs. 15-3, 15-4). This is
compartmentalized by numerous trabeculae, not a cylindrical channel, but a bowl-shaped
whereas in man trabeculae are poorly devel¬ cavity, which separates the capsule from the
oped and the cortex appears as a diffuse, cortical parenchyma. It is traversed by the
continuous mass. The trabeculae are promi¬ collagenous trabeculae and communicates at
nent in large lymph nodes, but in small nodes the hilus with the lumen of the efferent
they are thin and frequently interrupted. lymph vessels. Arising from the marginal

Figure 15-3. A, Capsule, marginal sinus, and part of two germinal centers in a mesenteric lymph node of a dog. B,
Medullary sinuses and cords from a mesenteric lymph node of a dog. A trabecula is seen on the right carrying an artery
and a vein; the vein receives tributaries from neighboring medullary cords. Hematoxylin-eosin.
452 * LYMPH NODES

Small lymphocytes Marginal sinus

Artery
Reticular cell
Afferent Blood vessel Capsule

Marginal sinus
Periphery
of nodule

Blood
vessel

Trabecula

Intermediate
sinus Blood vessel
Small vessel
lymphocytes Germinal center
Figure 15-4. Diagram of the marginal sinus in a lymph node of a dog. Hematoxylin-eosin-azure II. (After A. A.
Maximow.)

sinus are radially directed lymph channels with myriad surface protrusions. Also pres¬
called the intermediate or cortical sinuses, which ent are smaller cells, probably lymphocytes.
penetrate the cortical parenchyma, usually The framework of the sinus walls is a layer
following along the collagenous trabeculae. of reticular fibers, continuous with the paren¬
The compact appearance of the cortex is chymal reticulum. These fibers directly un¬
primarily due to the relatively small number derlie the sinus lining cells without any inter¬
and narrow lumen of the intermediate sin¬ vening basal lamina. The intraluminal
uses. These continue into the medulla as network of cell processes is supported by a
medullary sinuses—large, tortuous, irregular skeleton of reticular and elastic fibers an¬
channels, that branch and anastomose re¬ chored to the reticulum of the sinus walls
peatedly, thus fragmenting the lymphoid and suspended from the collagenous frame¬
parenchyma into a number of medullary cords work of the capsule and the trabeculae. The
(Figs. 15-3, 15-5). The sinuses of the med¬ fibers traversing the sinuses are not directly
ullary substance are confluent with the mar¬ exposed to the lymph, but are completely
ginal sinus at the hilus and form there a invested by the luminal stellate cells, often
plexus of tortuous vessels, which penetrate lying embedded in deep invaginations of the
the thickened capsule of this region and cell surface.
continue into the efferent lymphatics. There is no doubt that the wall of the
Seen with the scanning electron micro¬ sinuses is freely permeable to the constituents
scope (Fig. 15-6), the sinuses appear as tun¬ of the lymph and is continually crossed by
nels lined by a layer of attenuated cells, but wandering cells* which move freely between
their lumen is bridged by a meshwork of lymph and lymphoid parenchyma. However,
stellate cells, which are connected to each the nature and physiological properties of
other and to the opposite walls of the sinus the cells lining the walls of the sinuses and
via slender cell processes. Projecting from those traversing their lumens have long been
the sinus walls and luminal network of stellate subjects of controversy and are far from
elements are rounded macrophages, hirsute being settled satisfactorily. Traditionally,
 LYMPH NODES • 453

Medullary Medullary
Trabecula cord Arteriole sinus
Trabecula

Medullary sinus

Medul¬
lary
cord

Arteriole

Medullary
sinus

Trabeculae

Vein

Medullary Medullary
sinus
Figure 15-5. Diagram of medulla in a lymph node of a dog. Hematoxylin-eosin-azure II. (After A. A. Maximow.)

these cells were identified with the “reticular cells. The source of the macrophages is still
cells” of the lymphoid parenchyma and were unknown. Nor is it clear whether they are
thought to be capable of phagocytosis. At permanently associated with the endothelium
present, however, there are indications that (tissue macrophages), having migrated from
there are two distinct categories of lining the parenchyma into the sinuses, or are
cells: macrophages and flattened or stellate lymph-borne elements that secondarily took
endothelial cells. These latter have an incon¬ up residence in the sinus walls and acquired
spicuous complement of cell organelles. They phagocytic properties.
are connected to each other by specialized The relative proportions of macrophages
junctions and they take up only small and endothelial cells vary in different regions
amounts of particulate matter by endocytosis, of the node. The capsular wall of the mar¬
like the endothelium of blood vessels gener¬ ginal sinus and the wall of the intermediate
ally. Macrophages are inserted into the sinus sinuses adjacent to the collagenous trabeculae
wall or trapped in the meshes of the luminal are composed exclusively of flattened endo¬
network of tissue strands, apparently stuck thelial cells, whose outlines are easily stained
to the surface of the endothelial cells. They by treatment with silver nitrate. On the other
lack specialized intercellular junctions, do not hand, macrophages are especially numerous
invest the reticular fibers, and contain the in medullary sinuses, and there the outlines
pleomorphic complement of cell organelles of endothelial cells usually cannot be dem¬
and residual bodies typical of phagocytic onstrated by silver nitrate.
454 • LYMPH NODES

Figure 15-6. Scanning electron micrograph of the lumen of a sinus in a mesenteric lymph node of a dog. The spider
web of luminal stellate cells acts as a multitude of microscopic baffles that generate turbulence in the lymph and
facilitate the monitoring function of the macrophages deployed along the sinus walls. On left, two rounded cells, probably
lymphocytes, were trapped in the lumen of the sinus. (From Fujita, T., et al. Z. Zellforsch. 733:147, 1972.)

The organization of the lymph sinuses is been customary to classify certain regional
well suited to the filtering function of the differentiations in the cortical parenchyma
lymph node. The lymph entering the organ as primary lymphoid nodules, secondary nodules,
through the afferent vessels floods the mar¬ and diffuse lymphoid tissue. Secondary nodule
ginal sinus and slowly percolates through the is a term that has been used to describe either
intermediate and medullary sinuses, freely the germinal centers proper or the germinal
exchanging with the lymphoid parenchyma centers together with their cap of small lym¬
substances in solution, particulate matter, phocytes. Primary nodules and germinal cen¬
and cells. The system of sinuses functions as ters are usually located at the periphery of
a “trap” or “settling chamber,” with the intra¬ the node and together comprise the outer
luminal tissue strands acting as a multitude cortex, whereas the inner or deep cortex (also
of microscopic baffles that increase the sur¬ called the paracortical area) consists of dif¬
face exposed to the lymph, generate turbu¬ fuse lymphoid tissue (Fig. 15-7). No clear-
lence, and facilitate the monitoring function cut boundary exists between outer and deep
of the macrophages deployed along the sinus cortex, and the latter continues without de¬
walls. marcation into the medullary cords. Further¬
more, the relative proportions of the two
Cortex zones are quite variable in different lymph
nodes and in various functional states. This
The cortical parenchyma appears with the subdivision does, however, have considerable
light microscope as a dense mass of lymphoid physiological importance, because only the
cells, traversed in places by the collagenous deep cortex is populated by lymphocytes of
trabeculae and intermediate sinuses. It has the recirculating pool and it contains postcap-
 LYMPH NODES • 455

Figure 15-7. Portion of the auricular lymph node of a mouse. Notice the considerable thickness of the inner cortex.
Toluidine blue. (Courtesy of G. B. Schneider.)

illary venules with cuboidal endothelium, are polarized in such a way that their light
which represent the portal of entry of blood- regions, invested by the small lymphocyte
borne lymphocytes into the node. cap, point toward the marginal sinus that
Primary nodules are not clearly defined receives the incoming lymph (Fig. 15-8). In
morphological entities. In man and labora¬ the lymph nodes of the pig, these relation¬
tory rodents, they consist merely of the ships are reversed, with the germinal centers
lymphoid tissue intervening between the ger¬ pointing toward the central sinus, which in
minal centers of the outer cortex. Only in this species receives the incoming lymph. In
species such as the ox, whose lymph nodes germinal centers situated deep in the cortex,
possess a well-developed system of trabeculae the orientation of the light pole and lympho¬
and intermediate sinuses, does the paren¬ cytic cap is either random or toward a neigh¬
chyma of the outer cortex appear subdivided boring intermediate sinus. The structure of
into discrete, rounded aggregates, projecting germinal centers conforms to the general
slightly at the surface of the organ. The description provided in Chapter 13.
stroma of the primary nodules consists of a In the deep cortex, cells are more loosely
rather loose network of reticular fibers and packed than in the outer cortex. Small lym¬
associated reticular cells. The labyrinthine phocytes predominate, while large lympho¬
interstices of this reticulum are occupied by cytes, macrophages, and plasma cells are
small lymphocytes. Both large lymphocytes found only occasionally. Reticular fibers are
and macrophages are rare; mature plasma more abundant than in the outer cortex and
cells are usually lacking. increase additionally in number at the junc¬
Germinal centers occur in variable num¬ tion of the cortex with the medulla. Consid¬
bers in the outer cortex, are less commonly erable attention has been paid in the recent
found deeper in the cortex, and are only past to a type of cells normally found in the
exceptionally present in the medullary cords. resting lymph node, which are characterized
Those located at the lymph node periphery by euchromatic nucleus, pale cytoplasm, few
456 • LYMPH NODES

frequently, they form loops that continue

into other cords. The medullary cords are
not very prominent in resting lymph nodes.
They consist of a rich network of reticular
fibers and reticular cells, enclosing small lym¬
phocytes, mature plasma cells, and macro¬
phages.
The lymph node parenchyma contains var¬
iable, but usually small numbers of granulo¬
cytes and erythrocytes. The number of these
cellular elements may, however, be greatly
increased upon stimulation or in pathological
states.

Capsule and Trabeculae

The capsule of the lymph node consists of
dense collagenous fibers with a few fibroblasts
and, especially on its inner surface, networks
of thin elastic fibers. A few smooth muscle
cells are also found in the capsule around
the points of entry and exit of the afferent
and efferent lymph vessels. The outer aspect
of the capsule blends into the fat or loose
connective tissue surrounding the lymph
node, whereas its inner aspect is more sharply
defined and is lined by the endothelium of
Figure 15-8. Germinal center in the outer cortex of a the marginal sinus, except where it gives rise
mesenteric lymph node of a dog. Hematoxylin-eosin. to the cortical trabeculae. The trabeculae do
not represent complete septa, but cylindrical
or flattened beams of dense connective tissue;
they traverse the node completely en-
cell organelles, and numerous surface proc¬
sheathed by the intermediate and medullary
esses that interdigitate with those of the ad¬
lymph sinuses, and, near the hilus, they carry
joining lymphocytes. They are called interdi-
the major blood vessels.
gitating cells and are localized in the deep
cortex. In places, they contain granules that
have the same structure as those present in
Blood Vessels and Nerves
the cytoplasm of the Langerhans cells in the
epidermis. Furthermore, cells with numerous Almost all the blood vessels destined for
surface ruffles have been isolated from the the lymph node enter the organ through the
afferent lymph (veiled cells); they are con¬ hilus, with only occasional small ones enter¬
tained in the lumen of the sinuses or appear ing through the rest of the capsule (Fig. 15-
to be migrating across the endothelium of 1). The larger arterial branches initially run
the marginal sinus. Veiled cells may contain within the trabeculae, but they soon enter
Langerhans-type granules in their cytoplasm. the medullary cords and supply their capil¬
I heir relationships with interdigitating cells lary networks. Passing along the cords, the
and the dendritic cells of the germinal centers arteries reach the cortex, where they distrib¬
are unclear. ute to capillary plexuses of the diffuse cortical
parenchyma and around the germinal cen¬
Medulla ters. Special postcapillary venules with cuboidal
endothelium arise from these peripheral cap¬
The medullary cords consist of aggrega¬ illary plexuses and course radially through
tions of lymphoid tissue organized around the deep cortex to enter the medullary cords.
small blood vessels. The cords branch and Here they give rise to small veins lined with
anastomose freely with one another, and near normal endothelium. These in turn are tri¬
the hilus they terminate blindly or, more butaries of the larger venous channels that
 LYMPH NODES • 457

They have tall endothelial cells and no mus¬

cular coat, and their wall is traversed by great
numbers of blood-borne small lymphocytes,
which migrate into the lymph node paren¬
chyma either by perforating the endothelial
cells or by insinuating themselves into the
interendothelial clefts. Similar vessels occur
in certain segments of the vascular tree of
the tonsils, Peyer’s patches, and appendix.
The exact significance of the cuboidal endo¬
thelium is obscure, but it has been suggested
that the compliant endothelial cells adapt
their surface contours to the shape of the
wandering lymphocytes, thus limiting the
plasma loss that might otherwise be associ¬
ated with cell migration. Circulating lympho¬
cytes seem to be capable of specifically recog¬
nizing this segment of the vascular tree as
the portal of entry into the lymph node
parenchyma. The mechanism of this remark¬
able phenomenon is unknown, but it may
imply some sort of specific surface interaction
between lymphocytes and cuboidal endo¬
thelial cells. Experiments involving enzymatic
digestion suggest that carbohydrate moieties
of membrane glycoproteins are the essential
component of the recognition sites on the
surface of the lymphocytes. On the other
hand, immunoglobulins have been demon¬
Figure 15-9. Small lymphocytes migrating through the strated at the surface of the endothelial cells,
endothelium of a postcapillary venule in the popliteal and these might also be the basis for selective
lymph node of a rabbit. Toluidine blue. (Courtesy of C.
Compton.) capture of circulating lymphocytes.
Nerves enter the hilus of the node with the
blood vessels, forming perivascular plexuses.
accompany the major arterial trunks in the In the trabeculae and medullary cords, nerve
interior of the collagenous trabeculae. fibers are observed that are independent of
The postcapillary venules of the deep cor¬ the vessels, but in the cortex all nerves are
tex are of special interest (Figs. 15-9, 15—10). probably of vasomotor type.

Lumen of venule

Migrating
Migrating sma 11
sma 11 mphocyte
lymphoc
Figure 15-10. Diagram of lymphocyte
migration through the walls of the post¬
capillary venules. (After A. A. Maxi¬
Endothel ia I
mow.)
cell
nucleus

Granulocyte Small Endothelial

lymphocytes cell nucleus
458 • LYMPH NODES

Reticular cells

Sma! I
lymphocyte

Macrophage
with carmine
granules
Mitosis of
large
lymphocyte

Erythrocyte

Macrophage
containing
erythrocytes

macrophage with
carmine granules
Large lymphocyte
Figure 15-11. Medullary sinus of mesenteric lymph node of a rabbit that had repeated intravenous injections of lithium
carmine. Hematoxylin-eosin-azure II. (After A. A. Maximow.)

HISTOPHYSIOLOGY OF phages of the sinus walls remove them by

LYMPH NODES phagocytosis (Fig. 15—11). Lymph nodes,
however, represent a much less efficient bar¬
rier for lymph-borne cells and may even
The delicate walls of the lymphatic capil¬ facilitate dissemination of certain viruses.
laries are easily penetrated by macromole¬ Erythrocytes injected into the afferent lym¬
cules, particulate matter, and wandering cells phatic vessels are effectively retained by the
of the connective tissue. Furthermore, bac¬ node, although the filtering efficiency de¬
terial cells may cross the epidermis or the creases when very large numbers of red
epithelium of the mucous membranes lining blood cells are infused. On the other hand,
the body cavities. Escaping destruction by lymph nodes retain only a small proportion
local or blood-borne phagocytes, they may of lymph-borne cancer cells. Viruses that are
proliferate and release toxins. Both micro¬ capable of proliferating in the lymph node
organisms and their toxins easily gain access cells are quickly disseminated throughout the
to the lymph. Lymph nodes intercalated body, possibly carried by the recirculating
along the path of lymph drainage prevent lymphocytes.
entry into the bloodstream of potentially The function of the lymph nodes in the
harmful macromolecules, particulate matter, immunological defenses of the body is a
and bacteria carried by the lymph, and exert property of their lymphocyte and plasma cell
immunological surveillance on their antigenic populations, assisted by the macrophages in
determinants. both the preliminary phase of antigen rec¬
The filtering capacity of the lymph nodes, ognition and the terminal phase of antigen
l ecognized long ago by Virchow and substan¬ disposal. To initiate a primary immune re¬
tiated by experiments involving perfusion sponse, uncommitted T and B lymphocytes
with particulate matter and bacterial cells, is must be present in the resting lymph node,
based on both mechanical and biological whereas memory cells are required for a
mechanisms. The labyrinthine configuration secondary response. Upon interaction with
of the sinuses favors arrest of the particles antigens that elicit a humoral response, lym¬
suspended in the lymph, and the macro¬ phocytes are activated and undergo antigen-
 LYMPH NODES • 459

dependent proliferation and differentiation. thymus or the bone marrow. Neonatal thy¬
As a consequence, plasma cells appear in the mectomy is followed by a deficit of small
node and antibody is synthesized and re¬ lymphocytes in the deep cortex of lymph
leased into the efferent lymph, together with nodes, whereas outer cortex and medullary
lymphoid cells that propagate the response cords are unaffected; furthermore, experi¬
throughout the body. In cellular immune ments involving careful infusion of radioac¬
responses, lymphocyte activation and differ¬ tive label into the thymus show that thymic
entiation lead to the development and sys¬ lymphocytes specifically “home in” to the
temic dissemination of lymphokine-produc- deep cortex. Lymphocytes of bone marrow
ing and cytolytic lymphocytes. Although origin also appear to migrate continuously
these events are known to occur in the lymph into the resting lymph node; they do not
node, the precise location of the relevant cell seem to localize in specific regions of the
populations within the organ and the ultra- organ, but instead become distributed
structural details of their antigen-dependent throughout cortex and medulla. It is still a
differentiation are poorly understood. matter of conjecture whether or not this
In resting peripheral lymph nodes, such as direct inflow of lymphocytes from thymus
the popliteal node of the sheep, which has and bone marrow endows the lymph node
been studied in some detail, the afferent with a complement of newly formed, uncom¬
lymph contains very few cells. Among these mitted elements, capable of reacting to anti¬
are lymphocytes, macrophages, and occa¬ gens never experienced previously by the
sional granulocytes. The efferent lymph con¬ immune system. Also unknown is the life
tains 20 to 75 times more cells than the span of these cells and their contribution to
afferent lymph, and these are predominantly the recirculating pool.
small lymphocytes (98 per cent). Only a very Observations on the effects of thymectomy
small fraction of the small lymphocytes and the selective “homing” of both thymic
emerging with the efferent lymph actually and thoracic duct lymphocytes have given
arise from division of precursors in the node, rise to the concept that the parenchyma of
the vast majority (95 per cent) being derived the lymph nodes consists of a thymus-de¬
from the bloodstream. These lymphocytes pendent area, the deep cortex, and a bursa-
belong to the recirculating pool and enter dependent area, which includes the medul¬
the node through the cuboidal endothelium lary cords and the outer cortex with its ger¬
of the postcapillary venules in the deep cor¬ minal centers. This postulated compartmen-
tex. Blood-borne small lymphocytes are se¬ talization of the lymph node is easily
lectively trapped by these vessels, whereas understood in view of the facts that the deep
granulocytes and monocytes are specifically cortex is the major traffic area of the lymph
excluded. Most of the recirculating lympho¬ node and that the vast majority of the recir¬
cytes are thymus dependent and, upon en¬ culating lymphocytes are thymus dependent.
tering the node, they localize in the deep Less clear is the functional significance of the
cortex. Shortly thereafter, they migrate into outer cortex and medullary cords, regions
the sinus lymph and leave the organ through that seem to be inhabited by a population of
the efferent lymphatic vessels. The fate of lymphocytes that are either sessile or slug¬
the small component of the recirculating pool gishly migratory, and whose functional prop¬
that consists of B lymphocytes is poorly erties are still poorly understood.
understood. B cells recirculate at a slower The lymphocytopoietic activity of a quies¬
rate than T cells, either because they are cent lymph node seems to be slight, for 75
inherently sluggish or because they are some¬ per cent of the lymphocytes of the mesenteric
how separated from the main recirculatory nodes of the rat belong to the long-lived
pathway. Thoracic duct cells of animals con¬ variety. The only sites of sustained lympho¬
genitally or experimentally deprived of T cyte proliferation in the lymph nodes are the
lymphocytes cross the endothelium of the germinal centers, but the fate of the lympho¬
postcapillary venules and are initially local¬ cytes produced by these enigmatic structures
ized in the deep cortex. Subsequently, how¬ is still not known. Of the remaining cellular
ever, they “home in” to the outer cortex and elements of the resting node, the plasma cells
medullary cords. In the outer cortex, they located in the medulla are sessile, effete ele¬
take residence in the primary nodules and ments, which either arose locally as a conse¬
lymphocyte cap of the germinal centers. quence of a previous antigenic stimulation or
Lymph nodes are also likely to be seeded differentiated from ancestors seeded
with cells that have just emerged from the throughout the body from a distant focus of
460 * LYMPH NODES

immune activity. The macrophages of the and is retained in the intercellular clefts of
lymph node parenchyma are probably of the outer cortex. On days 2 and 3, granulo¬
monocytic origin, although definitive evi¬ cytes usually disappear and the large baso¬
dence on this point has not yet been obtained. philic or pyroninophilic cells proliferate and
In contrast to the ideas prevailing in the past, greatly increase in number. The lymph node
autoradiographic studies show that the turn¬ enlarges, and the deep cortex seems to
over of the reticular cells is extremely slow spread, invading the whole organ. Preexist¬
and not influenced by experimental proce¬ ing germinal centers usually disappear and
dures that deplete the lymphocyte population the medullary cords are greatly reduced in
of the node. Thus, the claim that they rep¬ length or are no longer evident. With the
resent primitive precursors of lymphocytes, electron microscope, the newly formed pop¬
plasma cells, or macrophages is devoid of ulation of basophilic cells appears to include
experimental foundation. Detailed and sys¬ lymphoblasts and transitional forms between
tematic studies on the ultrastructure and the lymphocytic and plasmocytic lines. The
functional properties of the reticular cells are lymphoblasts have a pale nucleus, a promi¬
still lacking. They probably are not a homo¬ nent nucleolus, and a profusion of cyto¬
geneous class of cells but may include fibro¬ plasmic polyribosomes. Cisternae of the gran¬
blasts responsible for the synthesis of the ular endoplasmic reticulum are only
reticular fibers, tissue macrophages, and den¬ exceptionally found iti these cells, and mito¬
dritic cells. The term dendritic cell encom¬ chondria are few, whereas the Golgi appa¬
passes a seemingly heterogeneous population ratus is well developed but lacking the dense
of lymph node constituents, which includes granules typical of this organelle in ceils of
the “follicular dendritic cells” of germinal the plasmocytic series. The transitional cells
centers, the interdigitating cells of the deep display increasing amounts of granular en¬
cortex, and the veiled cells of the afferent doplasmic reticulum, suggestive of a differ¬
lymph and lymphatic sinuses. All these cells entiation into immature plasma cells. Both
have irregular shape, with numerous branch¬ the lymphoblasts and the transitional ele¬
ing processes radiating in all directions. They ments have been shown to produce antibody.
are not phagocytic and have an inconspic¬ Later on, immature members of the plasma
uous complement of cell organelles, except cell family become more and more numer¬
for the occasional presence of granules iden¬ ous, with their eccentric nucleus, condensed
tical to those typical of the Langerhans cells chromatin, and large amounts of granular
of the epidermis. The dendritic cells of the endoplasmic reticulum. Antibody is actively
germinal centers have the property of retain¬ synthesized at this stage and appears in both
ing for long periods of time antigen-antibody efferent lymph and blood. Furthermore,
complexes in the labyrinth of clefts bounded well-circumscribed nests of small to large
by their surface processes. Interdigitating lymphocytes appear near the surface of the
cells do not carry receptors for antibody node. These probably represent early devel¬
molecules, but share with Langerhans cells opmental stages of new germinal centers.
the presence of histocompatibility (la) mole¬ Toward the end of the first week after the
cules on their surface. It is presently debated injection, the lymph node begins to reacquire
whether dendritic cells, including the Lan¬ its normal architecture. Numerous newly
gerhans cells of the epidermis, are a novel formed germinal centers have appeared in
cell type whose function is to present antigen the cortex; medullary cords are prominent
to T lymphocytes during the inductive phase again near the hilus and contain abundant
of the immune response. immature and mature plasma cells.
After administration of an antigen that During the second week after antigenic
elicits antibody production, the primary re¬ challenge, plasma cells begin to decrease in
sponse of the lymph node draining the site number and become exclusively confined to
of injection is characterized, during the first the medullary cords. Antigen is still present
day, by an increase in number of granulo¬ in the node, confined both to the residual
cytes in both the parenchyma and lymph bodies of medullary macrophages and to the
sinuses. Simultaneously, large and medium¬ intercellular clefts between the dendritic cells
sized basophilic mononuclear cells appear in of the germinal centers.
the deep cortex. The antigen can be dem¬ These events in the lymph node are accom¬
onstrated in phagocytic vacuoles of the mac¬ panied by characteristic changes in the effer¬
rophages lining the sinuses of the medulla ent lymph. During the first 24 hours that
 LYMPH NODES • 461

follow antigen administration, the output of lus, and a narrow rim of cytoplasm, filled by
cells from the node decreases. Blood lympho¬ parallel arrays of cisternae of the granular
cytes keep entering the node, but are re¬ endoplasmic reticulum.
tained in it by some unknown mechanism. During the secondary response, the lymph
From two to five days after antigen adminis¬ node undergoes changes that resemble those
tration, cell release by the node doubles, following the first exposure to antigen, but
because more lymphocytes are crossing from they occur earlier and are much more pro¬
the blood, but are no longer retained in the nounced.
node. The lymphocytes, however, which have When a lymph node is involved in a cellular
been stimulated by the antigen, do not ap¬ immune response, it undergoes morpholog¬
pear yet in the efferent lymph. After five ical changes that are not strikingly different
days, the output of cells declines, but the from those typical of a humoral response. If
newly emerging population contains a large skin is grafted from a donor onto an alloge¬
proportion of activated lymphocytes. These neic recipient (allograft or homograft) or if
appear as large lymphocytes, with euchro- delayed hypersensitivity is induced by appli¬
matic nucleus, prominent nucleoli, and abun¬ cation of a chemical sensitizing agent to the
dant cytoplasm very rich in polyribosomes. skin, the draining node enlarges markedly
They have only a few elements of the gran¬ and the deep cortex becomes very thick.
ular endoplasmic reticulum even though they Lymphoblasts with pale nucleus, prominent
are actively producing antibody; they are nucleoli, and abundant basophilic cytoplasm
motile and capable of incorporating DNA (large pyroninophilic cells) appear in the
precursors into their chromatin. The remain¬ deep cortex. With the electron microscope,
ing cells leaving the stimulated node include they display abundant cytoplasmic polyribo¬
antibody-producing small lymphocytes and somes, minimal amounts of granular endo¬
immature elements of the plasma ceil line. plasmic reticulum, a small Golgi apparatus,
The antibody-producing cells of the efferent and a few mitochondria. These are actively
lymph are thought to colonize successive dividing cells, and their number in the deep
lymph nodes along the chain and, by entering cortex increases rapidly, reaching a maxi¬
the bloodstream through the thoracic duct, mum toward the end of the first week. At
may propagate the immune response the beginning of the second week, the num¬
throughout the body. In fact, if the efferent ber of lymphoblasts declines rapidly and a
lymphatic vessel of a locally stimulated node second phase in the response becomes ap¬
is cannulated and its lymph drained off, parent: newly formed germinal centers ap¬
antibody fails to appear in the blood and the pear in the outer cortex, and immature and
dissemination of the immune response is pre¬ mature plasma cells become localized in the
vented. If the cells leaving a stimulated node medullary cords. At the same time, humoral
are recovered from the efferent lymph, la¬ antibody is detected in the blood.
beled radioactively in vitro, and reinfused After a slight time lag with respect to the
into an afferent lymphatic of a quiescent appearance of the lymphoblasts in the deep
node, they localize in the medullary cords, cortex of the draining node, lymphocytes and
proliferate, and finally differentiate into ma¬ macrophages begin to infiltrate the graft or
ture plasma cells. Antibody-producing cells the sensitized skin region. The peripheral
have also been demonstrated in the thoracic response reaches its peak and the graft is
duct lymph. The fate of these cells has been rejected when the number of lymphoblasts
studied after in vitro labeling and intrave¬ has already begun to decline in the draining
nous injection into a syngeneic animal. They node, but before antibody is produced in
are retained transiently in the lung, liver, and significant amounts.
spleen, but later they localize in the spleen, The mechanism of the cellular response is
lymph nodes, and gut-associated lymphoid poorly understood. Antigens from the homo¬
structures and especially in the lamina pro¬ graft or the site of application of the sensitiz¬
pria of the small intestine. ing agent may reach the regional lymph node
The antibody-producing cells appearing in through the afferent lymph. Small lympho¬
the thoracic duct lymph after local stimula¬ cytes in the deep cortex may react with the
tion of a lymph node seem to undergo fur¬ antigens and differentiate into proliferating
ther modulation as they enter the blood¬ lymphoblasts; these in turn produce lympho¬
stream. Here, they appear as elements with cytes of decreasing size, which emigrate from
a central nucleus, an inconspicuous nucleo¬ the regional node, circulate in the blood, and
462 • LYMPH NODES

are disseminated throughout the immune Borurn, K., and M. H. Claesson: Histology of the induc¬
tion phase of the primary immune response in
system. They invade the graft and destroy it
lymph nodes of germfree mice. Acta Pathol. Micro¬
or mediate the skin lesion typical of a delayed biol. Scand. [A] 79:561, 1971.
hypersensitivity reaction (contact dermatitis Brahim, F., and D. G. Osmond: The migration of
or erythematous papule). The antibody re¬ lymphocytes from bone marrow to popliteal lymph
sponse that accompanies the cellular immune nodes demonstrated by selective bone marrow la¬
beling with 3H-thymidine in vivo. Anat. Rec.
reaction seems to contribute but little to the
775:737, 1973.
rejection process, at least in laboratory ro¬ Caffery, R. W., N. B. Everett, and W. O. Rieke: Radioau¬
dents. tographic studies of reticular and blast cells in the
Neonatal thymectomy prevents the appear¬ hemopoietic tissues of the rat. Anat. Rec. 755:41,
ance of the lymphoblasts in the deep cortex 1966.
Cahill, R. N. P., H. Frost, and Z. Trnka: The effects of
in response to homologous transplantation
antigen on the migration of recirculating lympho¬
and prolongs the survival of the graft. Thus, cytes through single lymph nodes. J. Exp. Med.
the lymphoblasts appearing in the deep cor¬ 745:870, 1976.
tex are likely to be thymus dependent. Carr, I.: The fine structure of the mammalian lympho-
reticular system. Int. Rev. Cytol. 27:283, 1970.
Claesson, M. H., O. Jprgensen, and C. Ropke: Light and
Hemal Nodes electron microscopic studies of the paracortical post¬
capillary high-endothelial venules. Z. Zellforsch.
Even in normal lymph nodes, varying 779:195, 1971.
numbers of erythrocytes are found. These Clark, S. L., Jr.: The reticulum of lymph nodes in mice
studied with the electron microscope. Am. J. Anat.
have either entered the lymph from the af¬
779:217, 1962.
ferent vessels or come from the blood vessels Cohen, S., P. Vassalli, B. Benacerraf, and R. T. Mc-
of the node. Some of them pass with the Cluskey: The distribution of antigenic and nonan-
lymph into the efferent vessels, but most of tigenic compounds within draining lymph nodes.
them are engulfed by macrophages. Some Lab. Invest. 75:1143, 1966.
Conway, E. A.: Cyclic changes in lymphatic nodules.
nodes, however, called hemal nodes, are char¬
Anat. Rec. 69:487, 1937.
acterized by the exceptionally high content Dougherty, T. F., M. L. Berliner, G. L. Schneebell, and
of erythrocytes. They are most numerous D. L. Berliner: Hormonal control of lymphatic
and best defined in the ruminants (sheep); structure and function. In Bierman, H. R., ed.:
Leukopoiesis in Health and Disease. Ann. N. Y.
they probably do not occur in man. They
Acad. Sci. 775:511, 1964.
vary from minute bodies scarcely noticeable Downey, H.: The structure and origin of the lymph
to the size of a pea or larger, and are scat¬ sinuses of mammalian lymph nodes and their rela¬
tered along large blood vessels from the neck tions to endothelium and reticulum. Haematologica
to the pelvic inlet. They are also found near 5:431, 1922.
Downey, H., ed.: Handbook of Hematology. New York,
the kidneys and spleen, where they are be¬
Paul B. Hoeber, 1938.
lieved by some to be accessory spleens. Each Drinker, C. K., M. E. Field, and H. K. Ward: The
hemal node is covered by a dense capsule filtering capacity of lymph nodes. J. Exp. Med.
loosely connected to the surrounding tissue. 59:393, 1934.
Farr, A. G., Y. Cho, and P. P. H. DeBruyn: The structure
At the hilus a small artery and a large vein
of the sinus wall of the lymph node relative to its
enter and leave. The nodes are devoid of endocytic properties and transmural cell passage.
afferent lymphatics and have postcapillary Am. J. Anat. 757:265, 1980.
venules with walls infiltrated by migrating Fisher, B., and E. R. Fisher: Barrier function of lymph
lymphocytes. In the pig, a special type of node to tumor cells and erythrocytes. I. Normal
nodes. Cancer 20:1907, 1967.
hemal node has characteristics intermediate
Ford, W. L., and J. L. Cowans: The traffic of lympho¬
between the ordinary lymph node and the cytes. Semin. Hematol. 6:67, 1969.
typical hemal node. It has blood vessels as Forkert, P.-G., J. A. Thliveris, and F. D. Bertalanffy:
well as lymphatic vessels, and the contents of Structure of sinuses in the human lymph node. Cell
Tissue Res. 183:115, 1977.
both types of vessels mix in the sinuses. The
Fossum, S.: The architecture of rat lymph nodes. II.
functions of the hemal nodes are probably Lvmph node compartments. Scand. J. Immunol.
comparable with those of the spleen. 72:411, 1980.
Fossum, S.: The architecture of rat lymph nodes. IV.
Distribution of ferritin and colloidal carbon in the
draining lymph nodes after foot-pad injection.
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immunity. III. Distribution of iodinated antigens teric lymph node. Z. Zellforsch. 755:147, 1972.
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Aust. J. Exp. Biol. Med. Sci. 42:295, 1964. lymphocytes and thymocytes in the rat. I. The route
 LYMPH NODES • 463

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Gowans, J. L., and E. J. Knight: The route of recircu¬ Nossal, G. J. V., A. Abbot, and J. Mitchell: Antigens in
lation of lymphocytes in the rat. Proc. R. Soc. B immunity. XIV. Electron microscopic radioauto¬
759:257, 1964. graphic studies of antigen capture in the lymph
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Hall, J. G., B. Morris, G. D. Moreno, and M. C. Bessis: Nossal, G. J. V., G. L. Ada, and C. M. Austin: Antigens
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Harris, T. N., K. Hummeler, and S. Harris: Electron labelled antigens in the secondary response. Im¬
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Hay, J. B., M. J. Murphy, B. Morris, and M. C. Bessis: Thymus-dependent areas in the lymphoid organs
Quantitative studies on the proliferation and differ¬ of neonatally thymectomized mice. J. Exp. Med.
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J. Pathol. 66:1, 1972. Ropke, C., O. Jprgensen, and M. H. Claesson: Histo-
Hostetler, J. R., and G. A. Ackerman: Lymphopoiesis chemical studies of high-endothelial venules of
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lymphocytes through the endothelium of venules in munology 20:115, 1971.
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Lymph Node Cytology and Pathology. Berlin, Dendritic cells in the immune response: Character¬
Springer Verlag, 1969. istics and recommended nomenclature (a report
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Exp. Mol. Pathol. 4:155, 1965. structure and permeability of lymph node micro¬
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Ultrastructural analysis of antibody synthesis in cells 1976.
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lymph nodes. Cell. Immunol. 25:254, 1976. and Lymphoid Tissue. Cambridge, Harvard Uni¬
Nopajaroonsri, C., S. C. Luk, and G. T. Simon: Ultra¬ versity Press, 1956.
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65:1, 1971. •' lymphatic system in the opossum. Illinois Medical
Nopajaroonsri, C., and G. T. Simon: Phagocytosis of and Dentaf Monographs, Vol. 3, Nos. 1 and 2, 1940.
 16
SPLEEN
Elio Raviola

The spleen is an abdominal organ situated red pulp is due to the abundance of the
in the left hypochondrium beneath the dia¬ erythrocytes that fill the lumen of the sinuses
phragm; largely invested by visceral perito¬ and infiltrate the splenic cords.
neum, it is connected to the stomach, dia¬ The spleen, much like the lymph nodes,
phragm, and left kidney by peritoneal folds, has a collagenous capsule with inward exten¬
called gastrolienal, phrenicolienal, and lien- sions called trabeculae (Fig. 16—1). This cap¬
orenal ligaments. The lienorenal ligament sule is continuous with a delicate reticular
carries the splenic blood vessels, lymphatics, framework that occupies the rest of the in¬
and nerves. The spleen is a complex filter terior of the organ and holds in its meshes
interposed in the bloodstream. It is con¬ the free cells of the splenic tissue. The capsule
cerned with clearing the blood of particulate is thickened at the hilus of the organ, where
matter and effete cells, and with immune it is attached to the peritoneal ligaments and
defense against blood-borne antigens. In where arteries and nerves enter and veins
many vertebrates, but not in man, the spleen and lymphatic vessels leave the viscus.
is also involved in formation of erythrocytes, The structure of the spleen and the rela¬
granulocytes, and platelets, and in certain tionships between the red and white pulp
mammals it acts as a reservoir for mature depend on the distribution of the blood ves¬
erythrocytes that can be added to the circu¬ sels. They differ markedly in different ani¬
lation in response to unusual demands. The mal species and change in the course of
spleen contains a large amount of lymphoid immune responses or disturbances of blood
tissue and possesses a peculiar type of blood cell formation and destruction. Animal spe¬
vessel that allows the circulating blood to cies with a large blood volume (horse, rumi¬
come into contact with great numbers of nants, carnivores) have scanty white pulp and
macrophages. a robust connective tissue and muscular
framework. Species with a relatively small
blood volume (man, rabbit, laboratory ro¬
HISTOLOGICAL dents) have abundant white pulp, a less
prominent connective tissue framework, and
ORGANIZATION poorly developed smooth musculature. In
the splenic parenchyma, the white pulp is
On the freshly sectioned surface of the organized around the arteries and the red
spleen, elongate or rounded gray areas, 0.2 pulp fills the interstices among the venous
to 0.7 mm in diameter, are visible with the sinuses.
naked eye (Fig. 16—1). Together these com¬
pose the white pulp; they are scattered
White Pulp
throughout a soft, dark red mass, the red
pulp, which can easily be scraped from the The white pulp forms periarterial lym¬
cut surface of the organ. The white areas, phoid sheaths (PALS) about the arteries where
often called malpighian bodies, consist of dif¬ these leave the trabeculae to penetrate the
fuse and nodular lymphoid tissue. The red parenchyma (Figs. 16-1, 16-6). The periar¬
pulp consists of irregularly shaped blood ves¬ terial lymphoid sheaths follow peripherally
sels of large caliber (the venous sinuses) and along the vessels almost to the point where
die tissue occupying the spaces between them they break up into capillaries. In many places
(the splenic cords of Billroth). The color of the along their course the sheaths contain ger-
464
 SPLEEN 465

Penicillar
Capsule Trabecula Red pulp artery White pulp

Red pulp

Trabecula

Germinal Trabecula
center

WWwm
»-».<•» >i»vv ->ivv.

White
pulp
a
US!
S' Penicillar
artery
£S$.-.-.VV,';> •■*

rtf:'■■ £
gs% is
'■ ;■:■ -W&WlSk;^ :i
IMiJ 1 . ’

a
Trabecular
artery

Germinal center
Trabecular veins

Figure 16-1. Section of human spleen. A, Central arteries of the white pulp. (After A. A. Maximow.)

minal centers. Although both the periarterial cells form concentric layers that delimit the
lymphoid sheaths and the germinal centers lymphoid tissue from the surrounding red
consist of lymphoid tissue, they differ in their pulp. Near the central artery, a few elastic
physiological significance; the former, in fact, fibers are interspersed among the reticular
consist predominantly of lymphocytes be¬ fibers. The meshes of the reticular frame¬
longing to the recirculating pool, whereas the work are occupied by lymphocytes, predom¬
germinal centers are bursa-dependent struc¬ inantly belonging to the small and medium¬
tures, whose function is still poorly under¬ sized variety, in places associated with inter-
stood. digitating cells (see Chap. 13). Plasma cells
The periarterial lymphoid sheaths display and macrophages are only occasionally found
an organization similar to the deep cortex of but increase in number toward the periphery
the lymph nodes. They have a loose, irregu¬ of the sheath. Erythrocytes are rare, but they
lar framework of reticular fibers with associ¬ may occur at the boundary between white
ated reticular cells. At the periphery of the and red pulp. In the course of immune
sheath, the reticular fibers become circum¬ responses to blood-borne antigens, great
ferentially arranged, and flattened reticular numbers of large lymphocytes, lymphoblasts,
466 • SPLEEN

Figure 16-2. A, Drawing of red pulp of the human spleen. Venous sinuses filled with erythrocytes are separated from
each other by the pulp cords. Hematoxylin-eosin. (After W. Bloom.) B, Photomicrograph of a silver impregnation of the
reticular fibers of the spleen. The regularly spaced, circumferential ribs encircling the sinus endothelium are readily
distinguished from the randomly distributed reticular fibers of the surrounding splenic cords. (Preparation bv K.
Richardson.)
 SPLEEN • 467

and immature plasma cells appear in the in a finely filamentous matrix. They are com¬
periarterial lymphoid sheaths and soon be¬ pletely invested by stellate reticular cells (Fig.
come concentrated at their periphery. 16—4), which resemble fibroblasts in their
The germinal centers display the usual complement of organelles except for an un¬
architecture (see Chap. 13); they are eccen¬ common abundance of filaments in their cy¬
trically situated within the sheath and, when toplasm. Some of the reticular cells, as seen
fully developed, their light region and cap of with the light microscope, may actually rep¬
small lymphocytes are directed toward the resent tissue macrophages of monocytic ori¬
red pulp. Their number varies in different gin. The reticular fibers of the red pulp
animal species and tends to decrease pro¬ merge with ribs of basal lamina—like material
gressively with age. that support the sinus endothelium. Their
investing reticular cells are in turn anchored
Red Pulp to the sinus walls through footlike processes,
which are directed perpendicularly to the
The red pulp consists of a network of long axis of the sinuses.
branching and anastomosing, tortuous sin¬ The meshes of reticulum in the pulp cords
uses, separated from each other by highly are filled with great numbers of free cells,
cellular partitions, the splenic or pulp cords which include macrophages; all the cellular
(Figs. 16-2,16—3). The venous sinuses are elements circulating in the blood, including
discussed with the blood vessels of the spleen. great numbers of erythrocytes and platelets;
The splenic cords vary in thickness but typi¬ and a few plasma cells (Figs. 16-3,16-8).
cally form a spongy cellular mass supported With the light microscope the macrophages
by a framework of reticular fibers (Fig. 16— are readily recognized as large, rounded, or
2). The collagenous fibers of the trabeculae irregularly shaped cells, with a vesicular nu¬
continue directly into the reticular fibers of cleus and abundant cytoplasm. They often
the red pulp. As elsewhere in the body, the contain engulfed erythrocytes, neutrophils,
reticular fibers as seen with the electron mi¬ and platelets, or are loaded with masses of a
croscope consist of collagen fibrils embedded yellowish-brown pigment that stains for iron

Venous sinus
Endothelial
cell
Small of Erythro¬
Migrating lymphocyte lymphocytes sinus cytes Monocyte

Plasma cell

Reticular
cells
Macrophage

Small lymphocytes

Blast
Mitosis of medium
lymphocyte
Arteriole

Reticular cells
Macrophage

Migrating Monocyte
lymphocyte

Small lymphocyte

Small Free macrophage

Free macrophage
lymphocyte
Endothelial cell of sinus Free
macrophage

Figure 16-3. Cross section of splenic cord lying between two venous sinuses, from spleen of a rabbit injected with
lithium carmine and India ink. Hematoxylin-eosin-azure II stain. (After A. A. Maximow.)
468 • SPLEEN

Figure 16-4. Scanning electron micrograph of a splenic cord adjacent to a pulp vein in the dog. Spleen perfusion with
physiological salt solution has removed most of the free cells, thus exposing the three-dimensional network formed by
the reticular cells. (From Miyoshi, M., and T. Fujita. Arch. Histol. Jpn. 33:225, 1971.)

with the Prussian blue reaction and gives a the cords have a greater content of small
positive reaction for the lysosomal enzyme lymphocytes and plasma cells than the rest
acid phosphatase. This pigment represents of the red pulp. The marginal zone is the
the undigestible residues of phagocytized ma¬ region of the red pulp that receives the
terials, especially red blood cells. Its iron, in incoming arterial blood; thus, it is the site
the form of ferritin or hemosiderin, comes where blood-borne cells and particulate mat¬
from the degradation of hemoglobin. In ter first contact the splenic parenchyma.
many mammalian species (laboratory ro¬ Here, the lymphocytes of the recirculating
dents, hedgehog) and in the embryonic pool leave the blood of the sinuses to enter
spleen, the red pulp contains groups of ery- the periarterial lymphoid sheaths.
throblasts of various sizes, myeloblasts, mye¬
locytes, and megakaryocytes. In adult man,
Capsule and Trabeculae
these islands of hemopoietic tissue are lack¬
ing, but in certain infections, in some of the The capsule and the trabeculae of the
anemias, in leukemias, and in poisoning with spleen consist of dense connective tissue,
certain blood-destroying agents, they may smooth muscle cells, and elastic networks.
reappear, a condition described as myeloid The external surface of the capsule is covered
metaplasia. by a layer of flattened mesothelium, which is
Immediately peripheral to the white pulp part of the general peritoneum. In man,
is an 80- to 100-pm transitional region be¬ rabbits, and laboratory rodents, the capsule
tween lymphoid tissue and red pulp, called is rich in elastic fibers, especially in its deep
the marginal zone; it contains smaller venous layers, and in addition to typical fibroblasts it
sinuses, circumferentially oriented around contains a small number of stellate elements
the white pulp. In the marginal zone, the that display cytological characteristics inter¬
reticular fibers of the cords form a closely mediate between smooth muscle cells and
knit concentric network, and the meshes of fibroblasts, having filamentous cytoplasm and
 SPLEEN • 469

abundant glycogen. Smooth muscle cells are ters are embedded in the periarterial lymph¬
especially abundant in the splenic capsule of oid sheath, the artery is displaced to one side,
the horse, ruminants, and carnivores. thus losing its central position. It almost
The trabeculae are flattened or cylindrical never passes through the center of the nod¬
strands that carry arteries, veins, and lym¬ ules. The central artery is a muscular artery
phatics. They contain a larger number of with tall endothelial cells and one or two
elastic fibers than the capsule and varying layers of smooth muscle cells. Throughout
amounts of smooth muscle cells (Fig. 16-5). its course within the white pulp, the artery
Smooth muscle cells are sparse in the human gives off numerous collateral capillaries,
spleen. In those species in which muscle is which supply the lymphoid tissue of the
prominent, changes in the volume of the sheath. Initially, the capillary wall consists of
organ, either spontaneous or induced by in¬ tall endothelial cells, basal lamina, and an
jection of epinephrine, are due to contraction investment of pericytes; farther on, the en¬
of the smooth muscle in the capsule and dothelium becomes low and the pericytes
trabeculae, as well as to vasomotor changes disappear. Around the capillaries, the retic¬
in the amount of blood in the organ. ular meshwork of the white pulp is con¬
densed and contains a few elastic fibers.
Arteries These collateral capillaries, after coursing
through the white pulp, pass into the sur¬
The branches of the splenic artery enter rounding marginal zone; how they end is
the hilus and pass along the trabeculae, uncertain.
within which they branch repeatedly, becom¬ The central artery continues to branch and
ing smaller in diameter (Fig. 16-6). They are becomes thinner. On reaching a diameter of
muscular arteries of medium caliber and 40 to 50 pm, its lymphoid sheath appears
have a loose tunica adventitia surrounded by greatly reduced in thickness and the artery
the dense connective tissue of the trabeculae. suddenly branches into two to six vessels,
When the arterial branches have been re¬ called penicillar arteries (Latin, penicillus = a
duced by progressive dichotomous branching painter’s brush) or arteries of the red pulp.
to a diameter of approximately 0.2 mm, they These pursue a radiating course still invested
leave the trabeculae. At this point, the tunica by one or two layers of lymphocytes that
adventitia is replaced by a cylindrical sheath represent a greatly attenuated terminal ex¬
of lymphoid tissue, and the artery is desig¬ tension of the periarterial lymphoid sheaths.
nated the central artery. Where germinal cen¬ The penicillar arteries are 0.6 to 0.7 mm in

Red pulp
Smooth muscle cells

Elastic fibers

Smooth
Elastic muscle cells
fibers

Smooth
muscle cell
Smooth
muscle cell

Red pulp

Erythrocyte Lymphocytes Reticular cell

Figure 16-5. Portion of a trabecula from the spleen of a cat. Elastic fiber stain. (After A. A. Maximow.)
470 • SPLEEN
*

Venous sinus Capsule Trabecula Red pulp

Capillary
Sheathed
capillary — Capillary
Red pulp
Sheathed
capillary
Artery of pulp Artery of
pulp

Trabecular
vein
White
pulp

Artery leaving
trabecula

Trabecular vein

Trabecular vein
Trabecular vein
becula
Venous sinus

Trabecular artery Trabec¬ Trabecular artery

ular vein

Figure 16-6. Diagram of the vascular tree of the spleen. Capillaries are depicted as communicating with the venous
sinuses, according to the “closed circulation” theory. A, Central arteries. (After A. A. Maximow.)

length and have a tall endothelium resting filaments and rest on a discontinuous basal
on a continuous basal lamina, but they lack lamina. In man, the sheath is tubular and
an elastica interna. Their media consists of fairly thin (50 to 100 pm in length and 20 to
one layer of smooth muscle cells; they lack 30 pm in diameter), but in certain species it
an elastica externa and have a thin adventitia is very prominent and ellipsoidal or spherical
of collagenous and elastic fibers. in shape (pig, dog, and cat). Sheathed capil¬
Upon entering the red pulp, each penicil- laries are lacking in the spleen of laboratory
lar artery as a rule branches into two to three rodents. Not all capillaries arising from a
capillaries, which may exhibit a characteristic penicillar artery are sheathed, and most com¬
thickening of their walls, called the ellipsoidal monly only one sheath is associated with the
or Schweigger-Seidel sheath (Fig. 16-7). The terminal branches of a penicillar artery. Oc¬
capillaries are therefore named sheathed cap¬ casionally, multiple vessels (two to five) are
illaries. The endothelium of these vessels con¬ invested by a single sheath, and sometimes
sists of tall, fusiform cells arranged parallel two to three sheaths are arranged in series
to the long axis of the vessel. They are in along a single branch of a penicillar artery.
places connected by intercellular junctions The sheath consists of a closely knit network
and in other places separated by gaps of reticular fibers and cells. Most cells belong
through which blood cells can pass from the to two varieties, reticular cells and macro¬
lumen. They contain abundant intermediate phages, but elements of the circulating blood
 SPLEEN • 471

Medium-sized lymphocyte Capillary Reticular cells

Trabecula

Erythrocyte

Reticular
cells

Small
lym¬
phocytes
Heterophil
leukocyte

Erythrocytes

Sheath Capillary

Figure 16-7. Cross section of a Schweigger-Seidel sheath surrounding two capillaries in the spleen of a dog. Eosin-
azure stain. (After A. A. Maximow.)

are also present. Reticular cells are stellate in

Venous Sinuses and Veins
shape, are associated with the reticular fibers,
and, in addition to the usual organelles, con¬ The venous sinuses permeate the entire
tain abundant cytoplasmic filaments: inter¬ red pulp and are especially numerous around
mediate filaments coursing throughout the the white pulp (Fig. 16—8). These vessels are
cytoplasm and a feltwork of thin filaments called sinuses because they have a wide (12
on the inner surface of the plasmalemma. to 40 pm), irregular lumen, which varies in
Macrophages contain residual bodies and oc¬ size depending on the amount of blood in
casional phagocytized red blood cells. Upon the organ. The sinuses, even when only mod¬
intravenous injection of particulate matter erately distended, occupy more space than
they become avidly phagocytic; thus, there is the splenic cords between them. Venous sin¬
no doubt that they belong to the mononu¬ uses are missing in the mouse and cat spleen
clear phagocyte system. Red blood cells are and substituted for by conventional venules.
always present, in large or small numbers, Unlike true veins, the walls of the sinuses
among the cells of the sheath. There is gen¬ lack a muscular coat and display a unique
eral consensus that ellipsoidal sheaths func¬ arrangement of endothelium and basal lam¬
tion as filters that clear the blood of particu¬ ina. The endothelial cells are fusiform ele¬
late materials, yet the reasons for their ments, about 100 pm long, oriented parallel
location around arterial capillaries remain to the longitudinal axis of the sinus (Fig. 16-
unclear. 9). The central nuclear region of the cell
The sheathed capillaries continue as simple body is thick and tapers toward the ends;
capillaries that either do not divide or bifur¬ they are in contact with each other laterally
cate only once. Their mode of termination is but lack typical intercellular junctions. Micro-
unknown and will be discussed below after pinocytotic vesicles are plentiful on both the
the venous sinuses have been described. luminal and lateral surfaces, and the cyto-
472 • SPLEEN

Erythrocytes in diapedesis
Endothelial
cells Monocytes Venous sj
nus
Large lymphocyte
Migrating lymphocytes
Migrating lymphocytes
Plasma cell
Reticular cell
Erythrocytes
in diapedesis

Endothelial cell
Monocyte

Erythrocyte

Venous sinus

Lym¬
Venous phocytes
sinus

Eryth¬
rocytes

Small lym¬
phocyte Venous
sinus
Sheathed
capillary
Erythrocyte

Monocytes Plasma cell

Free macro¬
phage

Erythrocytes in cord
of Billroth Reticular cell
Monocytes Endothelial cel!
Free
macrophage j I Lymphocyte
Erythrocytes | Endothelial cell
Neutrophil leukocyte

Figure 18-8. Venous sinuses in the red pulp of the human spleen. Note the cuboidal shape of the cross-sectioned
endothelial cells. The cytoplasmic densities at the base of the cells (K) probably correspond to the condensations of
finely filamentous material revealed by the electron microscope. Eosin-azure stain. (After A. A. Maximow.)

plasm contains, in addition to the usual com¬ endothelial cells elsewhere in the body. Like
plement of organelles, two types of filaments, other endothelial cells, they display only a
both oriented parallel to the long axis of the limited capacity to take up particulate matter
cells. There are loosely packed intermediate injected into the bloodstream. Thus, the tra¬
filaments, free in the cytoplasmic matrix, and ditional interpretation of the sinus-lining cells
denser bands of finely filamentous material of the spleen as tissue macrophages is no
in the basal region of the cell. These latter longer tenable.
seem to run from one rib of the basal lamina Outside the endothelium, the wall of the
to the next, where they insert on the inner sinuses is supported by a system of circum¬
aspect of the plasma membrane; they are ferential ribs, about 1 pm in thickness, encir¬
probably responsible for the longitudinal cling the endothelial cells as the hoops em¬
basal striations of the endothelium seen in brace the staves of a barrel (Fig. 16—2). The
specimens stained with iron hematoxylin. Ex¬ ribs are spaced 2 to 5 pm apart and are
cept for their unusual shape, lack of inter¬ interconnected by relatively few thin, longi¬
cellular junctions, and abundance of fila¬ tudinal strands. At the light microscope level,
ments, the sinus-lining cells resemble they are observed to stain with silver impreg-
 SPLEEN • 473

Figure 16-9. Surface view of the endothelial cells of a venous sinus in the rabbit spleen as seen with the scanning
electron microscope. As the specimen was air-dried, cells have pulled apart, exposing the fenestrations of the basal
lamina. (From Miyoshi, M., et al. Arch. Histol. Jpn. 32:289, 1970.)

nation methods and with histochemical meth¬ to form the veins of the trabeculae; in turn,
ods for carbohydrates. In electron micro¬ these are drained by the veins at the hilus of
graphs, they appear to consist of finely the spleen, which are tributaries of the
filamentous material with a few embedded splenic vein.
collagen fibrils. Thus, in their fine structure,
they correspond to an unusually thick, fenes¬
Union of the Arteries with the Veins
trated basal lamina. The ribs are continuous
with the reticular fibers of the splenic cords In almost all other organs of the body, the
and are interposed between the endothelium arterial is joined to the venous system by a
on one side and the foot processes of the continuous capillary network, in which the
reticular cells of the cords on the other. vascular lumen is completely enclosed. In the
Cellular elements of the circulating blood can spleen, however, the connection of arterial
easily migrate through the sinus wall, tra¬ and venous systems may be different, and its
versing the interendothelial clefts and the details are still subject to dispute. There are
fenestrations of the basal lamina. Further¬ three main theories as to how blood gets
more, cordal macrophages are frequently from the arteries to the venous sinuses (Fig.
seen extending processes through the sinus 16—10). (1) The “open circulation” theory
walls into the lumen and they may also mi¬ holds that the capillaries open directly into
grate into the blood within the sinus. the spaces among the reticular cells of the
The venous sinuses empty into the veins of splenic cords, and that the blood gradually
the pulp, whose walls consist of elongated, filters into the venous sinuses. (2) The “closed
slender endothelial cells, a continuous basal circulation” theory holds that the capillaries
lamina, and a thin layer of smooth muscle. communicate directly with the lumen of the
They are supported externally by a conden¬ venous sinuses. (3) A compromise interpre¬
sation of the stroma of the red pulp and by tation holds that both types of circulation are
a few elastic fibers. The pulp veins coalesce present at the same time. One of the variants
474 • SPLEEN

Capillary —
Venous sinus

Germinal center -

White pulp--
Figure 16-10. Diagram to show
Venous sinus closed (1) and open (2) circulation
Pulp vein through the spleen.

Pulp cord
Trabecular vein

Trabecular artery

Trabecula

of this theory contends that a “closed” circu¬ the walls of the venous sinuses. According to
lation in a contracted spleen may become an the “open circulation” theory, these cells are
“open” circulation when the organ is dis¬ returning to the bloodstream from the extra-
tended. vascular spaces of the splenic .cords. Accord¬
The opposing theories are based on the ing to the “closed circulation” theory, both
following observations. (1) There are always plasma and cells of the blood are believed to
many erythrocytes scattered throughout the pass into the cordal spaces at the arterial end
tissue spaces of the splenic cords. Since in of the sinuses and to return to the circulation
most species there is little or no erythropoi- at their venous end, driven by a pressure
esis in the cords, it is concluded that the red gradient that may exist between the two ends
blood cells have come from the circulating of these blood vessels. (4) Studies with the
blood through gaps in the vascular segment electron microscope seem to favor the view
between arterioles and venous sinuses. Those that the circulation is open, for it has been
who maintain that the circulation is “closed” reported that arterial capillaries directly open
hold that the number of erythrocytes in the into the red pulp, whereas endothelial con¬
splenic cords is much smaller than it would tinuity between arterial and venous vessels
be if the capillaries opened directly into the has never been shown. (5) The problem of
pulp. They argue that if the capillaries were the circulation in the spleen would seem to
open, the red pulp would be completely filled be an ideal one for solution by direct obser¬
with blood, as in hemorrhages of the spleen. vation of the living organ. Unfortunately, the
(2) When the splenic arteries are injected techniques available for this are difficult, and
even at low pressures with dye solutions, the reports made on such studies are contra¬
India ink, or avian erythrocytes, the foreign dictory. According to one group, the circu¬
materials readily gain access to the tissue lation in the spleen appears to be closed,
spaces of the splenic cords, particularly in there is a marked intermittency of circula¬
the marginal zone. Only later do they reach tion, there is extensive filtering of the liquid
the venous sinuses. When the splenic vein is portion of the blood from the sinuses into
injected, the venous sinuses and the meshes the splenic cords, and erythrocytes normally
of the stroma can be filled easily, but the leave the sinuses by diapedesis.
arteries cannot. Those who favor a closed These conclusions are contradicted by an¬
circulation believe that this injection of the other group of observers who report the
red pulp by foreign materials is an artificial circulation to be open—i.e., without pre¬
situation resulting from the rupture of the formed connections between the arterial and
delicate vascular walls. (3) In every freshly venous systems, so that the blood from the
fixed spleen, granulocytes, lymphocytes, and terminations of the arterial tree percolates
erythrocytes can be found passing through between the reticular cells and macrophages
 SPLEEN • 475

of the splenic cords and finds its way through The destruction of aged, abnormal, or
openings in the wall of the sinuses. damaged blood cells and platelets takes place
in the meshes of the cords of the red pulp.
Lymphatic Vessels and Nerves How blood-borne elements and especially
erythrocytes, which lack motility, reach the
In man, lymphatic vessels are poorly de¬ tissue spaces of the cords is still a matter of
veloped and are found only in the capsule of controversy. According to the “open circula¬
the spleen and in the thickest trabeculae, tion” theory, the capillaries deliver the blood
particularly those in the vicinity of the hilus. directly to the pulp cords; this being true,
In some mammals, true lymphatic vessels the constituents of the plasma and the cells
follow the arteries of the white pulp. Net¬ of the blood can freely percolate through the
works of nerves that originate from the celiac interstitial spaces between cord macrophages
plexus and consist almost entirely of unmye¬ and reticular cells and finally reenter the
linated fibers accompany the splenic artery blood through the walls of the venous sin¬
and penetrate into the hilus of the spleen. In uses. In a “closed” vascular system, one must
the sheep and ox, these nerves form trunks postulate either that the pressure gradient
of considerable thickness. The nerve bundles between the arterial and venous ends of the
mainly follow the ramifications of the arteries sinuses drives plasma and erythrocytes into
and form networks that can be followed as the cord spaces and then back into the blood,
far as the central arteries of the white pulp or that rhythmic contractions of the sinus
and even along the branches of the penicillar endothelium squeeze the blood out of the
arteries. The terminal branches usually end vascular bed; this latter hypothesis implies
with button-like thickenings on the smooth the existence of a sphincteric mechanism at
muscles of the arteries and of the trabeculae. the venous end of the sinuses. With either
Apparently many branches penetrate into the theory, both plasma and blood cells establish
red as well as the white pulp, but their end¬ extensive contact with the macrophages of
ings here are not definitely established. the cords, which can then remove any un¬
desirable component. Unphagocytosed cells
can freely return to the intravascular com¬
partment through the fenestrations of the
basal lamina and the interendothelial clefts
HISTOPHYSIOLOGY of the sinuses. The precise role of the spleen
in removing aged erythrocytes, as well as the
The filtering function of the spleen de¬ extent to which this function is shared with
pends upon the abundant population of mac¬ bone marrow and liver, is poorly understood.
rophages of the splenic cords, which arise Splenectomy does not seem to affect the
from blood monocytes. Upon intravenous average life span of red blood cells signifi¬
injection, particulate matter or macro- cantly. There is no doubt, however, that the
molecular antigen first localize in the mac¬ spleen plays a major role in destroying path¬
rophages of the marginal zone and subse¬ ological or defective blood elements. When
quently spread to the phagocytes of the rest abnormal or experimentally damaged eryth¬
of the red pulp. Neither the endothelial cells rocytes are perfused through the spleen, they
of the sinuses nor the fibroblast-like reticular are retained by this organ, whereas normal
cells of the splenic cords contribute signifi¬ erythrocytes are not. Granulocytes damaged
cantly to clearing the blood of foreign mate¬ by endotoxin have also been shown to be
rial. Very little particulate matter enters the destroyed in the spleen. Moreover, splenec¬
white pulp, but in the presence of antibody, tomy is followed by the appearance in the
antigen may be trapped for long periods of bloodstream of defective erythrocytes con¬
time in the germinal centers. When lipid in taining remnants of the nucleus or cyto¬
the blood is increased in amount, the mac¬ plasmic organelles. The mechanism by which
rophages of the spleen, like the other phago¬ macrophages recognize old or abnormal
cytes of the body, have the capacity to remove blood cells is unknown. It has been postulated
it from the circulation. In this process, the that the immune system may react to changes
macrophages enlarge and become filled with in the erythrocyte surface and tag patholog¬
lipid droplets, thus acquiring a foamy ap¬ ical cells with opsonizing antibody. In normal
pearance. This phenomenon is observed in subjects, no lysis or fragmentation of red
diabetic hyperlipemia in man and in experi¬ blood cells is observed either in the lumen of
mental hypercholesterolemia of rabbits. the sinuses or in the cord spaces, and eryth-
476 • SPLEEN

rocytes seem to be phagocytized intact by blood that only a small fraction of the lym¬
macrophages. In pathological conditions, phocytes that leave the spleen via the veins
however, extracellular disintegration of red arise from division of precursors in that or¬
blood cells has been described. gan. A large fraction of splenic lymphocytes
Closely connected with erythrocyte de¬ belong to the recirculating pool and they are
struction by the macrophages is the function specifically localized in the periarterial
of the spleen in hemoglobin degradation and lymphoid sheaths. This has been shown by
iron metabolism. In the lysosomes of the experiments involving drainage of the tho¬
macrophages, the iron of hemoglobin is freed racic duct lymph and reinjection of thoracic
and stored by the cell as ferritin or hemosi¬ duct lymphocytes after labeling in vitro.
derin, readily available to the body for syn¬ Drainage of the thoracic duct lymph initially
thesis of new hemoglobin by bone marrow affects the central region of the periarterial
erythroblasts. The heme moiety of hemoglo¬ lymphoid sheaths; only after prolonged
bin is degraded by macrophages to bilirubin, drainage do the peripheral regions of the
which enters the plasma, where it binds to white pulp also become depleted of lympho¬
albumin. It is then captured by the liver, cytes. This finding has been interpreted as
conjugated to glucuronic acid, and secreted evidence favoring the idea that the rapidly
in the bile. recirculating T lymphocytes localize close to
In animal species in which the capsule and the central artery, whereas the sluggishly
trabeculae are rich in smooth muscle cells, migrating B cells assume a more peripheral
the spleen can act as a store for red blood position in the periarterial lymphoid sheaths.
cells; large numbers of them in fact can be After intravenous injection, labeled thoracic
retained in the red pulp and then given up duct cells, which are predominantly T lym¬
to the bloodstream when they are needed in phocytes, first localize in the marginal zone
the circulation. This may also occur experi¬ of the red pulp, but a few hours later they
mentally following injection of drugs, such as have migrated throughout the periarterial
epinephrine, that induce contraction of the lymphoid sheaths. Labeled B lymphocytes
splenic smooth muscle. The human spleen first appear in the marginal zone, then reside
has little storage capacity (about 30 to 40 ml for a few hours at the periphery of the
of erythrocytes), but it temporarily sequesters sheaths, and finally migrate to the cap of the
reticulocytes so that they can complete their germinal centers. Thus, the suggestion has
maturation; in addition, it traps a large frac¬ been advanced that the periarterial lymphoid
tion of blood platelets in a reserve pool avail¬ sheaths consist of a central, thymus-depend¬
able to meet physiological demands or emer¬ ent region and a peripheral, bursa-depend¬
gency conditions. ent region. In neonatally thymectomized ro¬
Although in the embryo the spleen con¬ dents, the periarterial lymphoid sheaths are
tains immature precursors of the circulating poorly populated with lymphocytes; thus, the
blood elements, the erythrocytes found in vast majority of their lymphocytes are rep¬
the red pulp in the normal, adult human are resented by T cells.
never formed there. In some pathological The transit time of the recirculating lym¬
conditions, especially myeloid leukemia, the phocytes through the spleen is very short and
red pulp of the spleen undergoes myeloid may be as brief as two hours. The pathway
metaplasia, after which a large number of followed by the lymphocytes in entering and
erythroblasts, megakaryocytes, and myelo¬ leaving the periarterial lymphoid sheaths is
cytes appear in the tissue, so that the red poorly understood. They probably enter the
pulp acquires a structure resembling that of cords of the marginal zone by crossing the
the red bone marrow. In many other adult walls of the venous sinuses and subsequently
mammals, some myelocytes and erythroblasts migrate into the white pulp. The route by
may be found normally in the red pulp, and which the lymphocytes reenter the blood is
megakaryocytes are consistently present in unknown; it has been suggested that they
the spleen of rats and mice. follow the lymphatic vessels that accompany
The spleen has great physiological impor¬ the central artery, but these vessels do not
tance in the immune response to bacteria, seem to be found consistently in the splenic
viruses, and foreign macromolecules that white pulp of all mammals.
have invaded the circulation. In an animal Upon introduction into the bloodstream of
not involved in an acute response to antigen, an antigen that elicits a humoral response,
it is evident from the small arteriovenous morphological changes are first seen in the
difference in lymphocyte counts on splenic periarterial lymphoid sheaths. One day after
 SPLEEN • 477

the injection, proliferating lymphoblasts ap¬ Blue, J., and L. Weiss: Species variation in the structure
pear, scattered throughout the sheaths. They and function of the marginal zone—an electron
microscope study of cat spleen. Am. J. Anat.
increase in number during the following one
161:169, 1981.
or two days and become more concentrated Blue, J., and L. Weiss: Electron microscopy of the red
toward the periphery of the sheaths. At the pulp of the dog spleen including vascular arrange¬
same time, antibody first appears in the ments, periarterial macrophage sheaths (ellipsoids),
and the contractile, innervated reticular meshwork.
bloodstream. Lymphoblasts also occur
Am. J. Anat. 161:189, 1981.
around the small arteries of the red pulp. Burke, J. S., and G. T. Simon: Electron microscopy of
These may have developed from the terminal the spleen. I. Anatomy and microcirculation. Am.
extensions of the periarterial lymphoid J. Pathol. 55:127, 1970.
sheaths that surround the penicillar arteries. Burke, J. S., and G. T. Simon: Electron microscopy of
the spleen. II. Phagocytosis of colloidal carbon. Am.
On days 4 to 6, an increasing number of
J. Pathol. 55:157, 1970.
immature plasma cells appear at the periph¬ Carr, I.: The fine structure of the mammalian lympho-
ery of the periarterial lymphoid sheaths and reticular system. Int. Rev. Cytol. 27:283, 1970.
along the penicillar arteries; mature plasma Chen, L-T., and L. Weiss: Electron microscopy of the
red pulp of human spleen. Am. J. Anat. 754:425,
cells are also found, but in very small num¬
1972.
bers. At this stage, morphological changes in Coons, A. H.: Some reactions of lymphoid tissues to
germinal centers are first seen; they contain stimulation by antigen. Harvey Lect. 55:113, 1959.
many proliferating lymphoblasts and macro¬ Edwards, V. D., and G. T. Simon: Ultrastructural aspects
phages loaded with debris of phagocytized of red cell destruction in the normal rat spleen. J.
Ultrastruct. Res. 55:187, 1970.
lymphocytes. At the end of the first week,
Ernstrom, U., and G. Sandberg: Migration of splenic
lymphoblasts and immature plasma cells be¬ lymphocytes. Acta Pathol. Microbiol. Scand. 72:379.
gin to decrease in number in the periarterial 1968.
lymphoid sheaths, and mature plasma cells Ford, W. L., and J. L. Gowans: The traffic of lympho¬
cytes. Semin. Hematol. 6:67, 1969.
are more numerous at the boundary between
Galindo, B., and T. Imaeda: Electron microscope study
white and red pulp and along the penicillar of the white pulp of the mouse spleen. Anat. Rec.
arteries. They also occur in the cords of the 143:399, 1962.
red pulp and not infrequently free in the Goldschneider, I., and D. D. McGregor: Migration of
sinus lumen. During the second week after lymphocytes and thymocytes in the rat. I. The route
of migration from blood to spleen and lymph nodes.
the introduction of the antigen, the structure J. Exp. Med. 727:155, 1968.
of the spleen reverts to normal, except for Hanna, M. G., Jr., and A. K. Szakal: Localization of 125I-
the germinal centers, which continue to re¬ labeled antigen in germinal centers of mouse spleen:
main prominent for about one month. histologic and ultrastructural autoradiographic
studies of the secondary immune reaction. J. Im¬
During the secondary response, the spleen
munol. 101:949, 1968.
undergoes changes that resemble those fol¬ Jacobsen, G.: Morphological-histochemical comparison
lowing the first exposure to antigen, but they of dog and cat splenic ellipsoid sheaths. Anat. Rec.
occur earlier and are much more dramatic. 769:105, 1971.
Jacobson, L. O., E. K. Marks, M. J. Robson, E. Gaston,
At the beginning of the response the spleen
and R. E. Zirkle: The effect of spleen protection on
is, per unit weight, the most active organ of mortality following x-irradiation. J. Lab. Clin. Med.
the body in antibody secretion, but it rapidly 54:1538, 1949.
falls off in production as the response be¬ Klemperer, P.: The Spleen. In Downey, H., ed.: Hand¬
comes propagated throughout the other pe¬ book of Hematology. New York, Paul B. Hoeber,
1938.
ripheral lymphoid tissues and organs of the
Knisely, M. H.: Spleen studies. I. Microscopic observa¬
body. tions of the circulatory system of living unstimulated
Inasmuch as macrophages and lympho¬ mammalian spleens. Anat. Rec. 65:23, 1936.
cytes are not restricted to the spleen, it is not Langevoort, H. L.: The histophysiology of the antibody
response. I. Histogenesis of the plasma cell reaction
surprising that the effects of splenectomy are
in rabbit spleen. Lab. Invest. 72:106, 1963.
transient and largely disappear as splenic Lewis, O. J.: The blood vessels of the adult mammalian
functions are assumed by other organs. spleen. J. Anat. 97:245, 1957.
MacKenzie, D. W., Jr., A. O. WTipple, and M. P.
Wintersteiner: Studies on the microscopic anatomy
and physiology of living transilluminated mammal¬
REFERENCES ian spleens. Am. J. Anat. 65:397, 1941.
McCuskey, R. S., and P. A. McCuskey: In vivo micros¬
Blue, J., and L. Weiss: Periarterial macrophage sheaths copy of the spleen. Bibl. Anat. 76:121, 1977.
(ellipsoids) in cat spleen-—an electron microscope Miyoshi, M., and T. Fujita: Stereo-hne structure of the
study. Am. J. Anat. 767:115, 1981. splenic red pulp. A combined scanning and trans¬
Blue, J., and L. Weiss: Vascular pathways in nonsinusal mission electron microscope study on dog and rat
red pulp—an electron microscope study of the cat spleen. Arch. Histol. Jpn. 55:225, 1971.
spleen. Am. J. Anat. 161:135, 1981. Miyoshi, M., T. Fujita, and J. Tokunaga: The red pulp
478 • SPLEEN

of the rabbit spleen studied under the scanning with special reference to sinus lining cells. Z. Zell¬
electron microscope. Arch. Histol. Jpn. 32:289, forsch. 97:300, 1968.
1970. Sussdorf, D. H., and L. R. Draper: The primary hemo¬
Mollier, S.: Uber den Bau der Capillaren Milzvenen lysin response in rabbits following shielding from x-
(Milzsinus). Arch. f. Mikros. Anat. 76:608, 1911. rays or x-irradiation of the spleen, appendix, liver
Movat, H. Z., and N. V. P. Fernando: The fine structure or hind legs. J. Infect. Dis. 99:129, 1956.
of lymphoid tissue. Exp. Mol. Pathol. 3:546, 1964. Szakal, A. K., and M. G. Hanna, Jr.: The ultrastructure
Movat, Id. Z., and N. V. P. Fernando: The fine structure of antigen localization and virus-like particles in
of the lymphoid tissue during antibody formation. mouse spleen germinal centers. Exp. Mol. Pathol.
Exp. Mol. Pathol. 4:155, 1965. 5:75, 1968.
Nieuwenhuis, P., and W. L. Ford: Comparative migra¬ Thiel, G. A., and H. Downey: The development of the
tion of B- and T-lymphocytes in the rat spleen and mammalian spleen, with special reference to its
lymph nodes. Cell. Immunol. 23:254, 1976. hematopoietic activity. Am. J. Anat. 25:279, 1921.
Peck, FI. M., and N. L. Hoerr: The intermediary circu¬ Weidenreich, F.: Das Gefasssystem der menschlichen
lation in the red pulp of the mouse spleen. Anat. Milz. Arch. Mikros. Anat. 55:247, 1901.
Rec. 769:447, 1951. Weiss, L.: An experimental study of the organization of
Pictet, R., L. Orci, W. G. Forssmann, and L. Girardier: the reticuloendothelial system in the red pulp of the
An electron microscope study of the perfusion-fixed spleen. J. Anat. 93:465, 1959.
spleen. I. The splenic circulation and the RES con¬ Weiss, L.: The structure of fine splenic arterial vessels
cept. Z. Zellforsch. 96:372, 1969. in relation to hemoconcentration and red cell de¬
Robinson, W.: The vascular mechanism of the spleen. struction. Am. J. Anat. 777:131, 1962.
Am. J. Pathol. 2:341, 19264 Weiss, L.: The structure of intermediate vascular path¬
Simon, G. T., and J. S. Burke: Electron microscopy of ways in the spleen of rabbits. Am. J. Anat. 773:51,
the spleen. IIE Erythroleukophagocytosis. Am. J. 1963.
Pathol. 55:451, 1970. Weiss, L.: The Cells and Tissues of the Immune System.
Snodgrass, M. J.: A study of some histochemical and Structure, Functions, Interactions. Englewood
phagocytic reactions of the sinus lining cells of the Cliffs, NJ, Prentice-Hall, 1972.
rabbit’s spleen. Anat. Rec. 767:353, 1968. Weiss, L..: The development of the primary vascular
Snook, T.: A comparative study of the vascular arrange¬ reticulum in the spleen of human fetuses (38 to 57
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1950. 1973.
Solnitzky, O.: The Schweigger-Seidel sheath (ellipsoid) Weiss, L., and M. Tavassoli: Anatomical hazards to the
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Sprent, J.: Circulating T and B lymphocytes of the Hematol. 7:372, 1970.
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Stutte, H. J.: Nature of human spleen red pulp cells
 17
HYPOPHYSIS

The hypophysis or pituitary is an endo¬ and the infundibular process. The relations of
crine gland located at the base of the brain. these components are depicted in Figure 17—
It is about 1 cm in length, 1 to 1.5 cm in 3. In many species, the pars intermedia is
width, and about 0.5 cm deep. It weighs closely adherent to the infundibular process
about 0.5 g in men and slightly more in to form the so-called posterior lobe, separated
women. Despite its small size it is one of the by a cleft from the pars distalis or anterior
most important organs in the body, produc¬ lobe. In man the cleft is largely obliterated
ing at least nine hormones and having many in late fetal and postnatal life, so that the
reciprocal relations with other endocrine anterior and posterior lobes are in continuity.
glands. It also has neural and vascular con¬ The subdivisions of the hypophysis and the
nections with the brain, to which it is attached accepted descriptive terminology are pre¬
by a slender stalk. By virtue of these connec¬ sented in tabular form in Figure 17—1.
tions, the hypophysis occupies a key position The hypophysis is lodged in a deep depres¬
in the interplay of the nervous system and sion in the sphenoid bone, the sella turcica,
the endocrine system—the two great inte¬ and is covered by a tough diaphragm, the
grating systems of the body. diaphragma sellae. This barrier between the
The hypophysis has two major subdivi¬ sella turcica and the intracranial cavity is
sions: the neurohypophysis, which develops as often incomplete, being penetrated by an
a process growing downward from the floor opening 5 mm or more in diameter around
of the diencephalon, and the adenohypophysis, the hypophyseal stalk. Some of the pia-arach-
which originates in the embryo as a dorsal noid may extend through this opening and
outpocketing of the roof of the embryonic occupy the narrow space between the dia¬
pharynx. There are three subdivisions of the phragm and the connective tissue capsule of
adenohypophysis: the pars distalis or anterior the gland. Elsewhere the dense collagenous
lobe, the pars infundibular is (pars tuberalis), and capsule is separated from the periosteum of
the pars intermedia. The neurohypophysis the sphenoid bone by a looser layer of con¬
generally is divided into three regions: the nective tissue containing numerous veins.
median eminence, a funnel-shaped extension This layer appears to be separate from the
of the tuber cinereum; the infundibular stalk; pia-arachnoid. In mammals other than man,

Pars distalis

Adenohypophysis Pars tuberalis

Pars intermedia

Pars nervosa
(Processus infundibuli)
Neurohypophysis
Infundibular stem
Infundibulum
Median eminence
of the tuber cinereum

Figure 17-1. Terminology of the divisions and subdivisions of the hypophysis. In addition, the pars intermedia and pars
nervosa together are sometimes called the posterior lobe, and the pars distalis and pars tuberalis are collectively called
the anterior lobe.
479
480 • HYPOPHYSIS

Figure 17-2. Schematic drawing of the hypophysis of an adult rhesus monkey, showing its relation to the sella turcica
of the sphenoid bone. Also depicted are the superior and inferior hypophyseal arteries (sha and iha) and the important
portal venules (pv) coursing down the infundibular stalk. The superior hypophyseal artery usually sends an ascending
branch (1) to the proximal part of the infundibular stalk and median eminence and a descending branch (2) coursing
distally. ar, Arachnoid membrane; ba, basilar artery; bv, basilar veins; d, dura; di, sellar diaphragm; Iv, lateral hypophyseal
veins; oc, optic chiasm; pc, posterior clinoid process; sas, subarachnoid space; sd, subdural space; v, dural vein; vpi,
veins of the infundibular process. (After Wislocki, G. B. Proc. Assoc. Res. Nerv. Ment. Dis. 77:48, 1936.)
 HYPOPHYSIS • 481

Figure 17-3. Diagram of midsagittai section of hypothal¬

amus and hypophysis of man to show relations of major
divisions and subdivisions of the gland to the hypothala¬
mus. (Modified from Tilney.)

c
Figure 17-4. Photomicrographs of anterior lobe of hypophysis of adult rats. A, Normal female rat. B, Castrated female
rat of same age. Hypertrophy of basophil cells ($), with enlargement of the Golgi complexes (here shown next to each
nucleus as a negative image and appearing as a clear halo), and reduction in acidophil (a) and chromophobe (C) cells
follow castration. Zenker-formol, Mallory-azan. x 1300. (Courtesy of I. Gersh.)
482 • HYPOPHYSIS

the diaphragma sellae is commonly incom¬ the staining affinities of the specific granules.
Historically these terms were reasonable and
plete.
served to distinguish two major classes of
chromophilic cells at a time when there were
only a few empirically developed staining
PARS DISTALIS combinations in routine use, and when the
great diversity of pituitary functions was not
yet appreciated. As time has passed, the num¬
The pars distalis or anterior lobe is the ber of hormones known to be secreted by the
largest subdivision of the hypophysis. It is adenohypophysis has increased to six. The
composed of glandular cells arranged in ir¬ effort to identify cell types to which synthesis
regular cords and clumps. These are inti¬ of each of these hormones could be attrib¬
mately related to an extensive system of thin- uted led to the development of numerous
walled sinusoids of the blood vascular system. staining methods. The terminological prob¬
The anterior lobe is largely enclosed by a lem has been greatly aggravated by the fact
dense collagenous capsule. The stroma of the that most of the staining procedures now
gland is not abundant, but some collagenous considered useful for the study of the ade¬
fibers, which accompany the superior hypo¬ nohypophysis do not make use of an acid
physeal arteries and the portal venules, pen¬ and a basic dye but involve mixtures of acid
etrate the anterior lobe at the pole adjacent dyes. With many of these methods, staining
to the pars tuberalis and fan out bilaterally, does not depend on the binding of a dye by
extending about a third of the way into the a tissue component of opposite charge, and
gland. There they become continuous with no conclusion as to the chemical nature of
reticular fibers that surround the cords of the granules can be drawn from their color
parenchymal cells and support the small in sections stained in this way. The color of
branches of the hypophyseal artery and the specific granules of the same cell type may
sinusoids. The sinusoids at the periphery of be red, orange, purple, or blue, depending
the gland continue into collecting venules on the combination of acid dyes used. With
that join an extensive venous plexus in the the trichrome staining methods, it has been
capsule. The endothelium lining the sinu¬ necessarv to establish the relation of the cell
soids was formerly considered to be phago¬ types to the traditional acidophilic, baso¬
cytic and was classified as a component of the philic, and chromophobic categories by com¬
reticuloendothelial system. This interpreta¬ parison of the same cells in consecutive sec¬
tion is not borne out by electron microscopy. tions stained with trichrome mixtures and
Uptake of particulate tracers in the gland is with hematoxylin and eosin.
confined to extravascular tissue macro¬ The most meaningful histochemical meth¬
phages. od for identification of cell categories is the
The glandular cells are classified as chro- periodic acid-Schiff (PAS) reaction, which
mophilic or chromophobic on the basis of their selectively stains the granules of basophils
avidity or lack of affinity for the dyes used because of their content of glycoprotein.
in routine staining of histological sections. Another approach of proven value for iden¬
The chromophilic cells were originally sub¬ tification of the cell of origin of various
divided into acidophilic cells or basophilic cells, hormones involves the use of immunohisto-
according to the tinctorial reactions of their chemical procedures. In these methods, an¬
specific granules in sections stained with eosin tibodies to a specific hormone are induced in
and alum-hematoxylin or with other combi¬ another species and are conjugated with flu¬
nations of an acidic and a basic dye. orescent dyes or with horseradish peroxidase.
It is important to realize that the terms These labeled antibodies are then reacted
acidophilic and basophilic as used by the with sections of hypophysis and the sites of
pituitary cytologist do not have the same the antigen in the tissue are localized by
connotation with respect to the chemistry of fluorescence microscopy or by the histochem¬
the cytoplasm that they generally have in ical method for peroxidase.
other fields of cytology. The basophilia of Electron microscopic studies have shown
the granules in the pituitary basophilic cell is that the specific granules of the cells in the
not to be confused with that attributable to adenohypophysis differ significantly in their
cytoplasmic ribonucleoprotein in other glan¬ size. Granule size and shape are therefore
dular cells. In the naming of the pituitary valuable criteria for distinguishing cell types
cells, acidophilic and basophilic refer only to in electron micrographs.
 HYPOPHYSIS • 483

Various systems of nomenclature based on structural features and immunohistochemical

Greek letter designations have been pro¬ staining properties.
posed to avoid the inconsistency involved in
continued use of acidophil and basophil, but
Mammotrophs (Prolactin Cells)
none of these has gained widespread accept¬
ance. The terminology now in general use is Acidophils of this type tend to be distrib¬
therefore a confusing mixture of terms in uted individually in the interior of the cell
which acidophil, basophil, and chromophobe cords. They secrete the hormone prolactin,
are used to designate three major classes of which stimulates the mammary gland. They
cells in the adenohypophysis, while various contain numerous small granules about 200
Greek letters or adjectives referring to dis¬ nm in diameter.
tinctive tinctorial reactions are used to iden¬ In the nonlactating, sexually mature fe¬
tify specific cell types within these classes. In male they are relatively small cells with a few
recent years it has become common practice short cisternae of reticulum near the cell
to substitute terms that identify the target periphery. In pregnancy they are stimulated
organ stimulated (e.g., thyrotroph, gonadotroph, by the elevated level of circulating estrogens,
corticotroph) or the hormone secreted (e.g., and undergo considerable hypertrophy. In
TSH cell, FSH/LH cell, ACTH cell). this condition they stain with azocarmine or
The description of the ultrastructure of erythrosin in trichrome stains, and they were
the cells that follows relies heavily upon stud¬ formerly called “pregnancy cells.” Their
ies of the rat, because comparable investiga¬ Golgi complex enlarges and the endoplasmic
tions of the human gland are few as yet. reticulum becomes more extensive, forming
Although there are interspecific differences, multiple layers parallel to the plasmalemma,
the general principles are the same. and the granules become larger, 550 to 600
nm in diameter and often irregular in outline
(Fig. 17-7).
ACIDOPHILS (ALPHA CELLS) The period of greatest activity of the mam¬
motrophs is during the postpartum prolactin
Acidophils in the human hypophysis are secretion necessary to initiate and maintain
most numerous in the posterolateral portions lactation. When suckling is terminated, lyso-
of the pars distalis. They are small rounded somes have an important role in the elimi¬
cells 14 to 19 pm in diameter, with a well- nation of excess secretory granules and the
developed juxtanuclear Golgi complex and hypertrophied cellular organelles involved in
small rod-shaped mitochondria. They stain the earlier period of active protein synthesis.
with eosin in routine histological sections, and Lysosomes fuse with the secretory granules
their granules are large enough to be re¬ to form autophagic vacuoles in which the
solved with the light microscope. Two cate¬ granules are degraded by hydrolytic enzymes
gories of acidophils can be distinguished by (Fig. 17—8). This method of disposal of secre¬
selective staining methods, and with greater tory product no longer needed is called cri-
precision by immunocytochemical proce¬ nophagy. Excess cytomembranes and ribo¬
dures using antibodies to specific hormones. somes are also enclosed in vacuoles and
degraded by autophagy until the mammo¬
Somatotrophs trophs have reverted to the relatively inactive
state characteristic of the normal cycling fe¬
These acidophils, occurring in groups male.
along the sinusoids, secrete growth hormone
(.somatotropin) and are therefore called soma¬
totrophs (STH cells). They contain very nu¬
BASOPHILS (BETA CELLS)
merous spherical granules 300 to 350 nm in
diameter (Figs. 17-5, 17-6), which are selec¬
tively stained by antibody against growth hor¬ This category of cells in the anterior pitui¬
mone. The cisternae of their well-developed tary stains poorly with hematoxylin and is
endoplasmic reticulum tend to be arranged less easily identified in routine preparations
parallel to the cell surface. than are the eosinophils. Basophils do stain
Human pituitary gigantism resulting from well with the aniline blue of Mallory’s tri¬
excess production of growth hormone is as¬ chrome stain and with resorcin-fuchsin. They
sociated with a tumor of the pars distalis are most easily distinguished from eosino¬
composed of acidophils having these ultra- phils by their pink staining with the PAS
484 • HYPOPHYSIS

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4 m A" - ■, . /■ ' - #
% ns®,,.* mi •* v - ': Jll,:#'. J1 fl|»s9,9.,/; ■ . / ji# f
' Jk*1 ' . ,,
*:.■ '"## ■ #;
# A
1 £Z** ^ ^ s ••%■•• m
kSM*** ."'i'll ' | ii 1 V1 'I 1 hi " '1 1 . 1 '. . '''''/ W i" i I, i ' i i 11 1 "|||| 1 lilll I I li
' , '9 .. ^ • j? : 49- , N.. ■ ,
V '"
! ,99y», «4 " 4;.f!;■ ..■■■' , skgf r ml ■ «s?s • :|«'“ , a.: ; 4V9 999/4
Bji ••,

P&.J&5 1
-- ** * #• aw--*i-r .#§
r $fr;jf£0# : : ;:#4Sx.,
4',^ ^9;..^;;^ 9--\P ; " MMWM
'- x^'f
:9>:.44v. IlfMiiiip ip®? 1 . |
••■ ' ,'v:. I - • a .a, vv

m
mm f Mammatrope '■• I '. , 'II *
4
iiij V
iSSi&SiSiiSSSS «SS ■■,•<■ aSsK;^ 4 2 | *& *a -.'.a.?- ,.'\ '« a.

Figure 17-5. Electron micrograph of an area of the pars distalis of rat hypophysis illustrating the fine structure and
relative size of the specific granules of a somatotrope, mammatrope, and corticotrope. (Micrograph from Nakayama, I.,
F. A. Nickerson, and F. R. Shelton. Lab. Invest. 27:169, 1969.)
 HYPOPHYSIS • 485

Figure 17-6. A typical somatotrope, showing numerous cisternae of endoplasmic reticulum, a well-developed Golgi
complex, and many specific granules about 350 nm in diameter. (Micrograph courtesy of M. Farquhar.)

Figure 17-7. Electron micrograph of a rat mammatrope. Notice the relatively large size and irregular shape of the
granules. A number of developing granules are associated with a large Golgi complex at lower right of figure. (Micrograph
courtesy of M. Farquhar and T. Kanaseki.)
486 • HYPOPHYSIS

In the human they are round or oval, are

distributed throughout the anteromedial
portion of the pars distalis, and commonly
invade a short distance into the neural lobe.
They contain granules that are intensely
stained with the PAS reaction. The granule
size 200 to 250 nm does not differ signifi¬
cantly from that of gonadotrophs and is not
a reliable criterion for identification. In ro¬
dents these cells are irregularly stellate in
shape, with cell processes that extend be¬
tween neighboring cells to end adjacent to
sinusoids. Their cytoplasm is of low density
with a rather sparse endoplasmic reticulum
(Fig. 17-5). The granules tend to be located
adjacent to the cell membrane. The small
granules are not easily resolved with the light
microscope and these cells were often mis-
identihed as chrorriophobes. The most de¬
pendable criterion for their identification is
Figure 17-8. Diagram illustrating the secretory pathway immunocytochemical staining with anti-
of a mammatrope. Small granules are formed in the Golgi ACTH or anti-LPH.
complex and subsequently fuse to form larger granules, It was long believed that there was one cell
often of irregular outline. During lactation, they are dis¬ type for each pituitary hormone. It is now
charged by exocytosis, but after the young are weaned,
excess granules fuse with lysosomes and are destroyed
known that the corticotrophs synthesize a
by autophagy. (After Smith, R. E., and M. G. Farquhar. J. 31,000-M.W. glycoprotein prohormone that
Cell Biol. 31:319, 1966.) includes the amino acid sequences of ACTH
and LPH. This molecule undergoes post-
reaction for carbohydrates. There are three translational cleavage to yield these two hor¬
distinct types of basophils. mones. In pig and rat, and possibly in other
species, the LPH is further processed to yield
beta endorphin, an opiate-like peptide with
Thyrotrophs (Beta Basophils, TSH
potent analgesic activity.
Cells)
Following adrenalectomy, corticotrophs
The cells that secrete thyroid stimulating hor¬ become more numerous and larger, and con¬
mone (TSH) are elongated or polygonal in tain greater numbers of granules. After pro¬
shape and arranged in clusters in the anter¬ longed administration of cortisol, they de¬
omedial portion of the anterior lobe. They crease in size and stain only faintly.
tend to be deeply situated in the cell cords,
not in contact with the sinusoids. They can Gonadotrophs (Delta Basophils,
be distinguished from other basophils by the Gonadotropes, FSH/LH Cells)
selective staining of their granules with al¬
dehyde thionin. In electron micrographs they These rounded cells are usually situated
have the smallest granules of any cell in the adjacent to sinusoids. They secrete two hor¬
pituitary, 140 to 160 nm in diameter. They mones, follicle stimulating hormone (FSH) and
are less dense than the granules of other luteinizing hormone (LH). They are PAS posi¬
basophils and tend to congregate at the pe¬ tive but do not stain with aldehyde fuchsin.
riphery of the cell. They stain immunocyto- In electron micrographs there is a prominent
chemically with anti-TSH antibody. juxtanuclear Golgi complex and a well-devel¬
Thyrotrophs hypertrophy following thy¬ oped endoplasmic reticulum with meander¬
roidectomy and atrophy after thyroxin ing cisternae that are often distended with a
administration. homogeneous content of low density (Fig.
17-9). The granules are spherical and vary
Corticotrophs (Corticotropes, over a wide range, 200 to 400 nm in diame¬
ACTH Cells) ter. Whether there are two gonadotrophs,
one secreting FSH and the other LH, has
These cells secrete adrenocorticotropic hor¬ long been a subject of controversy. Certainly
mone (ACTH) and lipotropic hormone (LPH). they exhibit considerable cytological varia-
 HYPOPHYSIS • 487

Figure 17-9. Gonadotrope with granules of relatively smaller size than the somatotrope (see Fig. 17-6) but displaying
considerable variability. The endoplasmic reticulum is typically distended with an amorphous material of low density.
(Micrograph courtesy of M. Farquhar.)

tion. Some are ovoid with a population of seems likely that many of the cells identified
small granules; others are angular or stellate as chromophobes with the light microscope
and contain only small granules, 200 to 220 are, in fact, partially degranulated chromo-
nm. Some stain immunocytochemically only phil cells.
with anti-FSH, others with anti-LH, and still Mitoses are relatively few in the anterior
others react to both antibodies. It remains lobe. For this reason, it was formerly thought
unsettled whether both hormones are found that the shifts in proportions of the three
in the same granule or in different popula¬ major cell categories were the result of trans¬
tions of granules. It seems likely that the formations of one cell type to another. In¬
varying cytological features observed repre¬ vestigations of these population shifts led to
sent different physiological states of the same proposals of several cell lineages based upon
cell type. observation of what were presumed to be
morphological transition stages. The most
widely accepted of these schemes considered
CHROMOPHOBES (RESERVE CELLS) the chromophobes to be a reserve population
of relatively undifferentiated cells capable of
Chromophobes are usually small cells lo¬ differentiating into either acidophils or ba¬
cated in groups in the interior of the cell sophils. It has become increasingly apparent
cords. They generally have less cytoplasm that the cells classified as chromophobes by
than the chromophilic cells but may rarely light microscopy are not a homogeneous pop¬
reach the dimensions of acidophils or baso¬ ulation. Some are evidently chromophils de¬
phils. Traditionally, chromophobes have granulated to the point at which their specific
been considered to be devoid of specific gran¬ nature is not detectable. There seems to be a
ules, but electron micrographs reveal rela¬ considerable degree of cytological specializa¬
tively few cells with no specific granules. The tion among the cells normally classified as
cells of the pars distalis are believed to have chromophobes. Some are said to have a Golgi
cyclic secretory activity, first accumulating apparatus characteristic of acidophils, while
and then releasing their specific granules. It in others the Golgi apparatus resembles that
488 • HYPOPHYSIS

Cell Type Staining reactions*

Electron microscopic
Hormones General Specific AF PAS description

Growth or somato¬ Acidophil Somatotrope — 350 nm. granules, cells

tropic hormone columnar and arranged
(STH) in groups on sinusoids

Lactogenic or luteo- Acidophil Mammotrope 600 nm. elliptical gran¬

tropic hormone or luteotrope ules, cells located indi¬
(LTH) vidually in interior of
cell cords

Thyrotropic Basophil Thyrotrope + + 140 nm. granules, cells

hormone (TSH) angular and not usually
located on sinusoids

Follicle stimulating Basophil FSH — + 200 nm. granules, cells

hormone (FSH) Gonadotrope located on sinusoids and
are usually rounded
»

Luteinizing Basophil LH + 200 nm. granules, cells

hormone (LH) Gonadotrope usually located on
sinusoids, rounded and
contain bizarre cyto¬
plasmic formations

Adrenocorticotropic Chromophobe Corticotrope 200-250 nm. granules,

hormone (ACTH) cells pale, stellate. Few
granules at cell
periphery

No specific hormone Chromophobe Acidophilic Few characteristic

chromophobe granules
Basophilic
chromophobe

*AF, Aldehyde fuchsin; PAS, periodic acid-Schiff.

Figure 17-10. Summary of current views of rat anterior pituitary cell types and their secretions. (Modified from McShan
W. H., and M. W. Hartley. Ergebn. Physiol. 56:264, 1965.)

of basophils. It is probable that many of the containing colloidal material of low electron
apparent chromophobes are already deter¬ density. In this form the cells have microvilli
mined and capable of differentiating into and occasional tufts of cilia on their luminal
only one of the chromophil types. surface. The apical plasma membrane has a
With the light microscope, chromophobes prominent surface coat and the cells are
were formerly estimated to make up 65 per joined by juxtaluminal junctional complexes.
cent of the cells in the pars distalis. It is now Unlike other epithelia, these cells have long
evident that the great majority of these were basal processes that extend outward between
degranulated secretory cells with so few re¬ the neighboring glandular cells. Their cyto¬
maining granules that their affinity for stains plasm contains the usual complement of or¬
was below a detectable level. If chromo¬ ganelles, occasional lipid droplets, numerous
phobes that are nonspecific stem cells exist at polyribosomes, and beta particles of glyco¬
all, they are evidently much less numerous gen. They may also occur as stellate cells that
than they were formerly thought to be. do not line follicles but extend branching
processes among the secretory cells. Similar
interstitial stellate cells form a loose mesh-
FOLLICULAR CELLS (STELLATE work throughout the pars intermedia. Their
CELLS) function is poorly understood. They are re¬
ported to be phagocytic in in vitro studies,
The principal nonsecretory cells of the surrounding and ingesting dead cells and
anterior pituitary are follicular cells, so named cellular debris. There is evidence that they
because they often adopt an epithelial mode have a similar scavenger function in vivo.
of association, lining small cystlike follicles The filaments in their processes react with
 HYPOPHYSIS • 489

antibody to gliofibrillar acid protein, and it pars intermedia and pars tuberalis. There is
has been suggested that they may have a as yet no Supporting physiological evidence
nurse cell function comparable with that of of neural control of secretion in the pars
the glial cells of the central nervous system. distalis.

NERVES BLOOD SUPPLY

The secretory activity of the pars distalis is The blood supply of the hypophysis is
controlled primarily by releasing hormones unusual and is intimately involved in the
generated in the hypothalamus and carried control of the secretory activity of the gland.
to it in the blood. Therefore, one would not Two inferior hypophyseal arteries from the in¬
expect to find nerves in the gland. There ternal carotid arborize within the capsule of
are, however, isolated reports of the occur¬ the gland, sending branches to the posterior
rence, in some species, of rare bundles of lobe and to a lesser extent to the sinusoids of
unmyelinated axons found in the perisinu- the anterior lobe. Several superior hypophyseal
soidal connective tissue and in the delicate arteries arise from the internal carotid artery
septa between clusters of secretory cells. The and posterior communicating artery of the
axons are described as having varicosities circle of Willis and anastomose freely in the
containing small clear vesicles and large region of the median eminence of the hypo¬
dense-cored vesicles. Some appear to termi¬ thalamus and base of the pituitary stalk (Figs.
nate in close association with somatotrophs. 17—2, 17-11). From these vessels, capillaries
It remains unclear whether these are func¬ comprising the so-called primary plexus extend
tionally significant or represent aberrant in¬ into the median eminence and are then re¬
vasion of the pars distalis by nerves from the turned to the surface, where they are col-

Figure 17-11. Drawing of a thick median sagittal section of a cat’s hypophysis after injection of the blood vascular
system with India ink. The main blood supply is via the superior hypophyseal artery (sha) and inferior hypophyseal
arteries (iha). The venous drainage is via systemic veins from the pars distalis (vpd) and the pars nervosa (vpi). Portal
veins arising in capillaries in the median eminence and pars tuberalis (pv) carry the neurohumoral releasing hormones
from the median eminence of the hypothalamus (me) to the pars distalis. sas, Subarachnoid space; sd, subdural space;
Viii, third ventricle; a7 and a2, branches of inferior hypophyseal artery; cs, capsular venous sinuses; int, pars intermedia.
(From Wislocki, G. B. Anat. Rec. 69:361, 1937.)
490 • HYPOPHYSIS

lected into veins that run downward around ling many different physiological processes.
the hypophyseal stalk to supply the sinusoids Its surgical removal results in cessation of
of the adenohypophysis below. The venules growth in young animals; atrophy of the
connecting capillaries in the median emi¬ gonads, thyroid gland, and adrenal cortex;
nence with the sinusoidal capillaries of the and disturbances of carbohydrate, protein,
anterior lobe constitute the hypophyseoportal and lipid metabolism. These profound and
system. The venous drainage of the hypo¬ potentially fatal effects of hypophysectomy
physis is chiefly through vessels that run in are the collective consequences of eliminating
the vascular layer of the capsule to the dia¬ the source of the hormones whose individual
phragm of the sella turcica and thence into functions are described below.
adjacent dural sinuses. Some venous blood
may also enter sinuses in the sphenoid bone. Growth Hormone (Somatotropin, GH,
It is firmly established that neurohumoral STH)
substances (releasing factors or hypophyseotropic
hormones) released by nerves in the median Growth hormone is a small protein con¬
eminence of the hypothalamus are carried in sisting of about 190 amino acids and no
the blood via the hypophyseoportal system to carbohydrate component. Unlike the other
the adenohypophysis, where they stimulate hormones of the anterior lobe, it does not
the cells to release their specific hormones. have a specific target organ but has a gener¬
alized effect upon cells throughout the body,
increasing their rate of protein synthesis.
HISTOPHYSIOLOGY OF THE PARS Other metabolic effects include increased
DISTALIS mobilization of fatty acids from adipose tissue
and a decreased rate of utilization of glucose.
The pars distalis or anterior lobe of the Its most conspicuous effect is upon the rate
hypophysis secretes at least six hormones that of growth of young animals. Absence of
stimulate several target organs—thyroid, ad¬ growth hormone results in pituitary dwarfism.
renal cortex, ovaries, testicles, and mammary Excess secretion leads to pituitary gigantism.
glands. It is an indispensable organ Control- Growth hormone produced in excess by a
pituitary tumor in adult life results in acro¬
megaly, a condition characterized by dispro¬
portionate thickening of the bones. Its effect
upon growth in stature in childhood is me¬
diated by smaller proteins, somatomedins,
which are synthesized in the liver and else¬
where in response to growth hormone. The
somatomedins in turn stimulate the prolif¬
eration of cartilage that is necessary for
growth in length of the long bones.

Prolactin
The lactogenic hormone prolactin is a pro¬
tein with a molecular weight of about 25,000
consisting of a single chain of 205 amino
acids. Its principal function is the promotion
of mammary gland development and lacta¬
tion. In pregnancy its concentration in the
blood rises progressively from the fifth week
until full term when it reaches levels ten times
that of the nonpregnant woman. This stim¬
ulates development of the mammary gland,
but the lactogenic effect of the hormone is
suppressed by high levels of estrogen and
progesterone until birth of the baby. There¬
Figure 17-12. Photomicrograph of anterior lobe of hy¬
pophysis of monkey injected intravenously with India after the precipitous fall of these ovarian
ink to show the irregular, richly anastomotic sinusoids, hormones allows the lactogenic effect of pro¬
x 165. (Courtesy of I. Gersh.) lactin to be expressed.
 HYPOPHYSIS • 491

In rodents prolactin also participates in Adrenocorticotropin

maintenance of the corpus luteum of preg¬ (Adrenocorticotropic Hormone,
nancy. Because of this function it has also ACTH)
been called luteotropin (LTH).
Adrenocorticotropin is a straight chained
polypeptide with 39 amino acids and a mo¬
Thyrotropin (Thyroid Stimulating lecular weight of about 4500. It stimulates
Hormones, TSH) the adrenal cortex to secrete cortisol. It is
Thyroid stimulating hormone is a glyco¬ produced in the corticotrophs by proteolytic
protein with a molecular weight of about cleavage of a larger precursor molecule, pro-
opiocortin or pro-opiomelanocortin. Another
2800. It appears to exert its effect exclusively
upon the thyroid gland where it promotes cleavage product, beta-lipotropic hormone
proteolysis of thyroglobulin and release of (LPH), is secreted with ACTH, but no pe¬
thyroid hormone into the blood. It also ripheral physiological effects of LPH have
causes hypertrophy of the thyroid cells and yet been identified. LPH may be further
cleaved to yield melanocyte stimulating hormone
increases their rate of synthesis of thyroid
(MSH), and beta endorphin. The physiolog¬
hormone.
ical role of these peptides is still unclear.

Gonadotropins: Follicle Stimulating

Hormone (FSH) and Luteinizing Occurrence of Pituitary Hormones in
Hormone (LH) Other Organs
These two hormones are produced by the It has long been assumed that the hor¬
cells of the anterior lobe called gonadotrophs. mones of the adenohypophysis were pro¬
As indicated earlier in this chapter, the ques¬ duced only by specific cells in that gland. As
tion is moot whether they are produced in sensitive radioimmunoassays for their detec¬
the same cell concurrently, or in a different tion and immunocytochemical methods for
phase of a secretory cycle, or by two sub¬ their localization have developed, all the hor¬
populations of the same cell type. mones of the anterior and intermediate lobe,
Follicle stimulating hormone (FSH) is a gly¬ except the gonadotropins, have been found
coprotein with a molecular weight of about within the central nervous system of several
30,000. In the female there is a cyclic increase species including humans. Although some
and decrease in the secretion of FSH each investigators insist that all such hormones are
month. A rising level stimulates the devel¬ of pituitary origin, evidence is accumulating
opment of several follicles in the ovary in to indicate that they are of neural origin.
preparation for ovulation of one, or occasion¬ This interpretation is supported by the find¬
ally two, at midcycle. In the male, FSH plays ing that the concentration of ACTH, a-MSH,
an important role in the initiation of sper¬ P endorphin, and growth hormone in certain
matogenesis at puberty. Its function in the areas of the brain remain unchanged or in¬
adult is less clear, but it has been shown to crease after hypophysectomy. Moreover,
act upon the Sertoli cells of the seminiferous brain slices or dispersed central nervous sys¬
epithelium to promote the synthesis of an tem cells maintained in culture secrete into
androgen-binding protein. the medium a greater amount of hormone
Luteinizing hormone (LH) is also a glycopro¬ than was present in the tissue when ex-
tein, having a molecular weight of about planted.
26,000. In the female it acts upon the ovary, There is now great interest in the possible
promoting secretion of estrogen by the de¬ functions of these molecules in the nervous
veloping follicles. It is necessary for matura¬ system. Effects of some of these peptides on
tion of the follicle, and a midcycle surge of learning and behavior of experimental ani¬
LH release triggers ovulation. The common mals have been reported, and it is speculated
oral contraceptives act by inhibiting this peak that they may function as neurotransmitters
of LH. After ovulation the hormone causes or may modulate the effects of classical neu¬
differentiation of lutein cells that form the rotransmitters on the receptors of specific
corpus luteum. sets of target neurons.
In the male, LH stimulates the interstitial ACTH immunoreactivity has also been de¬
cells of the testis to secrete testosterone, tected in certain cells in the gastrointestinal
which is essential for maintenance of sper¬ tract and in neoplasms of the lung that are
matogenesis. thought to arise from neuroepithelial bodies
492 • HYPOPHYSIS

in the bronchial epithelium. One explanation thalamohypophyseal portal system of blood

advanced for the synthesis of similar peptides vessels.
in neurons and endocrine cells postulates that Prodigious efforts to identify these sub¬
both are specialized ectodermal cells arising stances culminated in 1968 in the isolation
from the epiblast of the embryo and have from 300,000 sheep hypothalami of 1 mg of
similar potential because of their common a hypophyseotropic peptide designated thy¬
origin. rotropin releasing factor (TRF), now more com¬
monly called thyrotropin releasing hormone
(TRH)—a peptide of only three amino acids.
Histophysiological Correlations
This was the first of a series of similar efforts
The cell types believed to be responsible that led to isolation, sequencing, and synthe¬
for secretion of the hypophyseal hormones sis of other releasing hormones: gonadotropin
have already been identified. Elaboration of releasing hormone (GnRH), a decapeptide that
somatotropin and prolactin is attributed to causes release of both FSH and FH; and
two morphologically distinct types of acido¬ corticotropin releasing hormone (CRH), a 41
phil. The glycoprotein gonadotropic hor¬ amino acid polypeptide releasing ACTH. Hy¬
mones FSH and FH are assigned to the PAS- pothalamic inhibiting hormones have also
positive basophils. There is reason to believe been isolated and characterized: prolactin in¬
that basophils that also stain with aldehyde- hibiting hormone (PIH), which suppresses pro¬
fuchsin are responsible for secretion of thy¬ lactin secretion; and growth hormone inhibiting
rotropin. In addition to the histochemical hormone (GHIH), also known as somatostatin.
and biochemical evidence relating these hor¬ The latter is less specific in its action than the
mones to the basophils, there is experimental others, inhibiting secretion of glucagon, in¬
evidence based upon the negative feedback sulin, and other hormones of the gastrointes¬
mechanisms that operate in the regulation of tinal tract in addition to growth hormone.
hormone release. Endocrine glands that are Thus, nearly all the functions of the ade¬
under the direct control of the anterior lobe nohypophysis depend on peptide signals re¬
hormones usually exert a reciprocal inhibit¬ ceived from the hypothalamus in the portal
ing effect upon hypophyseal function via the blood. Secretion of TSH, FSH, FH, and
hypothalamus. Removal of the target organ ACTH occurs in response to releasing hor¬
therefore results in hypertrophy of those cells mones. Prolactin, on the other hand, seems
in the adenohypophysis responsible for elab¬ to be under tonic inhibition by the hypothal¬
oration of the corresponding tropic hor¬ amus. Therefore, its secretion by mamma-
mone. After castration, the rat hypophysis trophs is enhanced by pituitary transplanta¬
contains increased amounts of gonadotropic tion or explantation to tissue culture, while
hormones, and at the same time the basophils other cell types atrophy.
become markedly enlarged and vacuolated
in a characteristic way (castration cells, Fig. 17—
4). Thyroidectomy also results in an increase
in the percentage of another type of basophil, PARS INTERMEDIA
thyroidectomy cells.
In many mammals, the pars distalis is sep¬
Control of Anterior Lobe Function arated from the neurohypophysis by a cleft,
lined on the juxtaneural side by a multilay¬
If the pituitary is severed from the hypo¬ ered epithelium of basophilic cells making
thalamus and transplanted elsewhere in the up the pars intermedia. There is considerable
body, its cells remain viable but the rates of variation among species in the degree of
secretion of all the hormones except prolactin development of the pars intermedia. In rats
fall nearly to zero. However, if the anterior and mice, dogs, cats, and oxen it forms a
lobe is placed in tissue culture together with multilayered epithelium of basophilic cells.
fragments of the ventral hypothalamus, se¬ In marsupials it is reduced to a layer only
cretory function of the pituitary cells is better one or two cells deep, and in cetacea, sirenia,
maintained. These and other experiments and some birds it is absent. In the human
clearly indicated that the control of pituitary fetus it is represented by a typical stratified
function depended on the existence of secre¬ epithelium adjacent to the infundibular proc¬
tory products of hypothalamic neurons that ess and may constitute 3 per cent of the
are carried to the pars distalis in the hypo- glandular portion of the hypophysis, but in
 HYPOPHYSIS • 493

the adult it is no longer identifiable as a ilar ultrastructure and immunohistochemical

distinct layer. In the great majority of hu¬ properties may also be found scattered in
mans the hypophyseal cleft becomes discon¬ neighboring regions of the neural lobe.
tinuous in postnatal life and is represented Nonsecretory stellate cells extend long
in the adult by a zone of cysts (Rathke’s cysts). branching processes around and between the
These are often lined by ciliated epithelium glandular cells, forming a loose meshwork
and contain a colorless or pale yellow colloid throughout the pars intermedia. These cells
that varies in consistency but is often a highly react strongly with antibody to glial fibrillar
viscous fluid. With the disappearance of the acid protein, and their membranes exhibit
cleft, the epithelium of the pars intermedia Na + ,K + -ATPase activity. They are therefore
becomes discontinuous and the isolated cells similar to the astrocytes of the median em¬
or groups of cells that remain may extend inence and hypophyseal stalk and may con¬
some distance into the neural tissue of the ceivably play some role in regulation of the
infundibular process, where they are often secretory activity of the MSH cells.
overlooked in routine histological prepara¬ The pars intermedia is poorly vascularized
tions but can be detected immunohistochem- but richly innervated. Numerous axons con¬
ically. Thus, the pars intermedia of humans tain dense-cored vesicles 80 to 120 nm in
differs from that of most mammals in several diameter and are believed to be dopami¬
respects: the cleft is rarely complete; cysts nergic fibers originating in the rostral arcuate
are of common occurrence; and the remain¬ nucleus.
ing basophilic cells are few and may extend
into the neural lobe to a surprising degree.
In rodents such as the mouse, on the other HISTOPHYSIOLOGY OF THE PARS
hand, the pars intermedia constitutes 19 per INTERMEDIA
cent of the hypophysis. The following de¬
scription of its ultrastructure is based mainly The role of the hypophysis in the control
on studies of the rodent hypophysis. of pigmentation in fish and amphibia was
The principal cells of the pars intermedia discovered over 50 years ago. Removal of the
secrete melanocyte stimulating hormone (MSH). gland in frogs resulted in lightening of skin
They are large polygonal epithelial cells, rich color, and when the anterior and intermedi¬
in mitochondria and possessing a well-devel¬ ate lobes were then transplanted separately
oped rough endoplasmic reticulum and Golgi into tadpoles, only the transplants of the pars
complex. Numerous small secretory granules intermedia resulted in skin darkening. The
200 to 300 nm in diameter are distributed rapid color change induced by the hormone
throughout the cytoplasm. Some of these are in lower vertebrates is caused by centrifugal
electron dense and others are quite pale. dispersion of the pigment granules into the
Their variation in density is attributed to radiating processes of their melanophores.
partial extraction of their contents, which are It was long assumed that pigmentation in
not adequately preserved by osmium fixation. mammals was not under pituitary control,
Although the granules are inconspicuous at but the hormone of the intermediate lobe
the light microscope level, the cells stain with has been shown to induce synthesis of mela¬
the PAS reaction. The hormone secreted is a nin in melanoma cells grown in vitro. The
simple polypeptide, but the carbohydrate increased pigmentation that occurs in hu¬
staining of the granules is consistent with mans suffering from degeneration of the
biochemical evidence that the precursor is a adrenal cortex (Addison’s disease) is now
high-molecular-weight glycoprotein. attributed to release of excess MSH and
In addition to the MSH-secreting cells ACTH from the hypophysis. The darkening
there is a limited number of cells in the pars of the skin observed in human pregnancy
intermedia that have all the ultrastructural may also result from enhanced release of one
characteristics of the ACTH-secreting cells of or both of these hormones.
the pars distalis. They are smaller than the A highly basic melanocyte stimulating poly¬
MSH cells, are irregular in outline, and have peptide was isolated in 1954 from pig pitui¬
dense, 200-nm secretory granules located tary glands and called a-MSH. A slightly
mainly at the periphery adjacent to the acidic polypeptide was subsequently isolated
plasma membrane. These cells react with from the same source and designated (3-
anti-ACTH antibody as intensely as the cor- MSH. Analysis of the amino acid sequences
ticotrophs of the pars distalis. Cells with sim¬ and synthesis of a- and (3-MSH was soon
494 • HYPOPHYSIS

achieved; a-MSH contains 13 amino acids in stalk. The distinctive morphological charac¬
a sequence identical to that of one portion of teristic of the pars tuberalis is the longitudinal
the molecule of ACTH. (3-MSFI contains 18 arrangement of its cords of epithelial cells,
amino acids, of which seven are in a sequence which occupy the interstices between the lon¬
similar to a part of the ACTH molecule. The gitudinally oriented blood vessels.
cells of the pars intermedia have also been The pars tuberalis is the most highly vas¬
found to release significant amounts of a cularized subdivision of the hypophysis, be¬
corticotropin-like intermediate lobe peptide (CLIP) cause it is traversed by the major arterial
and of (3 endorphin, an opiate-like peptide. supply for the anterior lobe and the hypo-
It has now been found that melanotrophs thalamohypophyseal venous portal system.
of the pars intermedia, corticotrophs of the The pars tuberalis is separated from the
pars distalis, certain neurons of the hypo¬ infundibular stalk by a thin layer of connec¬
thalamus, and ceils in the placenta synthesize tive tissue continuous with the pia. On the
a high-molecular-weight parent molecule outside, the connective tissue is typical arach¬
variously called protropin, pro-opiocortin, or noidal membrane. Between these, the blood
pro-opiomelanocortin. Proteolytic cleavage of vessels and groups of epithelial cells are sup¬
this glycosylated common precursor molecule ported by reticular fibers.
gives rise to ACTH, lipotropic peptide hor¬ The epithelial cells of the pars tuberalis
mone (|3-LPH), melanotropin (a-MSH), and include undifferentiated cells and some small
endorphins. acidophilic and basophilic cells. The main
Different modes of post-translational proc¬ component is a cuboidal or columnar cell,
essing of the precursor molecule in the sev¬ which may reach 12 to 18 pm in size and
eral cell types yield different ratios of the contains numerous small granules or some¬
cleavage products. Some of these function as times fine colloid droplets. The mitochondria
active hormones and others may serve as are short rods, and numerous small lipid
potentiators or inhibitors of other hormones. droplets may be present. These are the only
As yet, little is known about the interactions cells in the adult hypophysis containing large
and target cells of these peptides. amounts of glycogen. The cells may be ar¬
Melanotropin, the principal product of the ranged to form follicle-like structures. Is¬
pars intermedia, acts primarily upon melano¬ lands of squamous epithelial cells may also
cytes, but a variety of nonpigmentary effects be present. Despite the occurrence of a pars
have been described. Systemic administration tuberalis in all vertebrates studied, the epi¬
of MSH to rats has been shown to affect thelial cells are not known to have any dis¬
avoidance responses and induce other behav¬ tinctive hormonal function.
ioral changes. It is also reported to improve
attention and vigilance in humans. The neu¬
rons in certain regions of the brain are im-
munoreactive to (3-LPH, a-MSH, endor¬ NEUROHYPOPHYSIS
phins, and a number of other peptides.
Optimal brain function may ultimately prove The neurohypophysis consists of the me¬
to depend on a delicate balance between the dian eminence of the tuber cinereum, the
amounts of various endogenous neuropep¬ infundibular stalk, and the infundibular
tides. process (Fig. 17—3). Much of its substance
consists of axons of neurons whose cell bodies
are located at a higher level in the hypothal¬
PARS INFUNDIBULARIS OR amus. Therefore, the organization and func¬
tion of the neural lobe cannot be adequately
TUBERALIS described without inclusion of the cell bodies
that are situated beyond its anatomical
Like the pars intermedia, the pars tuberalis boundaries.
constitutes only a small part of the hypo¬ The hypothalamus is the major neuroen¬
physis. Both are adjacent to and continuous docrine regulatory center of the brain.
with the anterior lobe. The pars tuberalis is Within it, two principal neurosecretory sys¬
25 to 60 pm thick and forms a sleeve around tems are recognized. In one, designated the
the stalk, the thickest portion being on its parvicellular system, axons extend from the cell
anterior surface (Fig. 17-3). It is frequently bodies to the median eminence, where they
incomplete on the posterior surface of the secrete the releasing and inhibiting hormones
 HYPOPHYSIS • 495

that control the levels of all the hormones of

the adenohypophysis. The other, the magno-
cellular neurosecretory system, consists of neu¬
ronal cell bodies located in the supraoptic
nucleus and paraventricular nucleus, and their
unmyelinated axons form the hypothalamohy-
pophyseal tract, which descends into, and
makes up the bulk of, the substance of the
neural lobe of the hypophysis. This system
secretes the hormones oxytocin and vasopressin.
In addition to the axons of hypothalamic
neurosecretory cells, the neural lobe contains
an intrinsic population of cells called pitui-
cytes, which do not appear to be secretory.
The cells of the hypothalamic nuclei are
large neurons with few processes, an eccen¬
tric nucleus, and abundant cytoplasm. The
endoplasmic reticulum tends to occur in par¬
allel cisternae at the periphery of the peri¬
karyon. There is a well-developed juxtanu-
clear Golgi complex, which forms small (120—
200 nm) neurosecretory granules. Neurotu¬
bules and neurofilaments converge upon the
axon and form an axial bundle in its center
along which the neurosecretory granules are
transported to the neural lobe at a rate of 1
to 4 mm an hour. Labeled precursors of the Figure 17-13. Photomicrograph of rat neurohypophysis
hormones injected intracisternally can be de¬ fixed by perfusion. The large clear areas are capillaries.
tected in the neural lobe in one to two hours. The dark rounded masses indicated by arrows are Herring
bodies. In electron micrographs these are resolved as
In histological preparations stained with
accumulations of neurosecretory material in dilatation of
chrome-alum hematoxylin, deeply stained nerve axons, x 950. (Courtesy of P. Orkand and S. L.
material is seen in aggregations of varying Palay.)
size throughout the infundibular process and
neural lobe. These were traditionally called Pituicytes
Herring bodies (Fig. 17—13). In electron micro¬
graphs they are found to be large aggrega¬ Pituicytes have slender processes that are
tions of the small secretory granules. The joined to processes of neighboring cells of
axons of the neurosecretory neurons are un¬ the same type to form a three-dimensional
usual in that they vary greatly in caliber and network ensheathing the neuronal elements.
have numerous dilatations along their length. In the human they are highly variable in size
It is in these that the bulk of the neurosecre¬ and shape and commonly contain lipochrome
tory material is stored. It is estimated that pigment granules and lipid droplets. Their
each axon has up to 4.4 x 102 of these cytoplasmic processes meander among
expansions along its course holding an aver¬ groups of preterminal secretory axons and
age of 2.2 X 103 neurosecretory granules. often intimately envelop their granule-filled
About 60 per cent of all neurosecretory ma¬ terminal expansions. The structural relation¬
terial resides in these dilatations, and about ship of pituicytes to the nerve fibers is thus
30 per cent in axon endings associated with similar to that of the neuroglial cells of the
the walls of fenestrated capillaries. Endings brain. They occupy 25 to 30 per cent of the
are distinguishable from other axon dilata¬ volume of the neural lobe. Their function is
tions by the presence of numerous small poorly understood. The processes of pitui¬
spherical vesicles in addition to the secretory cytes are connected by gap junctions, which
granules. These are similar in appearance to may provide for their metabolic coupling.
the synaptic vesicles of cholinergic nerve end¬ Cholinesterase has been localized in or on
ings but there is no evidence that they contain them, but no physiological interactions be¬
a neurotransmitter. Instead they are usually tween pituicytes and axons have yet been
interpreted as agents of membrane retrieval demonstrated. They are assigned no role in
after exocytosis of neurosecretory granules. the secretory process, but are generally be-
496 • HYPOPHYSIS

Neurosecretory
Endothelium granules
mMK'
/* *» *mWm
w:©* . ':>¥:V . ^ ■'• :W:X ■
mm
■lit
P 4;."*, • J?ar^« v<
g ■/T&V AWwM'kA ■''- / "•- & jp^V,- w ^
"^SU^VnSIl
A «k... M
m »
■ «—as ^mmm W ipfcCT iMr”^
a ■n i ^ :::' "" • »,■ • .-•• w ' . ' V• _
■:. VY/% fe ./ I . ‘ .« 4- Ip
< &•
P -# >A
■,<~*1 ililii %
j * *■••
.n ' ■<»”.•* . •’McsjPB5ai-19 -
f ‘ wx ■ . ..**2 H
j|V~ ' j';*' _* . ; *"- '-*-$gi 'I
:* viOf^v#V* ■■ dr-tfc*l
WiMWtflP '4 *» ■„•*-*
aeWfl f -llifehl?' > •'•• ■ ..• • ^~>i. * i
■lOff* ^ I B *■ * G a bBf JIa .•’ vNg-

• • w, # a v.a»u?>• _ - • . *• «tT -i >> • J

: >’ '“'” IS#8 ■ & j,,, 0,.1% j*jCBi&*y.' 5
iV-"4^'^4i «:BS
L_ ItfW »f'»^fw
' m- &m™rnm :': ’ Ca-Tu^
W*<A-■x:&2iTWm -w*- -■mw
-..r '■?#»« Wv • f ;-' -' ‘Ilf • • ' £ 2T |2l
wak*;«jE Aft2
m
fil ifflll
*. 'i *.•:- ymgs,*fr A
L x s.
S. A%, ... ■■ .. i .. ,-\ Wm -jS
\^C, gr
I I '%& *%
***?£*.
, - #> w®
- "■ X
fi^.....^llHmi y t* -fSSKs® * :
«• IraMF «' * JhMI^ \ ,.W ,M aflr? SiH| ISP 1m- - '>qm * Ip,. • |V ^
^ ,. m - #wU 2 w;y,
* v f7aTTB»iTI TiT n PStJP ?# \> caBSa^
.^L—— - --
: -:i^^^^Bftj:;;.,. it, ■dpfejt, * 1
Ji iSliliPlKY $-*m^ W
Figure 17-14. Electron micrograph of rat neurohypophysis, showing neurosecretory granules and small vesicles in the
axoplasm of fibers of the hypothalamo-hypophyseal tract ending in close relation to a capillary, x 22,000. (Courtesy of
P. Orkand and S. L. Palay.)

lieved to have a trophic and supportive func¬ and vasopressin are associated with proteins
tion and to maintain the appropriate ionic called neurophysins, which are immunologi-
composition of the extracellular fluid com¬ cally and biochemically distinct. During exo-
partment. cytosis of the neurosecretory granules, hor¬
mone and neurophysins are both released,
but the latter have no known physiological
HiSTOPHYSlOLOGY OF THE NEURAL effect.
LOBE

The two hormones of the neural lobe, Vasopressin

oxytocin and vasopressin (antidiuretic hormone,
ADH), are similar polypeptides both consist¬ Vasopressin acts upon vascular smooth
ing of nine amino acids and differing from muscle, causing constriction of the arterioles,
each other in only two amino acids. They thereby increasing peripheral resistance and
were formerly thought to be produced in raising blood pressure. Loss of blood is a
separate hypothalamic nuclei—oxytocin in potent stimulus for increased secretion of
the paraventricular nuclei and vasopressin in vasopressin. Severe hemorrhage may result
the supraoptic nucleus. Although they are in secretion of the hormone at 50 times the
synthesized in separate neurons, both types normal rate. Diminished blood volume is
are now known to be present in the supraop¬ sensed by pressure receptors in the carotid
tic and paraventricular nuclei. They are first artery and aorta that exert reflex control over
synthesized in the form of a larger precursor vasopressin secretion.
molecule, which is subsequently cleaved en¬ Vasopressin is also called antidiuretic hor¬
zymatically to yield active hormone. Oxytocin mone (ADH) because of its important role in
 HYPOPHYSIS • 497

Loss of water

Rise in serum tonicity

Vasopressin Thirst
release
Figure 17-15. Diagram of the role of vasopressin or
antidiuretic hormone of the neurohypophysis in regulation
of the concentration of body fluids. Interaction of the Increased tubular Increased water
neurohypophysis, the thirst center of the hypothalamus, reabsorption intake
and renal tubules results in maintenance of a constant of water
osmolarity of the body fluids. (After Leaf, A., and C. H.
Coggins. In Williams, R. H. Textbook of Endocrinology.
4th ed. Philadelphia, W. B. Saunders Co., 1968.) Dilution of body
fluid

Inhibition of Inhibition of thirst

vasopressin release

Loss of water

Pars nervosa

< "

Pars intermedia

Pars distal is

B
Figure 17-16. Photomicrograph of cross sections of rat hypophysis stained with paraldehyde-fuchsin. A, Hypophysis of
a normal control rat showing abundant, densely stained neurosecretory material in the neurohypophysis. B, Hypophysis
of a rat of the Brattleboro strain with hypothalamic diabetes insipidus. The neurohypophysis is unusually large but
contains very little neurosecretion (black), x 40. (After Sokol, H. W., and H. Valtin. Endocrinology 77:692, 1965.)
498 • HYPOPHYSIS

water conservation (Fig. 17—15). Its adminis¬ Brain

tration in minute amounts greatly decreases

excretion of water by the kidneys. The hor¬
mone increases the permeability of the col¬
lecting ducts to water and permits reabsorp¬
tion of most of the water in tubule fluid, by
an osmotic gradient existing between the lu¬
men and the interstitial fluid of the renal
papilla. The urine is concentrated and water
conserved. In the absence of ADH, the distal
tubules and collecting ducts are relatively
impermeable to water and concentration of
the urine does not occur. In dehydration or
osmotic stress induced by high intake of salt,
the neurons in the supraoptic and paraven¬
tricular nuclei send impulses along their ax¬
ons to the posterior lobe, releasing large
amounts of stored ADFI. If the osmotic stress
is of longer duration, the hypothalamic neu¬
rons enlarge and develop a more extensive
endoplasmic reticulum for enhanced synthe¬
sis of hormone.
In humans who have a tumor in the hy¬
pothalamus, the supraoptic and paraventric¬
Figure 17-17. Diagram illustrating the role of oxytocin in
ular nuclei may be invaded and destroyed. the suckling reflex. Stimulation of the nipples generates
Such patients are unable to secrete ADH and sensory impulses that pass to the brain via dorsal root
develop diabetes insipidus, a condition charac¬ ganglia. In the brain, these impulses are relayed to the
terized by constant thirst and drinking of hypothalamus, where they activate neurosecretory cells
whose processes extend into the pars nervosa of the
large amounts of water (polydipsia) and by
hypophysis. Stimulation of these cells results in release
excessive urination (polyuria). Much of our of the hormone oxytocin, which is carried in the blood to
understanding of the physiology of the the mammary gland, where it causes contraction of myo¬
hypothalamoneurohypophyseal system has epithelial cells, causing milk to be expressed.
been gained by study of a convenient animal
model of this disease—the Brattleboro rat. Oxytocin
This strain of rats inherit diabetes insipidus A principal target of this hormone is the
as a recessive trait. They excrete every day a myometrium of the pregnant uterus. It stim¬
volume of urine equivalent to 80 per cent of ulates contraction of uterine smooth muscle.
their body weight and drink correspondingly Its concentration in the blood increases dur¬
large amounts of water. Their posterior pi¬ ing the late stages of labor and it is believed
tuitary is unusually large but contains very to have a significant role in parturition.
little stainable neurosecretory material (Fig. Oxytocin is also responsible for milk ejec¬
17—16) and no immunocytochemically de¬ tion in the lactating mammary gland (Fig.
monstrable ADH. The failure of their hypo¬ 17—17). Stimulation of the nipple by the suck¬
thalamic neurons to synthesize the hormone ling infant sends afferent impulses to the
was long attributed to a simple deletion of brain stem and onward to neurons in the
the vasopressin gene. It has been found, supraoptic and paraventricular nuclei, which
however, that Brattleboro rats do synthesize respond by releasing oxytocin into the capil¬
vasopressin in their ovaries and adrenals. laries of the neurohypophysis. Blood-borne
Thus, it appears that their hypothalamic dis¬ oxytocin then stimulates contraction of myo¬
order is an inherited, tissue-specific, post- epithelial cells in the alveoli of the mammary
translational defect instead of a gene dele¬ gland, ejecting milk into the ducts. Milk be¬
tion. The occurrence of vasopressin in the gins to flow from the nipples about one
ovaries and adrenals of normal animals of minute after the onset of suckling.
several species is a relatively recent finding
and its physiological significance is not under¬ REFERENCES
stood. Immunoreactive vasopressin is also PARS DISTALIS
found in occasional tumors of the lungs and Baker, B. L. Functional cytology of the hypophyseal pars
gastrointestinal tract. distalis and pars intermedia. In Creep, R. O., and
 HYPOPHYSIS • 499

E. B. Astwood, eds.: Handbook of Physiology. Sect. Lerner, A. B., and Y. Takahashi: Hormonal control of
7, Vol. 4, part 1. Washington, DC, American Phys¬ melanin pigmentation. Recent Prog. Horm. Res.
iological Society, 1974. 72:203, 1956.
Duello, T. M., and N. S. Halmi: Ultrastructural immu- Stoeckel, M. E., G. Schmitt, and A. Porte: Fine structure
nocytolochemical localization of growth hormone and cytochemistry of the mammalian pars inter¬
and prolactin in human pituitaries. J. Clin. Endo¬ media. In Peptides of the Pars Intermedia. Ciba
crinol. 49:189, 1979. Foundation Symposium 81. London, Pitman Medi¬
Farquhar, M. G.: Processing of secretory products by cal, 1981.
cells of the anterior pituitary gland. Mem. Soc.
NE UROHYPOPHYSIS
Endocrinol. 79:79, 1971.
Farquhar, M. G., E. H. Skutelsky, and C. R. Hopkins: Baker, B. J., W. C. Dermody, and J. R. Reed: Distribu¬
Structure and function of the anterior pituitary and tion of gonadotropin releasing hormone in the rat
dispersed pituitary cells in in vitro studies. In Ante¬ brain as observed by immunocytochemistry. Endo¬
rior Pituitary. San Francisco, Academic Press, 1975. crinology 97:125, 1975.
Guillemin, R.: Control of adenohypophysial functions Barer, R., and K. Lederis: Ultrastructure of the rabbit
by peptides of the central nervous system. Harvey neurohypophysis with special reference to the re¬
Lect. Series 71, 1975-1976. lease of hormones. Zeitschr. Zellforsch. 75:201,
Herlant, M.: The cells of the adenohypophysis and their 1966.
functional signihcance. Int. Rev. Cytol. 17:299, Bargmann, W.: Neurosecretion. Int. Rev. Cytol. 79:183,
1964. 1966.
Kurosumi, K., and T. Kobayaski: Corticotrophs in the Bodian, D.: Cytological aspects of neurosecretion in
anterior pituitary glands of normal and adrenalec- opossum neurohypophysis. Bull. Johns Hopkins
tomized rats as revealed by electron microscopy. Hosp. 773:57, 1963.
Endocrinology 78:745, 1966. Bonner, T. I., and M. J. Brownstein: Vasopressin, tissue-
Moriarty, G. C.: Adenohypophysis: ultrastructural cy¬ specific defects and the Brattleboro rat. Nature
tochemistry. A review. J. Histochem. Cytochem. 310:17, 1984.
27:855, 1973. Dierickx, K., and F. Vandesande: Immunocytochemical
Moriarty, G. C.: Immunocytochemistry of the pituitary localization of the vasopressinergic and oxytocin-
glycoprotein hormones. T. Histochem. Cytochem. ergic neurons in the human hypothalamus. Cell
24:846, 1976. Tissue Res. 764:15, 1977.
Nakane, P.: Classification of anterior pituitary cell types Dierickx, K., and F. Vandesande: Immunocytochemical
with immunoenzyme histochemistry. J. Histochem. demonstration of separate vasopressin-neurophysin
Cytochem. 18:9, 1970. and oxytocin-neurophysin neurones in the human
Pelletier, G., F. Robert, and J. Hardy: Identification of hypothalamus. Cell Tissue Res. 796:203, 1979.
human pituitary cell types by immunoelectron mi¬ du Vigneaud, V.: Trail of sulfur research from insulin
croscopy. J. Clin. Endocrinol. Metab. 46:534, 1978. to oxytocin. Science 723:967, 1956.
Phifer, R. F., A. R. Midgley, and S. S. Spicer. Immuno- Gainer, H., Y. Same, and M. J. Brownstein: Biosynthesis
histologic and histologic evidence that follicle-stim¬ and axon transport of rat neurohypophyseal protein
ulating and luteinizing hormones are present in the and peptides. J. Cell Biol. 73:366, 1977.
same cell types in the human pars distalis. J. Clin. Morris, J- F.,J. J. Nordmann, and R. E. Dyball: Struc¬
Endocrinol. Metab. 36:125, 1973. ture-function correlation in mammalian neurosecre¬
Phifer, R. F., and S. S. Spicer. Immunohistochemical tion. Int. Rev. Exp. Pathol. 76:1, 1973.
and histologic demonstration of thyrotropic cells of Sawyer, W. H.: Vertebrate neurohypophyseal principles.
the human adenohypophysis. J. Clin. Endocrinol. Endocrinology 75:981, 1964.
Metab. 36:1210, 1973. Sokol, H. W., and H. Valtin: Morphology of the neu¬
Shiino, M., A. Arimura, A. V. Schally, and E. G. Rennels: rosecretory system in rats homozygous and hetero¬
Ultrastructural observations of granule extrusion zygous for hypothalamic diabetes insipidus (Brattle¬
from rat anterior pituitary cells after injection of boro strain). Endocrinology 77:692, 1965.
LH-releasing hormone. Z. Zellforsch. Mikrosk.
Anat. 128:152, 1972. BLOOD VESSELS
Shiino, M., M. G. Williams, and E. G. Rennels: Ultra-
structural observation of pituitary release of prolac¬ Bergland, R. M., and R. B. Page: Pituitary-brain vascular
tin in the rat by suckling stimulus. Endocrinology relations: a new paradigm. Science 264:18, 1979.
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laries of the anterior pituitary gland. Angiology
PARS INTERMEDIA
72:270, 1961.
Hadley, M. E., C. B. Heward, V. J. Hruby, T. K. Sawyer, Green, J. D.: The comparative anatomy of the portal
and Y. C. S. Yong: Biological actions of melanocyte- vascular system and of the innervation of the hy¬
stimulating hormone. In Peptides of the Pars Inter¬ pophysis. In Harris, G. W., and B. T. Donovan,
media. Ciba Foundation Symposium 81. London, eds.: The Pituitary Gland. Berkeley, CA, University
Pitman Medical, 1981. of California Press, 1966.
Jackson, S., J. Hope, F. Estivarex, and P. J. Lowry: Green, J. D., and G. W. Harris: The neurovascular link
Nature and control of peptide release from the pars between the neurohypophysis and adenohypo¬
intermedia. In Peptides and the Pars Intermedia. physis. J. Endocrinol. 5:136, 1949.
Ciba Foundation Symposium 81. London, Pitman Wislocki, G. B.: The vascular supply of the hypophysis
Medical, 1981. cerebri of the rhesus monkey and man. Proc. Assoc.
Lerner, A. B., and J. S. McGuire: Effects of alpha- and Res. Nerv. Ment. Dis. 7 7:48, 1938.
beta-melanocyte stimulating hormones on skin col¬
our in man. Nature 769:176, 1961.
 1ft
THE THYROID GLAND

The thyroid gland situated in the anterior The follicles are surrounded by an ex¬
part of the neck weighs 25 to 40 g. It consists tremely thin basal lamina, which usually is
of two lateral lobes connected by a narrow not resolved with the light microscope. With
isthmus, which crosses the trachea just below silver stains the follicles are seen to be en¬
the cricoid cartilage. In about one third of closed by a delicate network of reticular fi¬
the persons examined, a pyramidal lobe ex¬ bers. A close-meshed plexus of capillaries
tends upward from the isthmus near the left surrounds each follicle (Fig. 18—3). Between
lobe. the capillary nets of adjacent follicles are the
The gland is enclosed in a connective tissue blind terminations of lymphatic vessels, and
capsule that is continuous with the surround¬ in some rodent species lymphatics form ex¬
ing cervical fascia. This outer capsule is tensive perifollicular sinusoids. Numerous
loosely connected on its deep surface to an¬ nerve fibers accompany the blood vessels as
other layer of moderately dense connective they ramify among the follicles. These seem
tissue that is intimately adherent to the gland. to terminate mainly along the vessels, but in
This separation of the capsule into two layers some instances they appear to end in direct
creates a plane of cleavage between the two, contact with the base of the thyroid epithelial
which facilitates surgical access in subtotal cells. The nerves entering the thyroid are
thyroidectomy. postganglionic sympathetic fibers originating
The function of the thyroid is to elaborate, in the middle and superior cervical ganglia.
store, and release into the bloodstream thyroid There are also preganglionic parasympa¬
hormones, which are concerned with the reg¬ thetic fibers, and ganglion cells may occasion¬
ulation of metabolic rate. The thyroid differs ally be encountered within the thyroid. The
from other endocrine glands in that a mech¬ nerves to the thyroid are presumed to be
anism for extracellular storage of its hor¬ mainly vasomotor, inasmuch as transplanted
mones is highly developed, whereas in other thyroid tissue functions adequately, suggest¬
endocrine glands there are only rather lim¬ ing that an intact nerve supply is not neces¬
ited provisions for intracellular storage. sary for secretion.
The epithelial cells vary in height but are
commonly low cuboidal to squamous. In gen¬
eral, the epithelium tends to be squamous
HISTOLOGICAL ORGANIZATION when the gland is underactive and columnar
when it is overactive, but there are many
exceptions, and an accurate assessment of
The gland is composed of spherical, cyst¬ the functional activity of the gland cannot be
like follicles 0.02 to 0.9 mm in diameter, based on histological examination alone.
lined with a simple epithelium and containing The nucleus of the cells is spheroidal, cen¬
a gelatinous colloid (Figs. 18—1, 18-2). This trally situated, and poor in chromatin and
represents the stored product of secretory contains one or more nucleoli. The cytoplasm
activity by the epithelium lining the follicle. is basophilic; the mitochondria are thin rods
In the human there is great variability in the and the Golgi apparatus is usually supranu¬
size of the follicles, but the small predominate clear. Lipid droplets are common, and “clear
over the large. In other species the follicles droplets” have been described by various
are of more uniform size. In the rat and workers using the light microscope, and in¬
guinea pig, those at the periphery of the terpreted as intracellular globules of colloid.
gland are larger than those more centrally They stain with aniline blue and with the
situated. periodic acid—Schiff (PAS) reaction in much

500
 Figure 18-1. Photomicrograph of monkey thyroid showing variations in size of the follicles.

Figure 18-2. Photomicrograph of monkey thyroid at higher magnification to illustrate the homogeneity of the colloid and
the character of the epithelium.

501
502 • THE THYROID GLAND

Figure 18-3. Scanning electron micrograph of a monkey thyroid in which the blood vessels have been injected with
plastic and the tissue digested away. Each spherical follicle is surrounded by a dense network of capillaries. (Micrograph
from Fujita, H., and T. Murakami. Arch. Histol. Jpn. 36:181, 1974.)
 THE THYROID GLAND • 503

Mitochond ria violet absorption spectrophotometry, and by

the use of radioactive 1311 (Fig. 18-5). The
follicle cells were formerly believed to secrete
into the colloid a protease that split thyro¬
globulin into smaller molecules and liberated
the biologically active thyroxin and triiodo¬
thyronine. It is now widely accepted that the
proteases act within the thyroid cells upon
stored thyroglobulin taken up from the lu¬
men of the follicle.
In electron micrographs the follicles of the
human thyroid are found to be composed of
a single layer of low cuboidal cells surround¬
ing a homogeneous, moderately dense col¬
loid. A thin, continuous basal lamina about
50 nm thick surrounds the entire follicle. In
the interfollicular spaces are numerous cap¬
illaries of the fenestrated type, occasional
fibroblasts, and small bundles of collagen
fibrils. The follicle cells are joined laterally
by typical junctional complexes, and the free
surfaces bear a small number of short, irreg¬
ularly oriented microvilli. In follicles with
cuboidal epithelium, the microvilli are some¬
what more numerous. The basal plasma
Figure 18-4. Section through several follicles of human
thyroid. Aniline-acid fuchsin. (Courtesy of R. R. Bensley.) membrane is smoothly contoured and not
infolded. The relatively large nucleus is cen¬
trally placed and has an eccentric nucleolus.
the same way as the colloid in the lumen of The mitochondria are relatively few and uni¬
the follicle. Granules of varying size, located formly distributed. Their cristae are not es¬
mainly in the apical cytoplasm, give positive pecially numerous. The endoplasmic reticu-
staining reactions for acid phosphatase and
esterase and are therefore considered to be
lysosomes.
The unfixed colloid is optically homoge¬
neous, except for occasional desquamated
cells and rare macrophages. After fixation,
the colloid stains with either acid or basic
dyes, and with the trichrome stains it is not
uncommon for different follicles or even
different areas of the colloid in the same
follicle to be colored differently. Although
physiological significance has been erro¬
neously ascribed to this multiple staining, the
varied patterns observed appear to be due to
local differences in concentration of protein
that depend on the direction and rate of
penetration of fixative into the tissue block.
The colloid stains intensely with the PAS
reaction, because the thyroglobulin secreted by
the thyroid is a glycoprotein containing about
3 per cent carbohydrate. The thyroglobulin
of the colloid also contains various iodinated Figure 18-5. Low-power photomicrograph of autoradio¬
amino acids. Among these are thyroxin (tetra- graph of thyroid gland of rat previously injected with 1311.
Blackened areas represent sites of deposition of the
iodothyronine) and triiodothyronine, which are
radioactive material. There is great variability in the con¬
the thyroid hormones. The presence of these tent of the isotope in the several follicles. In a few places
compounds in the colloid has been demon¬ the epithelium is blackened. (Courtesy of C. P. Leblond,
strated by microchemical analysis, by ultra¬ D. Findlay, and S. Gross.)
504 • THE THYROID GLAND
\

lum varies in its degree of development. In chondria-rich cells, C cells, and ultimo branchial
the squamous cells there are only a few elon¬ cells. The term “parafollicular cell” was intro¬
gated cisternal profiles, but in the cuboidal duced to distinguish these cells from other
cells the reticulum is well developed (Fig. 18— interfollicular cells, some of which may be
6). The Golgi apparatus is in a supranuclear undifferentiated embryonal elements or con¬
or paranuclear position and is composed of nective tissue cells. They arise during embry¬
flattened or dilated saccules, vacuoles, and onic life from the last pair of pharyngeal
small vesicles. Small vesicles similar to those pouches. In fishes, amphibians, reptiles, and
of the Golgi apparatus are present in abun¬ birds, they form discrete epithelial cell masses
dance throughout the cytoplasm. Multivesi- called ultimo branchial bodies, located in the
cular bodies are also common. Membrane- neck or mediastinum. In mammals, they are
limited dense bodies 0.5 to 0.7 pm in diam¬ incorporated into tfie thyroid. They are often
eter are plentiful in the apical cytoplasm. larger than the principal cells, and in routine
These are lysosomes. histological preparations they stain less
In addition to the principal cells of the deeply. They can be selectively stained by the
thyroid follicles, there is another, smaller silver nitrate method of Cajal, which reveals
population of cells, present both in the follic¬ the presence of brown or black cytoplasmic
ular epithelium and in the interfollicular granules (Figs. 18—7, 18-8). The granules
spaces. These cells were hrst described by exhibit an affinity for aniline blue in tri¬
Baker (1877) and Hurthle (1894) and studied chrome stains. Cytochemically they are distin¬
in greater detail by Nonidez (1931) (Figs. 18— guished from follicular cells by their high
7, 18-8), but evidence permitting assignment level of activity of the mitochondrial enzyme
of a function to them was not forthcoming a-glycerophosphate dehydrogenase.
until 30 years later. They are variously des¬ Where the parafollicular cells are interca¬
ignated as parafollicular cells, light cells, mito¬ lated among the principal cells of the follicles,

Figure 18-6. Electron micrograph of the apical half of an epithelial cell from rat thyroid gland. The free surface of the
cell is provided with numerous short microvilli that project into the colloid of the follicle. The endoplasmic reticulum is
well developed and its cisternae are distended with an amorphous content of low density. The small dense granules
are lysosomes. (Micrograph courtesy of S. Wissig.)
 THE THYROID GLAND • 505

Figure 18-8. Drawing of an area occupied by small and

medium-sized follicles in the thyroid of an adult dog.
Numerous parafollicular cells are seen in clusters in the
interfollicular spaces. Cajal stain. (After Nonidez, J. F.
Anat. Rec. 53:339, 1932.)

ture. They are now adequately preserved by

aldehyde fixatives and appear as membrane-
limited, dense, spherical granules 0.1 to 0.4
pm in diameter (Fig. 18-9). The parafollic¬
ular cells elaborate and secrete calcitonin, a
hormone that lowers blood calcium concen¬
tration.

HISTOPHYSIOLOGY

The function of the thyroid gland is to

synthesize, store, and release hormones con¬
cerned with the regulation of metabolic rate
(thyroxin and triiodothyronine) and with
maintenance of blood calcium levels within
tolerable limits (calcitonin). The function re¬
lated to metabolic rate resides in the follicular
epithelial cells, whereas the calcium-regulat¬
ing action resides in the parafollicular cells.
D

Figure 18-7. Parafollicular cells in thyroid follicles. A, B,

Cat thyroid, Ehrlich’s hematoxylin; C, D, dog thyroid (35- THE PRINCIPAL CELLS
day-old puppy), Cajal silver nitrate method. (After Noni-
dez, J. F. Am. J. Anat. 49:479, 1932.)
The thyroid is the only endocrine gland
that stores its product extracellularly. The
they are close to the base of the epithelium. secretory process is therefore somewhat more
They never border directly on the lumen but complex than that in other glands. It involves
are separated from it by overarching proc¬ (1) synthesis of the large glycoprotein thyro-
esses of neighboring principal cells (Fig. 18— globulin (2) iodination of tyrosine molecules
9). The secretory granules are not easily that are important constituents of thyroglob-
preserved and were overlooked in the early ulin (3) release of thyroglobulin into the
descriptions of parafollicular cell ultrastruc- lumen of the follicle for storage (4) reabsorp-
506 • THE THYROID GLAND

Colloid

Follicular cell
... . 1 :: 'J

Parafollicular
cell

A
■* **

•% « ^

Figure 18-9. Electron micrograph of follicular and parafollicular cells of normal cat thyroid. The granular parafollicular
cells do not border on the lumen but are usually separated from the colloid by follicular ceils. When adequately fixed,
the parafollicular cells contain many dense spherical secretory granules. (Micrograph courtesy of S. Wissig.)

tion of thyroglobulin into the follicular epi¬ porated into the thyroglobulin. The synthesis
thelial cells (5) hydrolysis of thyroglobulin to of thyroglobulin and its iodination are ap¬
liberate thyroxin and triiodothyronine and parently independent, and while the sites of
(6) release of these hormones into the peri¬ synthesis of the glycoprotein are well known,
follicular capillaries and lymphatics. the site of its iodination has been a subject of
Synthesis of thyroglobulin involves the controversy—some believe that it takes place
same intracellular pathway described for in the cells, whereas others contend that it
other protein-secreting cells. Amino acids are occurs in the lumen of the follicle. After
assembled into polypeptides on ribosomes of iodine is actively transported from the blood
the endoplasmic reticulum, then transported into the cells, it is oxidized in the presence
to the Golgi complex. Since thyroglobulin is of hydrogen peroxide to a different ionic
a glycoprotein, the glycosyl transferases of the species. The oxidized iodide ion subsequently
Golgi apparatus no doubt play a role in iodinates the tyrosine residues of thyroglob¬
synthesis and conjugation of its carbohydrate ulin to form mono- and diiodotyrosine. Triio¬
components. From the Golgi complex, the dothyronine is formed when one molecule
product is transported in small vesicles to the each of monoiodotyrosine and diiodotyrosine
apical surface of the cell, where it is dis¬ are coupled. Thyroxin is formed when two
charged by exocytosis. molecules of diiodotyrosine are joined. Since
The thyroid has the capacity to concentrate both the initial oxidation of iodide and the
iodine to several hundred thousand times the subsequent iodination of tyrosine may be
concentration of this element in the blood catalyzed by the enzyme peroxidase, histo-
plasma. After an injection of inorganic io¬ chemical localization of this enzyme seemed
dide, 40 per cent of the circulating ion is a reasonable approach for determining the
concentrated in the thyroid gland within ten site of iodination. Peroxidase is found in the
minutes. The iodine is used to iodinate ty¬ perinuclear cisternae, endoplasmic reticu¬
rosine molecules that are ultimately incor¬ lum, inner lamellae of the Golgi, apical vesi-
 THE THYROID GLAND • 507

cles, and external surfaces of the microvilli and hence represent colloid taken up from
projecting into the colloid. The reaction the lumen, not newly synthesized thyroglob¬
product observed in the membranous cell ulin.
organelles is believed to represent continuous The hydrolysis of thyroglobulin, formerly
synthesis of peroxidase by the thyroid cells. attributed to proteases in the lumen of the
Peroxidase transported along this pathway is follicles, is now known to be a function of
probably incorporated into the cell surface lysosomes in the epithelial cells. Lysosomes
membrane. It is now believed that the prin¬ coalesce with the endocytosis vacuoles con¬
cipal site of iodination of thyroglobulin is taining reabsorbed colloid, and add to their
probably the microvillous border of the thy¬ contents cathepsins that hydrolyze thyroglob¬
roid cells, and possibly the apical vesicles. ulin. The tetraiodothyronine and triiodothy¬
This interpretation is in agreement with au¬ ronine liberated apparently diffuse into the
toradiographic studies localizing radioactive cytoplasmic matrix and through the cell base
iodine at early time intervals over the inter¬ to enter the blood. This phase of the process
face between the epithelium and the colloid. cannot be visualized with the electron micro¬
The uptake of colloid from the lumen of scope. As in other endocrine glands, the
the follicle has been studied mainly in animals capillaries of the thyroid are of the fenes¬
strongly stimulated to release hormone by trated type and offer little or no barrier to
administration of thyroid stimulating hor¬ passage of hormones to the blood. Whereas
mone (TSH) of the hypophysis (Fig. 18-10). the blood vascular system is undoubtedly the
Under these conditions, large droplets con¬ most important avenue of egress of thyroid
taining colloid appear in the apical cytoplasm. hormones because of the high rate of blood
By microinjection of ferritin into follicles flow, it has been shown that concentration of
before administration of TSH, it was shown hormone in the lymph draining the gland is
that the large colloid droplets which appear as much as 100 times greater than the con¬
in the cells after stimulation contain ferritin centration in venous blood. Thus, the lym-

Figure 18-10. Left side, Diagram depicting the normal ultrastructure of the thyroid cell secreting thyroglobulin into the
lumen of the follicle. Right side, The uptake of colloid by pinocytosis after stimulation with thyroid-stimulating hormone,
and the lysosomal degradation of thyroglobulin to release thyroxin. (From Fawcett, D. W., J. A. Long, and A. L. Jones.
Recent Prog. Horm. Res. 25:315, 1969.)
508 • THE THYROID GLAND

phatics must be considered a significant path¬ roids may secrete thyroxin at five to ten times
way for transport of hormone to the the normal rate. The patient suffers weight
circulating blood. loss, nervousness, weakness, rapid heart rate,
The activity of the thyroid is regulated by intolerance of heat, and tremor. The in¬
the thyrotropic hormone (TSH) of the ante¬ creased metabolic rate and associated symp¬
rior lobe of the hypophysis, and this in turn toms return temporarily to normal after
is controlled by thyrotropin releasing factor administration of iodine and of antithyroid
(TRF) of the hypothalamus. Lowered levels agents, such as thiouracil. The exact cause of
of thyroid hormone in the blood stimulates Graves’ disease is poorly understood, but the
the hypothalamus to secrete TRF, which re¬ blood of patients suffering from it often
sults in increased thyrotropin secretion. Ex¬ contains an immunoglobulin of the IgG type,
cess circulating thyroid hormone depresses which is a long-acting thyroid stimulator
thyrotropin secretion. Chronic hypersecre¬ (LATS). This substance may play a role in
tion of thyrotropin results in a highly vascular the pathogenesis of the disease.
gland with columnar follicular epithelial cells
and relatively little colloid. After hypophy-
sectomy, the thyroid is no longer capable of THE PARAFOLLICULAR CELLS
accumulating significant amounts of 131I, and •

only traces of thyroid hormone appear in the

blood. When it was first detected in the outflow
The most striking effect of thyroid secre¬ from a preparation of dog thyroid and para¬
tion is its control of the metabolic rate of the thyroid glands perfused with hypercalcemic
body. When a deficiency of thyroid hormone blood, calcitonin was believed to come from
occurs, the metabolic rate falls below normal; the parathyroids. It was later shown to be
when there is an excess, the metabolic rate present only in the thyroid, and several lines
rises above normal. When hypothyroidism be¬ of evidence associated it with the parafollic¬
gins in infancy and persists, it leads to cretin¬ ular cells: (1) sustained elevation of calcium
ism, a condition attended by stunting of phys¬ results in degranulation of these cells; (2)
ical and mental development. When extractable calcitonin correlates well with the
hypofunction begins in adulthood and per¬ presence and number of such cells; (3) hu¬
sists, it leads to myxedema, a disorder charac¬ man medullary carcinomas of the thyroid,
terized by a sallow, puffy appearance; dry, which are thought to arise from the parafol¬
sparse hair; lethargy; and slow cerebration. licular cells, contain 100 to 10,000 times as
In both conditions the basal metabolic rate is much calcitonin as normal thyroid tissue; (4)
reduced, and in both the symptoms may be fluorescent antibodies to calcitonin bind se¬
relieved through timely oral administration lectively to the parafollicular cells.
of dried thyroid gland. The hormone was extracted in pure form
Enlargement of the thyroid gland is called from pig thyroid glands by 1968, and it was
goiter, and any substance that causes enlarge¬ found to be a polypeptide consisting of a
ment of the thyroid is called a goitrogen. When single chain of 32 amino acids. The sequence
iodine is deficient in the diet, as it tends to of the amino acids has been worked out, and
be in a number of geographical regions, there the hormone has now been synthesized. Cal¬
is an excess production and accumulation of citonin of human thyroid origin differs
colloid, but in the absence of sufficient iodine, slightly from the porcine hormone.
relatively little active hormone is produced. Calcitonin appears to exert its effect by
This compensatory enlargement of the gland suppressing release of calcium into the blood
is called colloid goiter. Antithyroid substances from resorption of bone. Bone is constantly
occur naturally in some plants used for food. undergoing internal remodeling (see Chap¬
Excess consumption of these goitrogens may ter 9). Parathyroid hormone tends to alter
interfere with iodination in the thyroid and the balance between bone deposition and
result in enlargement. bone absorption so as to cause increased
A common form of hyperthyroidism in the resorption. The calcium released from bone
human is Graves’ disease or exophthalmic goiter. enters the blood, raising the blood calcium
The follicles become enlarged, with tall cells level. Parathyroid hormone acts upon osteo-
and papillary projections into the lumen. The cytes and osteoclasts, accelerating osteolysis.
mitochondria increase in number, the Golgi Calcitonin appears to have an opposite effect
apparatus hypertrophies, and colloid is di¬ upon these cells, decreasing bone resorption
minished or absent. Such hyperplastic thy¬ and thus lowering the blood calcium level.
 THE THYROID GLAND • 509

The role of calcitonin in normal human Isolation from thyroid gland and synthesis. 2. Phys¬
blood calcium regulation is less well estab¬ iological activity. Biochem. J. 53:645, 652, 1953.
lished than in experimental animals. How¬ Heimann, P.: Ultrastructure of the human thyroid. A
study of normal thyroid, untreated and treated toxic
ever, hypercalcitonism is thought to occur in
goiter. Acta Endocrinol. 53(Suppl. 110):5, 1966.
medullary carcinoma and in certain adeno¬ Hilfer, R. S.: Follicle formation in embryonic chick
mas of the human thyroid. thyroid. I. Early morphogenesis. J. Morphol.
775:135, 1964.
Hirsch, P. F., F. F. Gauthier, and P. L. Munson: Thyroid
hypocalcemic principle and recurrent laryngeal
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tion and purification of calcitonin. Nature 203:1927, in the thyroid follicle visualized by the “coated
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Copp, D. EL, E. C. Cameron, B. A. Cheney, A. G. F. Mitmaker: Elaboration of thyroglobulin in the thy¬
Davidson, and K. G. Henze: Evidence for calci¬ roid follicle. Endocrinology 74:333, 1964.
tonin—a new hormone from the parathyroid that Nonidez, J. F.: The origin of the parafollicular cell, a
lowers blood calcium. Endocrinology 70:637, 1962. second epithelial component of the thyroid gland
Daniel, P. M., Plaskett, L. G., and Pratt, O. E.: The of the dog. Am. J. Anat. 49:479, 1932.
lymphatic and venous pathways for the outflow of Nonidez, J. F.: Further observations on the parafollicular
thyroxine, iodoprotein and inorganic iodide from cells of the mammalian thyroid. Anat. Rec. 53:339,
the thyroid gland. J. Physiol. (Lond.) 188:25, 1967. 1932.
Degroot, L. J., and H. Niepomniszeze: Biosynthesis of Pearse, A. G. E.: The cytochemistry of the thyroid C
thyroid hormone: basic and clinical aspects. Metab¬ cells and their relationship to calcitonin. Proc. R.
olism 26:665, 1977. Soc. B 764:478, 1966.
Dempsey, E. W., and R. R. Peterson: Electron micro¬ Pitt-Rivers, R.: Mode of action of antithyroid com¬
scopic observations on the thyroid glands of normal, pounds. Physiol. Rev. 36:194, 1950.
hypophysectomized, cold-exposed and thiouracil- Pitt-Rivers, R., and W. R. Trotter, eds.: The Thyroid.
treated rats. Endocrinology 56:46, 1955. London, Butterworths, 1964.
Ekholm, R., and L. E. Ericson: Ultrastructure of the Seljelid, R.: Electron microscopic localization of acid
parafollicular cells of the rat J. Ultrastruct. Res. phosphatase in rat thyroid follicle cells after stimu¬
23:378, 1968. lation with thyrotropic hormone. J. Histochem. Cy-
Falck, B., B. Larsen, C. v. Mecklenburg, C. Rosengren, tochem. 73:687, 1965.
and K. Svenaeus: On the presence of a second Seljelid, R.: Endocytosis in thyroid follicle cells. II. A
specific cell system in mammalian thyroid gland. microinjection study of the origin of colloid drop¬
Acta Physiol. Scand. 62:491, 1964. lets. J. Ultrastruct. Res. 77:401, 1967.
Foster, G. V., A. Baghdiantz, M. A. Kumar, E. Slack, H. Seljelid, R., A. Reith, and N. F. Nakken: The early phase
A. Soliman, and I. MacIntyre: Thyroid origin of of endocytosis in the rat thyroid follicle cell. Lab.
calcitonin. Nature 202:1303, 1964. Invest. 23:595, 1970.
Foster, G. V., I. MacIntyre, and A. G. E. Pearse: Calci¬ Strum, J. M., and M. J. Karnovsky: Cytochemical local¬
tonin production and the mitochondrion-rich cells ization of endogenous peroxidase in thyroid follic¬
of the dog thyroid. Nature 203:1029, 1964. ular cells. J. Cell Biol. 44:655, 1970.
Fujita, H.: Outline of the fine structural aspects of Taurog, A.: Thyroid hormone synthesis and release. In
synthesis and release of thyroid hormone. Gunma Werner, S. C., and Ingbar, S. H. eds.: The Thyroid.
Symposium on Endocrinology 7:49, 1970. A Fundamental and Clinical Test. New York, Har¬
Fujita, H., and T. Murakami: Scanning electron micros¬ per 8c Row, 1978.
copy on the distribution of the minute blood vessels Turner, C. D., and J. T. Bagnara: General Endocrinol¬
of the dog, rat and Rhesus monkey. Arch. Histol. ogy. 5th ed. Philadelphia, W. B. Saunders Co., 1971.
Jpn. 36:181, 1974. Welzel, B. K., S. S. Spicer, and S. H. Wollman: Changes
Gittes, R. F., and G. L. Irvin: Thyroid and parathyroid in fine structure and acid phosphatase localization
roles in hypercalcemia: evidence for a thyrocalci¬ in rat thyroid cells following thyrotrophin adminis¬
tonin releasing factor. Science 148:1737, 1965. tration. J. Cell Biol. 25:593, 1965.
Gross, J., and R. Pitt-Rivers: 3:5:3'-Triiodothyronine. 1. Whur, P., A. Herscovics, and C. P. Leblond: Radioau-
510 • THE THYROID GLAND

tographic visualization of the incorporation of ga- Wissig, S. L.: The anatomy of secretion in the follicular
lactose-3H and mannose-3H by rat thyroids in vitro cells of the thyroid gland. II. The effect of acute
in relation to the stages of thyroglobulin synthesis. thyrotrophic hormone stimulation on the secretory
J. Cell Biol. 43:289, 1969. apparatus. J. Cell Biol. 16:93, 1963.
Wissig, S. L.: The anatomy of secretion in the follicular Wolman, S. H., S. S. Spicer, and M. S. Burstone: Local¬
cells of the thyroid gland; the fine structure of the ization of esterase and acid phosphatase in granules
gland in the normal rat. I. Biophys. Biochem. Cytol. and colloid droplets in rat thyroid epithelium. J.
7:419, 1960. Cell Biol. 21:191, 1964.
 PARA THYROID GLANDS

The parathyroid, glands are endocrine glands in man: principal cells and oxyphilic cells (Fig.
producing a hormone that is essential for 19-2).
maintaining the normal concentration of cal¬
cium in the blood and extracellular fluid of
The Principal Cells (Chief Cells)
the body. In most mammalian species their
removal results in a precipitous fall in blood The principal cells are polygonal and 7 to
calcium, leading to violent spasm of skeletal 10 |xm in diameter, with a centrally placed
muscle (tetany) and ultimately to death. In vesicular nucleus and a pale, slightly acido¬
the human there are usually four parathyroid philic cytoplasm that tends to shrink during
glands but accessory glands are common. fixation. There is a small juxtanuclear Golgi
They are small, yellow-brown, oval bodies apparatus and a number of elongated mito¬
adhering to the posterior surface of the thy¬ chondria. Coarse granular deposits of flu¬
roid gland. Their total weight varies from orescent lipofuscin pigment are often pres¬
0.05 to 0.3 g. They may range from 3 to 8 ent, and when the cells are appropriately
mm in length, 2 to 5 mm in width, and 0.5 stained, a considerable amount of glycogen
to 2 mm in thickness. Most of the glands are is found. In addition to these components,
associated with the middle third of the thy¬ small granules have been described that stain
roid, a smaller number with the inferior with iron-hematoxylin and exhibit argyro-
third. In 5 to 10 per cent of cases one or philia with the Bodian stain. These have been
more parathyroids are associated with the interpreted by some investigators as secretory
thymus and may therefore be deep in the granules.
anterior mediastinum. This association of the Electron microscopic studies reaffirm the
parathyroid glands with the thymus stems presence of all the components enumerated
from their common origin from the same above and reveal, in addition,, cisternal pro¬
pharyngeal pouch in the embryo. files of the granular endoplasmic reticulum,
Most parathyroid glands lie in the capsule sometimes aggregated in conspicuous paral¬
of the thyroid, but they may be embedded lel arrays. The argyrophilic granules at this
within the gland (Fig. 19-1). In either case, level of resolution have irregular outlines,
they are separated from the substance of are limited by a membrane, and have a dense
thyroid by a thin connective tissue capsule. granular content. They appear to arise in the
The capsular connective tissue extends into Golgi apparatus and tend to accumulate at
the parathyroid gland, and it is via these the periphery of the cell. A single abortive
trabeculae that the larger branches of blood cilium is often found projecting from the
vessels, nerves, and lymphatics enter. Be¬ principal cell into the narrow intercellular
tween the gland cells is a framework of retic¬ space.
ular fibers. These support the rich capillary A second category of principal cells is dis¬
network and the nerve fibers. The connective tinguished at the electron microscope level.
tissue stroma may contain numerous fat cells These have a smaller Golgi apparatus, few
and these may occupy half the volume of the dense “secretory” granules, and large lakes
gland in elderly individuals. of glycogen. These cells appear to be phys¬
The parenchyma of the parathyroid glands iologically less active than those described
consists of densely packed groups of cells, above and may outnumber them.
which may form a compact mass or may be
arranged as anastomosing cords, or less com¬ The Oxyphilic Cells
monly as follicles with a small amount of
colloidal material in the lumen. Two main The oxyphilic cells are greatly in the mi¬
types of epithelial cells have been described nority and occur singly or in small groups.
511
512 • PARATHYROID GLANDS

Artery

Parathyroid
gland

Connective
tissue

Thyroid gland

Figure 19-1. Photomicrograph of section of thyroid and parathyroid glands of Macacus rhesus, x 80.

They are distinctly larger than the principal gated mitochondria with numerous closely
cells. They have a small, darkly staining nu¬ spaced cristae. In the interstices among the
cleus and a strongly acidophilic cytoplasm. mitochondria are numerous glycogen parti¬
When stained by the aniline-acid fuchsin cles, but these do not form large masses as
method, they are found to have many more they do in the less active principal cells. The
mitochondria than the principal cells. This is Golgi apparatus is inconspicuous and the
borne out in electron micrographs, which endoplasmic reticulum sparse. The eosino¬
show a remarkable concentration of elon¬ philic granulation formerly described by light

Figure 19-2. Section through human parathyroid gland showing the small principal cells, often vacuolated, and the
large oxyphilic cells with fine purplish granules. Zenker-formol fixation. Mallory-azan stain, x 960.
 PARATHYROID GLANDS • 513

Figure 19-3. Photomicrograph of human parathyroid. Only one adipose cell is shown in this field, but such cells may
be very numerous. (Courtesy of S. I. Roth.)

microscopists must be attributed to the abun¬ apparatus also becomes smaller and more
dant mitochondria, for granules resembling compact. Both changes are suggestive of de¬
secretory granules are rarely encountered in creased functional activity. After two weeks
electron micrographs. the cells return to normal, both in size and
Another type of cell, intermediate between in morphology of the Golgi apparatus, sug¬
the oxyphilic and principal cells, has been gesting a resumption of normal secretory
described. It has a fine granular cytoplasm, activity. During the hypertrophy of the para¬
which stains faintly with acid dyes, and a thyroid glands in rickets, the Golgi apparatus
nucleus that is smaller and stains darker than is described as undergoing changes that in¬
that of the principal cells. Also, “water-clear” dicate great secretory activity in comparison
cells and “dark oxyphilic” cells have been with normal cells. It is not possible to extend
described, but it is not clear to what extent these conclusions to the human gland, for it
these latter types are to be attributed to has yet to be clearly shown which cells in man
vagaries of fixation. are equivalent to those in laboratory animals.
The parathyroid glands show certain
changes with increasing age: (1) an increase
in the amount of connective tissue, including
increased numbers of fat cells; (2) the oxy¬ HiSTOPHYSIOLOGY
philic cells are said to appear at the age of
4V2 to 7 years and to increase in number, 4 he function of parathyroid hormone is to
especially after puberty; (3) in the closely maintain the normal concentration of cal¬
packed masses of gland cells, some cords and cium in the extracellular fluid. It does this by
follicles appear in the 1-year-old infant, and acting directly upon the osteocytes and osteo¬
they increase thereafter; colloid accumula¬ clasts in bone, which is the principal store of
tion in the lumen of the follicles shows the calcium in the body. Its initial rapid effect is
same tendency. to increase the rate of release of calcium
When rats are given injections of a large from bone mineral. This appears to be a
dose of parathyroid extract, the cells of the result of stimulation of osteocytic osteolysis
parathyroid glands become smaller; the Golgi (Fig. 19—4). If the increase in circulating
514 • PARATHYROID GLANDS

Parathyroid graft Bone marrow

Figure 19-4. Section of parietal bone of rat 14 days after autogenous transplantation of parathyroid gland. The bone
beneath the graft has been nearly completely resorbed, x 50. (From a preparation by H. Chang.)

parathyroid hormone continues for some tional hexapeptide at one end. This proved
time, there is an acceleration of the rate of to be only an intermediate in the biosynthetic
internal remodeling of bone, which is a func¬ process, and a still larger precursor of 115
tion of the osteoclasts. amino acids, called pre-proparathyroid hormone,
Parathyroid hormone also acts directly was soon identified. This appears to be the
upon the kidney to minimize calcium loss in initial product resulting from transcription
the urine. Within a few minutes after admin¬ of the specific messenger RNA. A short
istration of the hormone to a parathyroidec- amino acid “signal” sequence serves to attach
tomized animal, there is an increase in excre¬ the polyribosomes to the membrane of the
tion of phosphate, sodium, and potassium in endoplasmic reticulum. As the nascent poly¬
the urine and a decrease in clearance of peptide chain extends through the mem¬
calcium. The hormone also influences the brane into the lumen, a specific peptidase
synthesis, in the intestine, of a form of vita¬ cleaves off the signal peptide, yielding pro¬
min D3 that increases the efficiency of calcium parathyroid hormone. This is channeled
absorption from dietary sources. through the reticulum to the Golgi complex
The secretion of parathyroid hormone is where the aminoterminal hexapeptide is re¬
under feedback control by the concentration moved to form the final product parathyroid
of calcium in the blood and extracellular hormone.
fluid. In response to a marked fall in the
blood calcium, the rate of hormone secretion
may reach 50 to 100 times the normal level.
The first biologically active extracts of the
parathyroid were prepared 70 years ago by
Collip. A highly purified fraction of the hor¬
mone was produced by Rasmussen in 1960
and its complete primary structure was de¬
termined by Brewer and Ronan in 1970. It
is a polypeptide of 84 amino acids. More
recent investigations have concentrated upon
elucidating its intracellular biosynthetic path¬
way and upon identifying its precursors.
Many proteins produced by cells are first
synthesized in the form of large precursor
molecules that are subsequently cleaved in-
tra- or extracellularly to yield the biologically
active product. Discovery of the precursors
of parathyroid hormone was somewhat de¬
layed because the very small size of the gland
made it difficult to obtain sufficient material
for analysis. One or two milligrams of a
precursor fraction were ultimately isolated
from several kilograms of bovine parathy¬
Figure 19-5. Photomicrograph of parathyroid gland of
roids and sequenced in 1972. This propara¬
monkey injected intravenously with India ink to show the
thyroid hormone consisted of the 84 amino acid extensively anastomotic capillary network in intimate con¬
chain of parathyroid hormone with an addi¬ tact with the gland cells, x 185. (Courtesy of I. Gersh.)
 PARATHYROID GLANDS • 515

There is relatively little storage of hormone Collip, J. B.: The extraction of a parathyroid hormone
in the parathyroid. The amount accumulated which will prevent or control tetany and which
intracellularly in bovine glands is estimated regulates the level of blood calcium. J. Biol. Chem.
63:395, 1925.
to be sufficient for only five to six hours of
Davis, R., and A. C. Enders: Light and electron micro¬
basal demand or IV2 hours under maximal scope studies on the parathyroid gland. In Creep,
stimulation. Increased demand for hormone R. O., and R. V. Talmage, eds.: The Parathyroids.
must be met by active synthesis, and the Springfield, IL, Charles C Thomas, 1961, p. 76.
DeRobertis, E.: The cytology of the parathyroid gland
intracellular mechanisms that couple secre¬
of rats injected with parathyroid extract. Anat. Rec.
tory activity to synthesis are poorly under¬ 78:473, 1940.
stood. Gaillard, P. J.: Parathyroid gland tissue and bone in
The precise mechanism by which parathy¬ vitro. Exp. Cell Res. 3(Suppl.): 154, 1955.
roid hormone exerts its effect upon bone and Gaillard, P. j.: Parathyroid and bone in tissue culture.
In Creep, R. O., and R. V. Talmage, eds.: The
kidney cells has yet to be completely worked
Parathyroids. Springfield, IL, Charles C Thomas,
out. There are strong indications, however, 1961, p. 20.
that activation of adenyl cyclase and elevated Grafflin, A. L.: Cytological evidence of secretory activity
intracellular concentrations of cyclic AMP are in the mammalian parathyroid. Endocrinology
26:857, 1940.
involved. The concentration of cyclic AMP in
Greep, R. O.: Parathyroid glands. In von Euler, U.S.,
the urine rises within minutes after adminis¬ and H. Heller, eds.: The Parathyroids. Comparative
tration of the hormone to intact animals. In Endocrinology. Vol. I. New York, Academic Press,
vitro, an increased activity of adenyl cyclase 1963, p. 235.
in kidney and bone can be demonstrated Greep, R. O., and R. V. Talmage, eds.: The Parathy¬
roids. Springfield, IL, Charles C Thomas, 1961.
within 15 seconds after adding parathyroid Habener, J. F. and J. T. Potts, Jr.: Biosynthesis of
hormone to the system. parathyroid hormone. Parts 1 and 2. N. Engl. J.
The parathyroids may give rise to tumors Med. 299:580, 635, 1978.
in which the cells produce excessive amounts Habener, J. F., D. Powell, T. M. Murray, G. P. Mayer,
and J. T. Potts, Jr.: Parathyroid hormone: secretion
of hormone, resulting in primary hyperparathy¬
and metabolism in vivo. Proc. Natl. Acad. Sci. U.S.A.
roidism. Such patients have high blood cal¬ 68:2896, 1971.
cium levels, rarefaction of their bones (osteitis Kemper, B., J. F. Habener, R. C. Mulligan, J. T. Potts,
fibrosa), low blood phosphate level, kidney and A. Rich: Pre-proparathyroid hormone: a direct
stones, and deposits of calcium in other soft translation product of parathyroid messenger RNA.
Proc. Natl. Acad. Sci. U.S.A. 71:3731, 1974.
tissues. In rickets, calcium and phosphate are Munger, B. L., and S. I. Roth: The cytology of the
not adequately absorbed from the diet, owing normal parathyroid glands of man and Virginia
to a deficiency of vitamin D. The resulting deer: a light and electron microscopic study with
low blood calcium results in a compensatory morphologic evidence of secretory activity, j. Cell
Biol. 16:379, 1963.
activation of the parathyroids called secondary
Potts, J. T., Jr., and L. J. Deftos: Parathyroid hormone,
hyperparathyroidism. Similarly, in severe kidney thyrocalcitonin, vitamin D and diseases of bone and
disease, there may be phosphate retention, bone mineral metabolism. In Bondy, P. K., ed.:
resulting in low blood calcium, and conse¬ Duncan’s Diseases of Metabolism. 6th ed. Philadel¬
phia, W. B. Saunders Co., 1969.
quent hypertrophy of the parathyroids.
Potts, J. T., Jr., T. Murray, M. Peacock, H. D. Niall, G.
W. Tregear, H. T. Keutmann, D. Powell, and L. J.
Deftos: Parathyroid hormone: sequence, synthesis,
immunoassay studies. Am. J. Med. 59:639, 1971.
REFERENCES
Rasmussen, H.: Chemistry of parathyroid hormone. In
Creep, R. O., and R. V. Talmage, eds.: The Para¬
Abe, M., and L. M. Sherwood: Regulation of parathyroid thyroids. Springfield, IL, Charles C Thomas, 1961,
secretion by adenyl cyclase. Biochem. Biophys. Res. p. 60.
Commun. 48:396, 1972. Rasmussen, H., and L. C. Craig: Isolation and charac¬
Aurbach, G. D., R. Marcus, J. Heersche, S. Marx, H. terization of bovine parathyroid hormone. J. Biol.
Niall, G. W. Tregear, H. T. Keutmann, and J. T. Chem. 236:759, 1961.
Potts, Jr.: Hormones and other factors regulating Roth, S. I.: Pathology of the parathyroids in hyperpara¬
calcium metabolism. In Robison, G. A., et al., eds.: thyroidism with a discussion of recent advances in
Cyclic AMP and Cell Function. Ann. N. Y. Acad. the anatomy and pathology of the parathyroid
Sci. 185:386, 1971. glands. Arch. Pathol. 73:492, 1962.
Barnicot, N. A.: The local action of the parathyroid and Trier, J. S.: The fine structure of the parathyroid gland.
other tissues on bone in intracerebral grafts. J. Anat. J. Biophys. Biochem. Cytol. 4:13, 1958.
52:233, 1948. Weymouth, R. J., and B. L. Baker: The presence of
Chang, H. Y.: Grafts of parathyroid and other tissues argyrophilic granules in the parenchymal cells of
to bone. Anat. Rec. 111:23, 1951. the parathyroid glands. Anat. Rec. 7 79:519, 1954.
 ADRENAL GLANDS AND
PARAGANGLIA

The paired adrenal or suprarenal glands are nucleoli. The cytoplasm is less abundant than
roughly triangular, flattened organs embed¬ in the cells of the other zones and is generally
ded in the retroperitoneal adipose tissue at acidophilic, but it contains some basophilic
the cranial pole of each kidney. They meas¬ material, which is usually disposed in clumps.
ure approximately 5 by 3 by less than 1 cm Lipid droplets are small and relatively scarce
and together weigh about 15 g. Both weight in this zone in most species but may be
and size may vary considerably, depending numerous in others. Mitochondria ^re fila¬
on the age and physiological condition of the mentous. The compact Golgi apparatus is
individual. The cut surface of the transected juxtanuclear and in some animals may be
gland presents a bright yellow cortex in its polarized toward the nearest vascular chan¬
outer part, with a reddish brown inner zone nel.
adjacent to the thin, gray medulla. At the electron microscopic level, the most
The adrenal glands comprise two distinct characteristic feature of the cytoplasm is its
endocrine organs that differ in their em- smooth-surfaced endoplasmic reticulum,
bryological origin, type of secretion, and which forms an anastomosing network of
function—the interrenal tissue and the chro¬ tubules throughout the cell body. Profiles of
maffin tissue. In mammals these are arranged granular endoplasmic reticulum are also
as cortex and medulla respectively, but in present in limited numbers, and there are
other vertebrate classes they may be inter¬ many polyribosomes free in the cytoplasmic
mingled in a variety of patterns or may be matrix. There is nothing unusual in the or¬
entirely dissociated. ganization of the Golgi complex or in the
centrioles that are associated with it. The
mitochondria as a rule have lamellar cristae
like those of most other organs. The plasma
THE ADRENAL CORTEX membrane is smoothly contoured over most
of the cell body but may have a few folds or
The cortex, which forms the bulk of the microvilli on the surface bordering on a peri¬
gland, has three distinguishable concentric vascular space and at the junctions where
zones—a thin, outer zona glomerulosa adjacent several cells meet.
to the capsule; a thick middle layer, the zona The zona fasciculata consists of polyhedral
fasciculata; and a moderately thick, inner zona cells considerably larger than those of the
reticularis contiguous with the medulla (Figs. glomerulosa and arranged in long cords dis¬
20-1, 20-2). In man these make up respec¬ posed radially with respect to the medulla
tively 15, 78, and 7 per cent of the total (Fig. 20—4). The cords are usually one cell
cortical volume. The transition from one thick and separated by sinusoidal blood ves¬
zone to another in histological sections is sels. The nucleus is central, and binucleate
gradual but may appear sharper in prepara¬ cells are common. The cytoplasm is generally
tions injected to show the vascular pattern. acidophilic but may contain basophilic
The zona glomerulosa consists of closely masses, particularly in the peripheral portion
packed clusters and arcades of columnar cells of the zone. The cells are crowded with lipid
that are continuous with the cell columns of droplets in the fresh condition, but after
the zona fasciculata (Fig. 20-3). The spherical treatment with the organic solvents used in
nuclei stain deeply and contain one or two preparation of routine histological sections,
516
 ADRENAL GLANDS AND PARAGANGLIA • 517

Capsule ^

Zona glomerulosa X

Zona fasciculata -J

Capsule

Zona glomerulosa

Zona reticularis -<

V Zona fasciculata

»'
*r _ 9
Medulla v
*
A , " # ' V 4 ' • * '
. ^

& £,
^ *-*
*' j Aft!? * #
... * i.,* *
.//V'8 \t ***>• '
pm
EsmiU’&OHLMa,N

Figure 20-1. Sections through the adrenal glands of a 6-month-old infant (left) and of a man (right). Mallory-azan stain,
x 110.
518 • ADRENAL GLANDS AND PARAGANGLIA

of the cortex, and in such places the zona

fasciculata is found immediately beneath the
cortex, but this is not common. There may
also be a thin transitional region between the
zona glomerulosa and zona fasciculata that is
relatively free of lipid droplets. Such a su¬
danophobic zona intermedia is particularly ob¬
vious in the rat adrenal.
In electron micrographs, ceils of the zona
fasciculata have a round nucleus, a promi¬
nent nucleolus, and clumps of heterochro¬
matin distributed at the periphery. In the
human the nuclear envelope has a distinct
internal fibrous lamina. The most conspicu¬
ous ultrastructural feature of the cells is a
very extensive smooth endoplasmic reticulum
in the form of branching and anastomosing
tubules (Figs. 20-5, 20-6, 20-7). This orga¬
nelle occupies 40 to 45 per cent of the cell
volume. Occasional parallel arrays of cister-
nae of rough endoplasmic reticulum are
found in the zona fasciculata of the human
adrenal but rarely in other species. The Golgi
complex is well developed and has multiple
stacks of cisternae and associated vesicles.
f* * , ..LV The mitochondria are numerous, occupy¬

BE dif» A »:>,
ing 26 to 36 per cent of the cell volume.
They are elongate in humans but generally
spherical in common laboratory rodents. The
^1/ TsT# ^r« matrix is of low density and the cristae are
m atypical in form. In the rat they appear to be
i>'* - . . I, &•*£&*■*
f spherical vesicles 60 to 70 nm in diameter,
«% It *.*“ : '.•**
TAT*/—{TT-.'.'.'®**
SP»'
* "di\ r ,7 „ Kf while in the human they are described as
■* tubulovesicular—short tubules with vesicular
H®* sdi 1 >■ t A *T^* A
dilatations along their length. In the hamster
^f*|,:^uy:s.^A*f!$ttV®'8^hS(?£P&%..1 they are slender tubules of uniform caliber.
JvT'^.§» 0 *4l&1 M Cristae of tubular form are also found in
f # .*•••*'■■. other steroid-secreting cells.
.V * • * ifiB * o Small dense bodies of irregular shape, as¬
fSgia« •>i.«' i fl /:-* ^ t'JiA44aPlo^& * sumed to be lysosomes, are present in small
e j c numbers throughout the cytoplasm, and mi¬
w f * - «* •
croperoxisomes have been identified with the
m ■ -r. 1 '
diaminobenzidene reaction. Deposits of lipo-
%*■■ &*■ ■ • /^*:.*;i‘* *v..,,
,=v.
; : 4»*f« : J-s, :
#.»*" -
chrome pigment are common in the human
* /y ^-.-vi • #. ’■ - .*[. A adrenal.
*'
..**-/ < ^ . Sj. -f^-L, ^
•> ■*; «s ■I ■ ■ ;'.' MM
. 0 . ^ *S Lipid droplets are usually present but there
mi
*♦ O #' ♦ • 9 **JB 0 « is considerable interspecific variation in their
,.*•/ t«< <*°> jl r*#5 abundance. They occupy 10 to 15 per cent
of the cell volume in the rat, and are numer¬
Figure 20-2. Photomicrograph of the full thickness of the
ous in the human and other primates but
adrenal cortex of a rhesus monkey, showing from the top
downward the zona glomerulosa, zona fasciculata, and usually absent in the ox and hamster.
zona reticularis. The relationships between cell ultrastruc¬
ture and function in adrenocortical cells is
better understood than in many other cell
the cytoplasm has a foamy appearance due types. The enzymes involved in biosynthesis
to the numerous clear vacuoles left by ex¬ of steroid hormones are located in the exten¬
traction of lipid (Fig. 20—4). In man, the zona sive smooth endoplasmic reticulum and in
glomerulosa may be absent in restricted areas the numerous mitochondria. The varying
Text continued on page 524
 ADRENAL GLANDS AND PARAGANGLIA 519

Figure 20-3. Photomicrograph of the monkey zona glomerulosa at higher magnification than in Figure 20-2, showing
the alveolar or glomerular arrangement of cells. These ceils have relatively few vacuoles, representing extracted lipid.
520 ADRENAL GLANDS AND PARAGANGLIA

Figure 20-4. Photomicrograph of the zona fasciculata. This zone consists of cords of cells filled with spherical vacuoles
resulting from extraction of the abundant lipid droplets characteristic of this zone.
 ADRENAL GLANDS AND PARAGANGLIA 521

Figure 20-5. Electron micrograph of a portion of a cell from the human fetal adrenal cortex, showing large spherical
mitochondria with tubular cristae, a few parallel arrays of cisternae of granular reticulum, and an extraordinary abundance
of smooth endoplasmic reticulum. (Micrograph courtesy of A. L. Jones and S. McNutt.)
522 • ADRENAL GLANDS AND PARAGANGLIA

Figure 20-6. Electron micrograph of a juxtanuclear area of a cortical cell from an adult human adrenal. The smooth
reticulum remains abundant, the mitochondria are variable in size and shape, and they generally have tubular cristae.
There is a marked tendency for accumulation of irregular dense masses of lipochrome pigment. (Micrograph courtesy
of J. Long.)
 ADRENAL GLANDS AND PARAGANGLIA 523

Figure 20-7. Electron micrograph of an area of cytoplasm of an adult adrenocortical cell at high magnification, illustrating
the tubular nature of the smooth reticulum and the fine structure of the mitochondria. (Micrograph courtesy of J. Long.)
524 • ADRENAL GLANDS AND PARAGANGLIA

abundance of lipid in different species is Others believe this requires modification of

correlated with the degree to which choles¬ the steroid molecule by sulfation or binding
terol, the precursor of steroid synthesis, is to a carrier molecule. Evidence has recently
taken up from the blood or is synthesized accumulated suggesting that some of the
from acetate in the smooth reticulum of the small dense bodies that have usually been
adrenocortical cells. Cholesterol stored in the interpreted as lysosomes or microperoxi¬
lipid droplets is released by cholesterol ester¬ somes are in fact secretory granules formed
ase and enters mitochondria, where it under¬ in the Golgi and released by exocytosis as
goes side-chain cleavage to yield pregneno¬ rapidly as they are formed. In support of
lone. Transferred from mitochondria to this interpretation is the observation that in
endoplasmic reticulum, pregnenolone is con¬ ACTH-stimulated cells that have been ex¬
verted to progesterone and then to 11-deoxy¬ posed to vinblastin to block the microtubule
corticosterone or 17-deoxycorticosterone. transport mechanism commonly associated
These intermediate products are converted with exocytosis, there is an increase in small
to the steroid hormones deoxycorticosterone and dense bodies in the cytoplasm and a tenfold
cortisol, respectively, by enzymes located in increase in intracellular corticosterone.
the mitochondria. The biosynthetic pathway In the zona reticularis, the regular parallel
thus involves transfer of precursor molecules arrangement of cel) cords gives way to an
and intermediate products to and fro be¬ anastomosing network of cell cords. The
tween mitochondria and smooth endoplasmic transition from the fascicuiata is gradual, the
reticulum. Little is known as to how these cells differing but little. The cytoplasm con¬
translocations occur. tains fewer lipid droplets. Toward the me¬
The stimulatory effect of adrenocortico¬ dulla there is a variable number of “light”
tropic hormone (ACTH) is mediated by cyclic and “dark” cells. The nuclei of the light cells
AMP. Upon binding of the hormone to re¬ are pale-staining; those of the dark cells are
ceptors in the membrane of the cells in the hyperchromatic and shrunken. The physio¬
zona fascicuiata, adenyl cyclase is activated to logical significance of these differences in
catalyze the formation of cyclic AMP from staining affinity is not known. In other or¬
ATP. This in turn activates cholesterol ester¬ gans, this appearance is often a fixation arti¬
ase, which hydrolyzes cholesterol ester stored fact. It is so common in the zona reticularis,
in lipid droplets, making free cholesterol however, that some interpret these cells as
available for steroid hormone biosynthesis. degenerative. The cells of this zone, particu¬
There is some evidence of ACTH enhance¬ larly the dark cells, contain large accumula¬
ment of transcription and translation, possi¬ tions of lipofuscin pigment. Apart from the
bly related to synthesis of carrier proteins presence of light and dark cells and the
involved in transfer of intermediate products greater amount of pigment, the cells of the
between organelles. zona reticularis resemble those of the fascic¬
The morphological correlates of prolonged uiata and, like them, have an abundance of
ACTH stimulation are depletion of lipid agranular reticulum.
droplets; increase in smooth endoplasmic re¬
ticulum; proliferation of mitochondria; and
enlargement of the Golgi complex. Unlike
many other glandular cells, steroid-secreting
THE ADRENAL MEDULLA
cells do not store their product in secretory
granules formed in the Golgi complex. The The boundary between the zona reticularis
role of the Golgi in steroid-secreting cells and medulla is usually irregular in the human
remains unclear, but it is speculated that it adult, with columns of cortical cells projecting
may be involved in sulfation of intermediate some distance into the medulla. In other
steroid molecules or their conjugation to car¬ animals, the boundary may be quite sharp.
rier proteins. The medulla is composed of large epithelioid
In the absence of unambiguous evidence cells arranged in rounded groups or short
of vesicular transport and exocytosis, the cords in intimate relation to blood capillaries
mechanism of hormone release by adreno¬ and venules (Fig. 20—8). When the tissue is
cortical cells is controversial. A majority of fixed in a solution containing potassium bi¬
investigators favor simple diffusion through chromate, these cells are seen to be filled with
the aqueous phase of the cytoplasm and fine brown granules. This browning of the
across the lipid bilayer of the cell membrane. cytoplasmic granules with chromium salts is
 ADRENAL GLANDS AND PARAGANGLIA • 525

Figure 20-8. Photomicrograph of the junction of the zona reticularis (above) with the clusters of large pale cells of the
adrenal medulla (below). Catecholamine granules ordinarily are not visible with the light microscope in the cells of the
medulla.

called the chromaffin reaction and is thought affinity for azocarmine and a positive acid
to result from the oxidation and polymeri¬ phosphatase reaction, and are not fluorescent
zation of the catecholamines epinephrine and or reactive with iodate or silver.
norepinephrine contained within the granules In electron micrographs, the most promi¬
of the cells. The medulla is colored green nent feature of the cells of the adrenal me¬
after treatment with ferric chloride—appar¬ dulla is the presence of large numbers of
ently for the same reason. There are other membrane-bounded dense granules 100 to
cells in the body that give a similar reaction, 300 nm in diameter (Fig. 20—9). The granules
notably some of the argentaffin cells of the are bounded by a membrane separated from
gastrointestinal tract and the mast cells, which the dense content by an electron-lucent gap.
are reactive because of their content of 5- When tissue is fixed in glutaraldehyde, two
hydroxytryptamine. The chromaffin cells of populations of cells are distinguishable on
the adrenal medulla, however, are derived the basis of the character of their granules.
from neuroectoderm, secrete catechol¬ Cells that store norepinephrine have gran¬
amines, and are innervated by preganglionic ules that contain a very electron-dense core,
sympathetic fibers. often eccentric in its location within the mem¬
In most species the application of a group brane-limited vesicle. Cells that store epi¬
of histochemical methods to the chromaffin nephrine have granules that are relatively
cells permits the identification of two types homogeneous and less electron dense. The
of cells, one containing norepinephrine and mitochondria are not remarkable. The cister-
the other epinephrine. The norepinephrine nae of the granular endoplasmic reticulum
storing cells are autofluorescent, give argen¬ form small parallel arrays. The juxtanuclear
taffin and potassium iodate reactions, exhibit Golgi apparatus frequently contains in its
a low affinity for azocarmine, and give a cisternae dense material interpreted as a pre¬
negative acid phosphatase reaction. The ep¬ cursor of the granules.
inephrine-storing cells have a high staining It has been established by cell fractionation
526 • ADRENAL GLANDS AND PARAGANGLIA

Figure 20-9. Electron micrographs of cells from the adrenal medulla of the cat, showing the abundant, membrane-
bounded, dense granules that are the sites of storage of catecholamines, x 9600 and x 13,600. (Courtesy or R.
Yates.)

and density gradient centrifugation that the macologically active substance reserpine may
hormones are contained within the secretory be the result of inhibition of the active trans¬
granules. The granules may contain as much port mechanism.
as 20 per cent by weight of the hormone. For some time there was a divergence of
Although they also contain a significant opiniop as to whether the granules of the
amount of protein, much of this is the soluble adrenal medulla are released by exocytosis,
protein chromogranin. The catecholamines do as are protein hormones, or whether the
not seem to be bound to a specific protein, smaller molecules of catecholamines are re¬
as are the hormones of the neurohypophysis leased from the granules within the cyto¬
and other neurosecretory cells. Therefore, it plasm and diffuse out of the cell. It now
is not entirely clear how such low-molecular- seems to have been well established experi¬
weight substances are retained in the gran¬ mentally that all or nearly all of the hormones
ules. According to one hypothesis, the hor¬ of the adrenal medulla are discharged by
mone forms high-molecular-weight aggre¬ exocytosis. Evidence for this resides not only
gates with adenosine triphosphate (ATP) and in morphological observations but in the
divalent cations, both of which are present in physiological finding that a perfusate of a
the granules. The active uptake of catechol¬ stimulated gland contains not only catechol¬
amine by the granules appears to be the amines but also soluble protein constituents
result of an active transport mechanism de¬ of the granules (chromogranin) and ATP, in
pendent on magnesium-activated ATPase in the same proportions that occur in a lysate
the limiting membrane. Thus, the ability of of isolated granules, whereas the insoluble
secretory granules in the adrenal medulla to constituents associated with the membranes
retain hormones in vivo probably depends of the granules are retained within the gland.
on (1) micelle formation between ATP and In addition to the chromaffin cells, sym¬
hormone and (2) an active transport mecha¬ pathetic ganglion cells occur singly or in small
nism in the membrane limiting the granules. groups in the adrenal medulla.
The depletion of catecholamines by the phar¬ The adrenal is enclosed by a thick capsule
 ADRENAL GLANDS AND PARAGANGLIA • 527

of collagenous connective tissue that extends

into the cortex to varying depths as trabecu¬
lae. Most of the rest of the supporting frame¬
work of the cortex consists of reticular fibers
that lie between the sinusoids and the cell
cords and penetrate to some extent between
the gland cells. Reticular fibers also enclose
the cell clusters in the medulla and support
the capillaries, veins, and nerves. Collagenous
fibers appear around the larger tributaries of
the veins and merge with the capsular con¬
nective tissue.

BLOOD SUPPLY AND

LYMPHATIC DRAINAGE OF
THE ADRENAL

The gland is richly supplied by a number

of arteries that enter at various points around
the periphery. Three principal groups are
recognized. The superior suprarenal arteries
arising from the inferior phrenic artery ap¬
pear to be the major source, but in addition
there are the middle suprarenals arising from
Figure 20-10. Schematic representation of the blood
the aorta and the inferior suprarenals, which supply of the adrenal, showing how blood from capsular
are branches of the renal artery. Arteries arterioles traverses the cortex and continues into capillar¬
from these several sources form a plexus in ies and venules in the medulla. This arrangement carries
blood rich in cortical steroids to the catecholamine-syn¬
the capsule. The cortical arteries arise from
thesizing cells in the medulla. (From Hamilton, W. J., ed.:
this capsular plexus and distribute to the Textbook of Human Anatomy. London, St. Martin’s Press,
anastomosing network of sinusoids sur¬ Macmillan and Co., 1957.)
rounding the cords of parenchymal cells in
the cortex (Fig. 20-10). The sinusoids of a
given region converge in the zona reticularis tant physiological consequences. The blood
upon a collecting vein at the corticomedullary reaching the medulla via the sinusoids has
junction. There is no venous system in the traversed the cortex and is rich in cortical
cortex. steroid hormones. There is evidence that
Some major arterial branches from the high concentration of glucocorticoids may be
capsule penetrate the connective tissue tra¬ required for induction and maintenance of
beculae and pass directly through the cortex, the enzyme phenylethanolamine-M-methyl
giving off few or no branches until they reach transferase, which is necessary for the syn¬
the medulla. In the medulla, they branch thesis of epinephrine. Thus, the adrenocor¬
repeatedly to form the rich capillary net tical steroids have a local downstream effect
around the clumps and cords of chromaffin on the medulla, as well as a general systemic
cells. The medulla thus has a dual blood effect. Indeed, whether a medullary cell se¬
supply—via the cortical sinusoids that anas¬ cretes norepinephrine or epinephrine may
tomose with its capillary bed across the cor¬ be determined by where it is situated with
ticomedullary junction, and via the medullary respect to steroid-rich blood from the cortex
arteries that course from the capsule directly that regulates the synthesis of the enzyme
to the medulla. The capillaries of the medulla necessary for conversion of norepinephrine
empty into the same venous system that to epinephrine.
drains the cortex. The multiple venules ulti¬ The cells lining the capillaries of the me¬
mately join to form the large central veins of dulla are typical endothelium. The nature of
the medulla, which emerge from the gland the lining of the sinusoids in the cortex is still
as the suprarenal vein. a subject of debate. These cells have been
This pattern of vascularization has impor- reported to take up colloidal vital dyes, such
528 • ADRENAL GLANDS AND PARAGANGLIA

as lithium carmine and trypan blue, and on HISTOPHYSIOLOGY OF THE

this basis they have been regarded as belong¬ ADRENAL CORTEX
ing to the reticuloendothelial system. Subse¬
quent studies suggest that these substances
simply adhere to the cell surface or that they The adrenal cortex is involved in a wide
are taken up by macrophages lying between variety of body functions, including mainte¬
the endothelium and the parenchymal cells. nance of fluid and electrolyte balance, main¬
Electron microscopic observations have failed tenance of carbohydrate balance, and main¬
to reveal any evidence of phagocytosis by the tenance of the normal function of certain
endothelial cells of the sinusoids. Except in cellular elements of the connective tissues. It
the regions occupied by the nucleus and cell is essential for life. Its removal or destruction
center, the endothelium is extremely atten¬ leads ultimately to death unless the patient is
uated and interrupted at intervals by small given exogenous adrenocortical hormones.
circular pores or fenestrae closed only by a The widely accepted zonal hypothesis of
very thin diaphragm. There is a continuous adrenal function affirms that different func¬
basal lamina supported at intervals by small tions can be ascribed to the morphologically
bundles of collagen fibrils corresponding to recognizable zones of the adrenal cortex.
the reticulum of light microscopy. The zona glomerulosa secretes hormones col¬
Lymphatics are limited to the capsule and lectively called miner aloe or tic oids (aldosterone
its cortical trabeculae, and to the connective and deoxycorticosterone), which are con¬
tissue around the large veins. cerned with fluid and electrolyte balance.
The zona fasciculata and zona reticularis se¬
crete hormones collectively referred to as
glucocorticoids (cortisol, cortisone, and corti¬
costerone), which are concerned with metab¬
NERVES OF THE ADRENAL olism of carbohydrates, proteins, and fat.
The concept of a separate function of the
The cells of the adrenal cortex apparently zona glomerulosa and the inner zones of the
are not innervated. Those of the zona fasci- cortex is supported by abundant experimen¬
culata are stimulated by circulating adreno¬ tal evidence. There is as yet no clear-cut
corticotropic hormone (ACTH) of the pitui¬ evidence of qualitative functional differences
tary; those of the glomerulosa are responsive between the zona fasciculata and zona retic¬
to changes in extracellular fluid volume or ularis.
changes in concentration of sodium and po¬ The most important hormone of the zona
tassium in the blood, possibly acting indi¬ glomerulosa is aldosterone. When released in
rectly via a hypothetical hormone of the dien¬ the body or administered exogenously, it has
cephalon, aldosterone-stimulating factor. three basic effects: it increases reabsorption
Preganglionic sympathetic nerve fibers of sodium by the tubules of the kidney; it
arising from cell bodies in the intermedio- increases the excretion of potassium by the
lateral column of the spinal cord in the lower kidney; and it appears to increase both the
thoracic and lumbar regions of the spinal movement of sodium into the cells of the
cord pass through the sympathetic chain and body and the associated transfer of potassium
via splanchnic nerves to the capsule of the out of the cells. If the secretion of this hor¬
adrenal, where they form a rich nerve plexus, mone is eliminated by disease or surgical
including a few sympathetic ganglion cells. removal of the adrenal, the sodium and chlo¬
From this plexus preganglionic nerves turn ride concentrations of the extracellular fluid
inward, traverse the cortex, and end in the decrease markedly, the volume of extracel¬
medulla around the cells that secrete epi¬ lular fluid is greatly diminished, and the
nephrine and norepinephrine. These cells patient goes into a shocklike state that is
are derived embryologically from the ner¬ followed by death within a few days, unless
vous system in early development and are treated by salt replacement or administration
analogous to postganglionic neurons. The of exogenous mineralocorticoids.
nerve terminals form typical synapses with As in other parts of the endocrine system,
the cells, in which the pre- and postsynaptic the secretion of aldosterone is carefully reg¬
membranes are separated by a cleft of 15 to ulated to maintain constancy of the internal
20 nm. The nerve terminals are cholinergic environment of'the cells of the body. If for
and contain multiple clear synaptic vesicles. some reason the extracellular sodium concen-
 ADRENAL GLANDS AND PARAGANGLIA • 529

tration falls, or if potassium rises, or extra¬ tions, such as pain, fear, and rage result in
cellular fluid volume is dangerously dimin¬ passage of nerve impulses to the hypothala¬
ished, this is reflected in the concentration in mus that initiate secretion of corticotropin
the blood and detected by centers in the releasing factor (CRF) into the hypophyseo-
midbrain. These, in turn, release a hormone portal vessels. Carried to the anterior lobe of
that stimulates the zona glomerulosa to re¬ the hypophysis, this promotes adrenocorti-
lease aldosterone, which acts upon the kidney cotropin release, which in turn causes dis¬
tubules to increase sodium absorption and charge of glucocorticoids from the adrenal.
decrease potassium resorption, thus correct¬ The resulting increase in blood amino acid
ing the imbalance. If increased demands are and glucose levels makes available to the cells
placed upon the homeostatic mechanisms for the energy-rich substrates that may be
maintaining electrolyte balance by giving an needed for combat, flight, or other responses
animal either an excess of potassium or a to stress.
sodium-deficient diet, there is a selective hy¬ The glucocorticoids also have effects upon
pertrophy of the zona glomerulosa and an inflammatory responses of the connective tis¬
increase in the abundance of smooth endo¬ sues and upon the immune system. The
plasmic reticulum in its cells. Conversely, mechanisms of these actions are poorly
injection of large doses of aldosterone or understood but they are widely utilized in
deoxycorticosterone results in atrophy of this the treatment of allergies, arthritis, rheu¬
zone. matic fever, and many other inflammatory
The glucocorticoids secreted by the two diseases. Cortisol somehow reduces the se¬
inner zones of the cortex are almost as im¬ verity of allergic reactions and suppresses the
portant in sustaining life as are the mineral- inflammation. Among other effects it causes
ocorticoids. Secretion or administration of destruction of lymphocytes, atrophy of
cortisol results in a great increase in the lymphoid tissue, and a decrease in level of
formation of glucose in the liver and its circulating eosinophils. Therefore, in surgical
intracellular storage as glycogen. Excess glu¬ grafting of organs, cortisol is often given to
cose is also released into the blood, producing suppress the rejection reaction.
a condition comparable with diabetes. The There is also a relationship between the
hormone also causes a decrease in the rate adrenal cortex and the reproductive system.
of protein synthesis and an increase in the Adrenalectomy is followed by loss of libido
rate of protein breakdown in the cells of the in the male and abnormal cycles in the fe¬
body. Therefore, the level of amino acids in male. Removal of the adrenal interrupts lac¬
the blood increases. Finally, cortisol acts upon tation. Whether or not these effects are me¬
adipose tissue, increasing both the rate at diated via the adenohypophysis is not clear.
which lipid is accumulated in the fat cells and It is known that under normal conditions,
the rate at which the lipids are mobilized small amounts of estrogens and androgens
from those cells. are produced in the two inner zones of the
The control of the secretion of glucocorti¬ cortex. In the pathological condition known
coids is quite independent of that of the as adrenogenital syndrome, the inner zones of
mineralocorticoids. The maintenance of the the cortex are hypertrophic, and high levels
inner zones of the cortex depends on secre¬ of circulating androgens may result in pre¬
tion of ACTH by the anterior lobe of the cocious puberty, increased hirsutism, and
hypophysis. Hypophysectomy results in a other manifestations of virilism.
marked atrophy of the zona fasciculata and Hyperadrenocorticism or Cushings disease is a
zona reticularis but has little effect on the condition in which the adrenal cortex is hy¬
zona glomerulosa. This atrophy of the inner perplastic, as a result of stimulation by pitui¬
zones can be prevented or reversed by injec¬ tary or other tumors producing excessive
tion of ACTH. Conversely, administration of amounts of ACTH. It is characterized by
large doses of cortisol to an intact animal obesity, particularly over the nape of the
suppresses hypophyseal secretion of ACTH neck; hirsutism; impotence or amenorrhea;
and results in atrophy of the inner zones of abdominal striae; and a characteristic round
the adrenal cortex. “moon face.” Hypoadrenocorticism or Addisons
The mechanisms of control of glucocorti¬ disease is due to destruction of the adrenal
coid secretion have already been presented cortex by tuberculosis or other infection of
in Chapter 3 but, briefly restated, many dif¬ the gland, resulting in chronic insufficiency
ferent kinds of alarming or stressful situa¬ of hormone production. It is characterized
530 • ADRENAL GLANDS AND PARAGANGLIA

by generalized weakness, weight loss, low vation of the blood pressure with very little
blood pressure, and abnormal pigmentation, effect upon heart rate or cardiac output. The
and leads ultimately to death if hormone effect of norepinephrine on the blood pres¬
replacement therapy is not undertaken. sure is not due to an action upon the heart¬
beat but is primarily a consequence of the
vasoconstriction it brings about in the periph¬
eral portion of the arterial system. Both epi¬
HISTOPHYSIOLOGY OF THE nephrine and norepinephrine cause lipolysis
ADRENAL MEDULLA and release of unesterified fatty acid from
adipose tissue isolated in vitro.
The adrenal medulla is not essential for Norepinephrine is not confined to the ad¬
life. Animals that have had their adrenal renal medulla but is present in the brain and
medullas removed can survive under ordi¬ in most of the innervated peripheral tissues,
nary circumstances but are unable to respond where it is localized mainly in tbe sympathetic
normally to emergency situations. The hor¬ nerve endings. It has been established as the
mones of the medulla, epinephrine and norep¬ principal transmitter substance of adrenergic
inephrine, are catecholamines, and unlike the neurons. Thus it is a neurohumor. Neurohu¬
steroid hormones of the cortex, they accu¬ mors such as norepinephrine, acetylcholine,
mulate in high concentration in the cells. The and serotinin are released by nerve cells,
granules of the cells are the site of storage of usually at their endings, and affect other
catecholamines. The two hormones are pro¬ neurons or muscles or glands. In general
duced in different proportions depending they act transiently and at very short range,
upon the species. Aggressive and predatory being destroyed enzymatically before they
animals tend to secrete large amounts of reach effective concentrations in the circula¬
norepinephrine, whereas the more timid and tion. The norepinephrine released into the
placid species produce relatively less. bloodstream by the adrenal medulla is some¬
Although the two hormones are very what exceptional among neurohumors in
closely related chemically, there are impor¬ that it does reach effective levels in the blood
tant qualitative and quantitative differences and acts at a distance. On the other hand,
in their physiological effects. the neurosecretory substances, such as the oxy¬
Epinephrine increases the heart rate and tocin and vasopressin of the posterior lobe of
cardiac output without significantly increas¬ the hypophysis, are long-acting products of
ing the blood pressure, and may increase the nerve cells that act at long range.
blood flow through some organs by as much Although the adrenomedullary hormones
as 100 per cent. It has effect on the cardio¬ are not essential for life, it appears that in
vascular system in very low concentrations. times of stress they do help to maintain
The denervated mammalian heart, for ex¬ homeostasis and to prepare tbe organism to
ample, is accelerated by as little as one part meet emergency situations. Epinephrine ac¬
epinephrine in 1.4 billion parts of fluid me¬ complishes this by elevating blood glucose
dium. Epinephrine also has a marked effect levels, increasing cardiac output, and redis¬
upon metabolism. It increases oxygen con¬ tributing blood within the circulation to en¬
sumption and basal metabolic rate. It elevates sure continuing rapid flow to those organs
the blood sugar level by mobilizing the car¬ vital for survival. Norepinephrine is less im¬
bohydrate stores of the liver; by promoting portant in these emergency adjustments, but,
the conversion of muscle glycogen into lactic as the mediator of adrenergic nerve impulses
acid, from which new carbohydrate can be throughout the body, it acts continuously on
made in the liver; and by causing the release the blood vessels of the normal animal to
of ACTH from the hypophysis, which in turn maintain blood pressure.
affects gluconeogenesis by stimulating secre¬ Hyperfunction of the adrenal medulla in
tion of glucocorticoids from the adrenal cor¬ man occurs with certain rare tumors of the
tex. The mobilization of glucose from the medulla or of extramedullary chromaffin tis¬
liver by epinephrine appears to result from sues. In such cases there may be attacks of
its activation of the enzyme phosphorylase, sweating, mydriasis, hypertension, and hy¬
which accelerates the first step in the break¬ perglycemia, terminating suddenly in death.
down of glycogen to glucose. The paroxysmal hypertension (acute high
Norepinephrine has relatively little effect blood pressure) of adrenomedullary tumors
upon metabolism but causes a marked ele¬ is decreased or abolished by intravenous
 ADRENAL GLANDS AND PARAGANGLIA • 531

administration of a series of compounds that of the common laboratory animals has a

have an epinephrine-inhibiting action. comparable fetal zone. In the mouse, the so-
The cells of the adrenal medulla are re¬ called X zone differentiates postnatally and
garded as modified postganglionic neurons, regresses at puberty in the male or at the
and their secretory activity seems to be time of the first pregnancy in the female.
largely, if not entirely, under nervous con¬ The hormonal factors controlling it appear
trol. Hormones of the medulla are increased to be different from those affecting the hu¬
in the blood of the adrenal veins after stim¬ man fetal zone.
ulation of the splanchnic nerves, and secre¬ The medulla arises from the ectodermal
tion is prevented by sectioning these nerves. neural crest tissue, which also gives rise to
After splanchnic nerve stimulation, about 75 sympathetic ganglion cells. Strands of these
per cent of the secretion is norepinephrine sympathochromafhn cells migrate ventrally
and 25 per cent epinephrine. Certain centers and penetrate the anlagen of the adrenal
in the posterior hypothalamus are known to cortex on its medial side to take up a central
relay impulses to the adrenal medulla by way position in the organ rudiment.
of the splanchnic nerves. In the intact animal,
certain kinds of emotional stimuli are espe¬
Cell Renewal and Regeneration in
cially effective in releasing norepinephrine,
the Adrenal Cortex
and other kinds of stimuli, such as pain or
hypoglycemia, promote the release of epi¬ There have long been divergent views as
nephrine. For this reason, it is believed that to the mode of growth and repair of the
the cells producing epinephrine and those adrenal cortex. It was formerly believed that
producing norepinephrine receive different the cells arose either from fibroblast-like cells
innervation and secrete their hormones in¬ in the capsule or by division of the cells in
dependently. the zona glomerulosa, and that they gradu¬
ally moved through the zona fasciculata and
degenerated in the zona reticularis. The dark
cells of this zone were interpreted as cells
HISTOGENESIS OF THE
undergoing regressive changes. During their
ADRENAL GLANDS migration, the cells were believed to go
through one cycle of secretion.
The cortex develops from the coelomic The bulk of the evidence now seems to
mesoderm on the medial side of the urogen¬ favor the view that, once formed, the cells of
ital ridge of the embryo. Mesothelial cells the glomerulosa and fasciculata do not move
near the cranial pole of the mesonephros in appreciably and that replacement and repair
8- to 10-mm human fetuses proliferate and take place as a result of local mitotic activity.
penetrate the subjacent, highly vascular mes¬ After injection of colchicine to arrest cell
enchyme. These cells ultimately form the fetal division at metaphase, mitotic figures are not
cortex. A second proliferation of the coelomic confined to any one region but are found
mesothelium taking place in 14-mm embryos throughout the cortex, the majority being in
later forms the definitive or permanent cortex. the zona fasciculata. Autoradiographic stud¬
The adrenal gland in the fetus is relatively ies after administration of tritiated thymidine
large, with the fetal zone composing about have produced equivocal results but, on the
80 per cent of the cortex. The cells of this whole, they provide little evidence of exten¬
zone are large and stain intensely with eosin. sive cell migration.
After birth the fetal cortex undergoes a rapid In experimental animals the adrenal cortex
degeneration and the permanent cortex en¬ has a considerable capacity for regeneration.
larges. Associated with these changes is a 50 If the gland is incised and all of the tissue in
per cent decline in the absolute weight of the its interior removed, leaving behind only the
adrenal during the first few postnatal weeks. capsule and a few adherent granulosal cells,
The adrenal in the fetus is functional and the whole cortex will be regenerated. The
under the control of ACTH secreted by the medulla is not restored. Studies of the ste¬
hypophysis. Monstrous anencephalic fetuses, roids elaborated during regeneration of the
which lack a normal hypophysis, have very cortex show that an adequate level of secre¬
small adrenal glands with no fetal zone. The tion of mineralocorticoids is established early,
physiological role of this zone during intra¬ although the secretion of glucocorticoids
uterine life is not well understood. Progress does not occur until one to two weeks later.
in this area is hampered by the fact that none Thus, in the regenerating gland, cells func-
532 ADRENAL GLANDS AND PARAGANGLIA

Figure 20-11. Cells of a paraganglion from the rabbit, showing chief cells, which contain numerous catecholamine¬
storing granules resembling those of the adrenal medulla. (Micrograph courtesy of R. Yates.)
 ADRENAL GLANDS AND PARAGANGLIA • 533

tionally similar to those of the normal zonae those of the adrenal medulla (Fig. 20-11).
fasciculata and reticularis differentiate from Histochemical methods indicate that these
cells of the zona glomerulosa. granules contain catecholamines, principally
if not exclusively norepinephrine. The sup¬
Phytogeny of the Adrenal Gland porting cells partially or completely surround
each of the chief cells. The nucleus of these
Chromaffin tissue that yields epinephrine¬ cells is elongated in section and frequently
like activity is present in the central nervous deeply infolded. The cytoplasm is devoid of
system of leeches and also in the mantles of secretory granules and is otherwise unre¬
certain molluscs. In cyclostomes and teleosts, markable.
the intervened bodies (which are homologous The paraganglia are richly vascularized by
with the cortex) are separate from the dis¬ capillaries lined by very attenuated endo¬
crete chromaffin bodies. In amphibians the thelial cells, occasionally exhibiting fenes¬
two components are in juxtaposition, or they trated regions. The chief cells are usually
may be intermingled; in reptiles and birds separated from the blood by a barrier con¬
they are commonly intermingled. The well- sisting of a thin intervening supporting cell,
known cortex and medulla relationship first two basal laminae, and the endothelium, but
appears in mammals, where this is the pre¬ there are areas in which the supporting cell
dominant form of organization. is absent and the chief cells are in close
relation to the capillary wall. These vascular
relationships are of some significance, for it
is not yet clear whether the paraganglion
THE PARAGANGLIA cells have an endocrine function, releasing
catecholamine into the blood, or whether the
The term paraganglia is used to describe chief cells are essentially interneurons syn-
small clusters of epithelioid cells that give a apsing on sympathetic neurons and exerting
chromaffin reaction. They are found rather an inhibitory effect on transmission in sym¬
widely scattered in the retroperitoneal tissue. pathetic ganglia.
Some of them are associated with sympathetic The parasympathetic paraganglia often do
ganglia, others with branches of the parasym¬ not give a conspicuous chromaffin reaction
pathetic nerves. The cells of the paraganglia and they were formerly classified by light
and those of the adrenal medulla are mor¬ microscopists as “achromaffin paraganglia”
phologically similar and of similar embryonic to distinguish them from those associated
origin. Together they make up the chromaffin with the sympathetic ganglia. It was even
system. In ordinary histological preparations suggested by some that they probably elabo¬
the cells of paraganglia appear pale or clear, rated acetylcholine. This distinction between
but they are stained by the chromaffin reac¬ chromaffin and achromaffin paraganglia re¬
tion. ceives no support from electron microscopic
These cells found in the ganglia and along studies and should probably be abandoned.
the nerves of the sympathetic nervous system The chief cells of vagal paraganglia contain
have now been studied electron microscopi¬ 50- to 200-nm electron-opaque granules that
cally and by histochemical procedures for the tend to be at the cell periphery or in its
identification of catecholamines. The para¬ processes. The supporting cells are somewhat
ganglia are surrounded by a rather thick less numerous than in sympathetic paragan¬
investment of collagenous connective tissue glia, and chief cells may adjoin one another
that extends inward between clumps of pa¬ and be attached by desmosomes. Nerve ter¬
renchymal cells. Two types of parenchymal minals apposed to the surface of the chief
cells are distinguishable, the chief cells and cells contain numerous clear synaptic vesicles.
supporting cells. The chief cells are irregular
in shape, with a single nucleus and well-
developed juxtanuclear Golgi complex. The
REFERENCES
endoplasmic reticulum is sparse, but occa¬
sional parallel associations of cisternae are
observed. Glycogen particles are present in ADRENAL CORTEX
small numbers scattered through the cyto¬ Baxter, J. D., and G. G. Rousseau: Glucocorticoid
plasmic matrix. In addition, there are nu¬ hormone action: an overview. In Baxter, J. D., and
merous membrane-limited, electron-opaque G. G. Rousseau, eds.: Glucocorticoid Hormone Ac¬
tion. Monogr. Endocrinol. 72:1, 1979.
granules 50 to 200 nm, closely resembling
534 • ADRENAL GLANDS AND PARAGANGLIA

Black, V. H., E. Robbins, E. McNamara, and T. Huima: Blaschko, H., G. Sayers, and A. D. Smith: Adrenal gland.
A correlated thin-section and freeze fracture anal¬ In Handbook of Physiology. Sect. 7, Endocrinology,
ysis of guinea pig adrenocortical cells. Am. J. Anat. Vol. 6. Washington, DC, American Physiological
756:453, 1979. Society, 1975.
Brown, M. S., P. T. Kovanen, and j. L. Goldstein: Coupland, R. E.: Electron microscopic observations on
Receptor mediated uptake of lipoprotein-choles¬ the structure of the rat adrenal medulla. I. Ultra¬
terol and its utilization for steroid synthesis in the structure and organization of chromaffin cells in
adrenal cortex. Recent Prog. Elorm. Res. 35:215, the normal adrenal medulla..J. Anat. 99:231, 1965.
1979. Landsberg, L., and J. B. Young: Catecholamines and
Gill, G. N.: ACTH regulation of the adrenal cortex. the adrenal medulla. In Bondy, P. K., and L. E.
Pharmacol. Ther. [B] 2:313, 1976. Rosenberg, eds.: Metabolic Control and Disease. 8th
Griffiths, K., and E. H. D. Cameron: Steroid biosynthetic ed. Philadelphia, W. B. Saunders Co., 1980, p. 1621.
pathways in the human adrenal. Adv. Steroid Smith, A. D., and H. Winkler: Catecholamines. In
Biochem. Pharmacol. 2:223, 1970. Blaschko, H., and E. Muschelli, eds.: Handbook
Lanman, J. T.: The fetal zone of the adrenal gland. of Experimental Pharmacology. Vol. 33. Berlin,
Medicine 52:389, 1953. Springer Verlag, 1972, p. 538.
Long, J. A.: Zonation of the mammalian adrenal cortex. Wurtman, R. J., and L. A. Pohorecky: Adrenal cortical
In Blaschko, H., G. Sayers, and A. D. Smith, eds.: control of epinephrine in health and disease. Adv.
Handbook of Physiology. Sect. 7, Endocrinology, Metab. Disord. 5:53, 1971.
Vol. 6. Washington, DC, American Physiological Yates, R. D.: A light and electron microscopic study
Society, 1975, p. 13. correlating the chromaffin reaction and granule
Long, J. A., and A. L. Jones: Observations on the fine ultrastructure in th^ adrenal medulla of the Syrian
structure of the adrenal cortex of man. Lab. Invest. hamster. Anat. Rec. 749:237, 1964.
77:355, 1967.
PARAGANGLIA AND PARA-AORTIC BODIES
Nussdorfer, G. G., G. Mazzocchi, and V. Meneghelli:
Cytophysiology of the adrenal zone fasciculata. Int. Brudin, T.: Catecholamines in the pre-aortic paraganglia
Rev. Cytol. 55:291, 1978. of fetal rabbits. Acta Physiol. Scand. 64:287, 1965.
Pohorecky, L. A., and R. J. Wurtman: Adrenocortical Chen, I., and R. D. Yates: Ultrastructural studies of
control of epinephrine synthesis. Pharmacol. Rev. vagal paraganglia in Syrian hamsters. Zeitschr. Zell¬
27:1, 1971. forsch. 766:309, 1970.
Coupland, R. E., and B. S. Weakley: Electron micro¬
ADRENAL MEDULLA scopic observations on the adrenal medulla and
Benedeczky, I., and A. D. Smith: Ultrastructural studies extra-adrenal chromaffin tissue in the post-natal
on the adrenal medulla of the hamster. Origin and rabbit. J. Anat. 766:213, 1970.
fats of secretory granules. Zeitschr. Zellforsch. Mascorro, J. A., and R. D. Yates: Microscopic observa¬
724:367, 1972. tions on abdominal sympathetic paraganglia. Tex.
Bennett, H. S.: Cytological manifestations of secretion Rep. Biol. Med. 26:59, 1970.
in the adrenal medulla of the cat. Am. J. Anat.
69:333, 1941.
 — 91
PINEAL GLAND

The pineal gland (epiphysis cerebri) is an en¬ rough endoplasmic reticulum and numerous
docrine gland that modulates the function of free polyribosomes. More abundant than the
the reproductive system. In seasonal breed¬ granular endoplasmic reticulum are tubular
ing species it responds to changes in day and vesicular elements of an atypical smooth
length and regulates gonadal activity. It is of endoplasmic reticulum (Fig. 21—6). Mito¬
less importance in species that breed contin¬ chondria are numerous in the perikaryon
uously. In the human brain it is a conical and quite variable in form. The Golgi com¬
gray body measuring 5 to 8 mm in length plex is moderately well developed and many
and 3 to 5 mm in its greatest width. It lies coated and smooth vesicles are associated
above the roof of the diencephalon at the with it. Some of these contain dense cores.
posterior extremity of the third ventricle. A In addition to the usual organelles, the
small ependyma-lined recess of the third ven¬ pinealocytes contain peculiar synaptic ribbons,
tricle extends into a short stalk, which joins which consist of a dense rod or lamella sur¬
the pineal body to the diencephalic roof (Fig. rounded by small vesicles that resemble the
21-1). synaptic vesicles of nerve endings. Similar
organelles are found in the photoreceptor
cells of the retina; in the organ of Corti in
the inner ear; and in certain other sensory
HISTOLOGICAL
organs where they are involved in synaptic
ORGANIZATION transmission. It has been proposed that the
pinealocytes of mammals originate from a
The organ is invested by pia mater, from sensory cell line that evolved from the pineal
which connective tissue septa containing photoreceptors of lower vertebrates. This
many blood vessels penetrate into the pineal interpretation fostered the belief that the
tissue and surround its clusters of epithelioid synaptic ribbons were phylogenetic relics with
cells. In sections stained with hematoxylin no functional significance. This view is no
and eosin, the pineal body is seen to consist longer espoused for it has been shown that
of cords of pale-staining epithelioid cells. there is a good correlation between the num¬
Their large nuclei are often irregularly in¬ ber of synaptic ribbons and the state of phys¬
folded or lobulated and have prominent nu¬ iological activity of the gland. Their function
cleoli. These cells, making up the bulk of the in the pineal is unknown. Where they occur
organ, are called pinealocytes. When appro¬ in sensory organs, synaptic ribbons are part
priately stained, the cytoplasm is moderately of the presynaptic complex and are thought
basophilic and often contains lipid droplets. to play a role in the transport of vesicles to
As described by del Rio-Hortega, who used the active sites of neurotransmitter release at
silver impregnation methods for their dem¬ the cell membrane. In the pinealocyte, they
onstration, human pinealocytes are cells with show no consistent relationship to other cel¬
long tortuous processes, which radiate from lular elements and may be found adjacent to
the cords toward the connective tissue septa other pinealocytes or glial cells, or in areas
where they end in bulbous swellings on or of the cell surface exposed to extracellular
near blood vessels (see Fig. 21—5). Such proc¬ spaces. They may be involved in transport
esses are visualized with some difficulty in and release of chemical mediators but they
routine preparations. do not appear to have a synaptic relationship
In electron micrographs the pinealocytes to other cells.
have nuclei of irregular outline with periph¬ Where synaptic ribbons are found in sen¬
erally distributed heterochromatin. The cy¬ sory organs they occur singly or in pairs, but
toplasm contains occasional short profiles of in pinealocytes they may be numerous. The
535
 536 • PINEAL GLAND

Group of cells with

little cytoplasm

Superior Cells with much cytoplasm

habenular
commissure

Ependyma
ineal recess "
Neuroglia Posterior end
of pineal body
Connection
with
posterior
commissure

Connective tissue sheath Group of cells with

cytoplasm
(pia mater)
little cytoplasm
Figure 21-1. Median section through pineal body of a newborn child. Blood vessels empty, x 32. A, Corpora arenacea
(sand granules) from the pineal body of a 69-year-old woman, x 160. (After Schaffer.)

pineal exhibits a circadian cycle of activity rier proteins called neuroepiphysins to distin¬
related to the periods of light and darkness. guish them from neurophysins, which are
In the dark phase of the cycle, when it is the carrier proteins of the neural lobe of the
most active, there may be a marked increase pituitary. When the vesicles are discharged
in the length of the synaptic rods or ribbons by exocytosis, the hormones become disso¬
and a two- to threefold increase in their ciated from the carrier protein and diffuse
number. They may assemble in sizable aggre¬ into the capillaries or the cerebrospinal fluid.
gations, called synaptic ribbon fields, containing The pineal body in humans and in cattle
20 or more of these organelles. contains extracellular concretions called cor¬
The pinealocytes arise from a neuroepi¬ pora arenacea or “brain sand” (Fig. 21-2).
thelium and have numerous gap junctions These are composed of calcium phosphates
that provide for electrical and metabolic cou¬ and carbonates in an organic matrix depos¬
pling of groups of cells. The cytoplasm often ited in concentric layers. Their significance
contains lipid droplets and deposits of lipo- and mode of formation are poorly under¬
chrome pigment, and is usually rich in mi¬ stood. They are not regarded as pathological
crotubules and intermediate filaments. The but rather as biproducts of secretory activity.
microtubules are randomly oriented in the It is suggested that the carrier proteins re¬
cell body but become associated in parallel leased during exocytosis bind calcium and
arrays in the cell processes. The shorter proc¬ become deposited in concentric layers
esses end blindly among the surrounding around portions of cellular debris that serve
cells, while the longer ones have bulbous as nucleation sites. The calcified corpora ar¬
terminal expansions in close proximity to enacea increase with age and are visible in
fenestrated blood capillaries or near epen¬ x-ray films or CAT scans of the head in 80
dymal cells lining the pineal recess. The end¬ per cent of individuals over 30 years of age.
ings may contain small dense-cored vesicles They are useful to the roentgenologist in the
that are believed to contain monoamines or diagnosis and localization of tumors or other
peptide hormones conjugated to specific car¬ expanding lesions in either hemisphere of
 PINEAL GLAND • 537

the brain, which may displace the pineal from whether these represent a cell type distinct
its normal midline position toward the op¬ from interstitial cells is not yet clear.
posite side.
The second cell type of the pineal is the
Histogenesis
interstitial cell. These cells occur in the peri¬
vascular areas and between the clusters of The pineal body first appears at about 36
pinealocytes. Their nuclei are elongated and days of gestation as a prominent thickening
stain more deeply than those of the paren¬ of the ependyma in the posterior part of the
chymal cells. The cytoplasm is somewhat roof of the diencephalon. Cells migrate from
more basophilic and drawn out into long this ependymal thickening to form a segre¬
processes. In electron micrographs the gran¬ gated mantle layer, whose cells tend to as¬
ular endoplasmic reticulum is well repre¬ sume a follicular arrangement that is gradu¬
sented, and free ribosomes are numerous. In ally transformed into the cordlike
addition, occasional deposits of glycogen are arrangement seen in later stages of develop¬
found. Rare microtubules are present, but ment. By the end of the sixth month, inter¬
these are overshadowed by a profusion of stitial and pineal parenchymal cells have dif¬
cytoplasmic filaments 5 to 6 nm in diameter ferentiated.
and of indeterminate length. They may occur It is generally believed that the pineal body
in large bundles or as single, randomly ori¬ increases in size until about 7 years of age.
ented filaments.
Some workers consider the interstitial cells,
Innervation
which represent about 5 per cent of the cells
in the pineal, to be glial elements. The abun¬ Nerves are found throughout the organ in
dance of filaments in their cytoplasm is in¬ silver-impregnated specimens. As the nerve
deed reminiscent of the fine structure of fibers penetrate into the organ, their myelin
astrocytes. The presence in rat pineal of sheaths terminate and the bare axons con¬
stellate cells that stain strongly with the acid tinue among the pinealocytes. Some of these
hematin method has been reported, but contain many small vesicles in the size range

Figure 21-2. Photomicrograph of human pineal, showing the characteristic concretions (pineal sand), x 200.
538 • PINEAL GLAND

of synaptic vesicles, suggesting that the HISTOPHYSIOLOGY

nerves have functional endings in close rela¬
tion to the parenchymal cells. The innerva¬ The pineal in mammals was long consid¬
tion appears to be exclusively via autonomic ered to be a vestigial organ of little functional
fibers originating in the superior cervical significance, but experimental investigations
sympathetic ganglion. of the past 20 years have firmly established
it as an endocrine gland whose primary func¬
tion-is the modulation of reproduction.
Pineal System of Lower Vertebrates The pineal secretes an indolamine hor¬
In contrast to the single, relatively solid mone, melatonin. The gland contains a higher
mass of tissue making up the pineal body in concentration of serotonin than any other
adult mammals, the organs in most lower organ. Serotonin is converted by the enzyme
vertebrates remain saccular throughout life serotonin TV-acetyltransferase to A-acetylser-
and are somewhat more complex. The pineal otonin, which in turn is converted to mela¬
system may consist of a single pineal sac (the tonin by hydroxyindole-O-methyltransferase.
intracranial epiphysis), as in most fishes and A unique feature of the pineal is that its
tailed amphibians, or it may be double. In
the latter case (primitive fishes, tailless am¬
phibians, and lizards) a second, parapineal
organ results either from elaboration of an
anterior end vesicle of the epiphysis, or from
a separate evagination from the diencephalic
roof situated more anteriorly. In frogs, the
parapineal component (the frontal organ)
comes to lie subepidermally and is discernible
externally on the median dorsal aspect of the
head. It is connected with the intracranial
epiphysis by a long pineal nerve. While many
nerve fibers and their endings are readily
demonstrable in pineal systems of lower ver¬
tebrates, there is no evidence that any are
sympathetic.
Electron microscopic examination of adult
saccular pineal systems reveals that, in most
lower vertebrates, the principal cell type is an
apparent photoreceptor. This cell closely re¬
sembles a retinal rod or cone, both in the
form of the membranous lamellated modi¬
fied flagellum that protrudes from the cell
apex into the pineal lumen and in the pres¬
ence of characteristic receptor synapses at
the cell base. Physiological data indicate that,
in these species, impulses course along pineal
tracts to surrounding brain regions in re¬
sponse to darkness or to light of various
wavelengths. The most elaborate pineal pho¬
toreceptor systems are found in certain liz¬
ards, such as the Tuatara (Sphenodon), in
which, in addition to a photoreceptor-type
intracranial epiphysis, the parapineal com¬
ponent (or parietal “eye”) is specialized to the
extent of possessing a distinct lens and a
retina composed of photoreceptor cells
backed by supportive cells containing pig¬ Figure 21-3. Section of pineal body of man stained with
hematoxylin and eosin, showing irregularly shaped cells
ment. Photoreceptor cells have not been dis¬ and their processes. Note the blood vessel in the center.
tinguished in mammalian pineal systems. Compare with Figure 21-5.
 PINEAL GLAND • 539

Figure 21-4. Photomicrograph of bovine pineal. The pinealocytes have large nuclei with prominent nucleoli. The outlines
of the stellate cells are not easily distinguished. Many densely stained processes are visible in this preparation, but it is
not possible to determine which belong to pinealocytes and which to interstitial cells. Iron hematoxylin stain, x 500.
(Courtesy of E. Anderson.)
540 • PINEAL GLAND

the evidence is not compelling. Various pep¬

tides normally found in the hypothalamo-
hypophyseal system have also been reported
in the pineal, but their localization and pineal
origin are disputed.
The pineal is involved in the reproductive
process to varying degree in different mam¬
malian species. The reproductive system of
the laboratory rat responds little and incon¬
sistently to experimental manipulation of the
pineal. This may be a consequence of being
inbred for many generations under unnatu¬
ral conditions that may have led to loss of
mechanisms essential for survival of a species
in the wild. The scientific community’s heavy
reliance upon the laboratory rat no doubt
contributed to slow progress in defining the
role of the pineal in reproduction. Many
species that inhabit temperate or arctic zones
are seasonal breeders, and it is important for
the young to be born in the spring to ensure
an appropriate ambient temperature and the
availability of food for their survival. At other
seasons of the year the gonads may regress
and only regain reproductive competence
with the approach of spring. In such animals
the critical environmental factor regulating
their seasonal pattern of reproduction is day
length (photoperiod). Information about the
photoperiod is transmitted to the reproduc¬
tive system via the pineal.
The photoreceptors for light detection are
in the retina. Information is carried from the
eyes over a neural pathway involving retino-
hypothalamic fibers, the suprachiasmatic nu¬
Figure 21-5. Specifically impregnated section of the clei, the hypothalamospinal fibers, and
pineal body of a young boy, showing interlobular tissue thence to the sympathetic nervous system and
(C) and vessel (D) with club-shaped processes of specific
via the cervical sympathetic ganglion to the
cells in its adventitia. Note parenchymatous cells and their
claviform processes bordering on C. (After del Rio-Hor- pineal. Interruption of this pathway severs
tega.) the link between photoperiod and pineal
function.
The influence of the pineal on reproduc¬
biosynthetic activity exhibits a diurnal rhyth- tion has been most thoroughly studied in the
micity correlated with the periods of daylight Syrian hamster, a species adopted as a labo¬
and darkness. Melatonin concentrations in ratory animal more recently than the rat.
the gland and in the blood increase during Either cutting the optic nerves or exposure
the dark phase of the diurnal cycle and there of intact hamsters to a short photoperiod
is evidence of seasonal variations. When ap¬ (less than 12.5 hours daily) for four weeks or
plied to the skin of amphibians, the hormone longer enhances melatonin secretion by the
causes aggregation of pigment granules in pineal and results in atrophy of the testes.
melanophores and results in blanching. Its Either pinealectomy or superior cervical gan-
effect is therefore opposite to that of mela¬ glionectomy prevents the gonadal atrophy
nocyte stimulating hormone of the pituitary. normally induced by shortening the photo¬
In mammals it has an antigonadotropic ac¬ period.
tion. In the annual cycle of seasonal breeders,
d he gland may also produce physiologi¬ the shortening of the day as autumn ap¬
cally active peptides. The most studied of proaches leads to enhanced melatonin secre¬
these is arginine vasotocin, which is reported tion by the pineal and atrophy of the gonads,
to have anti-FSH and anti-LH activity, but which remain inactive through the winter
Figure 21-6. Electron micrograph of a juxtanuclear area of pinealocyte cytoplasm, showing a centriole and part of the
Golgi complex, with associated vesicles with dense osmiophilic content. The cytoplasm generally contains abundant
small vesicles and numerous microtubules, x 30,000. (Courtesy of E. Anderson.)

>

CD
>
o
=3
T3
O
Q.
CD

CD
>
CD

Autumnal Vernal Summer

Winter
Equinox Solstice Equinox Solstice

i i i L
JULI AUG I SEP I OCT | NOV I DEC I JAN | FEB I MAR I APR I MAY I JUN I JUL

Size of
Testis
Figure 21-7. Schematic depiction of the relationships between the pineal gland and the annual reproductive cycle of a
seasonal breeding species such as the hamster. (From Reiter, R. J. Endocr. Rev. 7:109, 1980.) 54-1
542 • PINEAL GLAND

(Fig. 21-7). Although the pineal continues to Kitay, J. E, and M. D. Altschule: The Pineal Gland; A
Review of the Physiologic Literature. Cambridge,
secrete melatonin in this period, the neu¬
Harvard University Press, 1954.
roendocrine reproductive complex appears Knight, B. K., M. M. Hayes, and R. B. Symington: The
to become refractory to its suppressive effect, pineal gland—a synopsis of present knowledge with
and recrudescence of the reproductive or¬ particular emphasis on its possible role in control of
gans begins in the latter part of winter. This gonadotrophin functions. S. Afr. J. Anim. Sci.
3:143, 1973.
ensures that the animals will be sexually com¬ Kumado, K., and W. Mori: A morphological study of
petent when they emerge from hibernation the circadian cycle of the pineal gland of the rat.
in the spring. The refractory state subsides Cell Tissue Res. 182:565, 1977.
during the long days of the summer months Lerner, A. B., and J. D. Case: Pigment cell regulatory
factors. J. Invest. Dermatol. 32:221, 1959.
and the system again becomes responsive to
Lukaszyk, A., and R. J. Reiter: Histophysiologic evidence
the antigonadotropic action of the pineal. All for the secretion of polypeptides by the pineal gland.
the effects of the naturally occurring changes Am. J. Anat. 743:451, 1975.
in photoperiod can be duplicated in the lab¬ Matsushima, S., Y. Morisawa, I. Aida, and K. Abe:
oratory by administration of melatonin at the Circadian variations in pinealocytes of the Chinese
hamster Cricetus vriseus. Cell Tissue Res. 225*:231,
appropriate time and dosage. It is not yet
1983.
clear whether the pineal hormone brings Oksche, A.: Survey of the development and comparative
about reproductive incompetence by acting morphology of the pineal organ. Prog. Brain Res.
upon the hypothalamus or directly upon the 10:3, 1965.
gonads, and the mechanisms for develop¬ Quay, W. B.: Reduction of mammalian pineal weight
and lipid during continuous light. Gen. Comp. En¬
ment of, and recovery from, the refractory docrinol. 7:211, 1961.
state have yet to be elucidated. It is likely Quay, W. B.: Experimental and cytological studies of
that, with minor variations, the relationship pineal cells staining with acid hematin in the rat.
of the pineal to the control of the annual Acta Morphol. Neerl. Scand. 5:87, 1962.
Quay, W. B.: Circadian rhythm in rat serotonin and its
reproductive cycle in the hamster will be
modification by estrous cycles and photoperiod.
found to apply to other species living in Gen. Comp. Endocrinol. 3:473, 1963.
climatic zones where survival of the young Quay, W. B.: Retinal and pineal hydroxyindole-O-methyl
requires restriction of breeding to a favorable transferase activity in vertebrates. Life Sci. 4:983,
time of year. 1965.
Reiter, R. J.: Comparative physiology: pineal gland.
The degree of pineal participation in the
Annu. Rev. Physiol. 35:305, 1973.
reproduction of humans and other continu¬ Reiter, R. J.: Pineal control of seasonal reproductive
ous breeders is less clear. There is no doubt, rhythm in male golden hamsters exposed to natural
however, that the pineal exerts some degree daylight and temperature. Endocrinology 92:423,
1973.
of antigonadotropic influence in these species
Reiter, R. J.: The pineal and its hormones in the control
also. Pinealectomy or superior cervical gan- of reproduction in mammals. Endocr. Rev. 7:109,
glionectomy in young rats leads to enlarge¬ 1980.
ment of the reproductive organs and preco¬ Roth, W. D., R. J. Wurtman, and M. D. Altschule:
cious puberty. Conversely, sustained injection Morphologic changes in the pineal parenchymal
cells of rats exposed to continuous light or darkness.
of melatonin delays puberty. Young boys
Endocrinology 77:888, 1962.
with brain tumors that destroy the pineal also Soriano, F. M., H. A. Wether, and L. Volbrath: Corre¬
exhibit precocious puberty. lation of the number of pineal “synaptic” ribbons
and spherules with the level of serum melatonin
over a 24-hour period in male rabbits. Cell Tissue
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Ariens-Kappers, J., and J. P. Schade, eds.: Structure and changes in microtubules, synaptic ribbons and syn¬
function of the epiphysis cerebri. In Progress in aptic ribbon fields in the pinealocytes of the baboon
Brain Research. Vol. 10. Amsterdam, Elsevier Pub¬ Papio ursinus. Cell Tissue Res. 27 7:405, 1981.
lishing Co., 1965. Wartenberg, H.: The mammalian pineal organ: electron
Arstila, A. U.: Electron microscopic studies on the struc¬ microscopic studies on the fine structure of pineal¬
ture and histochemistry of the pineal gland of the ocytes, glial cells, and on the perivascular compo¬
rat. Neuroendocrinology (Suppl.) 2:1, 1967. nent. Z. Zellforsch. 86:74, 1968.
Axelrod, J., and H. Weissbach: Purification and prop¬ Wetterberg, L.: Melatonin in humans. Physiological and
erties of hydroxyindole-O-methyl transferase. J. clinical studies. J. Neural Transm. (Suppl.) 73:289,
Biol. Chem. 236:216, 1961. 1978.
Benson, B.: Current status of pineal peptides. Neuroen¬ Wolstenholme, G. E., and J. Knight, eds.: The Pineal
docrinology 24:241, 1977. Gland. Ciba Foundation Symposium. London,
Fiske, V. M., J. Pound, and K. Putnam: Effect of light Churchill, 1971.
on the weight of the pineal organ in hypophysec- Wurtman, R. J.: Effects of light and visual stimuli on
tomized, gonadectomized, adrenalectomized or endocrine function. In Ganong, W. F., and L. Mar¬
thiouracil-fed rats. Endocrinology 77:130, 1962. tini, eds.: Neuroendocrinology. New York, Aca¬
Kelly, D. E.: Pineal organs: photoreception, secretion demic Press, 1966.
and development. Am. Sci. 50:597, 1962.
 ^22
SKIN

The skin covers the surface of the body prints. It is well known that the complicated
and consists of two main layers, the surface patterns of ridges found on the fingers are
epithelium, or epidermis, and the subjacent subject to such marked variations that their
connective tissue layer, the corium or dermis impressions are a dependable means for
(Figs. 22—1, 22—2). Beneath the dermis is a identification of individuals. The same de¬
looser connective tissue layer, the superficial gree of variation holds for skin patterns in
fascia, or hypodermis, which in many places is other regions, but these are less commonly
largely transformed into subcutaneous adi¬ used.
pose tissue. The hypodermis is loosely con¬ The interface of the epidermis and the
nected to underlying deep fascia, aponeu¬ dermis is also uneven. A pattern of ridges
rosis, or periosteum. The skin is continuous and grooves on the deep surface of the epi¬
with several mucous membranes at mucocu¬ dermis fits a complementary pattern of cor¬
taneous junctions. Such junctions are found at rugations on the underlying dermis. The
the lips, nares, eyelids, vulva, prepuce, and projections of the dermis have traditionally
anus. been described as dermal papillae and those of
The skin is one of the largest of the organs, the epidermis as epidermal ridges, owing to
making up some 16 per cent of the body their respective appearances in vertical sec¬
weight. Its functions are several. It protects tions of skin. As will be seen later, these
the organism from injury and desiccation; it terms are not always accurately descriptive of
receives stimuli from the environment; it their three-dimensional configuration as seen
excretes various substances; and, in warm¬ in whole mounts.
blooded animals, it takes part in thermoreg¬ Although the boundary between the epi¬
ulation and maintenance of water balance. thelial and the connective tissue portions of
The subcutaneous adipose tissue has an im¬ the skin is sharp, the fibrous elements of the
portant role in fat metabolism. dermis merge with those of the hypodermis,
The specific functions of the skin depend so that there is no clear-cut boundary be¬
largely upon the properties of the epidermis. tween these layers.
This epithelium forms an uninterrupted cel¬
lular investment covering the entire outer
surface of the body, but it is also locally
specialized to form the various skin appen¬
THE EPIDERMIS
dages: hair, nails, and glands. Its cells produce
the fibrous protein keratin, which is essential The epidermis is a stratified squamous ep¬
to the protective function of the skin, and ithelium composed of cells of two distinct
melanin, the pigment that protects against lineages. Those comprising the bulk of the
ultraviolet irradiation. The epidermis gives epithelium undergo keratinization and form
rise to two main types of glands, one of which the dead superficial layers of the skin. They
produces the watery secretion sweat and the are derivatives of the ectoderm covering the
other the oily secretion sebum. embryo, and they constitute the keratinizing
The free surface of the skin is not smooth system. There are also cells in the deeper
but is marked by delicate grooves or flexure layers of the epidermis that do not keratinize
lines, which create patterns that vary from but are capable of producing the pigment
region to region. They are deeper on non- melanin. These are the melanocytes, which
hairy areas, such as knees and elbows, palms arise from the embryonic neural crest and
and soles. The most familiar of the surface invade the skin in the third to sixth months
patterns are those responsible for the finger¬ of intrauterine life. Collectively these cells
543
544 • SKIN

Hair follicle
Hair Duct of sweat gland

Epidermis

Sebaceous gland Dermis

Sweat gland

Subcutaneous
fat tissue

Blood vessel

Lymphatic vessel
Figure 22-1. Section through human thigh perpendicular to the surface of the skin. Blood vessels are injected and
appear black. Low magnification. (After A. A. Maximow.)
 SKIN • 545

Hair

-♦Epidermis

Arrector -Dermis

pili muscle

Hair
follicle

Hair
papilla
Subcutaneous
adipose
tissue

Blood
vessels

Galea
aponeurotica

Figure 22-2. Section of the skin of the scalp, x 15. (Courtesy of H. Mizoguchi.)

comprise the pigmentary system of the skin. In to 30 days, depending on the region of the
addition, there are two other cell types, not body and a number of other factors.
part of the keratinizing system. These are
the Langerhans cells and the Merkel cells. Epidermis of the Palms and Soles
The epidermis varies from 0.07 to 0.12
mm in thickness over most of the body, but The structural organization of the epider¬
it may reach a thickness of 0.8 mm on the mis can be studied to advantage in those
palms and 1.4 mm on the soles. In the fetus, areas of the skin where it attains its greatest
these sites are already appreciably thicker thickness—namely, the palm of the hand and
than other areas of skin, but continuous fric¬ the sole of the foot. In sections perpendicular
tion or pressure in postnatal life may cause to the surface (Figs. 22-3, 22-4), four prin¬
considerable additional thickening of the cipal layers can be distinguished—stratum ba-
outer layer of the epidermis on exposed areas sale, stratum spinosum, stratum granulosum, and
of the body surface. stratum corneum.
The superficial keratinized cells of the skin The stratum basale consists of a single layer
are continually exfoliated from the surface of cells resting upon the basal lamina and
and are replaced by cells that arise from underlying dermis. Its cells are cuboidal or
mitotic activity in the basal layer of the epi¬ low columnar. The nucleus is relatively large,
dermis. The cells produced there are dis¬ the cytoplasm basophilic. In electron micro¬
placed to successively higher levels by the graphs, there are a few profiles of rough
generation of new cells below them. As they endoplasmic reticulum, and a small Golgi
move upward, they elaborate keratin, which complex. The cytoplasmic matrix is rich in
accumulates in their interior until it largely ribosomes and contains abundant 10-nm fil¬
replaces all metabolically active cytoplasm. aments occurring singly or in conspicuous
The cell dies and its nucleus and other or¬ bundles. Desmosomes attach neighboring
ganelles disappear. It is finally shed as a cells, and hemidesmosomes are found along
flakelike, lifeless residue of a cell. This se¬ the membrane adhering to the basal lamina.
quence of changes, referred to as the cyto- Mitotic figures are common in this layer,
morphosis of the keratinocyte, takes from 15 formerly called the stratum germinativum be-
546 • SKIN

Stratum
disjunctum

Stratum corneum

Duct of
sweat gland

Stratum
lucidum
Stratum
granulosum

Tangential
sections
through
dermal
papillae
Blood
Stratum uessel
malpighii

Papillary
layer of
dermis

Reticular Duct of
layer of sweat gland
dermis

Figure 22-3. Section of human sole perpendicular to the free surface, x 100. (After A. A. Maximow.)
 SKIN • 547

Figure 22-4. Skin of the human finger tip, illustrating a very thick stratum corneum. Hematoxylin and eosin. x 65.

cause the proliferation of its cells is mainly shaped coarse granules that stain intensely
responsible for the continual renewal of the with basic dyes and'hematoxylin. These ker-
epidermis. As the cells generated here move atohyalin granules have no limiting membrane
up into the stratum spinosum, they assume a and consist of closely packed, dense, 2-nm
flattened, polyhedral form with their long subunits (Fig. 22—8). Tonofilaments may pass
axis parallel to the surface and their nucleus through them or be partially incorporated in
somewhat elongated in this direction. their periphery. The origin and chemical
The cells of the stratum spinosum are some¬ nature of keratohyalin granules have not
what basophilic and have the same comple¬ been clearly established. They were formerly
ment of organelles as their precursors. In believed to be associated with the process of
addition, electron micrographs show numer¬ keratinization, but this is questioned because
ous small granules with a distinctive substruc¬ they are not found in all keratinizing epithe-
ture, called lamellated granules or membrane¬ lia. They are absent in finger- and toenails,
coating granules. These ovoid, membrane- for example, but abundant in the epidermis
bounded granules are 0.1 to 0.5 |xm in di¬ forming the hooves of cattle.
ameter, containing parallel lamellae about 2 The lamellar bodies that first appear in the
nm thick, usually oriented transverse to the stratum spinosum increase in number in the
long axis. The lamellar structure consists of stratum granulosum, where they collect at
alternating electron-lucent and -dense layers. the cell periphery, and their contents are
Bundles of 10-nm filaments, corresponding secreted into the intercellular spaces. In the
to the tonofibrils seen with the light micro¬ granular layer, they occupy as much as 15
scope, are a prominent feature of these cells. per cent of the cytoplasmic volume. The
They extend into the spinous processes and lamellar bodies contain little phospholipid
terminate in the dense plaques of the des- but appear to be rich in glycolipids and
mosomes (Fig. 22—6). sterols. When released into the intercellular
The stratum granulosum consists of three to spaces, they form multiple membrane bilay¬
five layers of flattened cells whose distin¬ ers arranged in broad sheets between the
guishing feature is the presence of irregularly cells. Correlated with release of the contents
548 • SKIN

Nucleus

A •, \\ V'* % % I ***,>^11*15^

> few.g** s&« ^*«sa

- »**V3V *■ *

ate; -

£*«£
,1*#* % • , %* .J*

Papilla
Mitochondria
Figure 22-5. Section tangential to the surface, of the malpighian layer of epidermis of human palm, showing fibrils and
so-called “intercellular bridges” in cross section at x and in longitudinal section at y. The junction of the scalloped lower
surface of the epithelial cells with the dermis is at z.

plli

Figure 22-6. Electron micrograph of parts of three cells from the stratum germinativum of human epidermis, x 12,000.
(Courtesy of G. Odland.)
Figure 22-7. Electron micrograph of portions of two adjoining epidermal cells. The junction of the two cells runs
diagonally across the figure. Desmosomes attaching the apposed cell surfaces are indicated by the arrows. Bundles of
epidermal filaments run in various directions in the cytoplasm and terminate in desmosomes. x 50,000. (Courtesy of
G. Odland.)

Figure 22-8. Electron micrograph of cells of the stratum granulosum, running diagonally across the figure and containing
irregularly shaped keratohyalin granules. At upper left are several cell layers of the stratum corneum. x 12,000.
(Courtesy of G. Odland.)
549
550 • SKIN

of the lamellar bodies is an increase in the from the lamellar granules of the strata spi-
volume of the intercellular space from less nosum and granulosum.
than 1 per cent to 5 to 30 per cent of the Two additional layers are recognized by
total tissue volume. The release of the con¬ some histologists in the stratum corneum.
tent of the lamellar bodies appears to provide The stratum lucidum consists of a few layers
waterproofing intercellular lipids that are es¬ of closely compacted, highly refractile eosino¬
sential to the barrier function of the epider¬ philic cells on its deep surface. It appears in
mis. histological sections as a wavy clear stripe
The stratum corneum consists of many layers immediately above the stratum granulosum.
of flat, cornihed cells (Figs. 22—4, 22—10). The nucleus of the cells has already disap¬
The processes by which the cells were joined peared and keratinization of the cells is well
in the spiny layer are no longer present. The advanced. The outermost layers of the stra¬
cells lack a nucleus, and few remnants of tum corneum, where the fully keratinized,
organelles remain, nearly all of the cytoplasm lifeless cells are loosening and desquamating,
having been replaced by 8- to 10-nm fila¬ is sometimes referred to as the stratum dis-
ments of keratin embedded in an amorphous junctum. Electron micrographs reveal no dis¬
matrix. The plasma membrane appears tinctive cytological features of the cells in
thickened because of deposition of a layer of these regions, and their designation as sepa¬
dense nonkeratinous material on its inner rate layers of the stratum corneum probably
aspect, and is highly corrugated, its ridges serves no useful purpose.
interdigitating with the grooves of adjacent
cells. The abundant desmosomes present in Epidermis of the Body in General
the granular layer are still detectable but
greatly modified, with their two halves often On the rest of the body the epidermis is
widely separated. The intercellular spaces are much thinner and simpler in its structure
occupied by lipid-containing material derived (Figs. 22-11, 22—12). The stratum spinosum

Figure 22-9. Electron micrograph showing a portion of a cell of the granular layer (lower right) and several layers of
flattened cells of the stratum corneum (upper left). Area enclosed in rectangle is shown at higher magnification in Figure
22-10. Osmium fixation, x 22,500. (Courtesy of G. Odland.)
 SKIN • 551

Figure 22-10. Electron micrograph of the area of stratum corneum of human epidermis enclosed in the rectangle in
Figure 22-9. The cytoplasm of the flat keratinized cells appears devoid of organelles and seems to consist mainly of
closely packed, fine filaments embedded in a rather dense matrix. The desmosomes, indicated by arrows, have an
unusually thick, dense intermediate layer. The clear spaces between the cells are, in part, artifacts of specimen
preparation. Osmium fixation, x 62,000. (Courtesy of G. Odland.)

Stratum
corneum
Stratum
granulosum
Stratum
malpighii

Vessel
Papillary layer
of dermis

Vessel

Reticular layer
of dermis
Figure 22-11. Section through skin of the human shoulder, x 125. (After A. A. Maximow.)
552 • SKIN

Figure 22-12. Photomicrograph of skin of the abdomen. Compare the thickness of the stratum corneum with that of
the finger tip in Figure 22-4. x 60.

and stratum corneum are always present, genes and mRNAs. At different stages of
although the latter may be relatively thin. A their cytomorphosis, the keratinocytes pro¬
granular layer consisting of two or three duce different keratins. The cells in the basal
layers of cells is usually identifiable, but a layer contain only keratins of low molecular
definite stratum lucidum is seldom seen in weight, but as they move upward they pro¬
the thinner epidermis of the general body duce increasing amounts of the larger kera¬
surface. tins that make up the bulk of the cornified
The epidermis is entirely devoid of blood layer. In this layer, the keratins are also
vessels; it is nourished from capillaries in the extensively cross-linked by disulfide bonds.
underlying connective tissue by diffusion Another characteristic feature of keratin¬
through tissue fluid, which occupies an ex¬ ocytes late in their differentiation is the pres-
tensive system of intercellular spaces. Human
skin, unlike that of practically all other ver¬
tebrates, blisters after exposure to thermal
and certain chemical stimuli.

CELL TYPES OF THE EPIDERMIS

Keratinocytes
The principal cell type of the epidermis
and of other stratified squamous epithelia is
now called the keratinocyte, because of its ca¬
pacity to synthesize keratin. While keratin
filaments are a component of the cytoskele-
ton of most epithelia, it is most abundant in
the keratinocytes of the epidermis. As these
cells differentiate and move upward in the
Figure 22-13. Photomicrograph of a whole mount of a
epithelium, their content of keratins steadily sheet of epidermis from the thigh spread upon a slide and
increases, ultimately constituting 85 per cent viewed from the underside to illustrate the meianocytes.
of the total protein of the stratum corneum. The epidermis was separated from the dermis by treat¬
Keratin is not a single protein but a family ment of the excised skin with trypsin. The epidermal sheet
was then incubated in 1,3,4-dihydroxyphenylalanine
of polypeptides ranging from 40,000 to (DOPA), which selectively stains the melanocytes. Notice
70,000 M.W., which are products of different their branching process, x 300. (Courtesy of G. Szabo.)
 AREA AND NUMBER OF
NUMBER MELANOCYTES
SPECIES OF INDIVIDUALS (per mm.2 ± s.e.)

Man1
Caucasoid Thigh (35) 1000 ± 70
Mongoloid Thigh ( 3) 1290 ± 45
Negroid Thigh ( V) 1415 ± 255
Figure 22-14. Comparison of melanocyte numbers
Guinea Pis2
o
among color variants of mammals.
Black Ear ( 8) 920 ± 145
Red Ear ( 8) 865 ± 100

Mouse3
C57 Black Tail ( 4) 590 ± 65
DBL Dilute Tail ( 4) 590 ± 165

1Szabo (1969) and Szabo; Gerald et al. (1971).

2Billingham and Medawar (1953).
3Gerson and Szabo (1969).

Figure 22-15. Electron micrograph of a developing hu¬

man melanosome from the retina, showing the periodicity
in its structural framework. When the melanosome is fully
developed, its interior structure is obscured by the accu¬
mulated melanin. (Courtesy of A. Breathnach.)

Figure 22-16. Electron micrograph of a heavily pigmented keratinocyte from the stratum malpighii of human skin.
Whereas the melanosomes of melanocytes occur singly, those of keratinocytes are found in clusters of varying size
enclosed by a membrane. (Courtesy of G. Szabo.)

553
554 • SKIN

Figure 22-18. Section of human epidermis, showing gold

impregnated Langerhans cells at a high level in the
stratum malpighii. Gairn’s gold chloride technique. (After
Breathnach, A. S. Int. Rev. Cytol. 78:1, 1965.)

atively few organelles (Fig. 22—19). The ab¬

sence of desmosomes, melanosomes, and
bundles of tonohlaments serves to distinguish
them from the other cells of the epidermis.
Figure 22-17. Micrograph of a portion of a cell from the
stratum granulosum, showing dense keratohyalin gran¬
They contain numerous small vesicles, multi-
ules and several clusters of melanosomes. (Courtesy of vesicular bodies, and a few dense granules,
G. Szabo.) possibly lysosomes. Their most distinguishing
characteristic is the presence of peculiar,
ence of a dense layer (12 nm) of an insoluble membrane-bounded, rod-shaped granules
protein deposited on the inner aspect of the variously called Birbeck granules, Langerhans
plasma membrane. In the deposition of this cell granules, or vermiform granules. These are
layer, a soluble protein precursor called in- 15 to 50 nm long, 4 nm wide, with a centrally
volucrin is cross-linked by isopeptide bonds to placed linear density and faint striations ra¬
form an insoluble envelope. As in the case of diating from it to the enclosing membrane
the large keratins, involucrin is absent in the (Fig. 22-20).
basal layers but appears as the cells move Dendritic cells containing similar granules
toward the free surface of the epithelium. have been identified in lymph nodes, spleen,
and thymus. The Langerhans cells of the
skin have been shown to migrate from the
Langerhans Cells
skin to regional lymph nodes. The origin and
Throughout the epidermis, but particu¬ function of these cells has long been contro¬
larly in the upper layers of the stratum spi- versial. Evidence has recently accumulated
nosum, there are peculiar dendritic cells first indicating that they may participate in the
described by Langerhans in 1868 (Fig. 22— body’s immune responses. They have been
18). In routine hematoxylin and eosin prep¬ shown to possess surface la, Fc, and C3 re¬
arations, they have a dark-staining nucleus ceptors in common with macrophages. At
surrounded by a pale clear cytoplasm. When sites of allergic contact, cutaneous hypersen¬
stained by the gold chloride method, they sitivity lymphocytes are observed to gather
are blackened and their stellate or dendritic around Langerhans cells in close apposition
form is revealed. Their slender processes to their surface soon after antigenic chal¬
extend into the intercellular spaces among lenge. It is believed that Langerhans cells
the cells of the stratum spinosum and appear may be involved in presentation of antigen
to form an almost continuous network in the to lymphocytes and may participate in im-
epidermis. In electron micrographs, the nu¬ munoproliferative processes in the regional
cleus is highly irregular in outline, and the lymph nodes. The Langerhans cells are now
cytoplasm is of low density and contains rel- believed to be important agents in contact
Figure 22-19. Electron micrograph of a Langerhans cell surrounded by keratinocytes containing dense bundles of
filaments. The polymorphous appearance of the nucleus is typical. The stellate form of the cell is not evident here
because none of the processes is included in the plane of section. (Courtesy of G. Szabo.)

Figure 22-20. A small area of cytoplasm of a Langerhans cell, including one of the pair of centrioles, the Golgi complex,
and several vermiform granules (at heavy arrows). One of these is shown at higher magnification in the inset. The
dense granules in the cytoplasmic matrix are glycogen. (Courtesy of G. Szabo.)
555
556 • SKIN

allergic responses and other cell-mediated to these cells and appear to form expanded
reactions of the skin. endings applied to their surface. The nerve
endings contain no synaptic vesicles and are
presumed to be sensory. Because of their
Merkel Cell content of dense-cored vesicles and their in¬
The Merkel cell occurs in small numbers timate relationship to nerve terminals, the
in the basal portion of the epidermis. It bears Merkel cells are now considered to be para-
a superficial resemblance to the keratinocytes neurones involved in sensory reception.
to which it may be attached by desmosomes.
The nucleus is deeply invaginated (Figs. 22- Melanocyte
21, 22—22) and occasionally contains a pecu¬
liar inclusion consisting of a fascicle of The color of the skin is the resultant of
straight parallel filaments. The cytoplasm three components. The tissue has an inherent
contains bundles of filaments in the perinu¬ yellowish color, attributable in part to caro¬
clear zone and at the periphery but these are tene. The oxyhemoglobin in the underlying vas¬
less abundant than in the keratinocytes. A cular bed imparts a reddish hue, and shades
few transferred melanosomes may be pres¬ of brown to black are contributed by varying
ent. The most distinctive feature of the Mer¬ amounts of melanin. Of these three colored
kel cell is the presence in the cytoplasm of substances, only the melanin is produced in
80-nm dense-cored vesicles resembling those the skin. It is the product of specialized cells
found in cells of the adrenal medulla or with elaborately branching processes called
carotid body. However, it has not been estab¬ melanocytes, which are located in the basal
lished that these contain catecholamines. layer of the epidermis or in the underlying
The Merkel cells tend to be associated with connective tissue of the dermis. Although
areas where the dermis is especially well vas¬ melanin granules are also found in the ker¬
cularized and richly innervated. Unmyeli¬ atinocytes, they are formed only by the epi¬
nated axons are found in close relationship dermal melanocytes, for these cells alone pos-

Figure 22 21. Electron micrograph of the base of the human epidermis, showing a Merkel cell surrounded by
keratinocytes. Notice its pale cytoplasm and characteristic dense granules. (Courtesy of G. Szabo.)
 SKIN • 557

Figure 22-22. Portion of a Merkel cell from human gingival epithelium. The cytoplasm contains numerous dense
granules and filaments. The nucleoplasm may contain an unusual inclusion consisting of paracrystalline aggregations
of slender filamentous subunits (inset). (Micrograph courtesy of R. Winkelmann.)

sess the enzyme tyrosinase that is necessary basal epidermal cells varies between 1 to 4
for synthesis of the pigment. The fully and 1 to 10, depending on the region of the
formed melanin granules are transferred body. The melanocytes in the cheek and
from the melanocytes to the keratinocytes by forehead and in the genital, nasal, and oral
an unusual form of activity sometimes re¬ epithelium are about twice as numerous as
ferred to as cytocrine secretion. The melano¬ in other parts of the body surface. It is also
cytes are commonly located at the dermo- of interest that the number of melanocytes is
epidermal junction with their pigment-con¬ approximately the same in all human races.
taining processes extending for long dis¬ Racial differences in color are attributable to
tances upward into the interstices among the differences in the amount of pigment that
keratinocytes. They are not attached to the these cells produce and transfer to the kera¬
other cells by desmosomes, and in specimen tinocytes (Fig. 22-14).
preparation they may shrink away so that Melanin is formed in a specific cell particle,
they are surrounded by a clear space. Be¬ the melanosome?. In the human it is an elon¬
cause of their tendency to pass pigment to gated body with rounded ends, measuring
the keratinocytes, the melanocytes may ac¬ about 0.2 by 0.6 |xm, with a fibrillar or la¬
tually contain less melanin than the neigh¬ mellar internal structure exhibiting charac¬
boring epidermal cells, and their processes teristic periodic density variations along its
(or dendrites) are very difficult to identify in length in early stages of development. This
sections stained with hematoxylin and eosin. internal structure tends to be obscured by
They are best studied in whole mounts of accumulation of dense melanin in the mature
separated epidermis that have been treated melanosome. The size, shape, and internal
with 1,3,4-dihydroxyphenylalanine (DOPA) structure of the melanosomes vary with the
(Fig. 22—13). In such preparations, the mel¬ animal species and are characteristic of par¬
anocytes are blackened and appear as highly ticular genotypes within the same species. In
branched cells. The ratio of melanocytes to the human, however, melanosomes are uni-
558 • SKIN

formly elongated (Figs. 22—16, 22—17) except thicker than that of the adjacent skin and is
in red-haired individuals, in whom they tend more like that of a mucosa. They may have
to be spherical. Melanosomes are somewhat a thin, rudimentary, cornified layer. Nor¬
larger in the skin of Australoids, Negroids, mally they do not contain sweat glands, hair
and Mongoloids than they are in Caucasoids. follicles, hairs, or sebaceous glands, but are
Within the same individual they tend to be moistened by mucous glands situated within
larger in the hair follicles than in the skin. the body orifices. Since the keratinized layer
Lack of melanin in the epidermis of some is thin or may even be absent, the redness of
areas of the skin of animals may be due the blood in the underlying capillaries shows
either to absence of melanocytes or, as in through and gives the junction region a red
albinism, to the inability of the melanocytes to color as exemplified in the lips.
form pigmented melanosomes. In humans
the entire integument normally possesses
functioning melanocytes. Their activity is in¬
fluenced by hormones and by factors in the THE DERMIS
physical environment. During pregnancy, the
pigmentation of the areola of the nipples
increases. Freckles are intensified and some The thickness of the dermis cannot be
individuals develop cloasma, the so-called measured exactly, because it passes over into
“mask of pregnancy,” consisting of a pig¬ the subcutaneous layer without a sharp
mented area over the malar eminences and boundary. The average thickness is approxi¬
a brownish discoloration of the forehead. mately 1 to 2 mm; it is less on the eyelids
This gradually disappears after delivery. The and the prepuce (0.6 mm or less) but reaches
phenomenon of tanning on exposure to sun¬ a thickness of 3 mm or more on the soles
light results from an immediate darkening of and palms. On the ventral surface of the
the existing melanin and, after a few days, body and the appendages it is generally thin¬
an enhanced tyrosinase activity in the melan¬ ner than on the dorsal surface. It is thinner
ocytes that leads to the formation of new in women than in men.
melanin. The pigmentation of the skin is The outer surface of the dermis in contact
believed to protect the underlying tissues with the epidermis is usually uneven and is
against the potentially harmful effects of so¬ elevated into papillae that project into con¬
lar radiation. In the human, small melano¬ cavities between the ridges on the deep sur¬
somes often aggregate to form meianosome face of the epidermis. This sculptured sur¬
complexes within the keratinocytes (Fig. 22— face of the dermis is called the papillary layer,
16), whereas large melanosomes usually are and the deeper main portion of the dermis
individually distributed. A more effective is called the reticular layer. There is no distinct
protective layer against ultraviolet radiation boundarv between them.
results. The reticular layer consists of rather dense
Melanin is also found in the retinal pig¬ connective tissue. Its collagenous fibers form
ment epithelium of the eye (Fig. 22—15) and a feltwork with bundles running in various
in dermal melanocytes and melanophores of directions but, for the most part, more or
cold-blooded vertebrates. In the latter they less parallel to the surface. Occasional bun¬
constitute the pigmentary effector system re¬ dles are oriented almost perpendicular to the
sponsible for rapid changes of color for pur¬ majority. The papillary layer and its papillae
poses of camouflage and concealment. The consist of looser connective tissue with much
“ink” of squid and other cephalopods consists thinner collagenous bundles.
of small spherical melanosomes that are pro¬ The elastic fibers of the dermis form abun¬
duced in a special ink gland, stored in the dant, thick networks between the collagenous
ink sac, and squirted out to blacken the water bundles and are condensed about the hair
and conceal the animal threatened by a pre¬ follicles and the sebaceous and sweat glands.
dator. In the papillary layer they are much thinner
and form a continuous fine network in the
papillae beneath the epithelium. The cells of
MUCOCUTANEOUS JUNCTIONS the dermis are more abundant in the papil¬
lary than in the reticular layer and are similar
These are transitions between the mucous to those of the subcutaneous layer except for
membranes and skin. Their epithelium is the relative paucity of fat cells.
 SKIN • 559

A B C
Figure 22-23. The pattern formed at a dermoepidermal junction shows marked regional variations. The figures shown
here are views of the undersurface of separated sheets of epidermis stained with carmine. The light areas are the
depressions occupied in life by the dermal papillae. A, From the cheek; the undersurface of the epidermis is smooth,
except for the hair follicles. B, From the back. C, The breast. D, The elaborate pattern of concavities occupied by the
dermal papillae of a finger pad. (Courtesy of G. Szabo.)

Within the deep parts of the reticular layer At various levels of the dermis are the hair
in the areolae, penis, perineum, and scrotum, follicles and the sweat and sebaceous glands,
numerous smooth muscle cells form a loose which are epidermal derivatives extending
plexus. Such portions of the skin become down into the dermis. Blood vessels, nerves,
wrinkled during contraction of these muscles. and nerve endings are also abundant in this
Smooth muscle cells forming the so-called layer of the skin.
arrector pili muscles are also connected with
the hairs (Figs. 22-24, 22-25). In many places
in the skin of the face, cross-striated muscle Hypodermis
fibers terminate in the dermis. These are the
muscles of facial expression. They are responsi¬ The subcutaneous layer consists of loose
ble for smiling, frowning, and the voluntary connective tissue and is a deeper continuation
movement of the ears and scalp. These rep¬ of the dermis. Its collagenous and elastic
resent vestiges of a more extensive subcuta¬ fibers are directly continuous with those of
neous layer of muscle that is present in many the dermis and run in all directions but
mammals, called the panniculus carnosus. This mainly parallel to the surface of the skin.
layer is responsible for the voluntary move¬ Where the skin is flexible or freely movable,
ment of large segments of the integument, these fibers are few, but where it is closely
which can be observed when animals attempt attached to the underlying parts, as on the
to dislodge insects from their skin or to shake soles and palms, they are thick and numer¬
dry when they emerge from the water. The ous.
absence of this layer over most of the body Depending on the portion of the body and
in the human is disadvantageous in that, after the nutrition of the organism, varying num¬
wounds, the skin is likely to become immobile bers of fat cells develop in the subcutaneous
and bound down to the underlying structures layer. These are also found in clusters in the
because of shrinkage of scar tissue. Greater deep layers of the dermis. The fatty tissue of
disfigurement results than in other mammals the subcutaneous layer on the abdomen may
with more mobile skin. reach a thickness of 3 cm or more, but in the
560 SKIN

Stratum
corneum

Stratum
malpighii

Smooth muscle

Reticular layer

Sebaceous gland

Tangential section Hair shaft

of hair
Internal
root
sheath

Tangential section
Tangential
of hair section
of hair

Subcutaneous layer

Dermal sheath

External root
sheath
Hair matrix
Hair papilla

Figure 22-24. Section of the scalp of a man, showing the root of a hair in longitudinal section, x 32. (After Schaffer.)
 SKIN • 561

tends down into the dermis, where it is sur¬

rounded by connective tissue (Figs. 22-2, 22-
24, 22-25). The active follicle has a bulbous
terminal expansion with a concavity in its
underside occupied by a connective tissue
papilla (Figs. 22-24, 22-29). The papilla is
covered by epithelial matrix cells of hair and
root sheath. The cells on the dome of the
convexity form the hair root, which develops
into the hair shaft. The free end of the shaft
protrudes beyond the surface of the skin.
The hair is not a continuously growing
organ but has phases of growth that alternate
with periods of rest. The structure of the
hair follicle varies markedly according to the
stage of hair growth (Fig. 22-25). In the
resting hair (club hair), the follicle is relatively
short, its epithelium is more or less similar to
the surface epidermis, and the hair shaft is
firmly anchored into the follicle by fine fila¬
ments of keratin that penetrate between the
follicular cells. A cluster of dermal cells at¬
tached to the end of the follicular epithelium
is the remnant of the dermal papilla of the
growing hair and will again develop into a
typical dermal papilla at the next period of
Figure 22-25. Diagrammatic representation of an actively hair generation (Figs. 22T25, 22-30).
growing and a quiescent hair follicle and the accessory In a phase of growth the follicle elongates
structures. (Redrawn after Montagna, W. In Structure and and the epithelium again surrounds the der¬
Function of Skin. New York, Academic Press, 1956.)
mal papilla. The epithelial cells around the
papilla (the matrix) differentiate into several
eyelids and on the penis the subcutaneous types. (1) In certain types of coarse hairs, the
layer does not contain fat cells. central matrix cells on top of the convexity
The subcutaneous layer is penetrated of the papilla develop into the medulla of the
everywhere by large blood vessels and nerve hair shaft. The cells are large and vacuolated
trunks and contains many nerve endings. and eventually keratinize. This central part
of the hair shaft is not demonstrable in thin¬
ner hairs. (2) The next concentric layer of
matrix cells keratinize and develop into the
HAIRS cortex of the hair, the main constituent of the
shaft. Its cells are heavily keratinized and
The hairs are slender keratinous filaments tightly compacted, and they carry most of
that develop from the matrix cells of follicu¬ the pigment of the hair (Fig. 22-27). (3)
lar invaginations of the epidermal epithe¬ Peripheral to the matrix cells of the cortex
lium. They vary from several millimeters to lie those of the cuticle of the hair (Figs. 22-
over a meter in length and from 0.005 to 0.6 27, 22—28). These cells of the outermost layer
mm in thickness. They are distributed in are the most heavily keratinized and their
varying numbers (Fig. 22—40) and in variable imbrication (Fig. 22-26) prevents matting of
thickness and length on the whole surface of the erupted hairs. These three layers of cel¬
the skin, except on the palms, the soles, the lular components all undergo keratinization
sides of fingers and toes, the lateral surface in the so-called keratogenous zone of the folli¬
of the feet below the ankles, the lip, the glans cle, immediately above the dome of the der¬
penis, the prepuce, the clitoris, the labia mi¬ mal papilla, and form the solid hair shaft
nora, and the internal surface of the labia (Figs. 22-29, 22-31).
majora. The more peripheral concentric rows of
Each hair arises in a tubular invagination matrix cells produce the internal root sheath, a
of the epidermis, the hair follicle, which ex¬ transient structure surrounding the hair
Text continued on page 566
562 • SKIN

Figure 22-26. Scanning electron micrograph of a hair shaft emerging from a human scalp. (Micrograph by T. Fujita.)
 SKIN • 563

Figure 22-27 Figure 22-28

Figure 22-27. Electron micrograph of a mature black hair of a guinea pig in transverse section, showing a few melanin
granules in the concentrically arranged flattened cuticle cells, and a large number in the cortical cells in the interior of
the hair. No medullary cells are present at this level in the hair. For higher magnification of the area in the rectangle,
see Figure 22-28. (From Snell, R. J. Invest. Dermatol. 58:47, 1972.)

Figure 22-28. Higher magnification of the cuticle of a hair in an area similar to that in the rectangle in Figure 22-27.
The markedly flattened cuticle cells are separated with intercellular spaces filled with electron-dense amorphous material.
(From Snell, R. J. Invest. Dermatol. 58:47, 1972.)
564 . SKIN

v.

7 2 3 4 5 6 7 8 9

Figure 22-29. Longitudinal section through a hair from

the head of a 22-year-old man. 1, Medulla; 2, cortex;
3, hair cuticle; 4, inner sheath cuticle; 5, Huxley’s
layer; 6, Henle’s layer; 7, external root sheath; 8,
glassy membrane; 9, connective tissue of the hair
follicle; A, matrix; AW, external root sheath at the bulb;
P, papilla, x 350. (After Hoepke.)

AW
 SKIN • 565

A B
Figure 22-30. Unstained plastic sections of hair follicles. A, Active hair, showing large melanocytes and their processes
contributing pigment to the hair. B, Inactive or club hair, x 200. (Courtesy of R. Mitchell and G. Szabo.)

Connective tissue

Glassy
membrane

External
root sheath
Henle’s layer

Huxley’8
layer

Cuticle of
inner root
sheath
Cuticle of
hair

Cortical
substance
of hair

Figure 22-31. Cross section through a hair follicle in the skin of a pig embryo, at the level at which Henle’s layer is
completely cornified. x 375.
566 • SKIN

shaft below the level of the sebaceous glands, tive influence on the formation of the hair.
which is presumed to facilitate the movement If for any reason the dermal papilla is de¬
of the growing hair shaft. It consists of three stroyed in postnatal life, no hair is formed.
layers. The cuticle of the internal root sheath, It is noteworthy, too, that there is a greater
like the cuticle of the hair, consists of over¬ diversity of specialization and division of la¬
lapping thin scales with their free margins bor among the epidermal cells of the hair
directed toward the bottom of the follicle. matrix with respect to their fate in the process
Huxley’s layer consists of one to three layers of keratinization. In electron micrographs,
of cornihed cells, and Henle’s layer is a single the cells of the medulla, the cortex, and the
layer of elongated cells closely adherent to internal and external root sheaths can all be
the external sheath (Fig. 22—31). These three distinguished by characteristic differences in
layers form “trichohyalin” granules and ker¬ their granules and their mode of keratiniza¬
atinize, but they do not form a compact tion.
enduring structure, and they finally desqua¬ The pigmentation of the hair is attributable
mate at the level of the opening of the seba¬ to epidermal melanocytes located over the
ceous glands. tip of the dermal papilla, a site corresponding
The outermost layer of the follicle, the to their location in the base of the epidermis
outer root sheath, is basically similar to the generally (Fig. 22-30A). These melanocytes
unspecialized epidermal epithelium and is donate their pigment to the cells of the
continuous with it above. At the neck of the hair matrix and cortex. The melanosomes of
papilla it is one layer of flat cells. It becomes the hair are usually larger than those found
two-layered at the level of the middle third in the skin of the same individual. The me¬
of the papilla, and higher up it becomes lanocytes of the hair follicle function only at
stratified. the beginning of the growing phase of the
The glassy membrane, which is a part of the hair cycle, the onset of hair growth usually
dermis, separates the epithelial from the con¬ being heralded by increased melanogenic ac¬
nective tissue portion of the follicle. The tivity. In later stages of the growing phase or
latter portion is made up of two layers, a thin in the resting hair, melanocytes cannot be
internal layer formed by circular fibers and distinguished in the follicle.
an external, poorly outlined layer consisting In young rodents, hair growth is synchro¬
of longitudinal collagenous and elastic fibers. nized and spreads over the body in a wave
The hair matrix cells are analogous to the pattern. Later in life, however, this process
germinal cells of the epidermis insofar as the gives way to a mosaic pattern, hair growth
life cycle of each ends with formation of beginning in isolated islands here and there.
cornihed cells. However, the epidermis pro¬ In man the mosaic growth pattern prevails,
duces a relatively soft keratinous material and the duration of the growing and resting
that is steadily shed, whereas in the hair, the phases varies from one region to another. In
product of the matrix cells, is a hard, cohe¬ the case of scalp hair the growing phase is
sive, nonshedding, keratinous structure con¬ very long (several years), whereas the resting
sisting of cells that accumulate in numerous phase is of the order of three months.
concentric layers. Among mammals, humans are exceptional
Since the hair is not perpendicular to the in that their skin is not furry. It is by no
skin surface but inclined, it is very difficult means hairless, however (see Fig. 22—40 for
to find a perfect longitudinal section of a numbers of hair follicles per square centi¬
follicle that displays these concentric layers meter). In accordance with its relative paucity
well. The student will therefore have diffi¬ of hairs, the human epidermis is generally
culty in identifying all of the structures de¬ thicker than that of other mammals. The
scribed here. For this purpose the follicle architectural pattern of the dermoepidermal
needs to be reconstructed from serial sec¬ junction varies greatly from region to region.
tions. It is almost flat on the cheek, whereas deep
It is well to bear in mind certain differences dermal ridges occur on the soles and palms
between the keratinizing epidermis and the (Fig. 22-23).
keratinizing hair follicle. In the epidermis The human hairy coat also exhibits re¬
this process is general and continuous. In the gional differences in the competence of the
case of the hair follicle it is intermittent and hair follicles to respond to male sex hor¬
localized to a particular portion of the der¬ mones. At the onset of puberty in males, the
mis—the dermal papilla, which has an induc¬ areas of the mustache and the beard produce
 SKIN • 567

strongly pigmented thick hairs. The same The nail plate consists of closely compacted
areas in the female, although they contain scales, the dead residues of cornified epithe¬
the same number of hair follicles, continue lial cells so arranged that in section the nail
to produce fine hair. In other places, how¬ appears longitudinally striated. The nail wall
ever, such as the axillae and the pubic re¬ has the structure of skin, with all its layers.
gions, hair appears in both sexes at the onset Turning inward into the nail groove, it loses
of puberty. In males, there is often a char¬ its papillae, and the epidermis loses its cor¬
acteristic regression of the scalp hair with nified, clear, and granular layers. Under the
age, which varies in degree according to proximal fold, the stratum corneum spreads
genotype. In its extreme form, this male onto the free surface of the nail body as the
pattern of changing hair distribution pro¬ eponychium (Fig. 22-32). The stratum lucidum
gresses so far that all the hair follicles are lost and the stratum granulosum also reach far
(baldness) or only a few are left and produce inside the groove but do not continue along
very fine hair. the lower surface of the nail plate. On the
One or more sebaceous glands are associated surface of the nail bed, only the stratum
with each hair follicle. They discharge their basale and stratum spinosum of the epider¬
holocrine secretory product through a short mis are present.
duct into the upper portion of the follicular In the nail bed the dermis is directly fused
canal. with the periosteum of the phalanx. The
A band of smooth muscle cells, the hair surface of the dermis under the proximal
muscle or arrector pili muscle, is attached at edge of the nail is provided with rather low
one end to the papillary layer of the dermis papillae, but under the distal half of the
and at the other to the connective tissue lunula this surface is quite smooth. At the
sheath of the hair follicle (Figs. 22—24, 22- distal margin of the lunula, longitudinal, par¬
25). When this muscle contracts in response allel ridges project instead of papillae. The
to cold, fear, or anger, it moves the hair into
a more vertical position while depressing the
skin in the region of its attachment and Free edge of nail-—v Hyponychium
elevating the region immediately around the
hair. This is responsible for the erection of
hairs in animals and for the so-called “goose
flesh” in man.

NAILS
Malpighian
layer of nail bed
The nails are horny plates on the dorsal Nail plate
surfaces of the terminal phalanges of the Dermis of
nail bed
fingers and toes. The surface of the skin
Eponychium
covered by them is the nail bed. It is sur¬
rounded laterally and proximally by a fold Posterior nail iuall
of skin, the nail wall. The slit between the
Granular layer
wall and the bed is the nail groove. The Malpighian layer
proximal edge of the nail plate is the root of Stratum
the nail. The visible part of the nail plate, corneum Nail root
called the body of the nail, is surrounded by
the nail wall. The distal portion, becoming
free of the nail bed, extends forward and is
gradually worn off or is cut off. The nail is
semitransparent and permits the color of the
underlying tissue, rich in blood vessels, to
Phalanx
show through. Near the root, the nail has a Injected Posterior Nail matrix
whitish color. This crescentic portion, the uessel nail grooue
lunula, is usually covered by the proximal Figure 22-32. Longitudinal section of the nail of a new¬
portion of the nail fold. born infant. (After A. A. Maximow.)
568 • SKIN

boundary between the epithelium and the As new formation of the nail takes place
dermis of the nail bed is, therefore, scalloped in the matrix, the nail moves forward. Most
in a perpendicular section (Fig. 22—33), authors deny the participation of the epithe¬
whereas it is smooth in longitudinal sections. lium of the other portions of the nail bed in
Beyond the free edge of the nail the dermal the formation of the nail substance, believing
ridges are replaced by cylindrical papillae. that the nail simply glides forward over this
The epithelium of the nail bed distal to the region.
lunula retains the typical structure of the
basal layers of the epidermis. The epithelium
is thicker between the ridges of the dermis
than over them. The upper layer of cells, GLANDS
which touches the substance of the nail, is
separated from it in places by an even line,
The glands of the skin include the seba¬
while in others it is jagged. Under the free
ceous, sweat, and mammary glands. The last
edge of the nail the usual cornihed layer
will be described in Chapter 33.
again begins; it is thickened at this place and
is called hyponychium (Fig. 22—32).
The epithelium that lines the proximal Sebaceous Glands *

portion of the nail bed and corresponds

roughly in distribution with the lunula on The sebaceous glands are scattered over the
the upper surface is particularly thick distally entire integument except in the palms, the
and its upper portion gradually passes over soles, and the sides of the feet, where there
into the substance of the nail plate. Here the are no hairs. They vary from 0.2 to 2 mm in
new formation of the nail substance pro¬ diameter. They lie in the dermis, and their
ceeds; accordingly, this region of the epithe¬ excretory ducts open into the necks of hair
lium is called the nail matrix (Fig. 22—33). The follicles. When several glands are connected
cells of the deepest layer are low columnar with one hair, they lie at the same level. On
and mitoses can be observed frequently in the lips, around the corners of the mouth,
them. Above these are six to ten layers of on the glans penis and the internal fold of
polyhedral cells joined by three to 12 layers the prepuce, on the labia minora, and on the
of more flattened cells. This entire mass is mammary papilla, the sebaceous glands are
penetrated by parallel fibrils of a special “on- independent of hairs and open directly onto
ychogenic” substance. On passing into the the surface of the skin. To this category also
proximal edge of the nail plate, these cells belong the meibomian glands of the eyelids.
cornify and become homogeneous. The sebaceous glands in mucocutaneous

Nail plate
Lateral nail wall

Malpighian
layer of
nail bed
Derma I
Stratum papilla
corneum
of epidermis
Vessel
Stratum
lucidum

Stratum Dermis of
granulosum naii bed

Malpighian
layer of
epidermis

Figure 22-33. Cross section of the lateral edge of a nail and its surrounding parts. (After A. A. Maximow.)
 SKIN • 569

junctions are more superficial than those that that of many exocrine glands that have been
are associated with hairs. studied, in great detail.
The secretory portions of the sebaceous The eccrine sweat glands are simple,
glands are rounded sacs (alveoli). As a rule, coiled, tubular glands. Their secretory por¬
several adjacent alveoli form a mass like a tion is a tube convoluted into a ball, and the
bunch of grapes, and all of them open into duct is a slender unbranched tube (Fig. 22-
a short duct (Figs. 22-24, 22—25). A simple 35, 22—36). The bulk of the secretory portion
branched gland results. Much less frequently, is in the dermis, and the duct ascends
only one alveolus is present. In the meibo¬ through the epidermis to open at the skin
mian glands of the eyelids there is one long, surface. In response to stimulation by cholin¬
straight duct, from which a row of alveoli ergic nerves, the secretory portion forms a
project. precursor fluid with a composition similar to
The walls of the alveoli are formed by a an ultrafiltrate of blood plasma. Sodium and
basal lamina supported by a thin layer of excess water are reabsorbed in the duct, pro¬
fibrillar connective tissue. Along the internal ducing a hypotonic sweat released at the skin
surface is a single layer of thin cells with surface.
round nuclei. Toward the center of the al¬ The secretory coil is composed of three
veoli a few cells keratinize, but most of them cell types: clear cells, dark cells, and myoepithelial
become larger and polyhedral, gradually fill cells. The dark cells border nearly all of the
with fat droplets, and resemble multilocular luminal surface of the tubule and are either
fat cells. The nuclei gradually shrink and cuboidal in shape or pyramidal, with a broad
then disappear, and the cells break down into adluminal end and a narrower portion ex¬
fatty detritus. This is the oily secretion of the tending downward between the apices of the
gland, and it is secreted onto the hair and clear cells, which rest upon a thick basal
upon the surface of the epidermis. The ducts
of sebaceous glands are lined by stratified
squamous epithelium continuous with the
external root sheath of the hair and with the
malpighian layer of the epidermis.
In sebaceous glands, the secretion results
from the destruction of the epithelial cells
and is therefore of the holocrine type. It is
accompanied by a regenerative multiplication
of epithelial elements. In the body of the
gland, mitoses are rare in the cells lying on
the basal lamina. They are numerous, how¬
ever, in the cells close to the walls of the
ducts, whence the new cells move into the
secretory regions.
The so-called uropygial or preen glands of
birds, especially aquatic birds, are specialized
sebaceous glands. They produce oily material
that is spread with the heak over the surface
of the feathers to make them impervious to
water.

Eccrine Sweat Glands

The eccrine sweat glands have not received
investigative attention commensurate with
their physiological importance. The human
skin contains 3 to 4 x 106 eccrine sweat
glands distributed over nearly the entire sur¬
face of the body. Each weighs 30 to 40 pig
and their aggregate weight is roughly that of Figure 22-34. Drawing of a portion of a human sweat
a kidney. A human can perspire as much as gland. (From Krstic, R. V. Die Gewebe des Menschen
10 liters a day, a rate of secretion exceeding und der Saugetiere. Berlin, Springer Verlag, 1978.)
570 • SKIN

These cells are believed to secrete a mucoid

substance but its biochemical properties have
not been analyzed. Glycogen, fat droplets,
and pigment deposits may also occur in the
cytoplasm. The clear cells rest directly on the
basal lamina or on the myoepithelial cells.
Intercellular canaliculi lined with microvilli
occur between adjacent clear cells and pro¬
vide the only egress from these cells to the
lumen of the gland. The commissures of
canaliculi are closed by occluding junctions
between the apposed membranes of neigh¬
boring clear cells. These cells are rich in
mitochondria, but poor in endoplasmic retic¬
ulum. Their cytoplasm contains varying
amounts of glycogen. The basal plasma mem¬
brane is elaborately infolded as in other cell
types involved in secretion of water and elec¬
trolytes. Myoepithelial cells occur at intervals
between the clear cells and the basal lamina.
Their function is not clear, for the lumen of
the gland is collapsed in the resting gland
and contains no accumulated fluid that could
be expressed by their contraction. There is
no evidence of their pulsatile contraction in
the stimulated gland.
At the transition from the secretory por¬
tion of the gland, the tube narrows and its
lumen takes on a slitlike or star-shaped con¬
figuration in cross section. The glandular and
myoepithelial cells are replaced by a double
Figure 22-35. Sweat gland from the palmar surface of layer of cuboidal cells. The peripheral row
an index finger. The drawing was based on study of of cells have comparatively large nuclei and
sections and a teased preparation, x 45. (Slightly modi¬ abundant mitochondria (Fig. 22—39). The
fied from von Brunn.) adluminal cells have large, irregularly shaped
nuclei and relatively little cytoplasm. The
lamina. The dark cells do not reach the basal membranous organelles of the cytoplasm are
lamina. In electron micrographs they contain poorly developed. Immediately beneath their
abundant ribosomes and secretory granules luminal surface is a remarkable condensation
with glycoprotein staining characteristics. of filaments constituting a conspicuous ter-

Fat cell

Duct

Myoepithelial cell Terminal portion

Vein

Basement mem¬
brane
Connectiue tissue

Figure 22-36. Section of human sweat gland, x 120.

 SKIN • 571

minal web. This was formerly referred to as the hypothalamic nucleus are conducted over
a “cuticular border,” but this is erroneous, autonomic pathways through the brain stem
since the cuticle of such borders is outside of and spinal cord and to the periphery over
the plasmalemma whereas this specialization postganglionic sympathetic fibers that termi¬
is in the apical cytoplasm. nate in unmyelinated axons around the ec¬
As the ducts pass through the dermis and crine sweat glands. These are generally be¬
epidermis, they take a curved, loosely helical lieved to be cholinergic. However, a loose
course, so that in sections they tend to appear network of catecholamine-containing nerves
discontinuous at sites where they curve out has been demonstrated in the sweat glands
of the plane of section. In the epidermis they of the palm in monkeys, and it is quite
are surrounded by concentrically arranged possible that the sweat glands of the palms
keratinocytes. On the palms and soles and and soles of humans may also have adrener¬
volar surface of the fingers, the rows of ducts gic as well as cholinergic innervation. This
open on the epidermal ridges with funnel- may explain the localized “emotional sweat¬
shaped openings that are easily seen with a ing” that is generally confined to the palms,
magnifying glass. soles, axillae, and forehead under emotion¬
Histophysiology of Eccrine Glands. The ally charged circumstances that activate the
sweat glands play an important role in the adrenergic portions of the sympathetic ner¬
control of body temperature. Heat is pro¬ vous system.
duced continuously as a by-product of me¬
tabolism and is continually being lost to the Apocrine Sweat Glands
environment by radiation, conduction, and
evaporation. Of these, evaporation is perhaps A second type of sweat gland occurs in the
the most important and the only one subject axilla (Fig. 22-37), the mons pubis, the areola
to physiological control. When the environ¬ of the mammary gland, and the circumanal
ment is warmer than body temperature, area. These are called apocrine sweat glands
evaporation is the only way in which the body and are larger than the eccrine; their coiled
can dissipate heat. Under normal conditions secretory portion may reach 3 to 5 mm in
there is a continuous evaporative heat loss of diameter compared with 0.3 to 0.4 mm for
about 600 ml per day from the lungs and eccrine glands. They are located deep in the
skin over which there is no control. At high subcutaneous connective tissue and each
environmental temperatures, loss of heat by opens into a hair follicle. They arise during
evaporation can be regulated by varying the fetal life as an epithelial bud from the side
rate of sweating. The eccrine glands do not of the hair follicle, whereas the eccrine glands
function simultaneously or under the same develop from cords of epithelial cells that
conditions on all parts of the body. When the grow downward from the epidermal ridges.
human body is exposed to excessive heat, Eccrine sweat glands become functional in
sweating begins on the forehead and spreads the neonatal period, but secretory function
to the face, and then to the rest of the body. does not begin in apocrine sweat glands until
Finally, the palms and the soles will show puberty. The designation apocrine implies^
increased sweat production. Under nervous that a portion of the apical cytoplasm of the
strain, however, palms and soles may start to cells is shed in the secretory process. How¬
sweat first. ever, the mechanism of secretion of apocrine
It has been shown that the glandular por¬ glands has not been established to everyone’s
tion of the eccrine sweat gland excretes more satisfaction. The protein and electrolyte com¬
electrolytes than are finally found at the sur¬ position of apocrine sweat in humans has not
face of the skin. It is assumed that an absorp¬ been determined.
tion of electrolytes takes place in the duct In certain parts of the skin the sweat glands
portion of the gland. have a peculiar arrangement and function
Sweating is controlled by centers in the Such are the glands that produce cerumen
preoptic area of the hypothalamus that func¬ “wax” in the external auditory meatus. They
tion like a thermostat to regulate body tem¬ reach a considerable size and extend to the
perature. An increase in body temperature perichondrium. The secretory portions of
may increase the rate of discharge of heat- the ceruminous glands branch, and the ducts,
sensitive neurons in these centers by as much which sometimes also branch, may open to¬
as tenfold. This is accompanied by vasodila¬ gether with the ducts of the adjacent seba¬
tion and profuse sweating. Impulses from ceous glands into the hair sacs of the fine
572 • SKIN

Figure 22-37. Axillary glands

from a 37-year-old woman during
the premenstruum, a, Greatly en¬
larged glands that change with the
menstrual cycle, e, Glands that do
not change. Resorcin-fuchsin
stain for elastic fibers. Prepara¬
tion of Loescke. x 110. (After
Hoepke.)

hairs. In the terminal portions are highly nervated mainly by cholinergic nerves. The
developed smooth muscle cells; the glandular apocrine glands are connected with hair fol¬
cells located upon them are particularly rich licles; they begin to function at puberty, pro¬
in pigment granules containing lipid. ducing a more viscous secretion; and they
Moll’s glands of the margin of the eyelid are supplied by adrenergic nerves.
are also a special kind of sweat gland, with
terminal portions that do not form a ball but
are irregularly twisted and provided with a
BLOOD AND LYMPHATIC
wide lumen. The excretory ducts open onto
the free surface or into the hair sacs of the VESSELS
eyelashes.
The secretion of the sweat glands is not The arteries that supply the skin are lo¬
the same everywhere. True sweat, a trans¬ cated in the subcutaneous layer. Their
parent, watery liquid, is excreted mainly by branches, reaching upward, form a network
the small sweat glands, while a thicker secre¬ (rete cutaneum) parallel to the surface on the
tion of complex composition is produced by boundary line between the dermis and the
the apocrine glands of the axilla and about hypodermis. From one side of this network,
the anus. In women, the apocrine sweat branches are given off that nourish the sub¬
glands of the axilla show periodic changes cutaneous stratum with its fat cells, the sweat
with the menstrual cycle. These changes con¬ glands, and the deeper portions of the hair
sist mainly of enlargement of the epithelial follicles. From the other side of this network,
cells and of the lumens of the glands in the vessels enter the dermis. At the boundary
premenstrual period, followed by regressive between the papillary and reticular layers
changes during the period of menstruation. they form the denser, subpapillary network
The differences between the eccrine and or the rete subpapillare (Fig. 22—41). This
apocrine sweat glands are as follows. The gives off thin branches to the papillae. Each
eccrine glands have no connections with hair papilla has a single loop of capillary vessels
follicles; they function throughout life, pro¬ with an ascending arterial and descending
ducing a watery secretion; and they are in¬ venous limb.
 SKIN 573

Figure 22-38. Electron micrograph of a sector of the

secretory coil of a normal eccrine sweat gland. Mucigen-
ous, “dark” cells border the lumen, while “clear” serous
cells are more deeply situated and surround intercellular
canaliculi. The myoepithelia! cells form an incomplete
layer at the periphery of the tubule, x 3400. (Courtesy of
R. E. Ellis.)

Figure 22-39. Electron micrograph of a portion of the

wall of the coiled duct of an eccrine sweat gland. The
luminal margin of the superficial cells contains a concen¬
tration of filaments formerly described as a “cuticular
border.” The peripheral cells have elaborately convoluted
surfaces and contain many mitochondria, x 4000. (Cour¬
tesy of R. A. Ellis.)
574 • SKIN

AVERAGE NUMBERS, ± S.E. OF MEAN.

HAIR FOLLICLES SWEAT GLANDS MELANOCYTES

PER SQUARE PER SQUARE PER SQUARE

CENTIMETER CENTIMETER MILLIMETER

Face 700 ± 40 270 ± 25 2120 ± 90

Trunk 70 ± 10 175 ± 20 890 ± 70
Arm 65 ± 5 175 ± 15 1160 ± 40
Leg •55 ± 5 130 ± 10 1130 ± 60

Average 330 ± 20 212 ± 15 1560 ± 110

Figure 22-40. Regional anatomy of the human integu¬

ment. (After G. Szabo.)

The veins that collect the blood from the

capillaries in the papillae form the first net¬
work of thin veins immediately beneath the
papillae. Then follow three flat networks of
gradually enlarging veins on the boundary
line between the papillary and reticular lay¬
ers. In the middle section of the dermis and
also at the boundary between the dermis and
the subcutaneous tissue, the venous network
is on the same level as the arterial rete cuta-
neum. Into this network the veins of the
sebaceous and the sweat glands enter. From Figure 22-41. Distribution of blood vessels in the skin.
the deeper network the large, independent, (Modified slightly from von Brunn.)
subcutaneous veins pass, as well as the deep
veins accompanying the arteries.
There are direct connections between the vessels are not associated with the hairs or
arterial and venous circulation in the skin glands of the skin.
without intervening capillary networks.
These so-called arteriovenous anastomoses
play a vital role in thermoregulation in the
body. NERVES
Each hair follicle has its own blood vessels.
It is supplied with blood from three sources: The skin is the most extensive sense organ
a special small artery that gives off a capillary in the body, receiving stimuli from the exter¬
network into the papilla; the rete subpapil- nal environment, and it is therefore abun¬
lare toward the sides of the hair sac; and dantly supplied with sensory nerves. In ad¬
several other small arteries that form a dense dition, it contains nerves that supply blood
capillary network in the connective tissue vessels, sweat glands, and the arrector pili
layer of the follicle. muscles.
There is a dense network of capillaries Firmly entrenched in the literature of
outside the basal lamina of the sebaceous physiology is the concept that the skin pos¬
and, particularly, of the sweat glands. sesses four distinct types of sensory nerve
The skin is rich in lymphatic vessels. In the endings, each responding to a specific kind
papillary layer they form a dense meshwork of stimulus: touch, heat, cold, or pain. How¬
of lymphatic capillaries. They begin in the ever, histologists to date have been unable to
papillae as networks or blind outgrowths, identify four kinds of receptor organs to
which are always deeper than the blood ves¬ match the four sensory modalities.
sels. From this peripheral network, branches Two broad categories can be recognized:
pass to the deeper network, which lies on the (1) the corpuscular receptors and (2) free
boundary between the dermis and the hy- nerve endings. The specialized corpuscular
podermis; under the rete cutaneum it has endings include the Meissner, Merkel, paci¬
much wider meshes, and its vessels are pro¬ nian, and genital corpuscles, which are de¬
vided with valves. From the deeper network, scribed in the chapter on the nervous system
large subcutaneous lymphatic vessels origi¬ (Chapter 11). It will suffice here to state that
nate and follow the blood vessels. Lymphatic all of these possess non-neural cellular and
 SKIN • 575

extracellular components arranged in such a the lever-like action of the stiff, keratinized
way as to convey mechanical stimuli to the hair shaft whenever the free portion of the
associated nerve axons. The free nerve end¬ hair is touched. The hair and its associated
ings are simpler in structure and require nerve endings thus form a sensitive mechan-
special silver staining techniques or electron oreceptor. Other nerve fibers bypass the
microscopy for their demonstration. The skin deeper region of the hair and form papillary
on the palmar surface of the fingers is espe¬ endings around the orifice through which it
cially sensitive and has a high concentration emerges from the epidermis.
of both corpuscular and free sensory recep¬
tors. The skin there has about 80 dermal
papillae per square micrometer, and some 60
HISTOGENESIS OF THE SKIN
per cent of all papillae contain one or more
free nerve endings, resulting in up to 100 AND ITS ACCESSORIES
endings per (am2. These approach the epi¬
dermis vertically, providing a punctate distri¬ The epidermis develops from the ecto¬
bution ensuring sensitive point-to-point dis¬ derm, and the dermis arises from the mes¬
crimination. It is important to understand enchyme. The epidermis in the human em¬
that the term “free” nerve ending signifies bryo, during the first two months, is a double¬
only the absence of a corpuscular receptor layered epithelium. The basal layer, which
specialization; it does not imply a naked axon. lies on the mesenchyme, consists of cuboidal
The terminal cell of the Schwann sheath or cylindrical cells that multiply rapidly. The
sends multiple cytoplasmic processes that peripheral layer consists of flat cells that are
provide a thin covering for the axon and its constantly formed anew from the elements
branches to their blind ends in the dermal of the deeper layer. Beginning with the third
papillae. month, the epidermis becomes triple layered.
In addition to the vertically oriented pap¬ The new intermediate layer above the basal
illary endings, there is an extensive horizontal cells consists of polygonal cells, which in¬
nerve plexus underlying the dermal-epider¬ crease in number and develop the surface
mal junction. This network consists of rami¬ projections formerly interpreted as intercel¬
fying subunits arising from nonmyelinated lular bridges. At the end of the third month,
preterminal fibers. Because of their tuftlike in the peripheral portions of the intermediate
pattern of arborization, these units have been layer, cornification begins and leads to the
termed penicillate endings. In each, the termi¬ formation of the layers found in the adult.
nal cell of the Schwann sheath, located 2 to The cornified scales are desquamated and
6 jam beneath the epidermis, is highly form part of the vernix caseosa.
branched and supplies cytoplasmic sheaths The irregularities on the lower surface of
for single or multiple axons that fan out from the epidermis arise at the end of the third
the parent nerve fiber. Communicating month on the inner surfaces of the fingers,
branches from adjacent penicillate units per¬ palms, and soles as parallel ridges protruding
mit exchange of axons and establish horizon¬ into the dermis. From the beginning they
tal continuity of the subepidermal plexus. It show a characteristic pattern, and from them
is not clear which sensory modality is served sweat glands develop.
by these endings, but their overlapping and The regional specificity of the epidermis
ramification over relatively large areas could has been the subject of detailed embryologi-
not provide highly localized sensation. cal study. It has been shown that maintenance
Where the skin bears even fine hairs, these of the adult specificity of the epidermis de¬
are important adjuncts to tactile sensation. pends on the dermis. When the dermis and
The myelin sheath of nerve fibers in dermis epidermis are separated and epidermis from
terminates as they approach hair follicles, the ear is implanted on dermis from the sole,
and the branching axons that continue are thick epidermis will develop. If a composite
individually ensheathed in processes of the graft from sole epidermis is maintained with
terminal Schwann cell. They terminate in ear dermis, the originally thick epidermis
multiple fusiform endings applied to the root becomes thinner and hair follicles develop.
sheath of the sloping hair shaft on its under¬ The dermis and hypodermis consist during
side just below the sebaceous glands. In this the first six weeks of mesenchyme with wan¬
position, the spindle endings are strategically dering cells. From the second month on, the
placed to sense deformation resulting from collagenous fibers appear and elastic fibers
576 • SKIN

follow. In still later stages, the mesenchyme layers of epidermis that cover the plate even¬
divides into a peripheral dense layer with a tually desquamate.
compact arrangement of its elements—the The development of the eccrine sweat
dermis—and a deep loose layer, the future glands proceeds independently of that of the
subcutaneous layer. In the dermis, in turn, hairs. The first primordia appear during the
the peripheral papillary layer ditferentiates. fifth month on the palms and soles and the
Hair first appears in the eyebrows and on lower surface of the fingers. At first they are
the chin and upper lip, at the end of the similar to the primordia of the hairs. An
second month. At first, in the deep layer of epithelial shaft with a terminal thickening
the epidermis, a group of dividing cells ap¬ grows into the underlying connective tissue.
pear. These grow into the underlying con¬ But, unlike that around the hairs, the con¬
nective tissue and produce a gradually elon¬ nective tissue here does not condense about
gating epithelial cylinder. This is the the epithelium. The shaft gradually elongates
primordium of a hair follicle, the so-called and becomes cylindrical, and its lower por¬
“hair germ”; it is rounded on its end. Under tion curls in the form of a ball. Beginning in
the latter an accumulation of condensed con¬ the seventh month, an irregular lumen forms
nective tissue appears early. From it the hair in this lower portion, which constitutes the
papilla forms and protrudes into the epithe¬ secretory part; along the course of the future
lial mass of the bulb. The epithelial cells at excretory duct another lumen develops and
the surface of the connective tissue papilla later unites with the first one. In the secretory
represent the matrix of the future hair. The portion, the epithelium around the lumen
connective tissue that surrounds the bulb forms two layers, which differentiate into an
later forms the connective tissue portions of external layer of myoepithelial elements and
the hair follicle. On the surface of the epi¬ an internal layer of glandular cells.
thelial hair bulb, two projections arise. The Quantitative investigations have shown that
upper represents the primordium of the se¬ in the embryo the density of the skin appen¬
baceous gland; its central cells early undergo dages, regardless of whether they are hair or
a fatty transformation. The lower protuber¬ eccrine glands, is originally the same. A large
ance becomes the insertion of the arrector proportion of these appendages on the head,
pili muscle on the hair sac. In the mass of however, will become hairs and a large pro¬
epithelium that forms the hair primordium, portion of them on the rest of the body will
a layer of rapidly cornifying cells differen¬ become eccrine sweat glands. No hairs will
tiates into the layers of matrix, cortex, and be found on the palms and the soles. Due to
inner and outer root sheaths. The shaft of the differential rate of growth of the body
the new hair elongates, owing to the multi¬ surface, the original uniform density changes
plication of the cells of the matrix on the because no new hair follicles or eccrine sweat
summit of the papilla, and perforates the top glands form after the original population is
of the hollow cone of Henle’s sheath. The tip established. These appendages subsequently
of the hair moves upward, pierces the epi¬ become widely spaced in the trunk and in
dermis, and protrudes above the surface of the extremities, which grow to a surface area
the skin. about three times as great as that of the head.
The development of the nails begins in the In wound healing, usually no new hair folli¬
third month by the formation, on the back cles are formed.
of the terminal phalanx of each finger, of a
flat area, the primary nail field, which is sur¬
rounded by a fold of the skin. In the region
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of the nail the epithelium has three or four
layers. The true nail substance is laid down
GENERAL
during the fifth month in the portion of the
Montagna, W. W., and P. F. Parakkal: The Structure
nail bed near the proximal nail groove. Here
and Function of Skin. 3rd ed. New York, Academic
the deep layer of the epidermis is trans¬ Press, 1974.
formed into the nail matrix, and its cells Rothman, S.: Physiology and Biochemistry of the Skin.
become flat, adjoin one another closely, and Chicago, University of Chicago Press, 1954.
give rise to the true nail plate. In the begin¬ EPIDERMIS AND KERA TINIZA TION
ning it is still thin and is entirely buried in Allen, T. D., and C. S. Pottan: Desmosomal form, fate
the epidermis of the nail held or bed. It and function in mammalian epidermis. J. Ultra-
gradually moves in the distal direction. The struct Res. 57:94,1975.
 SKIN • 577

De Bersaques, J.: Keratin and its formation. Curr. Probl. Wolff, K., and E. Scheimer: Uptake, intracellular trans¬
Dermatol. 6:34, 1976. port and degradation of exogenous protein by Lan¬
Elias, P. M.: Epidermal lipids, membranes, and keratin- gerhans cells. J. Invest. Dermatol 54:37, 1970.
ization. Int. J. Dermatol. 26:1, 1981. Wolff, K., and R. K. Winkelmann: Quantitative studies
Elias, P. J., J. Goerke, and D. S. Friend: Permeability on the Langerhans cell population of guinea pig
barrier lipids: composition and influence on epider¬ epidermis. J. Invest. Dermatol 46:504, 1967.
mal structure. J. Invest. Dermatol. 69:535, 1977.
MERKEL CELL
Franke, W. W., E. Schmid, C. Grund, H. Muller, I.
Engelbrecht, R. Moll, J. Stadler, and E. D. Jarash: Hashimoto, K.: Fine structure of the Merkel cell in
Antibodies to high molecular weight polypeptides human oral mucosa. J. Invest. Dermatol 56:381,
of desmosomes: specihc localization of a class of 1972.
junctional proteins in cells and tissues. Differentia¬ Kurosumi, K., V. Kurosumi, and K. Inoue. Morpho¬
tion 26:217, 1981. logical and morphometric studies on the Merkel
Green, H.: The keratinocyte as differentiated cell type. cells and associated nerve terminals of normal and
Harvey Lect. 74:101, 1980. denervated skin. Arch. Histol. Jpn. 42:243, 1979.
Green, H., E. Fuchs, and F. Watt: Differentiated struc¬ Mihara, M., K. Hashimoto, K. Ueda, and M. Kumakiri:
tural components of the keratinocyte. Cold Spring The specialized junctions between Merkel cell and
Harbor Symp. Quant. Biol. 46:293, 1982. neurite: an electron microscopic study. J. Invest.
MacKenzie, I. C.: The ordered structure of mammalian Dermatol 73:325, 1979.
epidermis. In Maiback, H. I., and Rovee, D. T., Winkelmann, R. K.: The Merkel cell system and a
eds.: Epidermal Wound Healing. Chicago, Year comparison between it and the neurosecretory or
Book Medical Publishers, 1972, p. 5. APUD cell system. J. Invest. Dermatol 69:41, 1977.
Matoltsy, A. G.: Desmosomes, filaments, and keratohy-
MELANOCYTES
alin granules: their role in the stabilization and
keratinization of the epidermis. J. Invest. Dermatol. Billingham, R. E., and W. K. Silvers: The melanocytes
65:127, 1975. of mammals. Qt. Rev. Biol. 35:1, 1960.
Matoltsy, A. G., and M. Matoltsy: The chemical nature Drochmans, P.: On melanin granules. Int. Rev. Exp.
of keratohyalin granules of the epidermis. J. Cell Pathol. 2:357, 1963.
Biol. 47:593, 1970. Fitzpatrick, T. B., G. Szabo, M. Seiji, and W. C. Quevedo:
Matoltsy, A. G., and P. E. Parakkal: Membrane-coating Biology of the melanin pigmentary system. In
granules of keratinizing epithelia. J. Cell Biol. Fitzpatrick, T. B., et al., eds: Dermatology in Gen¬
24:297, 1965. eral Medicine 2nd ed. New York, McGraw-Hill Book
Menton, D. N.: A minimum surface mechanism to Co., 1979, p. 131.
account for the organization of cells into columns Snell, R. S.: An electron microscopic study of melanin
in the mammalian epidermis. Am. J. Anat. 145:1, in the hair and hair follicles. J. Invest. Dermatol
1976. 56:218, 1972.
Montagna, W., and W. C. Lobitz, eds.: The Epidermis. Szabo, G.: The regional anatomy of the human integu¬
New York, Academic Press, 1964. ment with special reference to the distribution of
Rice, R. H., and H. Green: Presence in human epidermal hair follicles, sweat glands and melanocytes. Philos.
cells of a soluble protein precursor of the cross- Trans. R. Soc. Lond. [B]. 252:447, 1967.
linked envelope. Activation of the cross linking by
DERMIS AND EPIDERMAL-DERMAL
calcium ions. Cell 76:681, 1979.
JUNCTION
Sun, T. T., and H. Green: Keratin filaments of cultured
human epidermal cells. Formation of intermolecular Briggaman, R. A.: Biochemical composition of the epi¬
disulhde bonds during terminal differentiation. J. dermal-dermal junction and other basement mem¬
Biol. Chem. 253:2053, 1978. branes. J. Invest. Dermatol. 76:1, 1982.
Briggaman, R. A., and C. E. Wheeler: The epidermal-
LANGERHANS CELLS dermal junction. J. Invest. Dermatol. 65:71, 1975.
Breathnach, A. S.: The cell of Langerhans. Int. Rev. Montagna, W., J. P. Bentley, and R. L. Robson. The
Cytol. 18:1, 1965. dermis. Adv. Biol. Skin 10, 1970.
Katz, S., K. Tamaki, and D. H. Sacks. Epidermal Lan¬
HAIR AND NAILS
gerhans cells are derived from cells originating in
the bone marrow. Nature (Lond.) 282:324, 1979. Ebling, F. J.: Hair. J. Invest. Dermatol 67:98, 1976.
Rowden, G.: Immunoelectron microscopic studies of Montagna, W., and R. L. Robson, eds.: Hair growth.
surface receptors and antigens of human Langer¬ Adv. Biol. Skin 9, 1969.
hans cells. Br. J. Dermatol. 97:593, 1977. Orwin, D. F.: The cytology and cytochemistry of the
Shelley, W. B., and J. Lennart: The Langerhans cell: its wool follicles. Int. Rev. Cytol. 66:331, 1979.
origin, nature, and function. Acta Dermatol. Wyatt, E. H., and J. M. Riggot: Scanning electron
(Stockh.) Suppl. 79:7, 1978. microscopy of hair. Observations on surface mor¬
Silberberg, I., R. L. Baer, and S. A. Rosenthal: The role phology with respect to site, sex, and age in man.
of Langerhans cells in allergic contact hypersensitiv¬ Br. J. Dermatol. 96:627, 1977.
ity. A review of findings in man and guinea pigs. J. Zaias, N. J., and Alvarez, J.: The formation of the
Invest. Dermatol. 66:210, 1976. primate nail plate. An autoradiographic study in
Stingl, G., S. I. Katz, L. Clement, I. Green, and E. M. the squirrel monkey. J. Invest. Dermatol. 57:120,
Shevack: Immunologic functions of la-bearing epi¬ 1968.
dermal Langerhans cells. J. Immunol. 121:2005,
SEBACEOUS GLANDS
1978.
Tew, J. G., J. Thorbecke, and R. M. Steinman: Dendritic Bell, M. A.: A comparative study of sebaceous gland
cells in immune response. J. Reticuloendothel. Soc. ultrastructure in sub-human primates. Anat. Rec.
31:371, 1982. 776:331, 1971.
578 • SKIN

Rupee, M.: Zur Ultrastruktur der Talgdriisenzelle. Arch. Robertshaw, D., C. R. Taylor, and L. M. Mazia: Sweating
Klin. Exp. Derm. 234:273, 1969. in primates: secretion by adrenal medulla during
exercise. Am. J. Physiol. 224:678, 1973.
SWEAT GLANDS Sato, K.: The physiology, pharmacology, and biochem¬
Cage, G. W., and R. L. Dobson: Sodium secredon and istry of the eccrine sweat gland. Rev. Physiol.
reabsorpdon in the human eccrine sweat gland. J. Biochem. Pharmacol. 79:51, 1977.
Clin. Invest. 44:1270, 1965. Sato, K., and R. L. Dobson: Regional and individual
Dobson, R. I.: The correlation of structure and function variations in the function of the human eccrine
in the human eccrine sweat gland. In Montagna, sweat gland. J. Invest. Dermatol. 54:443, 1970.
W., R. A. Ellis, and A. F. Silver, eds.: Advances in Schaumberg-Lever, G., and W. F. Lever: Secretion from
Biology of Skin. Vol. III. New York, Appleton- human apocrine glands: an electron microscopic
Century-Crofts, 1962, pp. 54—75. study. J. Invest. Dermatol. 64:38, 1975.
Dole, V. P., and J. H. Thaysen: Variation in the func¬ Szabo, G.: The number of eccrine sweat glands in human
tional'power of human sweat glands. J. Exp. Med. skin. In Montagna, W., R. Ellis, and A. Silver, eds.:
98:129, 1953. Advances in Biology of Skin. Vol. III. New York,
Ellis, R. A.: Eccrine sweat glands. In Jadassohn, J., ed.: Pergamon Press, 1962, pp. 1-5.
Handbuch der Haut und Geschlechtskrankheiten I. Terzakis, J. A.: The ultrastructure of monkey eccrine
Band. Normale und Pathologische Anatomie der sweat glands. Z. Zellforsch. 64:493, 1964.
Haut. Berlin-Heidelberg-New York, Springer, Lino, H., and W. Montagna: Catecholamine containing
1967. nerve terminals in the eccrine sweat glands of ma¬
Forstrom, L., M. E. Goldyne, and R. K. Winkelmann: caques. Cell Tissue Res. 158:\, 1975.
Ig E in human eccrine sweat. J. Invest. Dermatol.
64:156, 1975. BLOOD VESSELS AND INNERVATION
Haimovici, H.: Evidence for adrenergic sweating in man.
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Kuno, Y.: Human Perspiration. Springfield, II, Charles human dermal microcirculation. II The capillary
C Thomas, 1956. loops of the dermal papillae. J. Invest. Dermatol.
Lloyd, D. P. C.: Secretion and reabsorption in eccrine 68:44, 1977.
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Biology of Skin. Oxford—London—New York, Per- innervation and modalities of cutaneous sensibility.
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Montgomery, I., D. M. Jenkinson, and H. Y. Elder: The Odland, G. F.: The fine structure of cutaneous capillar¬
effects of thermal stimulation on the ultrastructure ies. In Montagna, W., and R. A. Ellis, eds.: Advances
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Biochem. Cytol. 77:385, 1961. Thomas, 1960.
 23
ORAL CA V/TY AND
ASSOC/A TED GLANDS

The oral cavity is the entrance to the long gastrointestinal tract, such as the salivary
tubular digestive system, which consists of the glands, liver, and pancreas. However, these
lips, mouth, pharynx, esophagus, stomach, remain connected by long ducts to the epi¬
small and large intestine, rectum, and anus. thelial surface from which they originated in
On its way through this tract, the food under¬ embryonic life.
goes mechanical fragmentation and chemical In the oral cavity, esophagus, and rectum,
digestion. Products of degradation of the the wall of the digestive tube is surrounded
food are absorbed through the wall of the by a layer of connective tissue that attaches it
intestine into the blood, which carries them to the adjacent structures. The stomach and
to the tissues for utilization or storage. The intestines, on the other hand, are suspended
undigested residue of the food is eliminated in the abdominal cavity by mesenteries, and
as feces. their surface is covered by a moist serous
The inner surface of the digestive tube is membrane, the serosa or peritoneum, which
lined with a mucous membrane or mucosa, permits these organs to slide freely over one
which consists of a superficial layer of epithe¬ another within the cavity during the peristal¬
lium regionally specialized for different tic movements of the digestive tract.
digestive functions; and a supporting layer The wall of the digestive tube is richly
of loose connective tissue, the lamina propria. provided with blood vessels that bring to it
The peripheral layers of the wall of the oxygen and the metabolites necessary to sus¬
digestive tube are smooth muscle making up tain its secretory activities. The veins and
the muscularis externa (Fig. 23—1). In most lymphatics also carry away the absorbed
parts of the tract, the outer limit of the products of digestion. In addition, the wall
mucosa is demarcated by a thin layer of of the digestive tract contains an intricate
smooth muscle, the muscularis mucosae. Be¬ system of sympathetic ganglia and nerve
tween it and the muscularis externa is a layer plexuses concerned with coordination of the
of loose connective tissue, the submucosa. Nu¬ movements of the digestive tube.
merous blood vessels, nerves, lymphatics, and
lymphoid nodules are to be found in this
layer. Where a muscularis mucosae is absent,
there is a gradual transition from the lamina THE ORAL CAVITY
propria to the submucosa.
The mucosa of the developing gastrointes¬
tinal tract evaginates to form folds and villi The epithelium of the mucous membrane
that project into the lumen and increase the in the mouth is of stratified squamous type,
surface area of the absorptive epithelium. It like that of the skin. However, in the human,
also invaginates to form tubular crypts or it normally does not undergo complete ker-
glands whose lining cells produce mucus, atinization. The nuclei of the cells of the
digestive enzymes, and hormones. The ma¬ superficial layers condense and become met-
jority of these outgrowths remain confined abolically inert, but do not disappear, and
to the thickness of the mucosa. Other evagin- the cell bodies do not reach the same degree
ations proliferate to such an extent during of flatness as in the epidermis. The superficial
embryonic development that they give rise to cells are continually exfoliating in great num¬
separate organs, the accessory glands of the bers and are found in the saliva. In some

579
580 • ORAL CAVITY AND ASSOCIATED GLANDS

Mesentery

Intestinal
\ Serosa Muscularis
villi externa

Plica circularis
Submucosal
gland

Submucosa
Excretory duct of large
gland outside of the
digestive tube

Mucosa

Cells of 1?
Auerbach’s plexus-11
Epithelial
1 ning

Lamina
propria
Mucosa^ la n<jjs
(crypts erkuhn)

Muscularis
mucosae
Circular muscle

Cel Is of Meissner’s plexus

Longitudina Lymphoid
muscle nodule
Figure 23-1. Schematic representation of the general features of organization in the gastrointestinal tract. The concentric
layers of serosa, muscularis, and mucosa are common to virtually all regions of the tract. In the upper half of the
drawing, the mucosa is depicted with glands and villi, as in the small intestine; in the lower half, it is shown with glands
only, as in the colon.

places they contain granules of keratohyalin, submucous plexus of large vessels, from
and glycogen is present in the superficial and which arise branches that form a second
middle layers of the epithelium. In many plexus in the lamina propria. This in turn
animal species, particularly ruminants, the sends capillaries into the papillae. The lym¬
epithelium of the oral cavity undergoes ex¬ phatics also show an arrangement similar to
tensive keratinization. that in the skin, and begin with blind capillary
The lamina propria, in most places, ex¬ outgrowths in the papillae.
tends into concavities in the deep surface of The oral mucous membrane is very sensi¬
the epithelium forming connective tissue pa¬ tive and is provided with many nerve endings
pillae similar to those associated with the derived from the sensory branches of the
epidermis of the skin. Their structure is, trigeminal nerve. On that portion covering
however, more delicate, and the collagenous the tongue it also contains the specific end
and elastic fibers are thinner than in the organs of the sense of taste. Under the lamina
dermis. In the posterior region of the oral propria, especially in the cheeks and on the
cavity the lamina propria contains many lym¬ soft palate, there is a loose submucosa, which
phocytes, which are often found migrating gradually merges with the denser connective
into and through the epithelium. tissue of the mucosa. In regions with a well-
The arrangement of the blood vessels is developed submucosa the mucous membrane
similar to that in the skin. There is a deep can be easily lifted into folds, whereas in
 ORAL CAVITY AND ASSOCIATED GLANDS • 581

those places against which the food is crushed distance onto its nasal surface. On this sur¬
and rubbed, as on the hard palate, there is face, at varying distances from its posterior
no submucosa, and the mucous membrane is margin, the stratified epithelium is replaced
firmly bound to the periosteum of the un¬ by pseudostratihed, ciliated, columnar epi¬
derlying bone. thelium, which rests on a thickened basal
The inner zone of the lip margin in the lamina. The lamina propria contains small
newborn is considerably thickened and pro¬ glands of the mixed type, but no adipose
vided with many high papillae and numerous tissue, and it is heavily infiltrated with lym¬
sebaceous glands that are not associated with phocytes. A dense layer of elastic fibers is
hairs (Fig. 23-2). These structural features found between the glands and the underlying
seem to facilitate the process of suckling. muscle. A submucosa is not present.
The soft palate consists of layers of striated Most of the structures in the oral cavity—
muscle and fibrous connective tissue covered the salivary glands, the lining of the palate,
with mucous membrane. On the oral surface the anterior two thirds of the tongue, and
the latter has the structure typical of the oral the vestibule of the nose—are derived from
cavity-a stratified squamous epithelium, with the embryonic ectoderm. The tooth enamel
high papillae, and associated mucous glands. is said to be a neural crest derivative, whereas
The glands are surrounded by adipose tissue the dentin and pulp originate from mesen¬
and are scattered in a loose submucous layer chyme. Endodermal derivatives include the
separated from the lamina propria by dense tonsils, the epithelium of the posterior third
elastic networks. This oral type of mucous of the tongue, the pharyngeal tonsils, and
membrane also covers the posterior margin the remainder of the gastrointestinal tract
of the soft palate and continues for some and its associated glands.

Blood vessel Thickened epi¬

thelium {for
sucking)

Orbicularis
oris muscle

Red
portion
of lip
Oral
epithe¬
lium

X
Hair follicle

Mucous Sebaceous
glands glands

Skin

Figure 23-2. Camera lucida drawing of sagittal section through lip of a newborn infant. Stained with hematoxylin,
x 10.
582 • ORAL CAVITY AND ASSOCIATED GLANDS

Palatine tonsil
Median epiglottic fold
Vallate Lingua
papilla
tonsil
Epiglottis
Rows of
filiform papillae

Fungiform papilla

Fungiform papilla

Fungiform Filiform
Vallate
papilla papillae papiMa
Lingual tonsil
Lingual
tonsil Palatine tonsil

Figure 23-3. Surface of dorsum and root of human tongue. (After Sappey, from Schumacher.)

THE TONGUE and foliate. The filiform papillae are arranged

in more or less distinct rows diverging to the
The tongue consists of interlacing bundles right and left from the middle line and par¬
of striated muscle that run in three planes allel to the V-shaped boundary between an¬
and cross one another at right angles. The terior and posterior regions of the tongue.
muscular mass is covered by a tightly adher¬ The fungiform papillae are scattered singly
ent mucous membrane. The dense lamina among the filiform papillae and are especially
propria is continuous with the interstitial con¬ numerous near the tip of the tongue. The
nective tissue of the muscle. A submucous circumvallate papillae, numbering only 10 to
layer is present only on the smooth ventral 12 in man, are distributed along the diverg¬
surface of the tongue. The dorsal surface is ing arms of the V-shaped boundary (Fig. 23-
covered in its anterior two thirds by a multi¬ 3). The paired foliate papillae are found on
tude of small excrescences—the lingual pa¬ the dorsolateral aspect of the posterior part
pillae—whereas in its posterior one third it of the tongue.
presents only irregular bulges of larger size. The filiform papillae are 2 to 3 mm long.
The boundary line between the two regions Their connective tissue core forms secondary
is V-shaped, with the opening of the angle papillae with pointed ends (Fig. 23—4). The
directed forward (Fig. 23-3). At the apex of epithelium covering these connective tissue
the angle is a small invagination, the foramen outgrowths also forms short processes, which
caecum. It is the rudiment of the thyroglossal taper to a point (Fig. 23—6). The superficial
duct, which in early embryonic stages con¬ squamous cells are transformed into hard
nects the thyroid gland primordium with the scales containing shrunken nuclei. The axial
epithelium of the oral cavity. parts of the scales at the point of the papilla
are connected with its solid axial core and
Papillae their lower edges project from the surface of
the papilla like the branches of a hr tree.
Four types of papillae are present on the When digestion is disturbed in illness, the
tongue: filiform, fungiform, circumvallate, normal shedding of these scales is delayed.
 ORAL CAVITY AND ASSOCIATED GLANDS • 583

Fungiform papillae

Filiform papillae

Cornified tips
Figure 23-4. Surface of dorsum of
tongue, drawn from a combined study
using the binocular microscope and Epithelium
from sections. The anterior cut surface
corresponds with the long axis of the Papillae
tongue—the tip of the tongue being to
the reader’s left, x 16. (After Braus.)
Lamina propria

Blood vessel

Vertical muscle
Taste bud Longitudinal muscle

Filiform papillae

Lenticular papillae
(ilingual follicles)

Lingual
tonsils

Epithelium

Vallate
papillae

Outer wall
of furrow
Circular
furrow
Taste
buds
Glands of Leuko¬
v. Fbner cytes
in the
epith¬
elium

Striated
muscles Glands of the mucosa Glandular duct
Figure 23-5. Surface of tongue at the border between the root and the dorsum, x 16. (After Braus.)
584 • ORAL CAVITY AND ASSOCIATED GLANDS

Figure 23-6. Scanning electron micrograph of the filiform papillae of rabbit tongue. (Micrograph courtesy of F. Fujita.)

They then accumulate, in layers mixed with that project into recesses in the underside of
bacteria, on the surface of the tongue, which the epithelium, which has a smooth free
becomes covered with a gray him—the surface (Figs. 23—4, 23-7). On many of the
“coated” tongue. fungiform papillae the epithelium associated
The fungiform papillae have a short, with the secondary papillae contains taste
slightly constricted stalk and a slightly flat¬ buds. Because the core is rich in blood vessels
tened hemispherical upper part. The con¬ and the epithelium is relatively thin, the fun¬
nective tissue core forms secondary papillae giform papillae have a distinct red color.

Secondary papilla
Keratohyalin
in filiform lar stroma with
papilla vessels

Connective tissue
of fungif orm papilla Figure 23-7. Perpendicular section
through a fungiform papilla, x 46.
(After Schaffer, from Schumacher.)

Striated muscle
 ORAL CAVITY AND ASSOCIATED GLANDS • 585

Stratified squa¬
mous epithelium

Lamina propria

Figure 23-8. Section through circum-

vallate papilla of Macacus rhesus.
Photomicrograph, x 42.
Taste bud

Gland of u. Ebner

The circumvallate papillae are recessed can often be seen aligned on the lateral
into the surface of the mucous membrane, surface of the papilla. A few may be present
and each is surrounded by a deep, circular in the outer wall of the groove surrounding
furrow. The connective tissue core forms the papilla. The number of taste buds in a
secondary papillae only on the upper surface. single papilla is subject to great variations,
The covering epithelium is smooth con¬ but it has been estimated to average 250.
toured, whereas that of the lateral surfaces Connected with the circumvallate papillae are
of the papillae contains many taste buds (Fig. glands of the serous type (.glands of von Ebner),
23-8). In vertical sections, 10 to 12 of them which are embedded deep in the underlying

Figure 23-9 Photomicrograph of the foliate papillae of a rabbit, showing the alternating ridges and deep clefts with
numerous taste buds on either side of the cleft, x 150. An area such as that enclosed in the rectangle is shown at
higher magnification in Figure 23-10.
586 • ORAL CAVITY AND ASSOCIATED GLANDS

muscular tissue and whose excretory ducts

open into the bottom of the circumferential
furrow.
In the human, foliate papillae are rudi¬
mentary, but in many animals they are the
site of the main aggregations of taste recep¬
tors (Fig. 23-9). The fully developed foliate
papillae in the rabbit, for example, are oval
bulgings on the mucous membrane, consist¬
ing of alternating parallel ridges and grooves.
The epithelium of the sides of the ridges
contains great numbers of taste buds. Small
serous glands open into the bottom of the
grooves between the folia.
Taste buds are also found on the glosso-
palatine arch, on the soft palate, on the
posterior surface of the epiglottis, and on the
posterior wall of the pharynx as far down as
the inferior edge of the cricoid cartilage.
The nodular bulges on the root of the
tongue are caused by lymphatic nodules,
forming the lingual tonsils (Fig. 23—3). On the Figure 23-10. Photomicrograph of taste buds from the
free surface of each lingual tonsil a small foliate papillae of a rabbit, x 450. See Figure 23-9 for
location. Arrows indicate the taste pores.
opening leads into a deep invagination, the
crypt, lined with stratified squamous epithe¬
lium. The crypt is surrounded by lymphoid the taste buds of the rabbit. Typical synapses
tissue. Innumerable lymphocytes infiltrate of nerves with the epithelial cells of taste
the epithelium and congregate in the lumen buds have been observed only on Type III
of the crypt, where they degenerate and form cells. Basal cells and peripheral cells have also
masses of detritus with the desquamated ep¬ been described, and these are believed to be
ithelial cells and bacteria. The lingual tonsils undifferentiated progenitor cells of the taste
are often associated with mucous glands buds.
embedded in the underlying muscle tissue. Both Type I and Type II cells have large
The ducts of the latter open either into the microvilli that projects into the taste pore,
crypt or onto the free surface. where they are embedded in a rather amor¬
phous extracellular substance. Material of
Taste Buds similar density and texture is found in mem¬
brane-bounded vesicles in the apical cyto¬
The taste buds are seen in sections under plasm of Type I cells. It is presumed to be a
low power as pale, oval bodies in the darker- secretory product of these cells. The taste
stained epithelium (Fig. 23-9). Their long hairs described by light microscopists were no
axis averages 72 pm. They extend from the doubt the clusters of coarse microvilli found
basal lamina almost to the surface. The epi¬ in the pore region in electron micrographs.
thelium over each taste bud is pierced by a Zonulae occludentes are present on the
small opening—the taste pore (Figs. 23-10, boundaries between cells near the free sur¬
23-11,23-12). face. The lateral surfaces of the cells below
Light microscopists distinguished two types the junctional complexes may be folded and
of cells among the constituents of taste buds, interdigitated. The cytoplasm of the lighter
the dark supporting cells (Type I) and the light Type II cells has a rather extensive smooth
neuroepithelial cells (Type II). Electron micro¬ endoplasmic reticulum, which is not observed
scopists have confirmed the existence of two in Type I cells.
cell types but are not able to assign a func¬ There are only four fundamental taste
tional role to them with any certainty and sensations: sweet, bitter, acid, and salty. It
have therefore adopted the designation Type has been shown by the application of sub¬
I and Type II to avoid functional implica¬ stances to individual fungiform papillae that
tions. A Type III cell structurally comparable they differ widely in their receptive proper¬
with the Type I cell has been identified in ties. Some do not give any taste sensations,
 ORAL CAVITY AND ASSOCIATED GLANDS • 587

Figure 23-11. Electron micrograph of the pore region of a rabbit taste bud. Notice the large microvilli on the sensory
cells, surrounded by a dense amorphous secretory product. For a surface view of the pore, see Figure 23-12.
(Micrograph courtesy of M. Coppe.)

whereas others give sensations of one or subepithelial plexus, from which fibers pen¬
more of the four taste qualities. No structural etrate the epithelium. Some terminate as in-
differences in the various taste buds have tergemmal fibers by free arborization between
been found, in spite of the differences in the taste buds. Others, the perigemmal fibers,
sensation mediated. There is, moreover, a closely envelop the taste buds; and still oth¬
general chemical sensitivity in regions of the ers, the intragemmal fibers, penetrate the taste
mouth where there are no taste buds. buds and end with small terminal enlarge¬
ments in intimate contact with certain of the
taste bud cells. The functional significance of
Nerves these different nerve endings is unknown.
The anterior two thirds of the tongue is
innervated by the lingual nerve, which con¬
tains fibers of general sensibility from the
fifth cranial nerve (trigeminal) and fibers of GLANDS OF THE ORAL
gustatory sensibility from the seventh cranial CAVITY
nerve (facialis). The latter enter the lingual
nerve from the chorda tympani. The poste¬ General Description
rior third of the tongue is innervated by the
glossopharyngeal nerve for both general and Numerous salivary glands open into the oral
gustatory sensibility. Taste buds of the epi¬ cavity. Many of them are small glands in the
glottis and lower pharynx are innervated by mucosa or submucosa and are named accord¬
the vagus. These nerve fibers are lightly mye¬ ing to their location. They seem to secrete
linated. They branch profusely under the continuously and furnish a liquid, the saliva,
basal lamina, lose their myelin, and form a which moistens the oral mucous membrane.
588 • ORAL CAVITY AND ASSOCIATED GLANDS

Figure 23-12. Scanning micrograph of the pore of a taste bud opening onto the surface of the epithelium. (Micrograph
courtesy of M. Coppe.)

In addition, there are three pairs of large from the oral cavity varies, depending on the
glands, which constitute the salivary glands degree of participation of the various salivary
proper. They are the parotid, the submandib¬ glands in its formation. But even the secre¬
ular (submaxillary), and the sublingual glands. tions of the same gland may change consid¬
They secrete only when mechanical, thermal, erably with variations in the stimuli acting
or chemical stimuli act upon the nerve end¬ upon the oral mucous membrane, as, for
ings in the oral mucous membrane, or as the instance, with different kinds of food.
result of certain psychic or olfactory stimuli. The salivary glands may be classified in
The saliva collected from the oral cavity is three categories, according to the type of
a mixture of the secretions of the various their secretory cells. The glands containing
salivary glands. It is a viscous, colorless, opal¬ only mucous cells elaborate a viscid secretion
escent liquid that contains water, mucopro- that consists almost exclusively of mucin. In
tein, immunoglobulins, carbohydrates, and a glands with only serous cells, the secretion is
number of inorganic components, including a watery liquid that lacks mucus but contains
calcium, phosphorus, sodium, potassium, salts, proteins, and the enzymes amylase, ly¬
magnesium, chloride, and traces of iron and sozyme, peroxidase, DNase, and RNase. In
iodine. Enzymes are also present, especially the mixed glands, containing serous and mu¬
amylase (ptyalin), which splits starch into cous cells, the secretion is a viscid liquid
smaller, water-soluble carbohydrates. Saliva containing mucin, salts, and enzymes.
always contains desquamated squamous epi¬ All glands of the oral cavity have a system
thelial cells and the so-called salivary corpus¬ of branching ducts. The secretory portions
cles. The latter originate in the lymphoid or acini of the pure mucous glands are usu¬
nodules of the tongue and in the tonsils, and ally long, branching tubules. In the pure
aie degenerating lymphocytes or granulo¬ serous and mixed glands, the secretory por¬
cytes. tions vary from simple oval acini to tubulo¬
The composition of the saliva collected acinar forms.
 ORAL CAVITY AND ASSOCIATED GLANDS • 589

The initial intralobular ducts are thin, tion of mucigen. The Golgi apparatus, mito¬
branched tubules called the necks, or interca¬ chondria, and rough endoplasmic reticulum
lated ducts. Branches of the next larger order, are located toward the cell base, below the
also intralobular, have a vertically striated mucigen granules. The amount of endo¬
epithelium. These segments are called striated plasmic reticulum varies with the phase of
ducts. Then follow the larger branches. the cell’s secretory cycle. The juxtaluminal
Among them, lobular, interlobular, and pri¬ region of the epithelium is provided with a
mary ducts may be distinguished in the large network of occluding junctions. Secretory
glands. canaliculi are absent. The lumen of the ter¬
minal portions of the glands is large and
usually filled with masses of mucin.
Mucous Cells
When the secretion has been discharged,
In the pure mucous glands, the cells are the cell collapses, and only a few granules of
arranged in a layer against the basal lamina mucigen may remain near its free surface.
and have an irregularly cuboidal form. In In this depleted condition the mucous cells
fresh state their cytoplasm contains many may be mistaken for serous cells, but the
pale droplets of mucigen, the antecedent of absence of intercellular secretory canaliculi
mucin. In fixed and stained sections, the always distinguishes mucous from serous
droplets of mucigen are usually extracted so glands. The demonstration of these canalic¬
that the cell body appears clear and contains uli, however, requires special staining meth¬
only a loose network of cytoplasm and traces ods or electron microscopy. With the initia¬
of precipitated mucigen. This material stains tion of the secretory cycle, the cell organelles
red with mucicarmine, or metachromatically undergo morphological changes correlated
with thionine. The nucleus is near the base with the cell’s activity. Rarely under physio¬
of the cell and usually appears angular and logical conditions do the mucous cells dis¬
compressed when there is a large accumula¬ charge all of their granules. The cells, as a

Figure 23-13. Electron micrograph of mucous acinar cells in a human labial salivary gland. (Micrograph courtesy of B.
Tandler.)
590 • ORAL CAVITY AND ASSOCIATED GLANDS
^ \

rule, do not show any signs of degeneration The serous cells in the various oral glands
and apparently recover completely from dis¬ have a similar microscopic structure even
charge of their secretion. Mitoses have occa¬ though they are not functionally identical
sionally been observed in them. (Figs. 23—17, 23-18). They are grouped to¬
gether under a general heading because his¬
tological methods are not able to demonstrate
Serous Cells
differences corresponding to the variations
These cells are roughly cuboidal and sur¬ in the nature of their secretions.
round a small tubular lumen. In an unfixed Salivary glands exhibit considerable inter¬
resting gland they contain numerous highly specific variation in their cytology. In some,
refractile secretion granules that accumulate including the human parotid gland, cells with
between the nucleus and the free surface. the appearance of serous cells have secretory
After the gland has secreted for a certain granules that stain with the periodic acid—
period, the serous cells diminish in size and Schiff (PAS) reaction for carbohydrates and
their few remaining granules are confined to contain sialomucin and sulfomucin in addi¬
the juxtaluminal cytoplasm. The Golgi com¬ tion to enzyme proteins. Such cells are
plex is usually found in an apical or para¬ termed seromucous cells.
nuclear position, and cisternae of endo¬
plasmic reticulum are abundant toward the Cells in the Mixed Glands
cell base. Mitochondria are scattered among
the cisternae at the base and in the apical The relative numbers of the two kinds of
cytoplasm as well. The luminal surface of the glandular cells in the mixed glands vary
serous cells has sparse microvilli, and inter¬ within wide limits. In some cases the serous
cellular canaliculi lined with microvilli are cells are far more numerous than the mucous
found between the cells. In human subman¬ cells, whereas in other glands the reverse is
dibular glands, the serous acinar cells have true. In still other instances, both cell types
numerous basal processes that interdigitate are present in about equal numbers. The
to form a complex basal labyrinth typical of mucous and serous cells line different parts
cells transporting fluid and electrolytes. of the terminal secretory portion of the

Basket cell
Serous cells
Secretory capillary
/ (intercellular)
Serous terminal
portion

Demi
lune

Processes
of basket cell f Basal lamina
Intercalated
Mucous cells of branched terminal portion
portion

Salivary duct

e c

Figure 23-14. Reconstruction of a terminal portion of a submandibular gland with its duct, b, Cross section of a purely
serous terminal portion, showing basal lamina; c, cross section through a purely mucous terminal portion; d, cross
section through an intercalated portion; e, cross section through a salivary duct. (Redrawn and modified after a
reconstruction by Vierling, from Braus.)
 ORAL CAVITY AND ASSOCIATED GLANDS • 591

Figure 23-15. Photomicrograph of the human subman¬

dibular gland, a mixed gland, showing serous acini at
lower left and mucous acini with serous demilunes at
upper right, x 475.

Serous cells Mucous

cells

Myoepithelial
cells

Figure 23-16. Drawing of a tubuloalveolar end piece in a mixed salivary gland showing mucous cells capped by a
serous demilune. Human submandibular gland. (From Krstic, R.V. Die Gewebe des Menschen und der Saugetiere.
Berlin, Springer Verlag, 1978.)
Figure 23-17. Electron micrograph of a serous acinus from human submandibular gland. (From Tandler, B., and R. A
Erlandson. Am. J. Anat. 735:419, 1972.)

Figure 23-18. Electron micrograph of serous cells from submandibular gland of rhesus monkey. Notice the nonhomo
geneity of the secretory granules, with dense and less dense regions. (Micrograph courtesy of A. Ichikawa.)
592
 ORAL CAVITY AND ASSOCIATED GLANDS • 593

gland. In those mixed glands in which the Ducts of the Glands

*
serous cells predominate, some of the termi¬
nal portions may be exclusively serous (Fig. The ducts of the glands of the oral cavity
23-15). In others a part of the secretory are of variable length and have a low cuboidal
portion is lined with mucous cells and a part epithelium. Between the lining cells and the
is lined with serous cells. In sections the basal lamina are scattered myoepithelial cells.
mucous portions can usually be recognized The epithelium of the necks may show vary¬
by the clear, empty appearance of their cy¬ ing degrees of transformation to mucous
toplasm, but they are identified more cer¬ cells.
tainly by their color after staining of the In the columnar epithelium of the striated
mucus with the PAS reaction. segments of the ducts, the lower parts of the
As a rule, the mucous cells are located cell bodies exhibit a parallel striation, attrib¬
nearer the ducts, whereas the serous cells are utable to vertical orientation of mitochondria
located at the end of the terminal secretory in slender compartments formed by infold¬
portion. It is quite probable that the mucous ing of the basal cell membrane (Fig. 23-19,
cells in mixed glands arise by differentiation 23—20, 23-21). The numerous infoldings of
of the cells in the smallest ducts. Sometimes the basal surface of the cells in the striated
ducts are not resolved with the light micro¬
single mucous cells are scattered among the
scope but are visible with the electron micro¬
unspecialized cells of the neck of the gland.
In other cases where the mucous transfor¬ scope, as they are in other epithelia in which
mation affects all the cells, the neck of the there is rapid transport of water and ions.
duct ceases to exist as such. If the mucous In the larger ducts the epithelium is col¬
cells are not numerous, the secretory portion umnar and pseudostratihed, and occasionally
contains goblet cells. Nearing the opening on
of the gland will show an irregular mixture
the mucous membrane, it becomes stratified
of the pale mucous and dark serous cells.
for a short distance and is then succeeded by
If the mucous cells predominate, the ser¬
stratified squamous epithelium.
ous cells are displaced to the terminal portion
or into saccular outpocketings of the acinus.
Flere they form small groups, which in sec¬ Glands Opening into the Vestibule
tions appear as darkly staining crescents
Scattered in the mucous membrane of the
(idemilunes of Giannuzzi) surrounding the ends
upper and lower lips are small labial glands
of the tubules of mucous cells (Figs. 23-14,
(Fig. 23—13). Similar glands associated with
23—15, 23—16). In the demilunes the serous
the mucous membrane of the cheeks are
cells are small and flattened and often seem
called buccal glands. Both of these are glands
to be entirely separated from the lumen by
of mixed type. The secretory portion some¬
the large mucous cells. However, there are
times contains only seromucinous cells, but
always secretory canaliculi that conduct the
in most cases these are confined to the blind
secretion through clefts between the mucous
ends of the glands, with the remainder lined
cells into the lumen (Fig. 23-16).
with mucous cells. Since the necks of the
glands are short and branch very little, the
Basal Myoepithelial Cells mucous secretory portions often pass directly
into striated ducts.
In all the glands of the oral cavity, the By far the largest salivary glands opening
epithelium in the terminal portion, as well as into the vestibule are the two parotid glands.
in the ducts, is provided with basal cells. They Situated subcutaneously on either side of the
lie between the glandular cells and the basal face just in front of the ear, they extend from
lamina and appear as slender, spindle-shaped the zygomatic arch above to below the angle
or stellate elements (Fig. 23—16). Usually, of the jaw. Each is connected to the vestibule
only their nuclei can be discerned in sections. by a long parotid duct (Stenson’s duct), which
When seen from the surface, they exhibit a emerges from the anterior border of the
stellate cell body with processes containing gland and courses forward and then through
many cytoplasmic filaments. In their ultra¬ the cheek to open into the mouth opposite
structure, they resemble smooth muscle cells, the second upper molar tooth. In the adult
and are presumed to be contractile and to human, the gland is largely serous in its
facilitate the movement of the secretion into histological appearance. However, in large
the ducts. They are considered to be myo¬ areas of the gland the cells have a positive
epithelial cells and resemble those of the staining reaction for carbohydrate and there¬
sweat glands and mammary gland. fore must be interpreted as seromucous.
594 • ORAL CAVITY AND ASSOCIATED GLANDS

Acinar cell

Figure 23-19. Diagrammatic representation of the fine structural characteristics of the various cell types in the mouse
submandibular gland. (Redrawn after U. Rutberg.)
 ORAL CAVITY AND ASSOCIATED GLANDS • 595

At the posterior end of these is another

group of small glands called the glossopalatine
glands.
In the sublingual glands, mucous cells are
far more numerous than in the submandib¬
ular glands. There are no acini formed ex¬
clusively of serous cells. For the most part
these are arranged in thick demilunes. The
gland consists of about 60 per cent mucous
cells and 30 per cent serous cells. The striated
ducts are few and short and are sometimes
represented only by small groups of basally
striated cells in the epithelium of the inter¬
lobular ducts. The glossopalatine glands are
pure mucous glands.

Glands of the Tongue

On either side of the midline near the tip
of the tongue is an anterior lingual gland
(gland of Blandin or Nuhn). The anterior
portion of this gland contains secretory tu¬
bules with seromucinous cells only. Its pos¬
terior part consists of mixed branching tu¬
bules that contain mucous cells and, on their
Figure 23-20. Photomicrograph of a striated duct from
the parotid gland of a marmoset. Notice the orientation of blind ends, thin demilunes of seromucinous
mitochondria parallel to the cell axis, giving the basal cells.
cytoplasm a vertically striated appearance. (Micrograph
courtesy of B. Tandler.)

Glands Opening on the Floor of the

Mouth
In the space between the mandible and the
muscles forming the floor of the mouth, on
either side, is a large submandibular gland. Its
duct (Whartons duct), about 5 cm. long, leaves
the deep surface of the gland and runs for¬
ward to open at the tip of the sublingual
papilla on the floor of the mouth adjacent to
the frenulum of the tongue. The secretory
portion of the gland is mainly seromucous,
but some parts are mucous, with serous cells
at the blind ends. Typical serous demilunes
(crescents of Giannuzzi) are less common in
the submandibular gland of the human than
in other species. Striated ducts are numerous
and long and have many branches.
Deep to the mucous membrane on the
underside of the tongue, there is a large
sublingual gland, 3 to 4 cm long, on either
side of the frenulum, and a number of
smaller sublingual glands (Fig. 23-22). The
ducts of the large glands open into the ducts
Figure 23-21. Electron micrograph of basal region of
of the submandibular gland. The small striated duct cells from cat submandibular gland. Notice
glands open separately along a fold of mu¬ the desmosomes joining the interdigitating processes of
cous membrane called the plica sublingualis. neighboring cells. (Micrograph courtesy of B. Tandler.)
596 • ORAL CAVITY AND ASSOCIATED GLANDS

A B
Figure 23-22. Lingual glands, situated among the bundles of striated muscle in rabbit tongue. A, Mucous glands. B,
Serous glands, x 300.

The posterior lingual glands comprise the networks surround the ducts and the termi¬
glands of von Ebner that open into the groove nal portions. The lymph vessels are relatively
around the circumvallate papillae and the scarce.
mucous glands of the root of the tongue. The long
secretory portions of von Ebner’s glands are Nerves
branching tubules that contain only serous
cells. Rarely these show a slight reaction for Each salivary gland is provided with sen¬
mucus. The system of ducts is poorly devel¬ sory nerve endings and two kinds of efferent
oped; isthmuses are absent. These glands secretory nerves, parasympathetic and sym¬
form a thin serous secretion that evidently pathetic. The parasympathetic preganglionic
serves to wash out the circumvallate groove fibers for the submandibular and sublingual
and the associated taste buds. glands run in the chorda tympani nerve to
The glands of the root of the tongue and the the submaxillary ganglion. The sympathetic
palatine glands are of the pure mucous variety. preganglionic fibers go to the superior cer¬
Short isthmuses have been found in the latter vical ganglion. From here the postganglionic
group. fibers follow along the carotid artery. The
vasodilators are believed to be included in
Blood and Lymphatic Vessels the chorda tympani, the vasoconstrictors in
the sympathetic nerves.
In the interstitial reticular connective tissue The parotid gland receives its secretory
of the salivary glands are fibroblasts and fibers from the glossopharyngeal nerve. In
macrophages, with fat cells scattered singly the interstitial tissue, along the course of its
or in small groups. Plasma cells are of com¬ blood vessels, are found plexuses of myeli¬
mon occurrence and, occasionally, small lym¬ nated (preganglionic and sensory) and non¬
phocytes are also found. The larger blood myelinated fibers, and groups of sympathetic
vessels follow the larger ducts. Loose capillary multipolar nerve cells close to the larger
 ORAL CAVITY AND ASSOCIATED GLANDS • 597

ducts. On the outer surface of the terminal tericidal properties. The principal function
portions, nonmyelinated fibers form a net¬ of the enzyme lysozyme, secreted by serous
work that sends small branches through the cells of salivary glands, is hydrolysis of bac¬
basal lamina. These branches form a second terial cell walls. Deprived of their walls, the
network, from which branches penetrate be¬ bacteria are more easily penetrated and killed
tween the glandular cells, ramify, and end by thiocyanate ions present in saliva.
on the surfaces of the glandular cells with The connective tissue of the stroma and
small, terminal thickenings. interlobular septa of the glands contain lym¬
Stimulation of the parasympathetic nerves phocytes and plasma cells. The latter produce
of the submandibular gland causes the secre¬ immunoglobulins, mainly IgA. This forms a
tion of an abundant thin saliva, rich in water complex with a secretory piece synthesized in
and salts but poor in organic substances. the serous cells of the acini, and the IgA-
Stimulation of the sympathetic nerve, on the secretory piece complex is released into the
contrary, yields a small quantity of thick sa¬ saliva where it is believed to participate in
liva, with a high content of organic sub¬ immunological defense against bacteria in the
stances. The mechanism of action of the oral cavity.
nerves upon the glandular cells and the role The salivary acini elaborate a primary secre¬
of the vasodilators in secretion are not tion consisting of mucus, enzymes, and ions.
known. The presence of different kinds of The ionic concentration of this fluid is not
nerve endings has not been proved. It is even significantly different from that of the extra¬
doubtful whether the secretory fibers in the cellular fluid, but as it flows through the ducts
chorda tympani and in the sympathetic nerve it is greatly modified by active transport of
are of different nature. certain ions by the epithelium lining the
After sectioning of the chorda tympani ducts. Bicarbonate ion is actively secreted by
nerve in the dog, the so-called “paralytic” duct cells near the acini that contain carbonic
secretion in the corresponding submandibu¬ anhydrase catalyzing the combination of
lar and retrolingual glands occurs. This se¬ water and carbon dioxide to form carbonic
cretion is accompanied by intense degenera¬ acid. The resulting bicarbonate ions are se¬
tion and atrophy of the gland cells, especially creted and chloride ions absorbed in this
of the mucous elements in the retrolingual region. Potassium ions are actively secreted,
gland. and sodium ions reabsorbed throughout the
duct system. As a result of these active trans¬
port processes and ion exchanges, the sodium
HISTOPHYSIOLOGY OF THE and chloride concentration of the saliva is
SALIVARY GLANDS only about one eighth that of blood plasma
while the potassium concentration is seven
The human salivary glands continually se¬ times, and bicarbonate three times, greater
crete at a basal rate of 0.5 to 1 ml per minute, than in plasma. Aldosterone, the adrenal
and this is increased manyfold during meals. hormone that controls absorption of sodium
The total daily flow is about 1200 ml. The and excretion of potassium in the kidney,
saliva serves to moisten and lubricate ingested also influences the electrolyte concentration
food, and its enzyme a-amylase initiates the of the saliva. Acting upon the salivary duct
digestive process by hydrolyzing starch to system, it greatly increases reabsorption of
soluble sugars. sodium and the excretion of potassium.
The saliva also plays an important role in
controlling the bacterial flora of the oral
cavity. The mouth is always inhabited by a
very large number and variety of pathogenic
TONSILS
bacteria. Some of these contribute to the
degradative processes of dental caries; others The aperture through which the oral cavity
are capable of initiating ulcers and severe communicates with the pharynx is called the
periodontal inflammation. They are held in fauces. In this region the mucous membrane
check by the continual secretion and swallow¬ of the digestive tract contains accumulations
ing of saliva, which dislodges residual food of lymphoid tissue. In addition to small infil¬
particles and bacteria and sweeps them on trations of lymphocytes, which may occur
down the alimentary tract. In addition to anywhere in this part of the mucous mem¬
these mechanical effects, the saliva has bac¬ brane, there are well-outlined organs of
598 • ORAL CAVITY AND ASSOCIATED GLANDS

lymphoid dssue. The surface epithelium in- tive of inflammation, which is very common
vaginates into them, and they are called ton¬ in tonsils. Occasionally islands of cartilage or
sils. The lingual tonsils have been described bone are found, which are probably late
above. sequelae of earlier pathological processes in
Between the glossopalatine and the phar- the tonsils. In the deeper portions of the
yngopalatine arches are the palatine tonsils, crypts, the limit between the epithelium and
two large oval accumulations of lymphoid the lymphoid tissue is obscured by an intense
tissue in the connective tissue beneath the infiltration of the epithelium with lympho¬
mucous membrane. The overlying epithe¬ cytes. The epithelial cells are pushed aside
lium invaginates to form 10 to 20 deep ton¬ and distorted, so that only a few recognizable
sillar crypts. The stratified squamous epithe¬ epithelial cells remain on the surface (Fig.
lium of the free surface overlies a thin layer 23—24). Plasma cells are common here.
of connective tissue. The crypts reach almost The lymphocytes and neutrophils that pass
to the connective tissue capsule and are of through the epithelium are found in the
simple or branching form (Fig. 23—23). saliva as the salivary corpuscles. They appear
The nodules with their prominent ger¬ there as degenerating vesicular elements with
minal centers are embedded in a diffuse mass a pyknotic nucleus surrounded by a clear
of lymphoid tissue 1 to 2 mm thick, and are vesicle containing granules that show brown¬
usually arranged in a single layer under the ian movement. The salivary corpuscles that
epithelium. The epithelial crypts with their originate from polymorphonuclear leuko¬
surrounding sheaths of lymphoid tissue are cytes are recognized by the remnants of their
partially separated from one another by thin specific granules and their polymorphous nu¬
partitions of loose connective tissue that ex¬ cleus.
tend inward from the capsule. In this con¬ The lumen of the crypts may contain large
nective tissue there are always numerous lym¬ accumulations of living and degenerated lym¬
phocytes of various sizes, mast cells, and phocytes mixed with desquamated squamous
plasma cells. The presence of large numbers epithelial cells, granular detritus, and micro¬
of polymorphonuclear leukocytes is indica- organisms. These masses may increase in size
and form cheesy plugs, which are ultimately
eliminated. If they remain for a long time,
they may calcify. The microorganisms are
sometimes the cause of inflammation and
suppuration, and, carried to other parts of
the body, they may be responsible for some
general infections.
Many small glands are connected with the
palatine tonsils. Their bodies are outside the
capsule, and their ducts open for the most
part on the free surface. Openings into the
crypts seem rare.
In the roof and posterior wall of the na¬
sopharynx is the unpaired pharyngeal tonsil.
The epithelium on the surface of this tonsil
is the same as in the rest of the respiratory
passages—pseudostratihed ciliated columnar
epithelium with many goblet cells. Small
patches of stratified squamous epithelium are
common, however. The epithelium is not
invaginated to form crypts like those of the
palatine tonsil but is plicated to form numer¬
ous surface folds. It is abundantly infiltrated
with lymphocytes, especially on the crests of
the folds. A 2-mm thick layer of diffuse and
Figure 23-23. Section through palatine tonsil of man, nodular lymphoid tissue is found under the
showing crypts penetrating the tonsil from the free surface,
and connective tissue septa (S) penetrating the lymphoid epithelium and participates in the formation
tissue from beneath, x 6. (Redrawn and modified from of the folds. The lymphoid tissue of the tonsil
Sobotta.) is separated from the surrounding parts by
 ORAL CAVITY AND ASSOCIATED GLANDS • 599

Vessel Connectiue tissue cell of lamina propria

Boundary
between con¬
nectiue tissue
and epithelium
Lymphocyte

Epithelium

Epithelial cell

Vessel

Neutrophilic
leukocyte

Plasma cell

Superficial
squamous epi¬
thelial cells
Mucus with
lymphocytes and
granulocytes in
lumen of crypt

Figure 23-24. Human palatine tonsil, showing infiltration of the epithelium of the crypt with lymphocytes, neutrophilic
(heterophilic) granular leukocytes, and plasma cells. Hematoxylin-eosin-azure stain, x 520. (After A. A. Maximow.)

a thin connective tissue capsule, which sends 15 or earlier, though the nodules on the root
thin partitions into the core of each fold. of the tongue persist longer. The pharyngeal
Outside the capsule are small glands of mixed tonsil is usually found in an atrophic condi¬
character. Their ducts—often markedly di¬ tion in the adult, with its ciliated epithelium
lated—traverse the lymphoid tissue and largely replaced by stratified squamous epi¬
empty into the furrows or on the folds. thelium.
Other small accumulations of lymphoid
tissue occur in the mucous membrane of the
pharynx, especially around the orifices of the THE PHARYNX
eustachian tubes, behind the pharyngopala-
tine arches, and in the posterior wall.
Unlike the lymph nodes, the tonsils do not The posterior continuation of the oral cav¬
have lymphatic sinuses, and lymph is not ity is the pharynx. In this region of the
filtered through them. However, plexuses of digestive tract, the respiratory passage and
blindly ending lymph capillaries surround the pathway for food merge and cross. The
their outer surface. upper part of the pharynx is the nasal, the
The tonsils generally reach their maximal middle the oral, and the lower the laryngeal
development in childhood. The involution of portion. In the upper part its structure re¬
the palatine tonsils begins about the age of sembles that of the respiratory system,
600 • ORAL CAVITY AND ASSOCIATED GLANDS

Opening of Leukocytes
duct of around orifice Muscle

Figure 23-25. Longitudinal section of the wall

of the pharynx of an 11-year-old girl, x 27.
(After Schaffer.)

Muscular layer

whereas in the lower part it corresponds columnar ciliated, with many goblet cells. On
more closely to the general plan of the diges¬ the lateral sides of the nasopharynx this cil¬
tive tube. iated epithelium continues downward be¬
Instead of a muscularis mucosae, the mu¬ yond the aperture of the eustachian tube.
cous membrane is provided with a thick, With age, the ciliated epithelium may be
dense, elastic layer. A loose submucous layer replaced by stratified squamous epithelium
is well developed only in the lateral wall of over large areas.
the nasopharynx and where the pharynx Glands of a pure mucous type are found
continues into the esophagus; here the elastic in those places lined with stratified squamous
layer becomes thinner. In all other places, epithelium (Fig. 23—25). They are always lo¬
the mucous membrane is directly adjacent to cated under the elastic layer, sometimes pen¬
the muscular wall, which consists of an inner etrating deep into the muscle. Glands of
longitudinal and an outer oblique or circular mixed type, similar to those of the dorsal
layer of striated muscle. The elastic layer surface of the soft palate, are confined to the
fuses with the interstitial tissue of the muscle regions covered with ciliated epithelium.
and sends strands of elastic fibers between
the muscular bundles. In the fornix it is fused
with the periosteum of the base of the skull.
The lamina propria mucosae consists of
REFERENCES
dense connective tissue containing fine elastic
TONGUE AND TASTE BUDS
networks. Those areas covered with stratified
squamous epithelium are provided with small Beidler, L. M., and R. L. Smallman: Renewal of cells
with the taste buds. J. Cell Biol. 27:263, 1965.
papillae. In the area covered with pseudo-
Farbman, A. I.: Fine structure of the taste bud. J.
stratihed ciliated columnar epithelium there Ultrastruct. Res. 72:328, 1965.
are no papillae. Hodgson, E. S.: Taste receptors. Sci. Am. 204:135, 1961.
The two lower sections of the pharynx and Murray, R. G., and S. Fujimoto: Fine structure of gus¬
a part of the nasal region have stratified tatory cells in the rabbit taste buds. J. Ultrastruct
Res. 27:444, 1969.
squamous epithelium. Toward the roof of
Oakley, B., and R. P. Benjamin: Neural mechanisms of
the pharynx its epithelium becomes stratified taste. Physiol. Rev. 46:173, 1966.
 ORAL CAVITY AND ASSOCIATED GLANDS • 601

Tucker, D., and J. Smith: The chemical senses. Annu. Shackleford, J., and C. E. Klapper: Structure and car¬
Rev. Psychol. 20:129, 1969. bohydrate histochemistry of mammalian salivary
Zotterman, Y.: Olefaction and Taste. New York, Mac¬ glands. Am. J. Anat. Ill:25, 1962.
millan Co., 1963. Spicer, S. G., and L. Warren: The histochemistry of
sialic acid containing mucoproteins. J. Histochem.
SALIVARY GLANDS
Cytochem. 8:135, 1960.
Amsterdam, A., I. Ohad, and M. Schramm: Dynamic Strum, J. M., and M. J. Karmovsky: Ultrastructural
changes in the ultrastructure of the acinar cell of localization of peroxidase in submaxillary acinar
the rat parotid gland during the secretory cycle. J. cells. J. Ultrastruct. Res. 16:320, 1970.
Cell Biol. 41:753, 1969. Tamarin, A., and L. Screenby: The rat submaxillary
Amsterdam, A., M. Schramm, I. Ohad, et ah: Concom¬ gland. A comparative study by light and electron
itant syntheses of membrane protein and exportable microscopy. J. Morphol. 117:295, 1965.
protein of the secretory granules in rat parotid Tandler, B.: Ultrastructure of human submaxillary
gland. J. Cell Biol. 40:187, 1971. gland. I. Architecture and histological relationships
Castle, J. D., J. D. Jamieson, and G. E. Palade: Radioau¬ of the secretory cells. Am. J. Anat. 111:287, 1962.
tographic analysis of the secretory process in the Tandler, B.: Ultrastructure of human submaxillary
parotid acinar cell of the rabbit. J. Cell Biol. 53:290, gland. IE The base of the striated duct cells. J.
1972. Ultrastruct Res. 9:165, 1963.
Junquiera, L. C., and F. F. de Moraes: Comparative Tandler, B., C. R. Denning, I. D. Mandel, and A. H.
aspects of the vertebrate major salivary glands bi¬ Kutscher: Ultrastructure of human labial salivary
ology. In Wohlfarth-Botterman, K. E., ed.: Functi- glands. III. Myoepithelium and ducts. J. Morphol.
onelle und Morphologische Organization der Zelle: 130:227, 1970.
Sekretion und Excretion. Heidelberg-Berlin-New Young, J. A.: Salivary secretion of inorganic electrolytes.
York, Springer Verlag, 1965. Int. Rev. Physiol. 19:1, 1979.
Eeeson, C. R.: Structure of the salivary glands. In Hand¬ Young, J. A., and E. W. van Lennep: Morphology and
book of Physiology. Vol. 2, Sect. 6. Washington, physiology of salivary myoepithelial cells. Int. Rev.
DC, American Physiological Society, 1967. Physiol. 72:105, 1977.
Mason, D. K., and D. M. Chisholm: Salivary Glands in
Health and Disease. Eondon, W. B. Saunders Co.,
1975.
 24
THE

The adult human has 32 teeth of which 16 The soft parts associated with the tooth are
are in the alveolar process of the maxilla and the pulp, which fills the pulp chamber; the
16 in the mandible. These permanent teeth periodontal membrane, which connects the ce-
are preceded by a set of 20 deciduous teeth mentum-covered surface of the root with the
that begin to erupt about seven months after alveolar bone; and the gingiva, that portion
birth and reach their full complement at 6 to of the oral mucous membrane surrounding
8 years of age. These are shed between the the tooth. In young0 persons the gingiva is
sixth and 13th year and are gradually re¬ attached to the enamel; with increasing age
placed by the permanent or succedaneous teeth. it gradually recedes from the enamel, and in
This process of replacement extends over a old people it is attached to the cementum.
period of about 12 years, and full dentition
is usually attained by the age of 18 with the
Dentin
eruption of the third molars. Each of the
several types of teeth in each set has a dis¬ Dentin makes up the bulk of the tooth. It
tinctive form adapted to its specific function. is formed by a layer of odontoblasts that lines
Thus, the incisors are specialized for cutting the pulp cavity. Dentin is yellowish in color
or shearing; the pointed canines for punctur¬ and semitranslucent in the fresh condition.
ing and holding; and the molars for crushing Although it resembles bone in structure and
and grinding. chemical composition, it is harder than com¬
All teeth consist of a crown projecting above pact bone. It consists of 20 per cent organic
the gum or gingiva and one or more tapering and 80 per cent inorganic matter. The or¬
roots that occupy conforming sockets or alveoli ganic part is 92 per cent collagen, and most
in the bone of the maxilla or of the mandible. of the inorganic components are in the form
The junctional region between the crown and of hydroxyapatite crystals. Upon decalcihca-
the root is called the neck or cervix. Incisors tion in acids, only the organic part remains
have a single root, lower molars two, and and the substance of the tooth becomes soft.
upper molars three. The tooth contains a Upon incineration, only the inorganic mate-
small central pulp chamber or cavity that cor¬
responds roughly in its shape to the outer
form of the tooth. This continues downward
into each root as a narrow canal that com¬ Enamel
municates with the periodontal membrane Dentin
through an apical foramen at the tip of the Pulp
root. gj..\ Gingiva

The hard portions of a tooth consist of r4- Alueolar bone

three different tissues: dentin, enamel, and
cementum (Fig. 24-1). The bulk of the tooth
is formed of the dentin, which surrounds the Periodontal
membrane
pulp chamber. It is thickest in the crown and
gradually tapers as it reaches the apex of the
Cementum
root. Its outer surface is covered, in the
region of the crown, by a layer of enamel,
which is thinnest in the cervical region. On
the root the tooth is covered by a thin layer
of cementum, which extends from the neck to
Figure 24-1. Diagram of sagittal section of adult human
the apical foramina.
lower first permanent molar. (Courtesy of I. Schour.)
602
 THE TEETH • 603

rial remains. The inorganic material is much portions they become narrower. In their out¬
the same as in bone, except that it is denser ward course from the pulp cavity, most of
and more insoluble. The organic part, like the dentinal tubules describe an S-shaped
other collagen-rich tissues, also contains gly- curve. The layer of dentin immediately sur¬
cosaminoglycan—protein complexes. rounding each tubule, the sheath of Neumann,
In a ground section passing through the differs from the rest of the dentin in its high
axis of an extracted tooth, the dentin has a refringence and denser staining in decalcified
radially striated appearance (Fig. 24—2). This specimens.
is attributable to the presence of innumerable Between the dentinal tubules are bundles
minute canals, the dentinal tubules, which ra¬ of collagenous fibrils corresponding to the
diate from the pulp cavity toward the periph¬ collagenous fibrils within bone. They are
ery. Apical cytoplasmic processes from the embedded in a ground substance consisting
formative cells, called odontoblasts, extend of glycosaminoglycans and protein. The
into these minute tubules. Near the pulp, course of the fibrillar bundles is, in general,
their diameter is 3 to 4 fjim, but in the outer parallel to the long axis of the tooth and
perpendicular to the dentinal tubules. Some
Schreger's lines investigators distinguish a peripheral layer,
of enamel the cover or mantle dentin, characterized by a
branching pattern of dentinal tubules, and a
Stripes of thicker inner layer, the circumpulpar dentin,
Retzius with thinner, straighter tubules.
The calcification of the developing dentin
is not always complete and uniform. The
mineral deposits that appear during devel¬
opment in the organic ground substance have
the form of spherical aggregations of apatite
crystals, which gradually gain in size and
Dentin Interglobular finally fuse. These nuclei of mineralization
space of Owen are found initially in rows or chains on and
within collagen fibrils. In incompletely calci¬
fied regions, between the calcified spheres,
there remain angular “interglobular” spaces
that contain only the organic matrix of the
dentin (Fig. 24-3). The dentinal tubules con¬
tinue uninterrupted through the spheres and
interglobular spaces. In a macerated tooth
from which all organic parts have been ex¬
Tomes' granular
tracted, the tubules, as well as the interglob¬
layer of dentin
ular spaces, are filled with air and appear
dark in transmitted light. In many normal
human teeth there are layers of large inter¬
globular spaces in the deeper parts of the
dentin of the crown. Mineralization in dentin
is not uniform, and as a result, curvilinear
Cell free lines of appositional growth are apparent.
cementum These are called the contour lines of Owen.
Immediately under the dentinocemental
junction in the root there is always a layer of
small interglobular spaces, the granular layer
of Tomes (Fig. 24—2).
In sections through a decalcified tooth,
Cellular
each dentinal tubule contains a slender cyto¬
cementum plasmic process, Tomes’ fiber, which in life
of root probably completely fills the lumen of the
tubule, but which in fixed preparations ap¬
Figure 24-2. Longitudinal ground section of human cus¬
pid. The top of the crown has been abraded, x 7. (After pears shrunken away from the wall. When
von Ebner, from Schaffer.) the tubules are seen in cross section, each
604 • THE TEETH

Matrix

Shrunken dentinal fiber

Margin of dentinal tubule
v X
* ' 4 ;

Figure 24-4. Tangential section through the root of a

molar of an ape. The margin of the dentinal tubule is also
called the sheath of Neumann. The dot in the center of
each dentinal tubule is a somewhat shrunken Tomes’
fiber, x 740. (After Schaffer.)

oped, enamel consists almost entirely of cal¬

cium salts in the form of large apatite crystals;
only 0.5 per cent is organic substance. The
protein matrix of enamel, secreted by cells
called ameloblasts, has been isolated, and ori¬
Figure 24-3. Dentinoenamel junction of a tooth of a man; ented samples have been subjected to x-ray
ground section. The enamel prisms appear as a fine, diffraction analysis. The protein was found
wavy striation. The interglobular spaces in the dentin are
to be in the cross-(3 configuration. Complete
black (air filled). Between these lacunae are the dentinal
tubules, x 80. (After Braus.) amino acid analysis revealed that one fourth
of the amino acid residues are proline. The
protein therefore cannot be either keratin or
small oval contains a dark dot (Fig. 24—4). collagen. Relatively high concentrations of
These Tomes’ fibers are the processes of the
odontoblasts. Dentin continues to be formed
very slowly throughout life, and the pulp iurface of tooth
cavity is therefore progressively narrowed itripes of Retzius
with advancing age.
The dentin is sensitive to touch, to cold, to
acid-containing foods, and the like. Only oc¬
casional nerve fibers penetrate for short dis¬
tances into the dentin. It has been suggested
that the odontoblastic processes may transmit
sensory stimulation to the pulp, which is
richly innervated.
In old age the dentinal tubules are often Enamel
obliterated through calcification. The dentin
then becomes more transparent. When the
dentin is exposed by extensive abrasion of
the enamel of the crown, or when the outside
of the tooth is irritated, a production of new
or “secondary” dentin of irregular structure
may often be observed on the wall of the
pulp chamber. 44iis may be so extensive as
to fill the chamber completely.
Enamel tuft

Enamel
Enamel is the hardest substance found in
the body. It is bluish white and transparent Figure 24-5. Portion of a ground cross section of crown
in thin-ground sections. When fully devel- of a human cuspid, x 80. (After Schaffer.)
 THE TEETH • 605

stand upright on the surface of the dentin,

usually with a pronounced inclination toward
the incisal or occlusal edge (Fig. 24—5). Be¬
tween the rods is “interprismatic substance,”
which has a substructure identical to that of
the rods but is oriented in a different direc¬
tion. Surrounding each rod is a clear area of
organic matrix called the enamel sheath or
prisynatic rod sheath (Fig. 24—6). Every rod runs
through the whole thickness of the enamel
layer. This, however, cannot be seen in sec¬
tions of the enamel, because the rods are
twisted and soon pass out of the plane of
Figure 24-6. Enamel rods of human tooth in cross section. In a ground preparation, the sub¬
section. The dark lines are the interprismatic substance stance of a rod in its longitudinal section
between the pale rods. Photomicrograph at high magni¬ seems homogeneous. However, after acid
fication. (Courtesy of B. Orban.)
acts upon such a section, a distinct cross
organic-bound phosphorus have been found striation appears in the rods; this indicates
in bovine enamel organ matrix. This may that the calcification probably proceeds layer
play a role in the initiation of enamel calcifi¬ by layer.
cation. After decalcihcation of a fully devel¬ Studies with the electron microscope show
oped tooth, the enamel, as a rule, is com¬ that the enamel rods or prisms and the inter¬
pletely dissolved. prismatic substance are both composed of
As seen with the light microscope, the apatite crystals and organic material. The
enamel consists of thin rods or prisms that relations of the crystals in the prisms and in

Figure 24-7. Slightly oblique section of undecalcified calf enamel, showing the roughly ovoid enamel prisms (A) and
the interprismatic substance (B). Note the remarkable orientation of the apatite crystals within the individual prisms, and
the different orientation of the crystals in the interprismatic substance. Note also the clear areas that define the prisms.
Embedded in methacrylate and sectioned with the diamond knife. Osmium fixation, x 18,000. (Courtesy of E. J. Daniel
and M. J. Glimcher.)
606 • THE TEETH

the interprismatic substance are clearly of the enamel they bend helically, and in the
shown in Figures 24—7 and 24—8. outer zone they again assume a direction
In the human tooth, most of the rods in perpendicular to the surface. In addition, the
cross section have the form of fluted semicir¬ rods show numerous small, wavy curves. On
cles. The convex surfaces of all rods face the the lateral surfaces of the crown the rods are
dentin, and their cross sections have a scale¬ arranged in zones that encircle the tooth in
like appearance (Fig. 24—6). The three-di¬ horizontal planes. The bends of the rods in
mensional configuration and arrangement of two neighboring zones cross one another. In
the enamel rods in human teeth is still con¬ axial, longitudinal ground sections, the cross¬
troversial. Some investigators report that in ing of groups of rods appears in reflected
electron micrographs the enamel rods in light as light and dark lines, more or less
cross section have a keyhole shape, and that perpendicular to the surface—the lines of
the asymmetrical projection is what others Schreger (Fig. 24—2).
call interprismatic substance. This form and In a cross section of the crown, the enamel
arrangement are explained by calcification shows concentric lines, which are brown in
beginning earlier on the side of the rods that transmitted light and colorless in reflected
lies nearest the dentin. This inner, harder light. In longitudinal, undecalcified axial sec¬
side is supposed to press into the softer side tions they are seen to run obliquely inward
of the adjacent rod, compressing it and leav¬ from the surface and toward the root. They
ing one or two groovelike impressions. are called the lines of Retzius or incremental
The exact course of the enamel rods is lines of Retzius and are connected with the
extremely complicated but seems to be per¬ circular striation on the surface of the crown
fectly adapted to the mechanical require¬ (Fig. 24-2). They are believed to result from
ments of the grinding and crushing of food. rhythmic deposition and mineralization of
Starting from the dentin, the rods run per¬ enamel matrix.
pendicular to its surface; in the middle zone The free surface of the enamel is covered

Figure 24-8. Higher magnification of an area of bovine dental enamel similar \o that in Figure 24-7, showing
longitudinally oriented prismatic crystals (A) with the interprismatic crystals (B) oriented approximately 30 degrees to
the direction of the crystals within the prism. Osmium fixation, x 100,000. (Courtesy of E. J. Daniel and M. J. Glimcher.)
 THE TEETH • 607

by two thin layers. The inner enamel cuticle Coarse collagenous bundles from the peri¬
(formerly called Nasmyth’s membrane) is odontal membrane penetrate the cementum.
about one micron thick, and appears to be These fibers, corresponding to Sharpey’s fi¬
the final product of the activity of the enamel¬ bers of bone, remain uncalcihed and in
forming ameloblasts before they disappear. ground sections of a tooth appear as empty
The outer is an acellular layer, probably canals.
derived from the keratinized remnants of the Not infrequently, epithelial cells are found
dental sac of the developing tooth. It is con¬ in cementum. These are thought to be rem¬
tinuous with the cementum covering the root nants of Hertwig’s epithelial root sheath.
and is similar in composition and histological Unlike the dentin, which may remain un¬
structure. This layer is tenaciously adherent changed even after the destruction of the
to the tooth and is distinct from the connec¬ pulp and the odontoblasts and after the “fill¬
tive tissue of the gingiva. It may persist for ing” of the pulp cavity, the cementum readily
some time after eruption of the tooth. undergoes necrosis when the periodontal
In an axial section of the tooth, the line of membrane is destroyed. On the other hand,
junction between the dentin and the enamel new layers of cementum may be deposited
(dentinoenamel junction) is uneven and scal¬ on the surface of the root. This deposition is
loped. Pointed or spindle-shaped processes called cementum hyperplasia when it becomes
of dentin penetrate the enamel. The spindle- excessive and is considered to be a reaction
shaped processes of the dentinal matrix pen¬ to chronic irritation.
etrating a short distance into the enamel are
called enamel spindles. Pulp
Local disturbances of the enamel during
development cause the so-called enamel lamel¬ The pulp occupies the pulp cavity of the
lae and tufts. These lamellae, usually found tooth and is the connective tissue that formed
in cervical enamel, are organic material ex¬ the dental papilla during embryonic devel¬
tending from the surface of the enamel to¬ opment. In the adult it has an abundant,
ward and sometimes into the dentin. The gelatinous, metachromatic ground substance
tufts extend from the dentinoenamel junc¬ similar to that of mucoid connective tissue.
tion into the enamel for one third of its It contains a multitude of thin collagenous
thickness. The tuftlike shape, however, is an fibrils running in all directions and not ag¬
optical illusion, due to the projection, into gregated into bundles. Elastic fibers are
one plane, of fibers lying in different planes. found only in the walls of the afferent vas¬
They are groups of poorly calcified, twisted culature.
enamel rods with abundant cementing sub¬ The odontoblasts are the cells of the pulp
stance between them. adjacent to the dentin. Beneath the layer of
odontoblasts is a relatively cell-free area, the
zone of Weil. With silver impregnation tech¬
Cementum
niques, bundles of reticular fibers (Korffs
The cementum covering most of the root is fibers) are found in this zone (Fig. 24—9).
coarsely fibrillar bone substance. The peri¬ These fibers, described by light microscopists,
odontal ligament attaches to it and to the pass from the pulp into interstices between
alveolar bone. Of all the dental hard tissues, the odontoblasts and their distal ends are
cementum is the closest to bone in physical incorporated in the dentinal matrix. Occa¬
and chemical characteristics. In the adult the sional cells are found in the zone of Weil,
organic matrix is elaborated by the cemento- and capillaries and nerves are plentiful. Ad¬
cytes embedded in the apical cementum. The jacent to this cell-poor zone, at the periphery
cervical portion of the cementum is acellular, of the pulp proper, is a cell-rich zone. Spin¬
whereas at the apex there is only a thin layer dle-shaped or stellate fibroblasts are the pre¬
of acellular cementum adjacent to the dentin. dominant cell type of the pulp. Other cells
The remainder of the cementum in this re¬ present in limited numbers include mesen¬
gion is cellular cementum. Canaliculi, haversian chymal cells, macrophages, lymphocytes,
systems, and blood vessels are normally ab¬ plasma cells, and eosinophils.
sent. The layer of cementum increases in The pulp continues into the narrow root
thickness with age, especially near the end of canal, where it surrounds the blood vessels
the root, and then haversian systems with and nerves, and continues through the open¬
blood vessels may appear. ings in the root apex into the periodontal
608 • THE TEETH

attachments. The fiber bundles of the peri¬

odontal membrane have a slightly wavy
course. When the tooth is not functioning,
they are slack and permit it to move slightly
on the application of stress. The ligament
adjacent to the cementum contains only ce-
mentoblasts and the usual complement of con¬
nective tissue cells. On the alveolar bone side
of the periodontal ligament, osteoblasts and
osteoclasts may be found.
Scattered in many places in the periodontal
membrane, especially near the surface of the
cementum, are blood and lymph vessels and
nerves embedded in a thin layer of loose
connective tissue, and small islands of epithe¬
lium. These islands are vestiges of the epi¬
thelial sheath of Hertwig, which will be de¬
scribed below in _ •
the section on the
histogenesis of teeth. The epithelial rests fre¬
quently degenerate and undergo calcifica¬
tion, giving rise to the cementicles.
Figure 24-9. Continuation of Korff’s fibers of the pulp into
the matrix of the dentin at (f). Photomicrograph, x 700. The Gingiva
(Courtesy of B. Orban.)
The gum or gingiva is that part of the
membrane. The pulp contains blood vessels mucous membrane that is firmly connected
and lymphatics, which enter and leave the with the periosteum at the crest of the alveo¬
pulp through the apical foramen. The cir¬ lar bone. It is a keratinized stratified squa¬
culation of the pulp consists of a system of mous epithelium with numerous connective
arterioles and capillaries close to the bases of tissue papillae projecting into its base. It is
the odontoblasts. These drain into small veins also linked to the surface of the tooth by the
more centrally situated in the pulp. Arterio¬ epithelial attachment ofi Gottlieb, which gradually
venous anastomoses are readily demonstrable recedes with advancing age, moving toward
in the pulp. Numerous bundles of myelinated the root.
nerve fibers, which arise from small cells in Although the gingiva generally has high
the gasserian ganglion, enter the pulp cavity papillae, the epithelial attachment to a tooth
through the canal of the root. They form a is devoid of papillae except when chronically
plexus in the pulp, from which arises a finer inflamed. Between the epithelium and the
plexus of nonmyelinated fibers in the periph¬ enamel there is a small furrow surrounding
eral layers. Nerve endings have been de¬ the crown, the gingival crevice, which is lined
scribed between the odontoblasts. by a nonkeratinizing stratified squamous ep¬
ithelium. The crevicular margin of the epithe¬
lial attachment is the junction between the
Periodontal Membrane
attached gingiva and the marginal gingiva,
The periodontal membrane, which also serves which surrounds the tooth like a collar about
as periosteum to the alveolar bone, furnishes 1 mm wide.
a firm fibrous attachment of the root to the The thinner portions of the gingival epi¬
bone. It differs from the usual periosteum thelium covering the papillae protrude
by the absence of elastic fibers. It consists, in slightly, giving a nubbled appearance to the
part, of thick collagenous bundles, that run otherwise smooth free surface. The crevicu¬
from the alveolar wall into the cementum of lar epithelium is devoid of papillae except
the roots. The orientation of the fibers varies when chronically inflamed. Three groups of
at different levels in the alveolus. From root collagenous fibers are associated with the
tip to neck, there are the cementoalveolar fibers, gingiva: the gingivodental group, the cuti-
which are variously designated as apical, cular group, and the transseptal group, the
oblique, or horizontal, and alveolar crest fibers. latter usually being associated with the fibers
The terms describe their direction or their of the periodontal ligament. Small aggregates
 THE TEETH • 609

of lymphocytes and plasma cells are usually several points budlike thickenings, the tooth
found in the lamina propria at the base of germs, that are the primordia of the teeth
the gingival crevice. (Fig. 24—10A,B). There are ten tooth germs
in each jaw, one for each deciduous tooth.
Alveolar Bone In each germ a group of epithelial cells
becomes conspicuous as the enamel knot or
As the teeth are formed, so is the bone cord, a temporary structure that later disap¬
that supports them, and it is to this bone that pears (Fig. 24-11). The cells of the mesen¬
the principal fibers of the periodontal liga¬ chyme under the enamel knot aggregate in
ment attach. This alveolar bone consists of a dense group to form the primordium of
cancellous bone between two layers of cortical the papilla (Figs. 24-10C, 24-11, 24-12). The
bone. The outer cortical plate is a continua¬ dental lamina then extends posteriorly be¬
tion of the cortical layer of the maxilla or yond the last deciduous tooth germ and
mandible. The inner cortical plate adjacent slowly forms germs of the permanent molars,
to the periodontal membrane is referred to which are not preceded by corresponding
by radiologists as the lamina dura. It sur¬ deciduous teeth.
rounds the roots of the teeth to form the Beginning with the 10th to 12th weeks, the
socket. The vessels and nerves to the teeth remainder of the dental lamina again pro¬
course through the alveolar bone to the apical duces solid epithelial buds—the germs for the
foramina of the roots where they enter into permanent teeth—one on the lingual side of
the pulp chamber. Considerable resorption each deciduous germ (Fig. 24-13). After the
of alveolar bone can occur after loss of per¬ formation of the permanent tooth germs, the
manent teeth or as a result of periodontitis dental lamina disappears. The germs of the
(inflammation of the supporting tissues of permanent teeth undergo the same transfor¬
the teeth). Alveolar bone is very labile and mations as do those of the deciduous teeth.
serves as a readily available source of calcium The papilla enlarges and invaginates into
to maintain blood levels. The cancellous tra¬ the base of the epithelial tooth germ (Figs.
beculae buttressed by the labial and lingual 24-10C,F>, 24-11, 24-12). The latter, while
cortical plates aid in support of the teeth still connected by an epithelial strand with
during mastication. the dental lamina, becomes bell shaped and
caps the convex upper surface of the papilla.
Thenceforth it is called the enamel organ,
HISTOGENESIS OF THE TEETH because in its further development it pro¬
duces the enamel. Both the papilla and the
The teeth are derivatives of the oral mu¬ enamel organ gradually gain in height, and
cous membrane. They may be considered to the latter soon acquires approximately the
be modified papillae of the epithelium that shape of the future tooth.
have been covered by a thick layer of calcified A concentric layer of connective tissue, the
material originating in part from the epithe¬ dental sac, develops around the tooth primor¬
lium and in part from the underlying con¬ dium and interrupts its epithelial connection
nective tissue. The most primitive toothlike with the oral cavity. Around the sac, and at
structures are the placoid scales found in the a certain distance from it, the maxillary or
integument of elasmobranch fish, where their mandibular bone develops (Fig. 24—12).
derivation from papillae of the epidermis is The peripheral cells of the enamel organ
quite evident. are arranged in a regular, radial fashion. On
In human embryos of the fifth week, the the convex outer surface of the enamel or¬
ectodermal epithelium lining the oral cavity gan, the cells remain small and cuboidal. On
presents a thickening along the edge of the its invaginated base, the cells of the inner
future upper and lower jaws. The thickening enamel epithelium become tall and columnar
consists of two parallel epithelial ridges that and differentiate into ameloblasts that elabo¬
extend into the subjacent mesenchyme. Of rate the enamel. In the interior of the enamel
these, the labial ridge later splits and forms organ a clear liquid accumulates between the
the space between lip and alveolar process of cell bodies, which remain connected by long
the jaw. The lingual ridge, nearer the tongue, processes. This tissue of epithelial origin thus
produces teeth and is called the dental lamina. acquires a reticular appearance like that of
The edge of the dental lamina extends into connective tissue and forms the stellate retic¬
the mesenchyme of the jaw and shows at ulum of the enamel pulp (Fig. 24—12).
610 • THE TEETH

A B c D
Bud stage Cap stage Bell stage Apposition and
Calcification of bone calcification of
enamel and dentin j
Y L ~v
.J
GROWTH CALCIFICATION

F G H
(Intra-osseous) (Into oral cavity)

v
■j
ERUPTION ATTRITION
Figure 24-10. Diagram of life cycle of a human deciduous incisor. The normal resorption of the root is not indicated.
Enamel and bone are drawn in black. (Redrawn and modified from Schour and Massler.)
 Epithelium on floor of mouth Epithelium of dental lamina

Neck of
enamel organ

Enamel cord Labial epithelium

External enamel
epithelium
Enamel pulp

Preformed
membrane

Dental papilla
Internal end of
dental lamina

Dental follicle
Internal enamel epithelium

Figure 24-11. Primordium of the right lower central incisor of a human fetus of 91 days, in sagittal section. Collagenous
fibers are black. Mallory’s connective tissue stain, x 80. (Redrawn and modified from Schaffer.)

Gum wall

Epithelial
pearl
Mi
St ratified squamous
epithelium of lip
Residue of
Enamel cap
permanent
dental lamina ■**— Dentin
Primordium of odontoblasts
Stratum intermedium
Internal enamel epithelium

Enamel pulp

Alveolar bone

VAX' j

Alveolar x. /’? \- / , > *--i tiki-Dental pulp

p.
External enamel
epithelium

Transition of external
into internal enamel
epithelium
Dental
follicle

Figure 24-12. Primordium of lower central incisor of a 5-month fetus, in sagittal section, x 30. (After Schaffer.)
Compare with photomicrographs in Figure 24-14.
611
612 • THE TEETH

blast process where they discharge their con¬

tent by exocytosis. The tropocollagen then
polymerizes extracellularly to form the
Enamel striated collagen fibers of the dentin matrix.
The odontoblasts are unusual among col¬
Dentin
lagen-secreting cells in their polarization. In
fibroblasts, procollagen is released anywhere
on the cell surface. Exocytosis in odontoblasts
is confined to the apical process. The secre¬
tory pathway is clearly demonstrated in ra¬
dioautographs. When radioactively labeled
precursors of collagen are administered, the
Enamel i grains first appear in autoradiographs over
thelium i
the endoplasmic reticulum of the odonto¬
blasts. At later time intervals they are concen¬
trated over the Golgi complex and then over
the cell apex. In a few hours they are located
predominantly over the predentin.
Dentin
Newly formed predentin around the ta¬
Stellate
reticulum pering apical processes of the odontoblasts is
unmineralized. It stains poorly in routine
histological sections and is apparently devoid
Hertwig’s
of glycoprotein, for it is not stained by the
PAS reaction. The transition from predentin
to dentin is abrupt. Beyond this line the
Figure 24-13. Diagram of deciduous tooth and the tooth matrix is PAS positive, and the collagen fibers
germ (below) of its corresponding permanent tooth. Note are larger and heavily encrusted with hy¬
the surrounding alveolar bone, x 5. (Redrawn from a droxyapatite crystals.
photograph by B. Orban.)
The dentin first appears as a thick limiting
layer between ameloblasts and odontoblasts,
sometimes called the membrana perforata. The
When the formation of the hard substances layer of dentin extends down the slopes of
of the tooth begins in fetuses of about 20 the dental papillae (Fig. 24—14). It gradually
weeks, the mesenchyme of the dental papilla grows thicker and is transformed into a solid
contains numerous blood vessels and a few cap of dentin by the apposition of new layers
reticular fibrils between its cells. The pulp on its concave surface. As the odontoblasts
cells adjacent to the layer of ameloblasts be¬ recede with the deposition of new dentin,
come transformed into odontoblasts (Figs. 24— thin processes of their apical cytoplasm re¬
12, 24-14). main in the deposited dentin as the odonto¬
The odontoblasts that form the dentin are blastic processes.
tall columnar cells with their base toward the When the dentin first appears, it is a soft
interior of the pulp and their tapering apical fibrillar substance. The fibrils are continua¬
processes embedded in a layer of predentin tions of the fibrils of the papilla. They are
(Figs. 24—15, 24—16). The long, slender ex¬ argyrophilic and are generally called Korffs
tensions of their apical processes continue fibers. They enter the dentin, spread out fan¬
into the dentinal tubules. The layer of odon¬ like, and are incorporated into the fibrillar
toblasts is epithelioid in appearance but the collagenous matrix of the dentin (Fig. 24-9).
cells are not in as close apposition laterally as Mantle dentin is formed first, followed by
in typical epithelium, and capillaries may formation of circumpulpar dentin.
penetrate between them nearly to the pre¬ In dentin formation, calcification follows
dentin. These cells have an extensive rough closely the deposition of the fibrillar organic
endoplasmic reticulum and a large supranu¬ matrix. During the whole process, however,
clear Golgi complex. Condensing vacuoles there is always a thin layer of uncalcihed
associated with the transface of the Golgi predentin adjacent to the odontoblasts (Fig.
contain filamentous material that has been 24-15).
identified immunocytochemically as procol¬ Almost immediately after the appearance
lagen. After their condensation the secretory of the first calcified dentin on the papilla, the
granules are distributed along the odonto¬ ameloblasts begin the elaboration of enamel
 THE TEETH • 613

IP j>>

Figure 24-14. Photomicrographs of a portion of a developing tooth, showing the enamel stained red and dentin stained
blue near the dental pulp and pink near the dentinoenamel junction. The columnar epithelium of ameloblasts is closely
applied to the enamel. Peripheral to this epithelium are the stellate cells of the enamel pulp. The dental pulp is limited
by an epithelial layer of odontoblasts depositing dentin. For further orientation, see Figure 24-12.
614 • THE TEETH

near the base (Fig. 24-18). They exhibit an

unusual segregation of organelles in that
most of the mitochondria are clustered be¬
tween the nucleus and the cell base. A long
Dentin cylindrical Golgi apparatus is situated in the
."...is |*s sift
aliUHMMli
If •»%' *
if ' C

Predentin

Figure 24-15. Photomicrograph from growing end of rat

incisor, showing the relationship of the odontoblasts to
the predentin and dentin. Notice the basal nuclei and the
apical processes projecting into the predentin. An area
such as that enclosed in the rectangle is shown at higher
magnification in Figure 24-16. (From Weinstock, M., and
C. P. Leblond. J. Cell Biol. 60:92, 1974.)

matrix. It is deposited layer by layer onto the

surface of the calcifying dentin (Fig. 24—14).
On the slopes of the papilla the height of the
ameloblasts decreases, and at the base of the
papilla they continue into the cuboidal outer
enamel epithelium.
As the mass of enamel increases, the ame¬
loblasts recede. Light microscopists believed
that the enamel matrix was a specialization
of the apical cytoplasm of the ameloblast and
therefore intracellular, and that the calcified
enamel prisms were essentially prolongations
of the columnar cells. This interpretation has
now been discarded, since a distinct cell mem¬
brane has now been observed in electron Figure 24-16. Electron micrograph of apical region of an
odontoblast, showing the appearance of the odontoblast
micrographs between the cytoplasm and the process and the fibrillar and amorphous components of
calcified matrix. The ameloblasts are tall col¬ the surrounding matrix of the predentin. (From Weinstock,
umnar cells with their elliptical nuclei located M., and C. P. Leblond. J. Cell Biol. 60:92, 1974.)
 THE TEETH • 615

Site
of
mineralization
DENTIN

distal

Odontoblast
process
PREDENTIN

proximal

Terminal
web

Apical r ER
Figure 24-17. Drawing depicting the uitrastructure of a
typical odontoblast from rat incisor. (From Weinstock, M.,
and C. P. Leblond. J. Cell Biol. 60:92, 1974.)

Golgi
ODONTOBLAST apparatus

Supranuclear
r ER

Nucleus
616 • THE TEETH

Growth
re g i o n s

ENAMEL
MATRIX

Secretory
APICAL granules
PROCESS

Terminal
web

Fibril

GOLGI Condensing
APPARATUS vacuoles

Figure 24-18. Photomicrograph of secretory amelo-

blasts in a region of enamel matrix secretion in rat
maxillary incisor. Notice the clustering of mitochondria rER
at the cell base, the basal location of the nuclei, and the
apical processes extending into the enamel matrix.
(From Weinstock, A., and C. P. Leblond. J. Cell Biol.
57:26. 1971.)

NUCLEUS

MITOCHONDRIA

Figure 24-19. Diagrammatic representation of the ultra¬

structure of a secretory ameloblast from rat maxillary
incisor. (From Weinstock, A., and C. P. Leblond. J Cell
Biol. 57:26, 1971.)
 THE TEETH • 617

membrane-limited secretory granules are

found in the Golgi region, the apical cyto¬
plasm, and the proximal portion of the apical
cell process (Figs. 24—19, 24-20). 4 he secre¬
tory granules contain glycoprotein constitu¬
ents of the enamel matrix. I hey are formed
in the Golgi, transported to the cell process,
and released there.
Calcification of the enamel matrix starts at
the periphery of each enamel prism and
proceeds toward its interior. When the or¬
ganic matrix has fully calcified, so little or¬
ganic material remains that the enamel is
completely dissolved when teeth are decalci¬
fied. That the calcification is seldom abso¬
lutely uniform has been mentioned. The rate
of deposition of enamel has been studied
using sodium fluoride, and of dentin, with
vital injections of alizarin. The daily thick¬
ening of dentin in the developing tooth is
about 4 pm, and unusual increments (neona¬
tal lines) appear in the enamel and dentin
formed in the deciduous teeth at the time of
birth.
When the definitive thickness and linear
extent of the enamel are reached in the neck
region of the tooth, the ameloblasts become
small cuboidal cells and then atrophy. Before
they disappear, they elaborate the enamel
cuticle, which covers the external surface of
the enamel of recently erupted teeth. At the
deep end of the enamel organ, the outer and
inner enamel epithelium form a fold, the
epithelial sheath of Hertwig.
The development of the root begins shortly
before the eruption of the tooth, continues
after the crown has emerged from within the
mucous membrane, and is not completed
until much later. The epithelial sheath of
Hertwig disappears when the root is com¬
pletely developed, but as mentioned previ¬
ously, remnants of this epithelial sheath are
Figure 24-20. E ectron micrograph of apical processes found in cementum and in the periodontal
(Tomes’ processes) of the ameloblasts and associated ligament.
enamel matrix from rat incisor. (From Weinstock, A., and When the germ of the permanent tooth
C. P. Leblond. J. Cell Biol. 51:26, 1971.)
begins to develop, its growth pressure causes
resorption of the bony partition between the
two teeth, then of the root, and eventually
supranuclear region, and numerous cisternae even of a part of the enamel of the deciduous
of rough endoplasmic reticulum are found tooth. Osteoclasts are prominent in this proc¬
in the supranuclear and apical regions of the ess of tooth destruction just as in the resorp¬
cytoplasm (Fig. 24-19). There is a diffuse tion of bone. The crown of the permanent
terminal web, and distal to this is a broad tooth moving upward gradually expels and
apical process, Tomes process, embedded in takes the place of the crown of the former
the enamel matrix. Numerous spherical deciduous tooth.
618 * THE TEETH

REFERENCES phological and biochemical considerations in struc¬

tural studies of the organic matrix of enamel. J.
Ultrastruct. Res. 75:281, 1965.
GENERAL
Kallenbach, E.: Fine structure of differentiating amelo-
Miles, A. E. W.: Structural and Chemical Organization blasts in the kitten. Am. J. Anat. 745:283, 1976.
of Teeth. Vols I & II. New York, Academic Press. Katz, E. P., J. Seyer, P. T. Levine, and M. J. Glimcher:
1967. The comparative biochemistry of the organic matrix
Schour, I., ed.: Noyes’ Oral Histology and Embryology. of developing enamel. Arch. Oral Biol. 74:533,
8th ed. Philadelphia, Lea & Febiger, 1960. 1969.
Sicher, H., ed.: Orban’s Oral Histology and Embryology. Kerebel, B., G. Daculsi, and L. M. Kerebel: Ultrastruc¬
6th ed. St. Louis, C. V. Mosby Co., 1971. tural studies of enamel crystallites. J. Dent. Res.
58(B): 844, 1979.
DENTIN
Leblond, C. P., and H. Warshawsky: Dynamics of enamel
Bernard, G. W.: Ultrastructural observations of initial formation in the rat incisor tooth. J. Dent. Res.
calcification in dentin and enamel. J. Ultrastruct. 58(B): 950, 1979.
Res. 41:1, 1972. Matthiessen, M. E., and P. Romert: Fine structure of the
Garant, P. R., G. Szabo, and J. Nalbandian: Fine struc¬ human secretory ameloblast. Scand. J. Dent. Res.
ture of the mouse odontoblast. Arch. Oral Biol. 86:67, 1978.
13:857, 1968. Smith, C. E.: Ameloblasts: secretory and resorptive func¬
Josephsen, K., and H. Warshawsky: Radioautography tions. J. Dent. Res. (B):695, 1979.
of rat incisor dentin as a continuous record of the Warshawsky, H.: The fine structure of secretory ame¬
incorporation of a single dose of HMabelled proline loblasts in rat incisors., Anat. Rec. 767:121, 1968.
and tyrosine. Am. J. Anat. 164:45, 1982. Weinstock, A., and C. P. Leblond: Elaboration of the
Munhoz, C. O. G., and C. P. Leblond: Deposition of matrix glycoprotein of enamel by the secretory
calcium phosphate into dentin and enamel as shown ameloblasts of the rat incisor as revealed by radioau¬
by radioautography of sections of incisor teeth fol¬ tography after galactose 3H injection. J. Cell Biol.
lowing injection of 45Ca into rats. Calcif. Tissue 57:26, 1971.
Res. 75:221, 1974.
Reith, E. J.: Collagen formation in developing molar PERIODONTAL TISSUES
teeth of rats. J. Ultrastruct. Res. 27:383, 1968. Beertsen, W., M. Brekelmans, and V. Everts: The site
Weinstock, A., and C. P. Leblond: Synthesis migration of collagen resorption in the periodontal ligament
and release of precursor collagen by odontoblasts of the rodent molar. Anat. Rec. 792:305, 1978.
as visualized by radioautography after (3H) proline Listgarten, M. A.: The ultrastructure of the human
administration. J. Cell Biol. 60:92, 1974. gingival epithelium. Am. j. Anat. 774:49, 1964.
ENAMEL Smuckler, H., and C. J. Dreyer: Principal fibers of the
periodontium, j. Periodont. Res. 4:19, 1969.
Frank, R. M.: Tooth enamel: current state of the art. J. Stern, L. B.: An electron microscopic study of the
Dent. Res. 58(B):684, 1979. cementum: Sharpey’s fibers and periodontal liga¬
Glimcher, M. J., L. C. Bonar, and E. J. Daniel: The ment in the rat incisor. Am. J. Anat. 775:377, 1964.
molecular structure of the protein matrix of bovine Susi, F. R.: Anchoring fibrils in the attachment of epi¬
dental enamel. J. Mol. Biol. 5:541, 1961. thelium to connective tissue in oral mucous mem¬
Glimcher, M. J., P. T. Levine, and L. C. Bonar: Mor¬ brane. J. Dent. Res. 48:144, 1969.
 THE ESOPHA GUS AND
STOMACH

ESOPHAGUS with the muscularis mucosae, forms numer¬

ous longitudinal folds, which result in an
The esophagus is a muscular tube 25 cm in irregular outline of the lumen in cross sec¬
length that conveys food rapidly from the tion. During the swallowing of food these
pharynx to the stomach. The greater part of folds are smoothed out. This is made possible
its length is intrathoracic but the terminal 2 by the elasticity of the connective tissue that
to 4 cm are below the diaphragm. Its wall forms the submucous layer.
includes all the layers characteristic of the The muscularis of the human esophagus is
digestive tube in general (Fig. 25-1). The 0.5 to 2.2 mm thick. In the upper quarter of
mucosa is 500 to 800 pm thick. The stratified the esophagus, both its outer and inner layers
squamous epithelium (Figs. 25-2, 25-3) con¬ consist of striated muscle. In the second
tinues from the pharynx into the esophagus. quarter, bundles of smooth muscle begin
At the junction of the esophagus with the gradually to replace the striated muscle, and
cardia of the stomach, there is an abrupt in the lower third only smooth muscle is
transition from stratified squamous to simple found. The relations between the two types
columnar epithelium (see Fig. 25-5). On of muscular tissue are subject to individual
macroscopic examination, the boundary line variations. The two layers of the muscularis
between the smooth, white mucous mem¬ are not as regularly circular and longitudinal,
brane of the esophagus and the pink surface respectively, as tfiey are elsewhere in the
of the gastric mucosa appears as a jagged alimentary tract. In the inner layer there are
line. many spiral or oblique bundles. The longi¬
In humans the flattened cells of the super¬ tudinal muscular bundles of the outer layer
ficial layers of the epithelium contain a small are also irregularly arranged in many places.
number of keratohyalin granules but do not The outer surface of the esophagus is con¬
undergo true cornihcation. The lamina pro¬ nected with the surrounding parts by a layer
pria consists of loose connective tissue with of loose connective tissue constituting the
relatively thin collagenous fibers and net¬ tunica adventitia.
works of fine elastic fibers. In addition to the
usual connective tissue cells, numerous lym¬
phocytes are scattered throughout the tissue. GLANDS OF THE ESOPHAGUS
Small lymphatic nodules are found around
the ducts of the esophageal mucous glands. Two kinds of small glands occur in the
At the level of the cricoid cartilage of the esophagus: esophageal glands proper and esoph¬
larynx the elastic layer of the pharynx gives ageal cardiac glands. The esopfiageal glands
way to the muscularis mucosae, which consists proper are small, compound glands with
of longitudinal smooth muscle fibers and thin richly branched tubuloalveolar secretory por¬
elastic networks. Near the stomach the mus¬ tions containing only mucous cells (Fig. 25—
cularis mucosae attains a thickness of 200 to 4). They are unevenly distributed in the sub¬
400 pm. mucous layer and can just be recognized with
The dense connective tissue of the submu¬ the naked eye as elongated white spots. The
cosa consists of collagenous and elastic fibers branches of the smallest ducts are short and
and small infiltrations of lymphocytes about fuse into a cystically dilated main duct, which
the glands. The submucous layer, together pierces the muscularis mucosae and opens

619
620 • THE ESOPHAGUS AND STOMACH

Mucous glands

Circular muscle

Longitudinal muscle

Mucous glands

Tunica muscularis

Lamina
propria
mucosae

Lumen

Lamina muscular
mucosae

Su bmucosa

Inner longitudina

Epithelium

Figure 25-1. Cross section from the middle third of the esophagus of a 28-year-old man. x 8. (After Sobotta.)

Figure 25-2. Esophageal stratified epithelium of a rhesus monkey.

 THE ESOPHAGUS AND STOMACH 621

g»*sr mm

?W%&- ■' *&**&&?*

mM

^sag*^-
Ji |

«p:%
^■k : >; • -f
’ Msl*
' %•*%'' $***? '’$*<?-

i. /_7‘ -3l v '- * I

vA a A
FJ&t4
-Vi-Si fsST (A- Stt/A
r irW>f> Ui^lm

ct>?

flW' Ia - * ■>A!#t§

<\

; ’Vi v

Figure 25-3. Electron micrograph of cells of the basal layer of rodent esophageal epithelium. Division of these cells is
responsible for continual renewal of the epithelium. They have abundant free ribosomes and occasional fascicles of
cytoplasmic filaments (at arrows). They are attached by well-developed desmosomes. One enclosed in rectangle at
upper right is shown at high magnification in the inset. Notice the prominent intermediate line in the interspace between
the two membranes. (Micrograph courtesy of S. McNutt.)
622 • THE ESOPHAGUS AND STOMACH

Figure 25-4. A, Photomicrograph of esophageal wall, showing the lumen and lining epithelium and the esophageal
glands in the submucosa, x 120. B, Esophageal glands at higher magnification, illustrating the dark pyknotic-appearing
nuclei displaced to the base of the cell by the accumulated mucigen in the apical region, x 300.

Esophageal
Gastric epithelium epithelium

Figure 25 5. Esophagogastric junction. Notice the abrupt transition from stratified squamous to simple columnar
epithelium (at arrow). Hematoxylin and eosin. x 375.
 THE ESOPHAGUS AND STOMACH • 623

through a small orifice. The epithelium in they have a mixed character. No esophageal
the smallest ducts is low columnar, whereas glands are found in rodents, horses, and cats.
that in the enlarged main duct is stratified
squamous epithelium. The mucous glands
may give rise to cysts of the mucous mem¬ HISTOPHYSIOLOGY OF THE
brane. ESOPHAGUS
The esophageal cardiac glands closely resem¬
ble the cardiac glands of the stomach. Two At the junction of the pharynx and esoph¬
groups of them can be distinguished. One is agus is a region of higher muscular tone
in the upper part of the esophagus at the referred to as the pharyngoesophageal
level between the cricoid cartilage and the sphincter, Similarly, the terminal few centi¬
fifth tracheal cartilage; the other is in the meters of the esophagus serve as a gastroe¬
lower part of the esophagus near the cardia. sophageal sphincter normally maintaining an
They show great individual variation and intraluminal pressure slightly higher than
sometimes are entirely absent. intragastric pressure. There is no anatomical
Unlike the esophageal glands proper, they thickening or local change in orientation of
are always confined to the lamina propria the musculature in these regions. Thus, they
mucosae. Their terminal portions are are physiological rather than anatomical
branched and convoluted tubules lined by sphincters. Nevertheless, the gastroesopha¬
columnar or cuboidal cells with a pale gran¬ geal sphincter is normally quite efficient in
ular cytoplasm, which sometimes gives a preventing reflux of gastric contents. How¬
staining reaction for mucin. The smallest ever, the esophageal hiatus in the diaphragm
ducts drain into a larger duct that is some¬ not infrequently fails to close completely
times cystically dilated and always opens on around the esophagus during embryonic de¬
the tip of a small papilla. Its columnar epi¬ velopment, resulting in a hiatus hernia
thelium often gives a staining reaction for through which a portion of the stomach may
mucin and resembles the mucous epithelium project into the thoracic cavity. This often
of the gastric foveolae. interferes with the normal sphincteric func¬
In the regions of esophageal mucous mem¬ tion of the terminal esophagus, permitting
brane that contain the upper and lower reflux of gastric contents. The esophageal
groups of cardiac glands, the stratified squa¬ epithelium is poorly equipped to resist the
mous epithelium may be supplanted in places acidity of gastric secretions, and the resulting
by a simple columnar epithelium resembling inflammatory response may cause difficulty
that in the gastric pits of the stomach mucosa. in swallowing and lead ultimately to devel¬
Seen with the naked eye, such patches may opment of fibrosis and stricture of the lower
be mistaken for erosions—that is, places de¬ esophagus.
nuded of epithelium. Sometimes these In swallowing, the tongue propels the food
patches lined with mucous epithelium are of back into the pharynx. This sets in motion a
considerable size and exhibit pitlike invagi¬ train of coordinated voluntary and involun¬
nations and tubular glands like those of the tary movements by pharyngeal and esopha¬
stomach; they may even contain typical zy¬ geal musculature. These involve closure of
mogenic and parietal cells. the glottis, elevation of the larynx, constric¬
The number and development of the tion of the pharynx, and reflex relaxation of
esophageal cardiac glands, as well as of the the pharyngeoesophageal sphincter. When
islands of gastric mucosa in the esophagus, the bolus of food enters the esophagus, the
are subject to great individual variation. Ac¬ local stimulus of distention initiates a peri¬
cording to some investigators, the presence staltic wave of contraction that is propagated
of this ectopic gastric epithelium is of some toward the stomach at a rate of 4 to 6 cm
importance in relation to the origin of cysts, per second. The gastroesophageal sphincter
ulcers, and carcinomas of the esophagus. relaxes transiently in anticipation of the ar¬
In many mammals, especially those that rival of the peristaltic wave, allowing the food
consume coarse vegetable food (rodents, to pass into the stomach.
ruminants, and equids), the stratified squa¬ The nerves to the esophagus are derived
mous epithelium of the esophagus undergoes from the vagus and the cervical and thoracic
keratinization. The esophageal glands are sympathetic trunks. They form a plexus of
present in most of the mammals, but instead fibers and clusters of cell bodies between the
of being purely mucous, as in the human, two layers of the muscular coat and a second
624 • THE ESOPHAGUS AND STOMACH

plexus in the submucosa. They are important

for coordination of movements involved in
swallowing. Disturbances of the neuromus¬
cular apparatus are fairly common in older
persons and individuals of nervous temper¬
ament, and may result in muscle spasm, dif¬
ficulty in swallowing, and severe substernal
pain.

Pyloric
STOMACH antrum

Figure 25-6. Drawing of the human stomach presenting

The stomach is an organ specialized for the terminology of its various regions.
%

storage and processing of food for subse¬

quent absorption in the intestine. Its storage greater and lesser curvatures. The stomach
function is minimal in the human and is best wall consists of the layers previously defined
exemplified in the multichambered stomach as characteristic of the entire digestive tract:
of ruminants, where food taken in during mucosa, submucosa, muscularis, and serosa.
grazing or browsing is temporarily stored in
the rumen and later returned to the oral cavity
for further mastication. It is then swallowed THE GASTRIC MUCOSA
again and proceeds successively through the
reticulum, omasum, and abomasum. In other The mucosa of the living stomach is a
mammals having a relatively simple, single- grayish pink in color with paler zones at the
chambered stomach, there is little or no stor¬ cardia and pylorus. In the empty contracted
age, but the wall of the stomach is capable of stomach, the mucosa forms numerous longi¬
considerable expansion during feeding with¬ tudinal folds or rugae. This plication is per¬
out significant increase in internal pressure. mitted by the loose consistency of the sub¬
In the human, it can accommodate up to 1 mucosa and results from contraction of the
liter. muscularis mucosae. In the full stomach, the
In the stomach, the semisolid material re¬ rugae flatten out and the mucosa is smooth
sulting from mastication is reduced to a fluid contoured to the naked eye. On closer in¬
by contractions of the muscular wall of the spection, a pattern of low elevations is created
organ and admixture with the acid and en¬ by shallow furrows that demarcate slightly
zymes secreted by its glandular mucosa. The bulging areas 2 to 6 mm in diameter (Fig.
content of the upper portion of the stomach 25-7). At low magnification, the surface of
may remain semisolid for some time after a each area is further subdivided by a system
meal, while that of the most distal portion is of intercommunicating shallow furrows, the
reduced to a pulplike fluid mass referred to gastric pits or foveolae. This convoluted pattern
as the chyme. When it has attained the neces¬ of ridges and foveolae is seen most clearly in
sary softness, it is passed on in small portions scanning electron micrographs of gastric mu¬
to the duodenum. Thus, the digestive func¬ cosa (Fig. 25—8). At this magnification, the
tion of the stomach is in part mechanical and convex apical surfaces of the individual epi¬
in part chemical. thelial cells can also be identified. In histolog¬
The area immediately surrounding the ical sections of the mucosa, the foveolae ap¬
opening from the esophagus into the stom¬ pear as funnel-shaped invaginations of the
ach is called the cardia. To the left of the surface epithelium continuous at their base
cardia a dome-shaped bulge above the level with numerous tubular gastric glands (Fig.
of the esophageogastric junction is called the 25-9). The columnar epithelium that covers
fundus. The capacious central portion is the the ridges and lines the foveolae is of uniform
corpus or body, and the more distal region of cellular composition throughout the gastric
transition to the duodenum is the pylorus (Fig. mucosa, but there are regional differences in
25—6). Gastroenterologists often incorporate the cell populations comprising the gastric
the fundus and corpus together under the glands, and this is reflected in significant
term fundic region. The convex left and con¬ regional differences in the nature of the
cave right margins are called respectively the gastric secretions.
 THE ESOPHAGUS AND STOMACH • 625

Surface epithelium

Gastric pits

Figure 25-7. Surface of gastric mucosa

of a man; drawing based on binocular
microscope view. The cut surfaces are
slightly diagrammatic. At left, the normal
distribution of the gastric glands; to right,
only a few are indicated. Glands, gray;
gastric pits, black, x 17. (After Braus.)

Lymphoid nodule

Gastric glands

Lamina propria mucosae

Submucosa i
Muscularis mucosae
i
Smooth muscle layers Serosa

Figure 25-8. Scanning electron micrograph of the luminal surface of the gastric mucosa. The cells are all the same
type—surface mucous cells. The convex apical surfaces of the individual epithelial cells are clearly seen. The convoluted
pattern of ridges and gastric pits, or foveolae, is also evident. (Micrograph courtesy of J. Riddell.)
626 • THE ESOPHAGUS AND STOMACH

Figure 25-9. Photomicrograph of the gastric mucosa of a macaque, showing the gastric glands opening into the gastric
pits, or foveolae. Hematoxylin and eosin. x 120.

Surface Epithelium stances and the potentially damaging acid

and enzymes of the gastric secretions.
The gastric pits and the ridges between At the electron microscopic level, there are
them are lined by a tall columnar epithelium numerous microvilli with a prominent gly-
that begins abruptly at the cardia under an cocalyx on the luminal surface, underlain by
overlying margin of the stratified squamous a narrow zone of cytoplasm rich in filaments
epithelium of the esophagus (Fig. 25-5). The forming a terminal web. Below this are mu¬
apical portion of the cells of the surface cous granules closely packed with little inter¬
epithelium is occupied by mucigen, the intra¬ vening cytoplasm (Fig. 25-10). The granules
cellular precursor of mucus. In routine his¬ are spherical, ovoid, or discoid and vary in
tological sections this portion of the cell may density from pale to completely electron-
be unstained, but after the periodic acid- opaque. In some species, they have dense
Schiff (PAS) reaction for carbohydrates it cores and a peripheral zone of lower density.
stains intensely. Although there is no obvious Material with a texture similar to that of the
morphological difference among surface mu¬ granules may be found in cisternae of the
cous cells, the characteristics of the mucus supranuclear Golgi complex. The paranu¬
exhibit regional variations when studied his- clear and basal cytoplasm contains cisternal
tochemically. The interfoveolar cells contain profiles of endoplasmic reticulum, numerous
neutral mucosubstance, while the cells deep mitochondria, and limited numbers of micro¬
in the foveolae contain acidic carbohydrates. tubules and filaments. The nucleus is often
Cells on the surface also contain more mu¬ irregular in outline.
cous granules than those deeper in the pits Adjacent cells are held together by juxta-
or isthmus of the glands. The mucus secreted luminal zonulae occludentes and adherentes,
plays an essential role in lubricating the sur¬ and desmosomes and gap junctions are
face epithelium and forming a barrier layer, sparsely distributed on the lateral cell bound¬
protecting it from injury by ingested sub¬ aries. Toward the base of the epithelium, the
 THE ESOPHAGUS AND STOMACH • 627

Figure 25-10. Electron micrograph of the apical portion of several surface mucous cells. The short microvilli have a
prominent glycocalyx and the surface of the epithelium is covered by a layer of mucus. (Courtesy of S. Ito.)

membranes often diverge, forming a con¬ cardiac glands varies from 5 to 30 mm in

spicuous intercellular cleft into which slender width in the human to as much as a third of
folds project from lateral surfaces of the cells. the stomach in pigs.
Surface mucous cells are continuously des¬
quamated into the lumen, and the entire Gastric Glands (Oxyntic Glands)
population is renewed about every three
days. Evidence of regeneration is seen only This glandular type is characteristic of the
in the depths of the foveolae and necks of mucosa throughout the fundus and corpus
the gastric glands where mitoses are frequent of the stomach, and makes the greatest con¬
in relatively undifferentiated cells that con¬ tribution to secretion of the gastric juice. The
tain few mucigen granules. The newly oxyntic glands (also called gastric glands or
formed cells are slowly displaced upward to fundic glands) are closely packed together and
replace cells lost at the surface. extend through the entire thickness of the
mucosa (0.3-1.5 mm). From one to seven
open through slightly constricted necks into
Cardiac Glands
the bottom of each foveola. They are 30 to
In a narrow zone of the cardia around the 50 pm in diameter with a very narrow lumen.
esophageogastric junction, the foveolae are Their blind distal ends are slightly expanded
relatively shallow, and each is continuous and coiled and may divide into two or three
below with several tortuous tubular cardiac terminal branches. There are estimated to be
glands. Unlike the glands in other segments 15 million oxyntic glands associated with 3.5
of the stomach, these consist mainly of mu¬ million gastric foveolae.
cous cells, with smaller numbers of undiffer¬ The oxyntic glands are composed of four
entiated cells and scattered endocrine cells. cell types: (1) mucous neck cells (2) chief cells
The endocrine cells are believed to secrete (also called zymogenic cells) (3) oxyntic or parietal
the hormone gastrin. The area occupied by cells and (4) endocrine cells (Fig. 25—13). For
628 THE ESOPHAGUS AND STOMACH

Figure 25-11. Micrograph of chief cells in a gastric gland of monkey stomach. The base of the cells is occupied by
closely spaced cisternae of rough endoplasmic reticulum, and the apex crowded with paie secretory granules containing
pepsinogen. (Courtesy of S. Ito.)
 THE ESOPHAGUS AND STOMACH • 629

Zymogen
Ergastop lasm granules

Figure 25-12. Drawing of the chief or zymogenic cell as seen with the electron microscope. (From Ito, S., and R. J.
Winchester. J. Cell Biol. 76:541, 1963.)

descriptive purposes, the gland is customarily with the PAS reaction, mucicarmine, or mu-
divided into three regions. At its confluence cihematein, the granules that fill the apical
with the foveola is the isthmus, containing cytoplasm are deeply colored, whereas those
surface mucous cells and oxyntic cells; fol¬ of the chief cells are unstained. There is
lowed by the neck, consisting mainly of mu¬ evidence that the mucus secreted by these
cous neck cells and oxyntic cells; and the base cells is somewhat different from that of the
or fundus, where chief cells are the predom¬ surface mucous cells.
inant cell type, with smaller numbers of ox¬ The mucous neck cells are deformed by
yntic and mucous neck cells. Scattered en¬ neighboring cells and therefore tend to be
docrine cells may be found in any segment quite irregular in shape; some have a wide
of the gland. Mitotic activity is largely con¬ base and narrow apex, others a broad apex
fined to the neck of the glands and it is and narrow base. The nuclei are at the bases
probable that the chief, oxyntic, and other of the cells and are often somewhat flattened.
cell types develop from a small population of Where the necks of the glands open into
relatively undifferentiated cells in this region. the bottoms of the foveolae, a series of tran¬
Mucous Neck Cells. These cells are rela¬ sitional forms between mucous neck cells and
tively few in number and are lodged between surface mucous cells can be identified. New
the parietal cells in the neck of the glands cells arise by transformation of the relatively
where the latter open into the gastric pits. undifferentiated cells found in the depths of
Deeper in the glands, they are abruptly suc¬ the foveolae and in the necks of the glands.
ceeded by the zymogenic cells. In fresh, un¬ In some gastric glands, mucous neck cells
stained preparations they are filled with pale, extend deeper into the gland and may be
transparent granules. These cells are easily found scattered singly among the zymogenic
overlooked or mistaken for zymogenic cells cells. This is especially prominent in glands
in preparations in which mucus either is not near the pyloric region. According to some,
preserved or is unstained. In sections stained the glands of a narrow intermediate zone
630 • THE esophagus and stomach

gland (Fig. 25-14). Their cytoplasm stains

intensely with eosin and phloxine. Their most
unusual feature is the presence of a mean¬
dering canalicular invagination of the apical
surface that may encircle the nucleus and
extend nearly to the basal lamina (Figs. 25—
15, 25—16). This was called the “intracellular
canaliculus” by light microscopists—a some¬
what misleading term. Although this channel
makes a deep incursion into the cell body, its
limiting membrane is continuous with the
plasmalemma at the cell apex where it opens
into the lumen of the gland. Thus, it is not a
closed intracellular canal within the cyto¬
plasm as the name implies, and the term
secretory canaliculus is probably preferable.
Like the apical surface of the cell, the canal¬
iculus is lined by mjcrovilli whose number
and length vary with the state of the cell’s
secretory activity.

Figure 25-13. Diagram of an oxyntic gland from the

corpus of a mammalian stomach. (From Ito, S. In Johnson,
L. R., ed.: Physiology of the Gastrointestinal Tract. New
York, Raven Press, 1981.)

may contain only mucous neck and parietal

ceils, and may be devoid of zymogenic cells.
Under the electron microscope, the lu¬
minal surfaces of the columnar mucous neck
cells are studded with short microvilli that
have a fuzzy glycocalyx. The lateral surfaces
of neighboring cells are interdigitated, par¬
ticularly toward the base of the cell. The
apical region of the cell contains numerous
dense granules of spheroid, ovoid, or discoid
form. Rod-shaped mitochondria of the usual
internal structure are scattered through the
cytoplasm. There is a sizable supranuclear
Golgi complex. Membranous profiles of en¬
doplasmic reticulum are present in small
numbers.
Oxyntic Cells (Parietal Cells). The most
conspicuous and distinctive cell type in the
gastric mucosa is the oxyntic or parietal cell. Figure 25-14. Drawing of a portion of a gastric gland,
These large pyramidal cells up to 25 pm in showing the parietal cells bulging outward, zymogenic
cells, and endocrine cells with small basal granules. (From
diameter have broad rounded bases that
Krstic, R. V. Die Gewebe des Menschen und Saugetiere.
often bulge outward at the periphery of the Berlin, Springer-Verlag, 1978.)
 THE ESOPHAGUS AND STOMACH • 631

Golg
Complex

Secretory
Canaliculus

Mitochondria

Basal Folds
Secretory
Canaliculus

Basement Lamina

Figure 25-15. Drawing of the ultrastructure of the gastric parietal cell or oxyntic cell. It is a large cell with very abundant
large mitochondria and an intracellular canaliculus lined with microvilli. (Drawing courtesy of S. Ito.)

Secretory granules are lacking and the canaliculus are subjects of continuing contro¬
Golgi complex, located in a paranuclear or versy among electron microscopists. It is clear
basal position, is less well developed than in that the appearance of this system varies
other glandular cells. The cytoplasm is greatly, depending on the state of activity of
crowded with large mitochondria that occupy the cell and the method of specimen prepa¬
up to 40 per cent of the cytoplasmic volume. ration. The descriptive term tubulovesicular
The large number of mitochondria and the system, now widely used, represents a compro¬
amplification of their internal surface by nu¬ mise among conflicting interpretations. Re¬
merous cristae are consistent with the very cent application of the technique of rapid¬
high energy requirement of their secretory freezing with liquid helium and freeze sub¬
function. Isolated parietal cells have a rate of stitution strongly suggests that the system is
oxygen consumption five times that of the predominantly tubular in the living state (Fig.
neighboring mucous cells. 25-17).
Another unique ultrastructural feature of Hydrochloric acid is first detected in the
the cell is the presence in the cytoplasm of fetal stomach at a time coinciding with the
an extensive system of membrane-limited tu¬ differentiation of parietal cells, and the
bules that do not have the properties typical amount of acid produced is positively corre¬
of the endoplasmic reticulum of other cells. lated with the number of these cells in the
Whether in the living cell these profiles are gastric mucosa. This and other indirect evi¬
predominantly tubular or vesicular, continu¬ dence have led to general agreement that it
ous or discontinuous, and whether they are is the parietal cells that are responsible for
closed or open to the lumen of the secretory the stomach’s remarkable capacity to secrete
632 • THE ESOPHAGUS AND STOMACH

P Lumen

Intracellular
canaliculus
■ .lii18 HfiSlii

Intracellular
Kdnaliylrus

Figure 25-16. Electron micrograph of a parietal cell of bat gastric mucosa. The extensive secretory canaliculus within
the limits of the cell is filled by large numbers of irregularly oriented microvilli, x 10,000. (Courtesy of S. Ito.)

hydrochloric acid. It is this function that is pelling morphological evidence for such a
implied in the alternative term “oxyntic” cell. translocation of preformed membrane is still
This cell type also secretes gastric intrinsic lacking. When active secretion ceases, the
factor, a substance required for absorption of oxyntic cells revert to their resting state and
vitamin B12 in the ileum. an extensive tubulovesicular system is recon¬
The internal organization of the oxyntic stituted.
cell undergoes striking changes in different Chief Cells (Zymogenic Cells). The chief
phases of secretory activity. In the nonsecret¬ cells form a cuboidal epithelium lining the
ing cell, microvilli on the surface and in the lower third or half of the gastric glands in
secretory canaliculus are short and sparse, the corpus of the stomach. They are absent
and the tubulovesicular system is extremely from cardiac glands, sparse in the glands of
developed. After stimulation of acid secre¬ the fundus, and rare in the pylorus. Chief
tion, there is a rapid increase in length and cells are difficult to preserve adequately in
number of microvilli, resulting in a fivefold routine histological preparations. When
increment in surface area. A concomitant and properly fixed, they contain numerous apical
equally dramatic reduction in the tubuloves- secretory granules and a strongly basophilic
icuiar membranes of the cytoplasm has fos¬ cytoplasm as in other protein-secreting zy¬
tered the belief that the tubulovesicular mem¬ mogenic cells. They synthesize and secrete
branes may fuse with the plasmalemma and, pepsinogen which is activated to form the
by exteriorization, contribute to the rapid digestive enzyme pepsin.
increase in microvillar surface area. Freeze- In electron micrographs, the chief cells
fracture studies demonstrate a similarity in have the ultrastructure characteristic of pro¬
pattern of intramembrane particles consis¬ tein-secreting glandular cells such as those of
tent with the hypothesis that tubulovesicular the pancreas—a well-developed supranuclear
membranes fuse with the plasmalemma at Golgi complex; abundant rough endoplasmic
the onset of induced acid secretion, but com¬ reticulum concentrated in the basal cyto-
 THE ESOPHAGUS AND STOMACH • 633

Figure 25-17. Electron micrograph of a portion of a parietal cell including a segment of the secretory canaliculus.
Notice the numerous elements of the tubulovesicular system. Specimen rapidly frozen with liquid helium and further
processed by freeze substitution in osmium tetroxide and acetone. (Courtesy of N. Sugai and A. Ichikawa.)

plasm; and numerous moderately dense, In the past 20 years, advances in biochem¬
spherical secretory granules in the apical cy¬ ical extraction methods and radioimmuno¬
toplasm. In the chief cells of humans, dense assay procedures have led to the identifica¬
bodies, apparently lysosomes, are commonly tion and purification of a large number of
found among the pepsinogen granules. The peptide hormones and biogenic amines. By
luminal surface bears sparse microvilli lim¬ using labeled antibodies to these substances,
ited by a plasma membrane with a thin gly- many of them have been localized to specific
cocalyx. single cells widely scattered throughout the
Although the chief cells are the main gastrointestinal tract. Whereas the silver-
source of gastric pepsin, studies with fluores¬ staining cells were formerly divided into two
cent-labeled antibodies to pepsinogen also categories, immunocytochemical methods
localize small amounts in the mucous neck have now identified nine different endocrine
cells of the gastric glands, but not in mucous or paracrine cell types in the stomach, and
cells of the pyloric region. the list continues to grow. All are small py¬
Endocrine Cells. Small granulated cells ramidal or ovoid cells lodged between the
scattered individually among the epithelial bases of the neighboring exocrine cells of the
cells of the gastric glands have been recog¬ epithelium. The narrow apex of some ex¬
nized for more than a century. They can be tends to the lumen; others are confined to
stained selectively with silver or chromium the base of the epithelium. All possess small,
salts and therefore were long referred to as membrane-limited granules usually concen¬
argentaffin or enterochromaffn cells. Although trated in their basal cytoplasm. Some types
these methods permitted recognition of two are readily identifiable in electron micro¬
or more categories of silver-reducing cells, graphs on the basis of differences in size or
they shed no light upon the chemical nature substructure of their granules. Those con¬
of their granules or their physiological sig¬ taining monoamines exhibit a characteristic
nificance. fluorescence. The most reliable means of
634 • THE ESOPHAGUS AND STOMACH

identification of the peptide-secreting cells ology is the G cell or gastrin cell. Its hormone
involves use of labeled specific antibody to is a potent stimulator of acid secretion by the
their respective secretory products. oxyntic cells, and is also thought to stimulate
Collectively these cells are now referred to mucosal cell proliferation. The cell is pyram¬
as the enteroendocrine cells. They are found idal in form, with a narrow apex extending
throughout the enteric tract. The terminol¬ to the lumen, and possessing a tuft of long
ogy of cell types, the morphological criteria microvilli. The granules are quite heteroge¬
for their identification, and the nature of neous, varying in size from 150 to 400 nm.
their secretions will be considered in greater Some appear to be vesicles with little content,
detail in the chapter on the intestines (Chap¬ others have dense cores, and still others are
ter 26). Three of the enteroendocrine cell of intermediate density (Figs. 25-18, 25—19).
types are largely confined to the gastric mu¬ Granules are concentrated at the cell base
cosa: theTC cell, secreting serotonin; the ECL and release their product into the lamina
cell, believed to release histamine; and the G propria by exocytosis.
cell, which secretes the hormone gastrin. The Although the enteroendocrine cells appear
EC cells contain small (~ 300 nm) oval or in tissue sections to be few and widely scat¬
crescentic granules with a dense core, and tered, the aggregate number in the gastric
have an apical extension of their cytoplasm mucosa is quite large. Gastrin cells alone are
to the lumen. The ECL cell contains larger estimated to number 5 X 105 per square
granules (up to 450 nm) with an eccentric centimeter of mucosa in dog, cat, and rat.
dense core of varying shape. It is found The presence of food distending the stomach
exclusively in the gastric mucosa. causes release of gastrin, which diffuses into
Of particular importance in gastric physi¬ the blood and is distributed to the glands of

Figure 25 18. Schematic representation of the ultrastructure of the gastric cell (G cell). As in other endocrine cells the
secretory granules are concentrated at the vascular pole. (From Ito, S. In Johnson, L. FT, ed.: Physioloqy of’ the
Gastrointestinal Tract. New York, Raven Press, 1981.) y
 THE ESOPHAGUS AND STOMACH • 635

the gastric mucosa, where it stimulates the Pyloric Glands

oxyntic cells and to a lesser extent the chief
The pyloric glands are found in the distal
cells. In response, the oxyntic cells increase
4 to 5 cm of the stomach. In this region, the
their rate of hydrochloric acid secretion as
foveolae are deeper than elsewhere in the
much as eightfold.
stomach, extending down into the mucous
membrane for half its thickness. The glands
here are also of the simple, branched tubular
type, but they branch more extensively, the
lumen is larger, and the tubules are coiled,
so that in perpendicular sections they are
seldom seen as continuous longitudinal struc¬
tures (Fig. 25—20). The principal cell type of
the pyloric glands has a pale cytoplasm with
an indistinct granulation. Secretory capillar¬
ies have been described between the cells.
The nucleus is often flattened against the
base of the cell. In sections stained with
hematoxylin and eosin, they are difficult to
distinguish from mucous neck cells or the
cells of the glands of Brunner in the duo¬
denum. Some investigators believe that the
pyloric glandular cells are identical with the
mucous neck cells, for both give similar stain¬
ing reactions for mucus. Cresyl violet and the
Giemsa mixture of dyes, however, seem to
stain them in a specific way. In the human
stomach, the pyloric glands in the region of
the sphincter may contain parietal cells. Gas¬
trin cells are also described in the pyloric
glands.

Gastric pits

Pyloric.
glands

Muscu laris
mucosae
S**.- '. -—7*)

k5V5- 'fj.
?) /t .v
'av.'••
Figure 25-19. Electron micrograph of one of the types of
granular argentaffin or endocrine cells. This kind is be¬
lieved to secrete the hormone gastrin. (Micrograph by S. Figure 25-20. Pyloric glands from human stomach.
Ito.) Slightly diagrammatic, x 75. (After Braus.)
636 • THE ESOPHAGUS AND STOMACH

Lamina Propria from the muscularis mucosae. It consists of

three layers of smooth muscle fibers: longi¬
Connective tissue of the lamina propria tudinal, circular, and oblique, but these are
occupies the narrow spaces between the not as clearly defined as in the muscularis of
glands and between their bases and the un¬ the intestine and their relationships are not
derlying muscularis mucosae. It forms larger easily described. The longitudinal fibers, di¬
accumulations only between the necks of the rectly beneath the serosa, are continuous with
glands and between the foveolae. It consists the longitudinal fibers of the esophageal wall.
of a delicate network of collagenous and They radiate from the cardia, are best devel¬
reticular fibrils and is almost devoid of elastic oped along the greater and lesser curvatures
elements. In addition to oval pale nuclei, of the stomach, and do not extend to the
which seem to belong to fibroblasts, the pylorus. This layer is incomplete, and longi¬
meshes of the fibrous network contain nu¬ tudinal fibers cannot be found in some areas
merous lymphocytes and some plasma cells, of the dorsal and ventral surface of the cor¬
eosinophils, and mast cells. Lymphoid cells pus.
with coarsely granular acidophilic inclusions, The circular fibers form a complete, uni¬
called Russell’s bodies, may be found between form layer over the whole stomach deep to
the epithelial cells of the glands. These may the longitudinal fibers. The circular muscle
develop under physiological conditions but is continuous with the corresponding layer in
are more common in pathological states. In the esophageal wall. It is thickest in the py¬
the lamina propria, especially in the pyloric lorus, where it forms an annular pyloric sphinc¬
region, strands of smooth muscle may be ter.
found and small accumulations of lymphoid The oblique fibers are deep to the circular
tissue occur normally. fibers and again do not form a complete
layer. They are most abundant at the cardia,
Muscularis Mucosae and sweep downward in broad bands parallel
to the lesser curvature, many of their number
Underlying the gastric glands is the mus¬ diverging toward the greater curvature and
cularis mucosae, consisting of an inner circular intermingling with the fibers of the circular
and outer longitudinal layer of smooth mus¬ muscle layer. Oblique fibers are generally
cle fibers. In some regions there may be an lacking along the lesser curvature of the
additional outer circular layer. Thin strands stomach.
of smooth muscle cells extend from the inner The work of the muscular components of
circular layer toward the surface in the lam¬ the stomach wall is regulated with great pre¬
ina propria between the glands. Their con¬ cision by autonomic nerve plexuses between
traction compresses the mucous membrane its layers. By varying smooth muscle tonus,
and may facilitate discharge of secretion from the stomach wall adapts itself to changes in
the glands. the volume of its contents with little or no
alteration of the pressure in its cavity. Emp¬
tying of the stomach depends on intermittent
SUBMUCOSA peristaltic waves of contraction that sweep
from the cardia toward the pylorus.
The connective tissue layer peripheral to
the muscularis mucosae is called the submu¬
cosa. It is denser than the lamina propria with SEROSA
more abundant collagenous fibers and nu¬
merous lymphocytes, eosinophils, and mast The outermost component of the stomach
cells. This layer contains the arterioles sup¬ wall is a thin layer of loose connective tissue
plying capillaries to the mucosa and a venous overlying the muscularis externa and covered
plexus that receives venules descending from on its outer aspect with mesothelium. It is
the mucosa. There is also a submucous net¬ continuous with the serous covering of the
work of lymphatics. Occasional adipose cells greater and lesser omentum.
are found in this layer.

BLOOD SUPPLY
MUSCULARIS EXTERNA
The arterial supply to the mucosa origi¬
The musculature of the stomach wall is nates from arterioles in the submucosa.
called the muscularis externa to distinguish it These give rise to numerous vessels of capil-
 THE ESOPHAGUS AND STOMACH • 637

Subsurface
capillary network Mucosal
i Maries
Surface
mucous cells

Gastric
mucosa

1 Muscularis
r mucosae

r Submucosa

Mucosal
venule

Submucosal
venule

Submucosal
arterioles
Figure 25-21. Diagramatic representation of the blood vascular supply of human gastric mucosa. The vascular
relationships of the gastric gland, marked with an asterisk, are shown in greater detail in Figure 25-22. (Redrawn after
Gannon, B. J., J. Browning, and J. E. McGuigan. Gastroenterology 86:866, 1984.)

lary size that ascend in the lamina propria cosal defense against potential damage by the
between the gastric glands (Fig. 25-21). acid content of the gastric lumen.
These capillaries are fenestrated and pass in
close proximity to the parietal cells in the
depths of the glands. They continue upward, CELL RENEWAL AND REPAIR
forming a rich capillary network immediately
underlying the surface epithelium. This su¬ The superficial portion of the gastric mu¬
perficial capillary bed is drained by venules cosa has a very rapid rate of cell renewal.
that descend directly to a submucous venous Studies carried out in laboratory rodents us¬
plexus. There appear to be no arteriovenous ing radiolabeled thymidine to identify divid¬
anastomoses in the gastric mucosa. ing cells have shown that the surface mucous
The flow of blood upward to the subsur¬ cells are completely renewed in about three
face capillary plexus past the parietal cells in days. The rate of renewal of the surface
the gastric glands encourages the speculation epithelium in human gastric mucosa is prob¬
that bicarbonate ion generated by actively ably somewhat slower than in small labora¬
secreting parietal cells would diffuse into the tory animals. Mitotic activity is largely con¬
fenestrated ascending capillaries and be fined to cells in the depths of the gastric
transported rapidly to the capillary bed lying foveolae and the mucous neck cells in the
beneath the surface epithelial cells (Fig. 25- isthmus of the gastric glands (Fig. 25—23).
22). There it would be favorably distributed Epithelial cells exfoliated at the surface are
to neutralize any back-diffusing hydrogen replaced by upward migration of cells from
ions from the lumen. It is tempting to believe this region of mitotic activity. The parietal
that the microvascular architecture of the cells and chief cells can be replaced by dif¬
stomach is arranged to provide maximal mu¬ ferentiation of mucous neck cells, but auto-
638 • THE ESOPHAGUS AND STOMACH

Acid flow damaged epithelium in the lower third of the

gastric pits is stimulated to migrate over the
vacated basal lamina of the superficial epithe¬
lium. Within 15 to 30 minutes, it is covered
with a continuous sheet of squamous or low
cuboidal cells, which then increase in height
and soon resume their secretory activity. This
capacity for epithelial migration provides a
rapid defense mechanism for prompt cover¬
age after chemical, thermal, and hyper¬
osmolar injuries that do not extensively dam¬
age the basal lamina. Very low pH in the
gastric lumen may destroy the basal lamina
and completely inhibit or greatly retard res¬
toration of epithelial continuity. Bicarbonate
or other antacids favor reconstitution by neu¬
tralizing luminal acid.

HISTOPHYSIOLOGY OF THE
STOMACH

The mechanical functions of storage, mix¬

ing of stomach contents, and emptying de¬
pend on the muscular wall. Peristaltic waves

Figure 25-22. Diagram illustrating the postulated vascular

transfer of bicarbonate ions, generated by active parietal
cells, to the subsurface capillary network where it may
protect the epithelium by neutralizing any back-diffusing
hydrogen ions from the lumen. (Redrawn after Gannon,
B. J., J. Browning, and J. E. McGuigan. Gastroenteroloqy
86:866, 1984.)

radiographic studies indicate that the cells in

the glands are relatively long-lived and are
renewed only very slowly.
In addition to the continual renewal of its
surface mucous cells, the gastric mucosa has
a remarkable capacity for reestablishment of
epithelial continuity after superficial injury.
In humans, the stomach is frequently in¬
sulted by aspirin (Fig. 25-24), strong alco¬
holic beverages, and other toxic substances
that cause superficial erosion of the mucosa.
Although extensive, irreversible damage and
exfoliation of the surface epithelium results,
there is rapid restoration of epithelial conti¬
nuity by migration of viable cells from the
depths of the foveolae. In experiments on
rodent gastric mucosa exposed to 20 mM
aspirin or 40 per cent ethanol (80 proof), the Figure 25-23. Autoradiograph of the gastric mucosa of a
resulting injury to the surface epithelium is mouse given three injections of tritiated thymidine over a
scarcely detectable after 30 minutes. The 12-hour period preceding fixation of the tissue. The dis¬
tribution of the black deposits of silver demonstrates that
time course of this repair is too short to
the principal site of mitoses is in the neck region of the
involve cell proliferation. Instead, the un¬ gastric glands, x 110. (Courtesy of A. J. Laaman.)
 THE ESOPHAGUS AND STOMACH • 639

Surface mucous ceils originate in the fundus and pass downward

over the stomach to the pylorus. Ten to 15
ml of gastric chyme enter the duodenum
with each peristaltic wave.
In species that are intermittent feeders, the
capacity of the stomach as a distensible res¬
ervoir for accumulation of food is consider¬
able. Although the luminal volume of the
empty stomach is only 50 to 75 ml, nearly
1000 ml can be swallowed before the intra¬
luminal pressure begins to rise. Distention
stimulates peristalsis.
The stimulation of gastric secretion is in
part neural and in part hormonal. Neural
stimulation of secretion travels via the vagus
nerves to the intrinsic nerve plexus, which in
turn innervates the gastric glands and mucus-
Swollen and vacuolated cells secreting glands. In addition the gastrin cells
(G cells) in the antral mucosa respond to
vagal stimulation by releasing the gastroin¬
testinal hormone gastrin. This hormone pro¬
motes pepsinogen secretion, stimulates gas¬
tric motility, and binds to receptors on the
oxyntic cells, stimulating acid secretion. Local
reflexes activated by distention of the stom¬
ach with food also result in gastrin release.
Substances ingested in the food, caffeine, and
low concentrations of alcohol act as secret¬
ogogues favoring the release of gastrin. The
activity of gastrin resides in four amino acids
at its C-terminal end. This tetrapeptide has
been synthesized and is used clinically to
stimulate acid secretion. Release of gastrin is
controlled by a feedback mechanism. When
the pH of the stomach contents reaches 2,
the stimulation of secretion by gastrin ceases,
and when the pH rises significantly at>ove
this level, gastrin is secreted again.
The secretory activity of the stomach is
considerable. The daily secretion of gastric
juice in a fasting human is from 500 to 1500
ml. At each meal, several hundred milliliters
of gastric juice is secreted. This clear, color¬
less fluid contains mucus, water, and electro¬
lytes and the proteolytic enzyme pepsin,
which is active only at the low pH that results
from hydrochloric acid secretion by the pa¬
rietal cells. The chief cells secrete pepsino¬
gen, 42,500 M.W., which upon exposure to
hydrochloric acid is converted to active pepsin,
Figure 25-24. The protective permeability barrier that M.W. 35,000. The enzyme is optimally active
protects the stomach wall from damage by the acidity of at pH 2, cleaving peptide bonds and facilitat¬
the gastric contents is broken down by aspirin or alcohol.
A, Normal surface mucous cells of mouse gastric mucosa. ing gastric digestion of proteins.
B, Mucosa 8 minutes after ora! administration of a solution The oxyntic or parietal cells secrete the
of 20 mM aspirin. The superficial cells are pale, their glycoprotein gastric intrinsic factor, which is
cytoplasm vacuolated, the chromatin marginated. C, Mu¬ essential for the absorption of vitamin B12 in
cosa 8 minutes after 20 mM aspirin and 1 mM HCI.
the ileum. Failure of absorption of vitamin
Damaged surface cells have exfoliated, exposing blood
vessels of the lamina propria. (From Hingson, R., and S. B12 leads to impairment of maturation of
Ito. Gastroenterology 61:156, 1971.) erythrocytes in the bone marrow, and results
640 • THE ESOPHAGUS AND STOMACH

in pernicious anemia. Patients with this disor¬ electron microscopy of normal human esophageal
epithelium. Virchows Archiv. [Cell Pathol.] 26:345,
der show atrophy of the oxyntic ceils in their
1978.
gastric glands and achlorhydria (deficiency Parakkal, P.: An electron microscopic study of the esoph¬
of HC1 secretion). ageal epithelium in the newborn and adult mouse.
The capacity to elaborate a secretion with Am. J. Anat. 727:175, 1967.
a pH ranging from 2 to as low as 0.9 is STOMACH
perhaps the most remarkable property of the Adair, H. M.: Epithelial repair in chronic gastric ulcers.
gastric mucosa. The concentration of hydro¬ Br. J. Exp. Pathol. 59:229, 1978.
gen ions may reach a level 2 million times Bensley, R. R.: The gastric glands. In Cowdry, E. V.,
greater than that of the blood. This is accom¬ ed.: Special Cytology. Vol. 1. 2nd ed. New York,
Paul Hoeber, 1932.
plished by the parietal cells. The precise
Forssmann, W. G., L. Orci, R. Pictet, A. E. Renold, and
mechanism is still a subject of some disagree¬ C. Rouiller: The endocrine cells in the epithelium
ment, but it is the prevailing view that chlo¬ of the gastrointestinal mucosa of the rat. J. Cell
ride ions are actively transported into the Biol. 40:692, 1969.
secretory canaliculus and that this in turn Gannon, B. J., J. Browning, and P. O’Brien: The micro-
vascular architecture of the glandular mucosa of rat
results in passive diffusion of potassium from
stomach, j. Anat. 735:677, 1982.
the cytoplasm into the lumen. Hydrogen and Gannon, B. J., J. Browning, and P. O’Brien: Mucosal
hydroxyl ions are dissociated in the cytoplasm microvascular architecture of the fundus and body
from water, and hydroxyl ions are actively of human stomach. Gastroenterology <56:866, 1984.
Goldstein, A. M. B., M. R. Brothers, and E. A. Davis:
secreted into the canaliculus in exchange for
Architecture of the superficial layer of the gastric
potassium, thus recovering most of the po¬ mucosa. J. Anat. 794:539, 1969.
tassium ions secreted with chloride. Hydro¬ Greider, M. EE, V. Steinberg, and J. E. McGuigan;
gen ions diffuse into the canaliculus, replac¬ Electron microscopic identification of the gastrin
ing the reabsorbed potassium. Water passes cell of the human antral mucosa by means of im-
munocytochemistry. Gastroenterology 63:572,
through the cell along an osmotic gradient
1972.
so that the content of the canaliculus is a Grossman, M. I.: Gastrin and its activities. Nature
solution of hydrochloric acid and potassium 228:1147, 1970.
chloride. In an essential concurrent reaction Helander, H. F.: Ultrastructure of fundus glands of the
in the parietal cell cytoplasm, carbonic an- mouse gastric mucosa. J. Ultrastruct Res. (Suppl.)
4:1, 1962.
hydrase catalyzes the combination of carbon Hingson, D. J., and S. Ito: Effect of aspirin and related
dioxide and water to form carbonic acid, compounds on the fine structure of mouse gastric
which dissociates to hydrogen and bicarbo¬ mucosa. Gastroenterology 67:156, 1971.
nate ions. Bicarbonate ion diffuses from the Hoedemaeker, P. J.,J. Abels, J. J. Wachters, A. Arends,
and N.O. Nieweg: Investigations about the site of
abluminal surface of the cell into the blood.
production of Castle’s gastric intrinsic factor. Lab.
The junctional complexes between the sur¬ Invest. 73:1394, 1964.
face mucosal cells normally provide an effec¬ Ito, S., and E. R. Lacy: Morphology of gastric mucosal
tive barrier preventing acid in the lumen of damage, defenses and restitution in the presence of
the stomach from diffusing back through the luminal ethanol. Gastroenterology <5<5:250, 1985.
Ito, S., and R. J. Winchester: The fine structure of the
epithelium. However, the “alkaline tide” of
gastric mucosa in the bat. J. Cell Biol. 76:541, 1963.
bicarbonate released into the lamina propria Leblond, C. P., and B. E. Walker: Renewal of cell
of the actively secreting gastric mucosa is populations. Physiol. Rev. 36:255, 1956.
believed to be beneficial in neutralizing any Lillibridge, C. B.: The fine structure of normal human
gastric mucosa. Gastroenterology 47:269, 1964.
back-diffusing hydrogen ions. This may be
Lipkin, M., P. Sherlock, and B. Bell: Cell proliferation
especially important when the efficiency of kinetics in the gastrointestinal tract of man. II. Cell
the barrier is greatly reduced by damage to renewal in stomach, ileum, colon, and rectum. Gas¬
the epithelium from imbibition of alcohol or troenterology 47:721, 1963.
other noxious substances. Samloff, I. M.: Pepsinogens, pepsins and pepsin inhibi¬
tors. Gastroenterology 60:586, 1971.
Siten, W., and S. Ito: Mechanisms for rapid epitheliali-
REFERENCES zation of the gastric mucosal surface. Annu. Rev.
Physiol. 47:217, 1985.
ESOPHAGUS Stevens, C. E., and C. P. Leblond: Renewal of the
mucous cells in the gastric mucosa of the rat. Anat.
Edwards, D. A. W. The esophagus. Gut 72:984, 1971. Rec. 775:231, 1953.
Hapwood, D., K. R. Logan, and 1. A. D. Bouchier: The
 26
/NTEST/NES

THE SMALL INTESTINE ileum, is situated in the lower abdomen. Al¬

though there are minor gross and micro¬
scopic differences between the three seg¬
The digestive process initiated in the stom¬
ments, they have the same basic organization.
ach is continued in the small intestine by the
As in other parts of the gastrointestinal
secretions of its intrinsic glands and those of
tract, the wall of the small intestine is made
its accessory glands—the liver and pancreas.
up of four concentric layers—the serosa, the
In the stomach, there is little or no absorption
muscularis, the submucosa, and the mucosa. Of
of nutrient materials released by digestion.
these, the mucosa is the most important in
Absorption is the principal function of the
relation to the digestive and absorptive func¬
small intestine and this takes place mainly in
tions.
its initial segments.
The small intestine is the portion of the
alimentary tract between the stomach and THE INTESTINAL MUCOSA
the large intestine. It is a tubular viscus 4 to
8 m in length grossly divisible into three To augment the efficiency of this physio¬
segments—the duodenum, jejunum, and ileum. logically important layer, there are a number
The duodenum is about 25 cm in length and of structural specializations that serve to in¬
is largely retroperitoneal, being firmly at¬ crease the area of surface exposed to the
tached to the dorsal body wall. The remain¬ lumen. The plicae circulares (valves of Kerck-
der of the small intestine is suspended from ring) are grossly visible crescentic folds that
the dorsal wall by a mesentery and is freely extend half to two thirds of the way around
movable. The proximal part of the free por¬ the lumen (Fig. 26-1). They are permanent
tion of the intestine is the jejunum, which structures involving both the mucosa and the
normally occupies the upper left portion of submucosa (Fig. 26—2). The larger plicae are
the abdominal cavity. The distal portion, the some 8 to 10 mm in height, 3 to 4 mm in

Openings of the intestinal glands Lymphoid nodule in the submucosa

Lamina propria

Muscularis
mucosae
Lymphoid nod¬
ule in the lamina
propria
Figure 26-1. Portion of small in¬ Submucosa
testine; drawn from sections using (iwith vessels)
a binocular microscope, x 17. Circular muscle
(After Braus.)

Longitudinal
muscle

Serosa

Value of
\ Kerchring
Villi
Lymphoid nodule

641
642 • INTESTINES

Figure 26-2. Drawing of a longitudinal section through the wall of the duodenum of an adult human, showing the plicae
circulares (valves of Kerckring), the villi, and submucosal glands of Brunner. (From Bargmann, W. Histologie und
Mikroskopische Anatomie des Menschen. 6th ed. Stuttgart, Georg Thieme Verlag, 1962.)

Figure 26 3. Whole mount of a human jejunal biopsy stained with PAS-Alcian blue, showing intestinal villi of both
finger-like and foliate form, x 37. (Photomicrograph from Pouisen, S. Scand. J. Gastroenterol. 72:235, 1977 )
 INTESTINES • 643

thickness, and up to 5 cm in length. They the fresh condition. Their number varies
are absent from the first portion of the duo¬ from 10 to 40 per square millimeter. They
denum, but begin about 5 cm distal to the are most numerous in the duodenum and
pylorus and reach their greatest development proximal jejunum.
in the last part of the duodenum and proxi¬ The surface of the epithelium is increased
mal portion of the jejunum. From there on¬ not only by villi, but also by invagination to
ward, they gradually diminish in size and form tubular glands between the bases of the
number and are seldom found beyond the villi, called crypts of Lieberkuhn (Fig. 26—5).
middle of the ileum. These are simple tubular glands 320 to 450
A second and more effective means of pm long, which extend downward nearly to
augmenting the surface area of the mucosa the thin layer of smooth muscle comprising
is the presence of enormous numbers of the muscularis mucosae. The spaces between
intestinal villi (Figs. 26—3, 26—4). These are the intestinal glands are occupied by the loose
minute, hngerlike projections of the mucosa connective tissue constituting the lamina pro¬
having a length of 0.5 to 1.5 mm, depending pria.
on the degree of distention of the intestinal
wall and the state of contraction of smooth Intestinal Epithelium
muscle fibers in their own interior. They
cover the entire surface of the mucosa and The free surface of the mucosa is covered
give it a characteristic velvety appearance in by a simple columnar epithelium in which

Figure 26-4. Thin section of monkey jejunal mucosa, showing the long villi and relatively short crypts. Visible in the
core of each villus is the lacteal, a blind-ending lymphatic capillary through which absorbed lipid and other nutrients are
transported to a submucosal lymphatic plexus, and thence via mesenteric lymphatic vessels and the thoracic duct to
the bloodstream. (Courtesy of Madera, J., from Neutra, M. In Weiss, L., and Greep, R. O., eds.: Histology. 5th ed. New
York, Elsevier, 1982.)
644 • INTESTINES

Villus

Crypts of
Figure 26-5. Photomicrograph of a histolog¬
lieberkuhn ical section of the duodenum of a macaque.
The villi, crypts of Lieberkuhn, and Brunner’s
glands are shown. A duct of Brunner’s gland
can be seen penetrating the muscularis mu¬
cosae to empty into one of the crypts. He¬
matoxylin and eosin. x 110.

Muscularis
Mucosae

Brunner's
Glands

three types of cells can be distinguished— In electron micrographs, the striated or

absorptive cells, goblet cells, and endocrine cells brush border of the absorptive cells is made
(argentaffin cells). up of large numbers of closely packed par¬
Absorptive Cells. The absorptive cells are allel microvilli that amplify the surface ex¬
columnar in form and 20 to 26 pm in height, posed to the lumen about 30-fold. The mi¬
with an ovoid nucleus situated in the lower crovilli are 1 to 1.4 pm in length, and about
part of the cell. As seen with the light micro¬ 80 nm in diameter (Figs. 26—7, 26—8). The
scope, the free surface has a prominent plasmalemma enclosing the microvilli has the
striated border (Fig. 26-7) and beneath this usual unit membrane structure but is unusual
is a clear zone usually devoid of organelles in having a nap of delicate branching fila¬
but occupied by the terminal web. This may ments that radiate from its outer leaflet, giv¬
exhibit birefringence when examined with ing the membrane a fuzzy appearance. These
Jthe polarizing microscope and it can be selec¬ filaments are longer and more numerous at
tively stained by a method that employs the tips of the villi than on their sides (Fig.
tannic acid, phosphomolybdic acid, and the 26—9). The intermingling of the filamentous
dye amido black. The cytoplasm below the excrescences of the microvillus tips forms a
terminal web is rich in mitochondria and continuous surface coat on the striated border
there is a well-developed supranuclear Golgi that varies from 0.1 to 0.5 pm in thickness,
apparatus. depending on the species (Fig. 26—8). This
 INTESTINES • 645

Lymphocytes Lacteal Fibroblast

cells border cej|s Wandering

cells
Figure 26-6. Longitudinal section of a villus from cat jejunum. Hematoxylin and eosin. x 600.

Figure 26—7. Electron micrograph of portions of two intestinal epithelial cells of the hamster, showing the striated
border, the terminal web, and the junctional complex, which corresponds to the terminal bar seen with the light
microscope, x 25,000. (Courtesy of E. Strauss.)
646 • INTESTINES

Figure 26-8. The plasma membrane on the microvilli of the intestinal brush border has a prominent surface coat
consisting of polysaccharide chains of integral glycoproteins. (Micrograph courtesy of S. Ito.)

surface coat is well developed in the human. border and its surface coat, which were not
It is regarded as an integral part of the cell resolved with the light microscope, have now
surface. It is glycoprotein in nature and is been shown to be of great physiological im¬
extremely resistant to both proteolytic and portance in the digestive and absorptive func¬
mucolytic agents. It is believed to have a tion of the small intestine.
protective role, but it is likely that it also In the interior of each microvillus is a
participates actively in the digestive process. bundle of thin straight filaments that run
Intraluminal digestive enzymes such as pan¬ longitudinally in an otherwise homogeneous
creatic amylase and proteases are believed to fine-textured cytoplasmic matrix (Fig. 26—
be adsorbed onto the very large surface pre¬ 10). The filaments extend downward from
sented by the filaments of the surface coat. the microvilli into the terminal web, which is
Thus, some of the digestive processes previ¬ resolved in electron micrographs as a felt-
ously thought to occur only in the lumen may work of exceedingly fine filaments oriented
also take place on the microvillous surface. (Fig. 26—11), for the most part, parallel to
Isolated intestinal striated borders hydrolyze the brush border. At the sides of the cell, the
disaccharides and polypeptides to monosac¬ filaments of the terminal web merge with
charides and amino acids. Therefore, the those associated with the zonula adherens.
disaccharidases and peptidases involved in The filaments in the core of the microvilli
the terminal digestion of carbohydrates and have been shown to form arrowhead com¬
proteins are believed to be localized in the plexes with heavy meromyosin and are there¬
membrane of the microvilli. In addition to fore identified as actin. There are about 20
its enzymatic activities, it has been suggested actin filaments in each microvillus of the
that the surface coat of the jejunum may brush border, and these filament bundles,
have specific receptors or binding sites for together with the intermediate filaments,
substances that are selectively absorbed in myosin, and associated proteins of the ter¬
this portion of the intestine and not in other minal web, form one of the most highly
regions. Thus, the membrane of the brush organized cytoskeletal apparatuses thus far
 INTESTINES • 647

ular endoplasmic reticulum are also found in

the supranuclear cytoplasm but they are
more abundant toward the base. The Golgi
complex has the usual arrays of parallel cis-
ternae and associated vesicles. It shows no
morphological evidences of activity except
during lipid absorption.
The lateral surfaces of the absorptive cells
are in close apposition in the upper part of
the epithelium and may have occasional slen¬
der interdigitating processes. In phases of
active absorption the lateral cell membranes
of neighboring cells diverge toward the base
of the epithelium, and chylomicra may accu¬
mulate in these widened intercellular clefts.
A juxtaluminal junctional complex bars ac¬
cess to the intercellular cleft from the lumen.
The presence of occluding junctions com¬
pletely encircling each cell near the lumen
ensures that material being absorbed from
the lumen must traverse the plasma mem¬
brane of the brush border, the apical cyto¬
plasm, and the lateral cell membrane to gain
access to the intercellular spaces of the epi¬
thelium.
Figure 26-9. Electron micrograph of the tips of several
microvilli from cat ileum, showing the branching protein-
polysaccharide (mucopolysaccharide) filaments attached
at one end to the outer leaflet of the unit membrane (at
arrows), x 120,000. (Courtesy of S. Ito, after Fawcett, D.
W. J. Histochem. Cytochem. 73:75, 1965.)

described. Until recently it was thought that

the actin filaments of the microvillous cores
interacted with myosin in the terminal web,
resulting in shortening of the microvilli. It is
generally agreed that isolated brush borders
contract in vitro in the presence of Ca+ + ions
and ATP; it is now believed, however, that
lateral contraction of the terminal web in¬
creases the convexity of the cell apex and
tends to spread the tips of the microvilli
apart, but the microvilli do not shorten. What
role, if any, this contractility of the cytoskel-
eton plays in intestinal absorption is not
known.
Below the terminal web, the cytoplasm con¬
tains occasional lysosomes and numerous
elongate mitochondria of orthodox internal
structure. Branching profiles of smooth en¬
doplasmic reticulum are plentiful in this re¬
gion. The membranes of this organelle con¬
tain enzymes essential for synthesis of Figure 26-10. Electron micrograph of cross sections of
triglycerides from fatty acids and monogly¬ microvilli from the brush border of an intestinal epithelial
cell. The bundle of actin filaments in the core of the
cerides. The smooth endoplasmic reticulum microvillus is clearly shown. Prepared by quick freezing
therefore plays an important role in intestinal in liquid helium and freeze substitution. (Micrograph cour¬
absorption of fat. Cisternal profiles of gran¬ tesy of A. Ichikawa.)
648 • INTESTINES

H0

Ml

jaHK*:

Figure 26-11. Brush border of an intestinal absorptive cell prepared by quick-freezing, deep-etching^and rotary
shadowing. Bundles of actin filaments can be seen extending from the cores of the microvilli downward into the
meshwork of filaments making up the terminal web. (Micrograph courtesy of N. Hirokawa and J. Heuser.)

The plasma membrane at the cell base is cytoplasm around the theca. A highly devel¬
closely applied to a continuous basal lamina oped Golgi complex is situated between the
that extends across the intercellular spaces at nucleus and the mucigen droplets in the
the base of the epithelium. Absorbed material theca. The individual droplets appear to orig¬
accumulating between cells must traverse this inate in the Golgi complex and move up into
barrier to reach the capillaries and lymphatics the theca. Each is enveloped by a delicate
of the intestinal villus. membrane, which is often disrupted in prep¬
Goblet Cell. The goblet cells are irregularly aration of the specimen. The basal and lateral
scattered among the absorptive cells (Fig. 26- plasma membranes are smooth-contoured
6). They are described in some detail in except for a few lateral interdigitations. The
Chapter 3, as examples of unicellular glands. goblet cells are attached to the neighboring
Their name derives from their fancied re¬ absorptive cells by juxtaluminal junctional
semblance to a wine glass. Their apical re¬ complexes. Sparse microvilli may be present
gion, called the theca, is distended with mu- on the free surface. Their length and number
cigen droplets, while the base of the cell is is influenced by the degree of distention of
relatively free of secretory material and the theca with mucigen. The tendency of
forms a slender stem or stalk. The nucleus mucigen droplets to swell in specimen prep¬
tends to be flattened and the surrounding aration has made it difficult to study the
cytoplasm strongly basophilic. The organelles mechanism of their release, but the mem¬
are difficult to study with the light micro¬ branes of the droplets appear to fuse with
scope in the mature goblet cell because of each other and with the plasmalemma, per¬
their close crowding. mitting the mucus to flow out while maintain¬
In electron micrographs, cisternae of gran¬ ing the integrity of the cell surface.
ular endoplasmic reticulum are arranged The secretory product of the goblet cell,
more or less parallel to the base and the mucus, is a viscous material with the consis¬
lateral surfaces of the cell. A few cisternae tency of raw egg white. It serves to lubricate
may continue upward into the thin layer of and protect the surface of the epithelium.
 INTESTINES • 649

Chemical analyses indicate that goblet cell many years, they were also called argentaffin
mucus is a very large glycoprotein of molec¬ cells. It was subsequently found that if tissue
ular weight 2 x 106, which is acidic owing to sections were treated with a reducing agent
the presence on the molecule of both termi¬ before being exposed to silver nitrate, a
nal sialic acid residues and sulfate esters such greater number of these basal granular cells
as A-acetylglucosamine-6-sulfate. It stains could be demonstrated. Cells stainable by this
brilliantly with the periodic acid-Schiff (PAS) method were termed argyrophilic cells. The
reaction for carbohydrates, with Alcian blue, differences in number of argyrophil and ar¬
toluidine blue, and other thiazine dyes that gentaffin cells, and the fact that some gran¬
stain acidic glycoproteins. There appear to ulated cells exhibited fluorescence in ultravi¬
be minor qualitative differences between the olet light while others did not, gradually led
mucus in the crypts and that on the villi, and to the realization that not all the enteroen¬
also differences along the length of the intes¬ docrine cells were the same. Although they
tine. The content of acidic carbohydrate side have a number of morphological features in
chains is generally higher in the goblet cells common, they have proved to be a highly
of the colon than in those of the small intes¬ heterogeneous cell population. In recent
tine. years, correlation of electron microscopic ob¬
The synthetic activities of goblet cells in¬ servations with immunocytochemical staining
clude the synthesis of protein from amino has produced a surge of new information
acid precursors; the formation of polysaccha¬ about their secretory products and their
rides from mono- and disaccharides; the link¬ probable function. More than 16 different
ing of oligo- and polysaccharides to proteins; endocrine cell types have now been described
and the incorporation of sulfate into acid in the gastrointestinal tract (Fig. 26-12).
polysaccharides. Autoradiographic studies For students to learn the identifying char¬
show that protein synthesis is associated with acteristics of all the enteroendocrine cell
the abundant granular endoplasmic reticu¬ types would be beyond the call of duty, but
lum at the cell base, whereas synthesis of they should be familiar with the structural
polysaccharide and its sulfation take place in features these cells have in common and
the Golgi complex. should be aware of the range of hormones
Enteroendocrine Cells (Basal Granular secreted by the enteroendocrine cells as a
Cells). More than a century ago, Heidenhain group.
(1870) described small cells near the base of At the light microscope level, they are
the gastrointestinal epithelium that possessed ovoid or pyramidal in the stomach and intes¬
staining reactions similar to those of the chro¬ tinal crypts, and more columnar on the epi¬
maffin cells in the adrenal medulla. These thelium of the villi. The bulk of the cell body
later came to be called the enterochromaffin is in the lower half of the epithelium, but a
cells. Their basal location in the epithelium narrow apical region usually extends to the
and the presence of secretory granules con¬ lumen and has a brush border. The nucleus
centrated between the nucleus and the basal is round and generally poor in heterochro¬
lamina suggested that they were endocrine matin. The cytoplasm is pale in relation to
cells liberating their secretion into the lamina the surrounding cells. The secretory granules
propria, rather than into the intestinal lu¬ vary in size in the different cell types and are
men. concentrated in the basal cytoplasm.
They occur singly and are widely scattered In electron micrographs, the microvilli of
throughout the epithelial lining of the gas¬ the enteroendocrine cells are often longer
trointestinal tract. They are present in mod¬ and thicker than those of adjacent absorptive
erate numbers in the stomach, are common cells—a finding that has suggested their pos¬
in the duodenum, and are more sparse in sible function as chemoreceptors. The cyto¬
the jejunum and ileum, where they occur plasm is relatively electron lucent. The cyto¬
both on the villi and in the crypts. They are plasmic organelles do not differ significantly
present in limited numbers throughout the in size or form compared with those of other
colon and are also found in the biliary tract epithelial cells. The rough endoplasmic retic¬
and the ducts of the pancreas. ulum varies somewhat in amount from one
In addition to their ability to bind alkaline cell type to another, but in no case is it highly
bichromates (chromaffinity), many of these developed. A few lysosomes are usually pres¬
cells also precipitate silver salts in the absence ent. Morphological identification of the sev¬
of a reducing agent (argentaffinity). For eral types of enteroendocrine cells depends
650 • INTESTINES

CELL SECRETION [ LOCALIZATION PRODUCT

TYPE GRANULES | Pancreas Stomach Intestines

1 Islets Glucagon, Glicentin

A 250 nm

350 Islets
B ®<#
Insulin

—r——”— Jejunum
0 W*^ Fundic
D 350 Islets 1 leum Somatostatin
Pyloric Colon
Fundic Jejunum
Dl 160 Islets Pyloric Ileum Unknown
Colon
Jejunum
Fundic Serotonin
EC 300 Islets ileum
Pyloric Various peptides
%5f Colon

Fundic Histamine
ECL 450

G 300 Pyloric I Duodenum 1 Gastrin

% • ©•
I Jejunum Cholecystokinin
1 250
1 Ileum
•v«
Jejunum I Gastric inhibitory
350
Ileum I peptide

Jejunum
Glucagon- like
400 Ileum
immunoreactivity
Colon

Jejunum
Mo I leum
Motilin

300 I leum Neurotensin

N
• #•
Fundic
•••
• •• •••
••
120
Pyloric
Jejunum I Unknown

pp few.
© © q© O
180 Islets Fundic
Pyloric
Colon
Pancreatic
polypeptide

Jejunum
200 Secretin
I leum

C-terminal gastrin
Jejunum
TG immunoreactivity

Fundic
300 Unknown
Pyloric

Figure 26-12. Summary of the enteroendocrine cell types thus far described, including their nomenclature, their
distribution, the ultrastructure of their granules, and their amine or peptide products. (Modified after Grube, H., and G.
Forssmann. Horm. Metab. Res. 11:603, 1979.)
 INTESTINES • 651

mainly on differences in size, shape, electron The enteroendocrine cells have thus been
density, and substructure of their secretory shown to be the source of the classical gas¬
granules complemented by immunocyto- trointestinal hormones long studied by
chemical staining of the granules with fluo¬ physiologists (gastrin, secretin, and chole-
rescein-labeled antibodies specific for their cystokinin-pancreozymin), and several other
amine or polypeptide products. biologically active peptides and monoamines
A majority of the cells traditionally de¬ with less well-established functions. Rather
scribed as argentaffin cells by histologists are puzzling is the fact that several polypeptide
now called EC cells. They are found in the hormones found in the endocrine cells of the
fundic and antral regions of the stomach and gut (gastrin, cholecystokinin-pancreozymin,
in the intestinal epithelium of all species so vasoactive intestinal peptide, and motilin)
far studied. Their cytoplasmic granules have have also been localized in certain cells of the
elongated cores of high electron density. In central nervous system. And conversely,
general, those in the stomach are smaller some polypeptide hormones originally iso¬
(200 nm) than those in the intestine (300- lated from the central nervous system (sub¬
350 nm). They synthesize and secrete sero¬ stance P, somatostatin, neurotensin) have also
tonin (5-hydroxytryptamine). been found to occur in enteric or pancreatic
A second category of enteroendocrine cells endocrine cells. The physiological or onto¬
found in the fundus and antrum of the genetic significance of such a diffuse distri¬
stomach is the D cell, so named because of its bution of isolated endocrine cells in organ
resemblance to the delta cell of the endocrine systems of such disparate function awaits
pancreas. Its round granules (350 nm) have clarification.
a core of finely granular texture and medium The majority of enteroendocrine cell types
electron density. They secrete somatostatin. A can take up and decarboxylate precursors of
subtype of similar distribution, designated biogenic monoamines and therefore are in¬
Dj, has smaller granules (160 nm) and is cluded as members of the so-called APUD
believed to produce vasoactive intestinal poly¬ cell series. They are also components of the
peptide. diffuse neuroendocrine system (see Chapter
In the human duodenal mucosa and in the 3).
stomach of other species are the A cells, so
called because of their resemblance to the Crypts of Lieberkuhn
alpha cells of the pancreatic islets. Their
granules (250 nm) have a core of high elec¬ The epithelium covering the villi continues
tron density surrounded by a narrow clear into the intestinal glands or crypts of Lieber¬
space beneath their limiting membrane. They kuhn (Fig. 26—5). The upper halves of the
are the source of enteric glucagon. walls of the crypts are lined with low colum¬
Occurring in large numbers in the gastric nar epithelium containing absorptive cells,
antrum of all species studied are the G cells. goblet cells, and a few basal granular cells.
Their granules (200-250 nm) are round or In the lower halves of the crypts the cells are
slightly angular with cores of variable density less clearly differentiated and there are nu¬
and heterogeneous texture. They secrete the merous mitoses. It is here that new cells are
hormone gastrin. formed to continually replace those that are
The L cells, generally confined to the intes¬ exfoliated at the tips of the villi. If tritiated
tinal mucosa, have quite large secretory gran¬ thymidine is given to an animal, it can be
ules (400 nm) with round or slightly angular shown by biopsies at later time intervals that
cores of high electron density closely sur¬ the label incorporated into nuclei of dividing
rounded by the limiting membrane. Their cells in the bottom of the crypts of Lieber-
product is still a subject of debate. kiihn gradually moves upward onto the villi.
5 cells are restricted to the intestinal mu¬ About a day after administration of thymi¬
cosa. Their granules are among the smallest dine the labeled cells are on the sides of the
found in enteroendocrine cells (200 nm) with villi, and by the fifth day they are being
round cores of medium to high electron exfoliated at the villus tips. Thus, billions of
density. There is general agreement that they cells are being shed every day from the hu¬
produce the hormone secretin. I cells occur in man gastrointestinal tract and are being re¬
the duodenal mucosa and have granules (250 placed by upward displacement of cells from
nm) intermediate in size between the L and localized regions of cell proliferation. In the
S cells. They are probably the source of small intestine, the proliferative activity is
cholecystokinin. confined to the crypts of Lieberkuhn.
652 • INTESTINES

Discovery of this rapid turnover of cells in in both longitudinal and transverse sections
the lining of the intestine has altered our of the glands. At early time intervals after
interpretations of the physiology of some of administration of tritiated thymidine, labeled
the cell types. For example, it was formerly fibroblasts are found in autoradiographs
thought that goblet cells might accumulate around the lower third of the crypts. At
secretions, discharge, and then refill, and that longer time intervals, these labeled cells are
this cycle was repeated many times. It is now localized around the necks of the glands. If
realized that the life span of intestinal goblet the changing distribution of the labeled epi¬
cells is only three or four days—the time thelial cells and labeled pericryptal cells are
taken for them to differentiate in the crypts, compared, it is evident that the two cell types
move up onto the villi, and be exfoliated at migrate in synchrony. Electron micrographs
the tip. Thus, it is probable that goblet cells show that there is a progressive increase in
secrete continuously and normally pass degree of cytological differentiation of the
through only one secretory cycle. This con¬ fibroblasts as they move upward. The contin¬
sists of an initial phase in the crypts when uous renewal and upward migration of peri¬
rate of synthesis of mucus exceeds discharge cryptal cells constitutes an exception to the
and mucin accumulates; an intermediate generalization that fibroblasts in the adult are
phase when the cells are in the upper part a relatively stable .population proliferating
of the crypts and lower half of the villi, only in response to injury.
synthesis and discharge are approximately in Paneth Cell. In addition to the cell types
equilibrium, and the cells appear engorged already described, there is a type of cell
with mucus; and a final phase when rate of occurring in small groups only in the depths
discharge exceeds synthesis as the cells ap¬ of the crypts of Lieberkiihn—the Paneth cells
proach the tips of the villi and appear de¬ (Fig. 26—13). They are pyramidal in form,
pleted. Although this is the normal course of with a round or oval nucleus situated near
events, it can be modified by irritants that the base, and conspicuous secretory granules
cause expulsion of mucus. If mustard oil is in the apical cytoplasm. They have the cyto¬
administered locally to experimental animals, logical characteristics of cells actively secret¬
the goblet cells expel nearly all of their mucus ing protein. The cytoplasm at the base is
stores. Under these conditions, they then are basophilic, and in electron micrographs con¬
found to initiate a new cycle of accumulation. tains parallel arrays of cisternae of rough
The discovery of the continual upward
migration of epithelial cells immediately
posed a number of puzzling morphogenetic
problems. Do the cells move with respect to
a relatively stationary basal lamina, or does
the epithelium as a whole move with respect
to the underlying lamina propria? The an¬
swers to these questions remain in doubt, but
valuable new insight into the problem has
been gained from autoradiographic studies
that reveal that there is also a continuous
renewal and upward migration of the peri-
cryptal fibroblasts. Much of the relevant ex¬
perimental work has been carried out on the
crypts of the colon but the findings are be¬
lieved to apply also to those of the small
intestine and other parts of the alimentary
mucosa.
The crypts are surrounded by a sheath of
flattened fibroblasts closely applied to the
base of the epithelium and by a highly or¬
dered reticulum laid down with the majority
of its small bundles of unit fibers of collagen
arranged circumferentially around the crypt.
Figure 26-13. Drawing of a crypt of Lieberkuhn, illustrat¬
The pericryptal fibroblasts are flattened squa¬
ing the Paneth cells at the base of the crypt. Higher up in
mous elements that present fusiform profiles the crypt are four argentaffin cells. Hematoxylin and eosin.
 INTESTINES • 653

endoplasmic redculum. The lumen of the digestion of the chitinous exoskeleton of the
cisternae not infrequently has a dense con¬ ingested ants, but there is no solid evidence
tent, which may occasionally be precipitated for this.
in crystalline form. The Golgi complex is Paneth cells evidently secrete continuously,
prominent, as in other active glandular cells, but the rate of secretion is enhanced by
and often contains the formative stages of feeding or by administration of pilocarpine.
secretory granules. The granules are homo¬ Despite decades of study, the functional role
geneous in density in humans and in a num¬ of Paneth cells is still not known. Research
ber of other species. In the mouse, they have on the chemical nature of the secretory prod¬
a dense core and lighter peripheral zone (Fig. uct of Paneth cells has been hampered by
26-14). The periphery consists of an acid inability to obtain their secretion free of con¬
glycoprotein, whereas the core appears to be tamination by constituents of other cell types.
a neutral glycoprotein. The granules usually Immunocytochemical studies have revealed
stain with acid dyes such as eosin or orange the presence of lysozyme in their well-devel¬
G. oped phagolysosomal system. Lysozyme is a
Paneth cells are a stable population with highly charged cationic protein capable of
little or no turnover. They do not incorporate digesting the cell walls of certain bacteria.
tritiated thymidine and they are not observed Under some circumstances Paneth cells have
in mitosis. They are abundant in humans, been observed to phagocytose and digest cer¬
monkeys, mice, rats, guinea pigs, and rumi¬ tain intestinal flagellates and spirilliform mi¬
nants, but are absent from the intestines of croorganisms commonly found in the crypts
dogs, cats, pigs, and raccoons. They are ex¬ of Lieberkiihn of rats. This has led to the
traordinarily abundant in the intestinal suggestion that they may play a role in reg¬
glands of the South American anteater—a ulating the microbial flora of the intestinal
finding that encouraged the speculation that glands. This leaves unexplained their obvious
in this species they might be involved in structural specialization as secretory cells, and

Figure 26-14. Electron micrograph of an area of cytoplasm from a Paneth cell, showing the abundance of granular
reticulum and the heterogeneity of the secretory granules. (Micrograph from Staley, M., and J. Trier. Am. J. Anat.
117:365, 1965.)
654 • INTESTINES

it is clear that the principal function of the Lamina Propria

Paneth cells is yet to be discovered.
The lamina propria of the intestinal mucosa
consists of loose areolar tissue that occupies
Cell Turnover and Renewal the spaces between the crypts of Lieberkuhn
and forms the cores of the intestinal villi
The epithelial lining of the intestinal tract
(Figs. 26-4, 26-5). It contains many free cells
is continuously being renewed by prolifera¬
enmeshed in a stroma of argyrophilic fibers.
tion of undifferentiated cells in the crypts,
Associated with the fibers are fixed cells re¬
migration upward onto the villi, and exfol¬
sembling the reticular cells of lymphoid or¬
iation of effete or dying cells at the villus tips.
gans. The fine collagen fibrils of the stroma
This whole process of cell renewal is referred
are more concentrated adjacent to the basal
to as the cell turnover of the epithelium, and
lamina of the epithelium. In addition there
its duration is the cell turnover time. The mu¬
are delicate networks of elastic fibers sur¬
cosa of the jejunum has the fastest rate of
rounding the crypts and extending into the
turnover of any tissue in the body.
villi. Thin strands of smooth muscle extend
The cells at the base of the crypts of Lie-
from the muscularis mucosae into the core
berkuhn have high activities of the enzymes
of each villus where they are oriented parallel
involved in nucleic acid synthesis and do not
to the central lacteal—a minute terminal
have the long interphase (G0) characteristic
branch of the submucous lymphatic plexus
of more slowly proliferating tissues. Synthesis
that ends blindly near the tip of each villus
of DNA occurs in a period of six to 11 hours
(S). The premitotic (G2) and postmitotic (GJ and provides an important channel for the
transport of absorbed lipid. Periodic contrac¬
stages of the mitotic cycle are brief and the
mitotic phase (M) takes about one hour. The tions of the smooth muscle in the villus core
complete cell cycle occupies 10 to 17 hours empty the lacteal, and propel lymph and
in rodents and about 24 hours in humans. absorbed nutrients toward the mesenteric
The intestinal epithelium is completely re¬ lymph nodes and thoracic duct.
placed in two to three days in laboratory Large numbers of lymphocytes, plasma
rodents and three to six days in humans. cells, eosinophils, and macrophages are
In the colon, villi are lacking but the pat¬ found in the lamina propria. The most nu¬
tern of renewal is similar. The proliferative merous are the lymphocytes, a reserve of
zone in the crypts is somewhat more exten¬ immunocompetent cells of which many are
sive, and cells are extruded at the mucosal capable of differentiating into antibody-pro¬
surface between crypts. Cell division and mi¬ ducing plasma cells. Lymphocytes also pene¬
gration are slightly slower, resulting in a trate into the intercellular clefts of the epi¬
turnover time of four to eight days in hu¬ thelium. The concentration of intraepithelial
mans. lymphocytes increases along the intestinal
The intestine has a remarkable capacity to tract and reaches its highest in the large
adapt to changing conditions of alimentation intestine. It was formerly thought that lym¬
and to surgical removal of large segments. phocytes by the millions migrated through
Starvation or protein deficiency results in the epithelium into the lumen and were lost
atrophy of both muscular and mucosal com¬ in the elimination of the intestinal contents.
ponents of the small intestine. The mitotic Radioautographic studies with tritiated thy¬
cycle in the crypts is prolonged and migration midine provide no support for this concept.
is slowed. These changes are reversed by Over 95 per cent of the labeled lymphocytes
feeding. Food intake above normal levels are located in the basal third of the epithe¬
results in hypertrophy of the villi and in¬ lium and there is no convincing evidence that
creased absorption of nutrients. After sur¬ they enter the lumen.
gical excision of a segment of bowel, there is
a compensatory increase in the height of the
Lymphoid Nodules
villi and depth of the crypts in the remaining
small intestine. This is accompanied by In addition to the large population of wan¬
growth in the muscularis and in the length dering lymphocytes, the lamina propria of
of the bowel. The hyperplastic response is the small intestine contains numerous solitary
proportional to the length of intestine re¬ lymphoid nodules, varying in diameter from
sected. Much less is known about the capacity 0.4 to 3 mm (Fig. 26—15). These dense ag¬
of the colon to adapt. gregations of lymphocytes are scattered all
 INTESTINES • 655

Figure 26-15. Photomicrograph of cat ileum, illustrating intestinal villi. Crypts of Lieberkuhn and submucosal lymphoid
nodules, or Peyer’s patches, are shown.

along the small intestine but are more abun¬ ment of the mesentery and are recognizable
dant and larger in its distal part. In the ileum, grossly as elongated thickened areas. Their
they may be found in the plicae circulares or long diameter is 12 to 20 mm, and the short
between them. The smaller nodules occupy 8 to 12 mm. In sections, they consist of dense
only that portion of the mucosa above the lymphatic tissue with large germinal centers
muscularis mucosae. The larger ones occupy in their interior. Their periphery is demar¬
the whole thickness of the mucosa, bulging cated by a thin layer of reticular fibers. In
at the surface and extending down through the vicinity of aggregated nodules, the lamina
the muscularis mucosae into the submucosa. propria and submucosa are always heavily
They are visible to the naked eye as bulging infiltrated with lymphocytes. In old age, the
round or oval areas devoid of overlying villi. solitary nodules and Peyer’s patches undergo
In sections, crypts are also lacking in the considerable involution.
attenuated epithelium overlying the nodules.
With scanning electron microscopy, a distinc¬ Secretory Immune System
tive cell type (M cell) has been identified in of the Intestine
the epithelium over large lymphoid nodules.
These cells stretched over the dome of the We swallow thousands of strains of bacteria
nodule are squamous, and their microvilli and viruses. Indeed, a large fraction of the
are coarser and more widely separated than intestinal contents consists of bacteria. For¬
those in the brush border of the neighboring tunately, only a small percentage of those
absorptive cells (Figs. 26—16, 26-17). taken in are pathogenic. The mucosal surface
Groups of nodules massed together are of the intestinal tract nevertheless presents
called aggregated nodules or Peyer’s patches. As an enormous area to be protected against
a rule they occur in the ileum, but occasion¬ invasion. A very important component of its
ally they may be encountered elsewhere. defense is the so-called secretory immune system,
From 30 to 40 can usually be found scattered which produces a special class of antibodies
along the ileum. They are always located on that restrain bacterial proliferation, neutral¬
the intestinal wall opposite the line of attach¬ ize viruses, and prevent penetration of enter-
656 • INTESTINES

Figure 26-16. Inset presents for orientation a low-power scanning micrograph of a lymphoid nodule in rat intestine
covered by an attenuated layer of epithelium and surrounded by villi. The area in the small square is shown at higher
magnification in the main figure. The polygonal outlines of the flattened cells is evident and certain of them have a
surface texture different from the majority (at arrows). This difference is seen more clearly in Figure 26-17 (a higher
magnification of area enclosed in rectangle). (Micrograph from Owen, R., and A. Jones. Gastroenterology 66:189,
1974.)

otoxins through the epithelium. The synthe¬ juxtaposition of macrophages to lymphocytes

sis of these antibodies and their delivery to and plasma cells in the lamina propria. Mac¬
the surface of the mucosa involves the coop¬ rophages are now believed to process anti¬
eration of the lymphocytes, plasma cells, and gens in some way that increases their effec¬
macrophages of the lamina propria with the tiveness in inducing an immune response
epithelium. (see Chapter 13). Weak antigens may be
As noted in earlier chapters, there are five hundreds of times more immunogenic when
or more classes of immune globulins in the associated with macrophages than they are in
human of which the most abundant and best the free state. Although macrophages may
understood is IgG, the antibody mediating phagocytize and destroy antigen, it is postu¬
general humoral immunity. A second impor¬ lated that some persists on their surface and
tant class is IgA, the so-called secretory immu¬ is liberated and presented to neighboring
noglobulin found in the serum and in the immunocompetent lymphocytes in a more
secretions of the parotid, submandibular, effective form. Thus, macrophages actively
mammary, and lacrimal glands and the tra¬ promote the immune response by priming
cheobronchial glands, as well as those of the the host for a secondary reaction to the
gastrointestinal tract. When the cells of the antigen.
lamina propria are surveyed with fluorescein- Antigens from the intestinal lumen that
labeled antisera specifically reacting with IgG cross the mucosal barrier interact with cells
and with IgA, the great majority of the in the lymphoid nodules of the lamina pro¬
plasma cells are found to be producing IgA. pria, which are precommitted to IgA pro¬
The mean population density of IgA reactive duction. The M cells overlying lymphoid
plasma cells is reported to be 180,000 per nodules are believed to be specialized for
mm3 as against 18,000 for IgG. uptake and transcellular movement of anti¬
Attention has been drawn to the frequent gen. When the underlying lymphoblasts have
 INTESTINES • 657

Figure 26-17. Scanning micrograph of the area shown in Figure 26-16. Two of the cells bear loosely packed microvilli
that are very much broader than those composing the brush border of the surrounding cells. (Micrograph from Owen,
R., and A. Jones. Gastroenterology 66:189, 1974.)

interacted with the antigen, they migrate to situated to interact with antigens, enterotox-
the mesenteric lymph nodes, and after fur¬ ins, and bacteria to prevent their attachment
ther maturation there, they enter the sys¬ to the cell membrane—an effect termed im¬
temic circulation. From the circulation, they mune exclusion.
“home” back to the intestine and become The mystery that long enshrouded the
widely distributed as free cells in the lamina function of the free cells and aggregations of
propria (Fig. 26—18). There they differen¬ lymphoid tissue in the lamina propria has
tiate into plasma cells and produce specific now been largely stripped away by advances
IgA antibody to the absorbed antigen. The in immunology, which have provided com¬
IgA that they produce is transported through pelling evidence that they constitute an effi¬
the overlying epithelium bound to a special cient local immune system with the primary
glycoprotein carrier called the secretory com¬ function of supporting the epithelium in its
ponent. The secretory component contributed role as a barrier to the penetration of path¬
by the epithelium not only serves as a carrier, ogenic organisms and toxins from the exter¬
but is believed to protect the IgA against the nal environment.
cells’ lysosomal system and against subse¬ It is interesting to note that in the contin¬
quent destruction by other enzymes on the uing “arms race” between microbes and
intestinal surface. This remarkable coopera¬ higher organisms, a few species of bacteria,
tion of IgA-producing plasma cells with epi¬ including Neisseria gonorrhoeae, have evolved
thelial cells producing secretory component as a counter weapon, IgA proteases—enzymes
appears to have evolved to ensure formation that readily cleave IgA immunoglobulin. No
of antibody molecules capable of coexisting other substrate for their activity has yet been
with the proteolytic enzymes in the intestinal identified. These proteases may play an im¬
lumen. The secretory antibodies released at portant part in the pathogenesis of certain
the free surface are retained in the glycocalyx human diseases by interfering with the im¬
of the epithelium where they are strategically munological defenses mediated by IgA.
658 • INTESTINES

Duodenal Glands (Submucosal

Glands of Brunner)
Brunner’s glands are usually encountered
first in the region of the pyloric sphincter,
but in the human they may extend a few
centimeters into the pyloric antrum. They
diminish in size and number along the duo¬
denum and usually terminate in its distal
third. In rare instances they may extend into
the proximal part of the jejunum. Brunner’s
glands tend to occupy the submucosa in the
plicae circulares and are separated from one
another by short, gland-free intervals (Figs.
26-2, 26-19).
The terminal secretory portions of the
glands consist of richly branched and coiled
tubules arranged in lobules 0.5 to 1.0 mm in
diameter (Fig. 26-5). The ducts ascend
through the muscuiaris mucosae to open into
a crypt of Lieberkuhn.
Examined with the electron microscope,
the secretory cells of the submucosal glands
share the ultrastructural features of zymo¬
genic and mucus-secreting cells. They have
numerous mitochondria and abundant gran¬
Figure 26-18. Schematic representation of the pathways
ular reticulum at the cell base. Their dense
involved in the secretory immune system. Lymphocytes secretory granules bear a superficial resem¬
in Peyer’s patches exposed to antigens from the gut blance to those of pancreatic zymogen cells.
lumen migrate to regional lymph nodes and thence to the The Golgi complex is unusually large and is
bloodstream. Circulating in the blood they home into the
believed to be the site of synthesis of the
lamina propria of the gut and there develop into plasma
cells secreting IgA, which acquires a secretory piece in carbohydrate moiety of the secretory product
the epithelium and is released into the gut lumen. (Re¬ and of its conjugation with the protein moiety
drawn after Walker, W. A., and K. J. Isselbacher. N. Engl. synthesized in the granular endoplasmic re¬
J. Med. 297:767, 1977.) ticulum.
The secretion is a clear, viscous, and dis¬
tinctly alkaline fluid (pH 8.2 to 9.3). Its prin¬
Muscuiaris Mucosae cipal function is to protect the duodenal
The muscuiaris mucosae averages 38 [xm mucosa against the erosive effects of the acid
in thickness, and consists of networks of elas¬ gastric juice. Its mucoid nature, its alkalinity,
tic fibers and thin inner circular and outer and possibly the buffering capacity of its
longitudinal layers of smooth muscle. Its con¬ bicarbonate content make it well suited to
traction increases the height of folds in the this role.
mucosa. It is usually contracted in fixed spec¬ Another possible function has recently
imens exaggerating the irregularity in the been suggested. Urogastrone, a polypeptide
outline of the lumen. It is conceivable that excreted in human urine, is a powerful in¬
such changes in the surface topography of hibitor of gastric acid secretion, and also
the mucosa occurring in vivo may play an stimulates epithelial proliferation. The cells
ancillary role in mixing the content of the of origin, storage, and secretion of urogas¬
gut. trone have long eluded detection. Applying
fluorescein-labeled antiurogastrone antibody
to a large variety of human tissues, specific
Submucosa
staining was observed only in the cells of
The submucosa consists of moderately Brunner’s glands and in the duct cells of the
dense connective tissue rich in elastic fibers. submandibular salivary gland. It is now spec¬
It may also contain occasional clusters of ulated that in addition to their other secre¬
adipose cells. In the duodenum, but not else¬ tory functions, Brunner’s glands may release
where, it is largely occupied by the glands of urogastrone into the intestinal lumen, where
B runner. its growth-stimulating properties may play an
 INTESTINES • 659

Figure 26-19. Photomicrograph of duodenum from a man who had committed suicide by drinking formalin. The mucosa
is unusually well preserved; the muscularis shows considerable shrinkage, x 20. (Courtesy of H. Mizoguchi.)

important role in the rapid turnover of the ward. The peristaltic waves are propagated
epithelial cells lining the intestine. A relation¬ for short distances along the intestine and
ship between urogastrone and the acid inhib¬ then die out, to be followed a few minutes
itory hormone enter ogastr one, said to be lib¬ later by another wave. Several waves may be
erated by the duodenum, remains to be in progress at the same time in successive
established. segments of the small intestine but it is rare
for a single peristaltic wave to pass along its
entire length. In addition to these traveling
Muscularis waves of contraction, there are segmental
movements, consisting of alternate constriction
The muscular coat of the small intestine and relaxation of short segments. These do
consists of an outer longitudinal and inner not advance the contents toward the large
circular layer of smooth muscle. Between intestine but result in to-and-fro movement
these layers is the sympathetic myenteric nerve that serves to agitate and mix the material in
plexus. Some strands of smooth muscle cells the lumen.
pass from one layer into the other. The In the terminal portion of the ileum the
smooth muscle cells have usually been re¬ muscularis is somewhat thickened, forming
garded as a static cell population, but auto¬ an ileocecal sphincter. It remains partially con¬
radiographic studies have now demonstrated tracted, delaying emptying of the contents
a slow rate of cell replication distributed into the cecum; After a meal there is a reflex
throughout the external muscle layer. There activation of ileal peristalsis and a relaxation
are differences in rate of replication in dif¬ of the ileocecal sphincter, permitting advance
ferent portions of the alimentary tract. of its contents into the large intestine. At the
The muscularis is responsible for peristalsis, ileocecal orifice, folds of mucosa project into
a wavelike contraction that travels along the the cecum. These act as a bicuspid valve,
intestine at the rate of a few centimeters a closing when the cecum fills and preventing
second and propels the luminal contents on¬ reflux into the ileum.
660 • INTESTINES

acellular detritus. It is difficult to make a

Serosa
clear distinction between the normal struc¬
The outermost layer of the gut wall, called ture and certain pathological conditions in
the serosa, consists of a continuous sheet o^ this organ. Villi are absent. The crypts of
squamous epithelial cells, the mesothelium, sep¬ Lieberkuhn radiating from the lumen have
arated from the underlying muscularis by a an irregular shape and variable length and
very thin layer of loose connective tissue. are largely embedded in the lymphoid tissue.
Most of the gastrointestinal tract is suspended The epithelium of the glands contains only a
from the dorsal wall of the abdomen by few goblet cells and consists mostly of col¬
mesenteries. Along the site of attachment of umnar cells with a striated border. The zone
the mesentery the serosa of the intestine is of mitotically active undifferentiated cells in
continuous with the two apposed leaves of the crypts is shorter than in the small intes¬
the mesentery, and at its base these, in turn, tine. In addition to occasional Paneth cells,
are continuous with the serous lining of the enteroendocrine cells are regularly present
abdominal cavity. Thus, the inner aspect of in the depths of the crypts and in smaller
the abdominal wall and the surface of the numbers in the upper parts of the glands.
organs suspended from it are covered by a They are much more plentiful than in the
continuous layer of mesothelium, usually re¬ small intestine, and6may number five to ten
ferred to as the peritoneum. That portion in a single gland in section.
lining the cavity is called the parietal perito¬ The lymphatic tissue of the appendix, like
neum and that covering the organs is the that of the tonsils, often shows chronic in¬
visceral peritoneum. The transudation of fluid flammatory changes. The muscularis mu¬
from underlying capillaries moistens the cosae of the appendix is poorly developed.
smooth, serous surfaces and facilitates fric¬ The submucosa forms a thick layer with
tion-free sliding of the loops of the intestine blood vessels and nerves and occasional fat
over one another during peristalsis. Bacterial lobules. The muscularis externa is reduced
contamination of the abdominal cavity due in thickness, but the two usual layers are
to perforating lesions of the gut wall results always identifiable. The serous coat is similar
in peritonitis, a severe inflammatory process to that covering the rest of the intestines.
that is often fatal.

THE CECUM AND COLON

THE LARGE INTESTINE The mucosa of the large intestine does not
form folds comparable with the plicae circu¬
The large intestine includes in succession lars, and villi cease beyond the ileocecal
the cecum; the ascending, transverse, and de¬ valve. The interior of the colon therefore has
scending colon; the sigmoid colon', the rectum', a smooth surface when examined with the
and the anus. The cecum is a blind-ending naked eye, but may be somewhat irregular
pouch continuous above with the ascending in outline in sections due to contraction of
colon. At the junction between the two, the the muscularis (Fig. 26—21). At low magnifi¬
ileum enters its medial side. The appendix is cation one can see the openings of innumer¬
a vermiform tubular appendage projecting able crypts or glands of Lieberkuhn (Fig. 26—
from the cecum posteromedial to the ileoce¬ 22). In histological sections these are straight
cal valve. tubular glands about 0.5 mm in length, some¬
what longer than their counterparts in the
small intestine. In the rectum they attain a
THE APPENDIX length of 0.7 mm. They differ from the
glands of the small intestine in the absence
The appendix arises from the blind end of of Paneth cells and in the greater abundance
the cecum and ranges in length from 2 to 18 of goblet cells (Figs. 26—23, 26—24). Entero¬
cm. Its wall has all of the layers typical of the endocrine cells are present in small numbers.
intestine but it is thickened by an extensive Although the goblet cells are the most con¬
development of lymphoid tissue, which forms spicuous element, the majority of the cells in
an almost continuous layer of large and small the middle and upper portions of the crypts
lymphatic nodules (Fig. 26-20). The small are columnar absorptive cells, and these rep¬
lumen has an angular outline in cross section resent the principal cell type in the epithe¬
and often contains masses of dead cells and lium at the surface of the mucosa. As in the
 ,Tunica muscularis externa

Serosa

Centers of

lymphatic
nodules

Crypts of

Lieberkuh n

Mesentery
Lumen with feces

Tunica submucosa '''

Figure 26-20. Cross section of appendix from a 23-year-old man. x 22. (After Sobotta.)

Mucosal
glands

K~:

mmik/
;w
* ' v
psS,|§#ii-
Wm
Figure 26-21. Drawing of a longitudinal section of the wall of the human colon. (From Bargmann, W. Histologie und
Mikroskopische Anatomie des Menschen. 6th ed. Stuttgart, Georg Thieme Verlag, 1962.)
661
Figure 26-22. Scanning electron micrograph of the surface of the descending colon of a monkey (Macaca mulatta)
showing the regular array of openings of the crypts. (From Specian, R., and M. R. Neutra, Am. J. Anat. 160:461, 1981).

Figure 26-23. Sections of the crypts of the colon of a macaque. A, Vertical section of the mucosa, showing the
columnar cells and goblet cells, x 550. B, Horizontal section of the crypts, showing the radial disposition of goblet cells
around the lumen and the cellular lamina propria between crypts. Periodic acid-Schiff reaction and hematoxylin.
Photomicrograph, x 425.
662
 INTESTINES - 663

the crypt and onto the surface. There, they

slough off into the lumen from extrusion
zones situated about midway between the
openings of neighboring crypts. The life span
of most of the cell population is about six
days, but the endocrine cells appear to be
exceptional, having a life span measured in
weeks rather than days. This implies that
they must migrate independently and at a
different rate from that of the surrounding
cells. Little is known about the mechanism of
epithelial migration—whether they move
over the basal lamina or whether it also
moves. It is clear from autoradiographic
studies that the subepithelial fibroblasts that
ensheath the intestinal crypts proliferate at
the base and move upward with the epithelial
cells.
The lamina propria of the colon is similar
in structure to that of the small intestine.
Scattered lymphoid nodules are always pres¬
ent and may extend deep into the submucosa.
The muscularis mucosae is well developed,
consists of longitudinal and circular fibers,
and may send slender fascicles of muscle cells
toward the surface mucosa. The submucosa
presents no unusual features.
The muscularis of the large intestine dif¬
fers in its organization from that of the small
intestine. Instead of forming a continuous
layer of uniform thickness, the longitudinal
fibers are aggregated into three evenly
spaced longitudinal bands, called the taenia
coli. Between the taenia, longitudinal smooth
muscle fibers form a very thin, and often
discontinuous, layer. The inner circular layer
of the muscularis is similar to that of the
small intestine. In the living the taenia are in
a state of partial contraction, which causes
the intervening portions of the wall to bulge
outward, forming sacculations termed haus-
trae. These are conspicuous in the ascending,
transverse, and descending colon and sig¬
moid flexure, but in the rectum the muscu¬
laris externa again becomes a continuous
layer of uniform thickness.
The serosa of the colon is unusual in hav¬
ing local accumulations of adipose cells be¬
Figure 26-24 Photomicrograph of the junction of the
neath the mesothelium that form pendulous
upper portion of a crypt with the surface epithelium of
human rectal mucosa. In the goblet cells at the surface protuberances called the appendices epiploicae.
the theca is smaller and the “stem” longer than in those
of the upper crypt. (Photomicrograph from Neutra, M. Lab.
Invest. 36:535, 1977.) THE RECTUM AND ANUS

small intestine, the epithelium is constantly The rectum is about 12 cm long and ex¬
being renewed. Undifferentiated cells in tends from the sigmoid colon to the pelvic
depths of the crypts divide and their progeny diaphragm. It is slightly dilated in its lower
differentiate into columnar, goblet, and en- portion to form the rectal ampulla. There are
teroendocrine cells, which slowly move up two or three transversely oriented crescentic
664 • INTESTINES

folds of the mucosa above the rectal ampulla. digest food, reducing the carbohydrates, fats,
The mucosa in the rectum is similar to that and protein to molecules that can be ab¬
of the colon but its crypts are somewhat sorbed by the mucosa. Each day some 8 or 9
longer (0.7 mm). liters of water; 100 g of fat; 50 to 100 g of
The rectum narrows rather abruptly at the amino acids; and several hundred grams of
lower end of the ampulla and continues as carbohydrate are absorbed from the small
the anal canal, about 4 cm in length. 4 he intestine.
mucosa here exhibits longitudinal folds, the Most physiologically important processes
rectal columns of Morgagni. The crypts of Lie- in living organisms take place at cell surfaces
berkuhn in this region suddenly become and at interfaces between membrane-limited
short and then disappear along an irregular intracellular compartments. The rate of met¬
line about 2 cm above the anal opening, and abolic activity per unit area of surface prob¬
there is an abrupt transition from simple ably cannot be increased above a certain limit.
columnar to stratified squamous epithelium. Therefore, at all levels of organization there
Here, at the level of the external muscular are architectural devices for increasing sur¬
sphincter of the anus, the lining of the canal face area without increasing the overall size
has the histological appearance of skin with of the organism. This principle of design is
typical sebaceous glands and special, large, dramatically exemplified in the structure of
circumanal apocrine glands. The lamina pro¬ the alimentary tract (Fig. 26—25). The ab¬
pria here contains a plexus of large veins sorptive surface is increased first by elonga¬
that, when abnormally distended and vari¬ tion and convolution of the tubular intestine.
cose, protrude at the anus as hemorrhoids. The mucosal surface is increased by plication
The circular layer of smooth muscle of the to form the plicae circulares. At higher mag¬
anal canal is considerably thickened, forming nification, the surface is found to be ampli¬
the internal anal sphincter. Distal to this is a fied further by about 40 intestinal villi per
circumferential annulus of striated muscle, square millimeter. At the level of the electron
the external anal sphincter. microscope, the surface of every absorptive
cell in the epithelium covering the villi is
augmented another 20-fold by several
HISTOPHYSIOLOGY hundred microvilli. At the molecular level,
OF THE INTESTINE the polysaccharide chains of membrane gly¬
coproteins forming the surface coat are
The function of the intestinal tract and its highly branched filaments, adding further to
accessory glands, the liver and pancreas, is to the enormous surface exposed to the intes-

Figure 26-25. Drawings illustrating the several levels of amplification of the absorptive surface of the intestine. A,
Lengthening and convolution of the gut. B, Plicae circulares. C, Villi. D, Microvilli on the cells. E, Polysaccharides of the
integral glycoproteins of the microvillar membrane. (From Fawcett, D. The Cell. 2nd ed. Philadelphia, W. B. Saunders
Co. 1981.)
 INTESTINES • 665

tinal contents. Within the cell, convolution is cannot be followed by microscopy. Fat lends
again seen in the configuration of the endo¬ itself best to morphological study because
plasmic reticulum, and plication to increase lipid can be preserved in the tissue and in¬
the surface is evident in the cristae of the tensely stained by fixatives containing os-
mitochondria. And these are but a few of
Nature’s stratagems for increasing the effi¬
ciency of the metabolic machinery with min¬
imal increase in body mass.
The secretions of the liver and pancreas
that are delivered into the duodenum at the
ampulla of Vater are essential to the digestion
of the chyme in the small intestine. The bile
from the liver and gallbladder, together with
the mechanical mixing action of peristalsis,
reduces ingested fat to a fine emulsion of
triglycerides. The pancreatic juice contrib¬
utes lipolytic, proteolytic, and carbohydrate
splitting enzymes. The intestinal mucosa it¬
self contributes intestinal juice, or succus en-
tericus.
The secretion of the crypts of Lieberklihn
was formerly believed to contain several
digestive enzymes, but it has become clear
from biochemical studies on isolated brush
borders that some of the enzymes believed
to be secreted into the lumen are instead
incorporated in the membrane of the brush
border of the absorptive cells. Among these
are leucine aminopeptidase, and the enzymes
that hydrolyze disaccharides—sucrase, which
cleaves sucrose to glucose and fructose; lac¬
tase, cleaving lactose to glucose and galactose;
and maltase, that hydrolyzes maltose, derived
from starch, to glucose. These enzymes resid¬
ing in the membrane and surface coat of the
microvilli reduce dietary carbohydrates in the
gut lumen to hexoses that are actively trans¬
ported, together with sodium ions, into the
absorptive cells by carrier protein present in
the brush border. Thus, the brush border is
not only a device for increasing surface area
for absorption but also the site of the enzymes
involved in the terminal steps in digestion of
carbohydrates and proteins, and it possesses
the carriers necessary for active transport of
glucose and amino acids into the cell.
A genetic defect resulting in absence of
one of the enzymes in the brush border is
the basis for an uncommon disease that pre¬
viously was poorly understood. Some infants
cannot tolerate milk, and feeding results in
bloating and copious diarrhea. These symp¬
toms have been traced to congenital absence
of the single enzyme lactase from the intes¬ Figure 26-26. Electron micrograph of rat intestinal cell.
tinal brush border. If lactose is eliminated Lipid accumulates in sizable droplets within the smooth
reticulum, but very little is seen in inpocketings of the cell
from the diet, these infants do very well. surface or in vesicles traversing the terminal web. Al¬
Intraluminal digestion of most food re¬ though pinocytosis undoubtedly occurs to some extent it
duces it to units of molecular size whose path is not the principal mechanism for absorption of lipid.
of absorption through the intestinal mucosa (Courtesy of S. L. Palay and J. P. Revel.)
666 • INTESTINES

mium tetroxide (Figs. 26—26, 26—27). Dietary ide forms numerous visible droplets in the
fat, consisting mainly of triglycerides, is hy¬ lumen of the reticulum in the apical portion
drolyzed by pancreatic lipase in the intestinal of the cell (Fig. 26-26). From there the lipid
lumen to free fatty acids and monoglycerides. appears to be transported to the Golgi com¬
These products combine with bile salts to plex for further processing that converts it
form minute micelles about 2 nm in diameter into chylomicra, the complex glycolipoprotein
(Fig. 26—28). When myriads of these come droplets that are transported via the lacteals
into contact with the microvilli of the in¬ and intestinal lymphatics to the bloodstream.
testinal absorptive cells, the fatty acids and The precise role of the Golgi complex in lipid
monoglycerides diffuse across the plasma absorption has not been established, but it
membrane and accumulate in the apical cy¬ seems likely that the carbohydrate moiety of
toplasm. The membranes of the smooth en¬ the chylomicra is added there to the triglyc¬
doplasmic reticulum located there contain eride synthesized in the smooth reticulum.
the enzymes for resynthesis of triglycerides The chylomicron also acquires in its passage
from fatty acids and monoglycerides. During through the Golgi complex a membranous
lipid absorption, the resynthesized triglycer¬ investment with properties that enable it to

Figure 26 27. Schematic representation of the fine structure of an intestinal absorptive cel! (A) in the fasting state and
(B) after a lipid-rich meal. Area in rectangle is depicted at higher magnification in Figure 26—27. (Redrawn after Cardell
R., S. Badenhausen, and K. R. Porter. J. Cell Biol. 34:123, 1967.)
 INTESTINES • 667

coalesce with the lateral plasma membrane

of the columnar cell. Vesicles of Golgi origin
thus discharge chylomicra into the intercel¬
lular cleft between adjacent absorptive cells
(Figs. 26—29, 26—30). From there they move
across the basal lamina of the epithelium and
into the lymphatic capillaries in the lamina
propria of the intestinal villi.
The absorptive cells of the intestine are
able to respond rapidly and to modify their
internal structure to adapt it to the require¬
ments of lipid absorption. In the fasting state,
there are some profiles of granular endo¬
plasmic reticulum and a limited amount of
smooth reticulum in the apical cytoplasm,
and a relatively quiescent supranuclear Golgi
complex. Ingestion of lipid stimulates the
formation of a more extensive smooth retic¬
ulum to provide for resynthesis of triglycer¬
ide and its transport to the Golgi complex.
The rapid turnover of Golgi membranes dur¬
ing lipid absorption and transport also re¬
quires accelerated synthesis of new mem¬
brane to replace that lost to the cell surface
in discharge of chylomicra. If protein synthe¬
sis is blocked by administration of puromycin,
membrane replacement in smooth reticulum
and Golgi complex is prevented, as is synthe¬
sis of protein constituents of the chylomicra.
Lipid transport is therefore inhibited and
large amounts of lipid accumulate in the
cytoplasm of the absorptive cells.
An important part of the absorptive mech¬
anism is the active movement of the intestinal
villi. These can be observed in a living animal
if a loop of intestine is split open and the
surface of the mucosa is observed with a
binocular microscope. A villus is seen to sud¬
denly shorten to about half its length with an
appreciable increase in its thickness, and it
slowly extends again to its original length.
Each villus contracts about six times a minute.
During the contraction, its volume is reduced
and the contents of the central lacteal are
Triglyceride
✓ .Fatty acid forwarded to the submucous lymphatic
A Bile salt plexus. Contraction occurs as a result of
T Monoglyceride
shortening of the longitudinally oriented
Diglyceride s Protein
strands of smooth muscle in the core of the
villus. The contraction of the villi is believed
<p Lipase
to be under the nervous control of Meissner’s
Figure 26-28. Schematic representation of events occur¬
ring in a small area of the apex of an intestinal absorptive submucous plexus. Direct mechanical stimu¬
cell (see Fig. 26-27). An emulsion of fine droplets of lipid lation of the base of a villus with a bristle also
in the lumen is broken down by pancreatic lipase to fatty calls forth a contraction, and the stimulus
acids and monoglycerides. These diffuse into the microvilli radiates from the affected villus to the sur¬
and apical cytoplasm, where they are esterified to form
rounding villi.
triglycerides in the smooth endoplasmic reticulum. (Re¬
drawn after Cardell, R., S. Badenhausen, and K. R. Porter. In addition to the absorption of water and
J. Cell Biol. 34:123, 1967.) nutrients, the small intestine has an essential
668 • INTESTINES

Figure 26-29. Electron micrograph of the boundary between two rat intestinal epithelial cells during lipid absorption.
The absorbed lipid has been discharged through the lateral cell surfaces and is seen to have accumulated here as
aggregations of chylomicrons in the intercellular spaces, x 30,000. (Courtesy of S. L. Palay and J. P. Revel.)

function in absorption and conservation of gested vegetable matter. The bacterial flora
ions. About 6 g of sodium is taken in daily of the intestine in humans digest only small
in the diet and 20 to 30 g are secreted into amounts of cellulose, but in herbivorous
the lumen in the succus entericus, but little mammals cellulose is an important source of
or none is normally lost in feces. Sodium is nutrients whose digestion depends on the
constantly reabsorbed in the intestine. It dif¬ bacteria of the gastrointestinal tract. In hu¬
fuses through the apical membrane of the mans the major contribution of the intestinal
absorptive cells and is actively transported flora is the production of vitamin B12, needed
through the lateral membranes, creating a for hemopoiesis, and vitamin K, which is
standing gradient in the intercellular clefts essential for maintenance of the clotting
that draws water through the epithelium and mechanism of the blood.
into the capillaries of the villi. Chloride ions
diffuse along the electrochemical gradient
following the sodium. The importance of this
reabsorption of ions becomes evident in chol¬
BLOOD VESSELS OF THE
era and other forms of severe diarrhea in GASTROINTESTINAL TRACT
which the sodium reserves of the body can
be reduced to lethal levels within several The arrangements of the blood and lymph
hours. vessels in the wall of the stomach and in the
Absorption of water and electrolytes con¬ wall of the intestine are basically similar.
tinues in the colon. About 1 liter of chyme Because the important differences depend
passes through the ileocecal valve daily, but mainly on the presence of villi, the small
normally less than 100 ml is lost in the feces. intestine shows significant peculiarities.
The bulk of the feces consists of dead bacteria In the stomach, the arteries arise from the
and undigestible fibrous constituents of in¬ two arterial arches along the lesser and
 INTESTINES • 669

cularis externa into the serosa. In the stom¬

ach the veins of the submucosal plexus are
provided with valves and a relatively thick
muscular coat.
In the small intestine, the submucous ar¬
terial plexus gives off two kinds of branches
that run toward the mucosa. Some of these
arteries ramify on the inner surface of the
muscularis mucosae and break up into cap¬
illary networks that surround the crypts of
Lieberkuhn in the same way they surround
the glands of the stomach. Other arteries are
especially destined for the villi, each villus
receiving one or sometimes several such small
arteries. These vessels enter the base of the
villus and form a dense capillary network
immediately under its epithelium (Fig. 26-
32). Near the tip of the villus, one or two
small veins arise from the superficial capillary
network and run downward, to anastomose
with the glandular venous plexus, and then
pass on into the submucosa, where they join
the veins of the submucosal plexus. These
veins in the intestine have no valves. How¬
ever, their continuations, which pass through
the muscularis externa with the arteries, are
provided with valves. Valves disappear again
in the collecting veins of the mesentery.
Figure 26-30. Electron micrograph of chylomicrons fixed
in osmium tetroxide, embedded and sectioned. They
range from 110 mm to 700 mm. (From Jones, A., and M.
Price. J. Histochem. Cytochem. 76:366, 1968. By permis¬
sion of the Histochemical Society, Inc.)
LYMPH VESSELS OF THE
GASTROINTESTINAL TRACT

greater curvatures and are distributed to the In the stomach, the lymphatics begin as an
ventral and dorsal surfaces. In the intestine, extensive system of large lymphatic capillar¬
the arteries reach one side in the mesentery. ies in the superficial layer of the mucosa
They run in the serosa and break up into between the glands. They are always situated
large branches that penetrate the muscularis more deeply than the blood capillaries. They
externa and enter the submucosal layer, anastomose everywhere throughout the mu¬
where they form a large plexus (Fig. 26—31). cosa. They surround the glandular tubules
In the stomach and colon the plexus gives and take a downward course to the inner
off branches directed toward the surface. surface of the mucosa, where they form a
Some of these break up into capillaries sup¬ plexus of fine lymphatic vessels. Branches of
plying the muscularis mucosae; others form the plexus pierce the muscularis mucosae and
capillary networks throughout the mucosa form, in the submucosa, a plexus of lym¬
and surrounding the glands. The capillary phatics that is provided with valves. From the
net is especially prominent around the fo- submucosal plexus, larger lymphatics run
veolae of the gastric mucosa. through the muscularis externa. Here they
From the superficial, periglandular capil¬ receive numerous tributaries from the lym¬
lary networks, veins of considerable caliber phatic plexus in the muscular coat and then
arise. They form a venous plexus between follow the blood vessels into the retroperito¬
the bottoms of the glands and the muscularis neal tissues. In the wall of the colon the
mucosae. From this plexus, branches run into lymphatics show a similar arrangement.
the submucosa and form a venous plexus. The lymphatic vessels are important in the
From this submucosal plexus, the large veins absorption of fat from the small intestine.
follow the arteries and pass through the mus¬ During digestion, all their ramifications be-
670 * INTESTINES

Submucous plexus
, t_
\ T mucosa
ikLmuscularis
mucosae Villus
submucosa 'layer
rc u I a r
ICyMC muscle
Longitudinal
r til musc*e
0m

Crypt
layer

}Mm
Vein •Sub
Artery

CM

Vei n } LM
Artery

C. , .
Intermuscular /
Mucous plexus
r
. .
V
plexus / i_onqjflidinal muscle
Circular muscle
fCsubmucosa

Lmuscularis
mucosae
Tmucosa

Mm

Submucous
plexus
Lymphatic vessel

Figure 26-31. Diagrams of distribution of blood vessels (A and B) and of lymphatics (C and D) in the small intestine of
the dog. B and D are drawn on a larger scale to show details. CM, Circular muscle; Cr, crypt; F, follicle; LM, longitudinal
muscle; Mm, lamina muscularis mucosae; PF, perifollicular plexus; Smp, submucous plexus; Sub, tunica submucosa;
V, villus. (Redrawn and slightly modified from Mall.)
 INTESTINES • 671

larger lymphatics. The latter also receives

tributaries from the dense network of large,
thin-walled lymphatic capillaries, which
closely surround the surface of the solitary
and aggregated follicles. The large lymphat¬
ics that run from the submucosal plexus
through the muscularis externa into the mes¬
entery receive additional branches from a
dense, tangential plexus located between the
circular and longitudinal layers of the mus¬
cularis externa.

NERVES OF THE INTESTINAL

TRACT

The gastrointestinal tract is innervated by

the autonomic nervous system, which consists
of three subdivisions: sympathetic, parasym¬
pathetic, and enteric. All three play a role in
regulating intestinal activity but the enteric is
B
the most important. The sympathetic and
Figure 26-32. Two villi of rat intestine injected with India
parasympathetic nerves to the gastrointes¬
ink in gelatin. The villi in this species are thin, leaflike
structures whose broad surface is presented in this figure. tinal tract constitute its extrinsic nerve supply
In the duodenum (B) they are larger and more richly and exert their influence on digestive func¬
vascularized than they are farther along in the jejunum tion through the intrinsic enteric nervous sys¬
(A). The large surface area presented by the villi and their tem, which consists of nerve cell bodies and
rich network of capillaries favors absorption of nutrients,
x 100. their processes located within the wall of the
tract. The extrinsic nerves are preganglionic
fibers from the vagus nerve and postgan¬
come filled with milky white lymph—a fine glionic sympathetic fibers that arise mainly in
emulsion of neutral fats. This white lymph, the celiac ganglion. They are distributed to
drained from the intestine, is called chyle, and the intestine with the blood vessels via the
the lymphatics that carry it away from the mesentery. The enteric nervous system con¬
epithelium are called lacteals. sists of ganglia and interconnecting bundles
In the small intestine the most conspicuous of nerve fibers that form extensive plexuses
parts of the lymphatic system are the central in various layers of the digestive tube and
lacteals in the core of the villi. Each conical extend throughout its length from the lower
villus has one lacteal, which occupies an axial esophagus to the internal anal sphincter. The
position and ends blindly near the tip. The enteric system contains 107 to 108 neurons
broader villi of the duodenum may contain including sensory neurons, interneurons,
two or perhaps more lacteals that intercom¬ and motor neurons, which form reflex path¬
municate. The lumen of these lacteals, when ways that are intrinsic to the digestive tract.
distended, is considerably larger than that of Unlike other subdivisions of the autonomic
the blood capillaries. The wall consists of thin nervous system, the enteric system is largely
endothelial cells and is everywhere connected autonomous and can carry out many of its
with the argyrophilic reticulum and sur¬ functions without input from the central ner¬
rounded by thin, longitudinal strands of vous system. Although the sympathetic and
smooth muscle. parasympathetic nerves do have some effect
The central lacteals at the base of the villi upon the digestive tract, cutting the extrinsic
anastomose with the lymphatic capillaries be¬ nerves results in surprisingly little impair¬
tween the glands. They also form a plexus ment of digestive function. If the intestine is
on the inner surface of the muscularis mu¬ detached from the mesentery and immersed
cosae. Branches of this plexus, provided with in warm physiological salt solution, it will
valves, pierce the muscularis mucosae and show normal peristaltic movements when the
form, in the submucosa, a loose plexus of mucosa is stimulated by introduction of ma-
672 INTESTINES

;i.

..
"% '
.
, /; /,«ji

Autonomic Nerve

Plexus

WSmm.

F fbroblasts

Nerve ,

Plexus

Figure 26-33. Photomicrograph of a whole mount of the longitudinal muscle coat of rabbit intestine impregnated with
silver to show the nerve bundles of Auerbach’s plexus, x 300. (After Richardson, K. C. J. Anat. 94:451, 1960. Labelinq
added.)
 INTESTINES 673

Figure 26-34. Scanning electron micrograph of the myenteric plexus of rat intestine. The overlying longitudinal muscle
and connective tissue has been removed by dissection and enzymatic digestion. (Micrograph from Fujiwara, T., and Y.
Uehara. J. Electron Microsc. 29:397, 1980.)
674 • INTESTINES

bodies and bundles of unmyelinated axons

that connect the ganglia to form a continuous
network (Fig. 26-34). The cells of the ganglia
are of two principal morphological types.
One is a multipolar cell (Figs. 26-35, 26-36)
with short dendrites in contact with the bod¬
ies of similar cells in the same ganglion, while
the axon can be traced for a considerable
distance to sites of synaptic contact with cells
of the second type in neighboring ganglia.
Cells of the second type are far more numer¬
ous and more variable in form. The dendrites
are diffuse receptive endings related to cells
of the first type, or the same type, in the
same or in other ganglia. The axon enters
one of the fiber bundles associated with the
ganglion, and its branches terminate in the
circular or longitudinal muscle layers. Thus,
cells of the first type appear to be associative
while those of the second type are motor. A
third cell type, termed the interstitial cell, has
short branching processes that intermingle
with those of the other cell types. It does not
contain demonstrable neurofibrils and may
be a form of glial cell.
Most of the unmyelinated fibers in the
ganglia and in the internodal strands of the
plexus are processes of enteric neurons. The
remainder are axons of extrinsic vagal or
sympathetic origins. The vagal fibers termi¬
Figure 26-35. Ganglion of the myenteric plexus of cat nate as perikaryal arborizations on ganglion
jejunum containing bipolar and multipolar neurons. Modi¬ cells of the second type. The sympathetic
fied Bielschowsky stain. (Photomicrograph courtesy of G. fibers do not seem to have synaptic relation¬
Schofield.)
ships with the nerve cells of the ganglia, but
are thought to terminate in the muscularis
and on blood vessels.
terial into the lumen. Thus, intestinal move¬ A deep muscular plexus (plexus muscularis pro¬
ments are determined by local neuromuscu¬ fundus) is situated on the mucosal aspect of
lar mechanisms that are modulated only by the circular muscle layer. It is devoid of
input through the extrinsic nerves. ganglia and consists of thin anastomosing
The most superficial neural elements in nerve bundles with their prevailing orienta¬
the gut wall form a subserous plexus, which tion parallel to the muscle fibers. Branches
generally lacks ganglia and consists of a loose from this plexus penetrate into the muscle
network of fine nerve fibers that connect the layer and some are connected with the myen¬
extrinsic nerves with the more deeply situ¬ teric plexus.
ated intrinsic plexuses. The majority of the The submucous plexus (Meissner’s plexus)
nerves coursing from the mesentery to the (plexus entericus interna) is a network of ganglia
deeper enteric plexuses traverse the longitu¬ and interconnecting nerve bundles within the
dinal muscle layer near the mesenteric at¬ connective tissue of the submucosa. Its fibers
tachment. innervate the muscularis mucosae and
The most conspicuous enteric plexus is smooth muscle strands in the cores of the
found between the longitudinal and circular intestinal villi. Fibers from the submucous
layers of the muscularis (Figs. 26-33, 26-34). plexus also form a mucosal plexus situated in
This myenteric plexus (Auerbach’s plexus) the lamina propria and extending compo¬
(plexus entericus externa) consists of ganglia nents between the intestinal glands and into
containing three to 50 or more nerve cell the villi.
 INTESTINES • 675

Figure 26-36. Scanning micrograph of a multipolar neuron in the myenteric plexus of a newborn rat. (Micrograph
courtesy of T. Komuro and Y. Uehara.)
676 • INTESTINES

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a variety of biologically active peptides in Grube, D., and W. G. Forssmann: Morphology and
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clude substance P, somatostatin, enkephalins, vas¬ tab. Res. 7 7:603, 1979.
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trastructural identification of a new cell type—the
rotensin. It is not known whether these serve
N-cell as the source of neurotensin in the gut
as neurotransmitters or neuromodulators. mucosa. Cell Tissue Res. 184:445, 1977.
 INTESTINES • 677

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Pearse, A. G. E., J. M. Polak, and S. R. Bloom: The ing linolenic acid, monoolein, and bile salt. J. Lipid
newer gut hormones. Cellular sources, physiology, Res. 7:307, 1966.
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72:746, 1977. CELL TURNOVER AND RENEWAL
Chang, W. W. L., and C. P. Leblond: Renewal of the
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IgA and IgM: a hypothetical model. In Immunology 1971.
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Owen, R. L., and A. L. Jones: Epithelial cell specializa¬ Am. J. Anat. 769:113, 1981.
tion with human Peyer’s patches: an ultrastructural Leblond, C. P., and B. Messier: Renewal of chief cells
study of intestinal lymphoid follicles. Gastroenter¬ and goblet cells in the small intestine as shown by
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Owen, R. L., and P. Nemanic: Antigen processing struc¬ into mice. Anat. Rec. 752:247, 1958.
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SUBMUCOSAL GLANDS OF BRUNNER
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LARGE INTESTINE
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 v;

THE LIVER AND

GALLBLADDER

LIVER protein components of the blood plasma, and

it exercises an important degree of control
over the general metabolism by virtue of its
The liver is the largest gland in the body, capacity to store carbohydrates as glycogen
weighing about 1500 g in the adult. It func¬ and to release glucose to maintain the normal
tions both as an exocrine gland, secreting bile concentration of glucose in the blood.
through a system of bile ducts into the duo¬
denum, and as an endocrine gland, synthesiz¬
ing a variety of substances that are released HISTOLOGICAL ORGANIZATION OF
directly into the bloodstream. To appreciate THE LIVER
the importance of the liver and to correlate
its structure with its functions requires an The liver is composed of epithelial cells
understanding of its blood supply and its arranged in plates or laminae that are inter¬
strategic location in the circulation. It receives connected to form a continuous tridimen¬
a large volume of venous blood from the sional lattice. The laminae are disposed ra¬
intestinal tract via the portal vein and a small dially with respect to terminal branches of
volume of arterial blood via the hepatic artery. the hepatic veins, which traditionally have
It is drained by the hepatic veins into the been designated as central veins because of
inferior vena cava near the heart. The liver their location in the centers of prismatic units
is thus interposed between the intestinal tract of liver parenchyma that constitute the liver
and the general circulation. It therefore re¬ lobules (Figs. 27-1, 27-2). The radially dis¬
ceives, in the portal blood, all of the material posed plates of liver cells are exposed on
absorbed from the intestinal tract except the either side to the blood flowing in a parallel
bulk of the lipid, which is transported in the system of vascular channels, the hepatic sinu¬
chyle via the mesenteric lymphatics to the soids. The radially oriented sinusoids closely
thoracic duct. The absorbed products of conform to the broad surfaces of the cellular
digestion are taken up and metabolized in laminae and intercommunicate through fe¬
the liver or are transformed there and re¬ nestrations in them to form a labyrinthine
turned to the blood for storage or utilization system of thin-walled vessels intimately re¬
elsewhere. The liver may also receive toxic lated to a very large surface area of liver
substances from the intestine or from the parenchyma.
general circulation, and is capable of degrad¬
ing them by oxidation or hydroxylation or The Liver Lobule
detoxifying them by conjugation. The prod¬
ucts of their degradation or their harmless In the pig and a few other species, a well-
conjugates are then excreted in the bile. The defined layer of connective tissue clearly de¬
bile is a complex fluid that can be regarded marcates the lobules. However, in most mam¬
as a secretion in that it plays an important mals, including the human, there is no
role in digestion, but it can also be regarded boundary between the lobules, the hepatic
as a vehicle for excretion to the extent that it parenchyma appearing quite continuous.
carries detoxified waste and potentially harm¬ The radial pattern of the cellular plates and
ful materials to the intestine for ultimate sinusoids is such that one can, nevertheless,
elimination. recognize the units of structure and assign
The liver also synthesizes several important imaginary boundaries to the lobule by relying
679
680 • THE LIVER AND GALLBLADDER

Interlobular
vessels, etc.

Branch of
Bile portal
duct vein
Branch of
hepatic artery

"-- Central veins of

I two lobules
II
II
I
I
II
I
I
I
l

l
I

Interlobular septa
('G/isson’s capsule) Hepatic
vein

Figure 27-1. Wax reconstruction (by A. Vierling) of a lobule of the liver of a pig. A portion of the lobule has been cut
away to show the bile capillaries and sinusoids, x 400. (After Braus.)

upon the regularly distributed central veins The traditional lobule as defined above is
and portal canals as landmarks (Fig. 27-2). not comparable with the lobules of most
The lobules in sections are typically hex¬ glands, which are centered on the ducts that
agonal, with the corners of the polygon each drain them. The liver lobule as just presented
occupied by a portal canal. This latter struc¬ is conceptually convenient, however, for as a
ture consists of a small branch of the portal result of differential deposition of glycogen
vein and one of the hepatic artery, as well as or fat, the hepatic tissue of which it is com¬
a bile ductule, enclosed in a common invest¬ posed frequently exhibits microscopically dis¬
ment of connective tissue (Fig. 27-3). Blood tinguishable zones, concentric around the
enters the hepatic sinusoids from small central vein. Moreover, in pathological con¬
branches of the hepatic artery and portal ditions, necrosis may selectively involve the
vein, flows through the lobule centripetally, central or the peripheral zone, depending
and leaves via the central vein (Figs. 27-4 upon the nature of the disease process.
27-5). The lobular pattern appears to develop as
 Branch of
hepatic artery

Bile ducts

Inter-
lobular
veins

Branches of
hepatic

a rtery

Interlobular
vein

Artery Bile Interlobular Liver cell plates Bile ducts Interlobular veins
ducts connective tissue
Figure 27-2. Portion of liver from a 22-year-old man. Two complete lobules are surrounded by portions of other lobules,
x 70. (After Sobotta.)

Figure 27-3. Photomicrograph of liver parenchyma at the periphery of a traditional lobule, showing a typical portal triad,
consisting of branches of the hepatic artery and portal vein, and a small bile duct.
681
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Figure 27-4. Diagram of hepatic structure. (From Gray’s Anatomy. London, Longman. After Prof. H. Elias.)
 THE LIVER AND GALLBLADDER • 683

Sinusoids center of the lobule, and the branches of the

hepatic vein were said to be situated around
its periphery. The lobule defined in this way
is called a portal lobule. In such a lobule, the
bile would drain toward a duct located with
the vascular supply in the center of the lob¬
ule, as is the case in most other glands.
In some respects this is a more satisfactory
way of interpreting the architecture of the
liver than the classical lobule, but it has been
argued that the portal lobule is not the small¬
est unit of functional organization of the
liver. A variant of the portal lobule has been
proposed by Rappaport and his colleagues,
who consider the functional unit to be a mass
of parenchymal tissue associated with the fine
terminal branches of the portal vein, hepatic
artery, and bile duct. These branches leave
the portal canals at intervals, coursing per¬
pendicular to the canals and to the central
vein, and run along the side of the hexagon
Figure 27-5. Diagrammatic representation of the radial
disposition of the liver cell plates and sinusoids around
that forms the section of the classical lobule.
the terminal hepatic venule or central vein, showing the The associated mass of hepatic tissue is
centripetal flow of blood from branches of the hepatic smaller than either of the lobules proposed
artery and portal vein, and the centrifugal flow of bile earlier and is composed of parts of two ad¬
(small arrows) to the small bile duct in the portal space.
jacent classical lobules (Fig. 27—6). It is called
(Redrawn and modified from Ham, A. W.: Textbook of
Histology. 5th ed. Philadelphia, J. B. Lippincott Co., 1965.)
Terminoi HepaticV.

a consequence of the hydrodynamics of the

blood flow through the liver. From this point
of view the liver may be considered as a
tough sac filled with fluid, in which is sus¬
pended a plastic spongework of liver tissue.
In the flow of fluid through the liver, the
terminal branches of the portal vein are
sources and the radicles of the hepatic vein
are sinks (Figs. 27—4, 27—5). The flow from
the one to the other is thought to determine
the radial pattern of the sinusoids character¬
istic of the lobule. It follows also that the cells
nearest the branches of the portal vein re¬
ceive blood first and therefore have first call
upon the nutrient and oxygen content of the
portal blood. As the latter diminish in the
passage of the blood from the periphery
toward the center of the lobule, a gradient
of metabolic activity is established, which is
expressed in the morphologically detectable
zonation of the lobule. Figure 27-6. Diagram illustrating the functional unit of
liver parenchyma (the acinus) according to Rappaport et
Some histologists have objected to the clas¬
al. It consists of the parenchyma centered around the
sical definition of the liver lobule because it terminal branches of the hepatic artery and portal vein. It
is inconsistent with the lobular organization should be noted that the cells in Zone 1 nearest these
typical of other exocrine glands. In an effort vessels have first call upon the incoming oxygen and
nutrients, while the cells of Zone 2 are less favored and
to make the liver conform to the same gen¬
those of Zone 3 near the terminal hepatic venules are
eral plan, Mall proposed an alternative con¬ least favorably situated. (Redrawn after Rappaport, A. M.,
cept of liver lobulation according to which A. J. Borowy, W. M. Lougheed, and W. N. Lolto. Anat.
the portal canal was considered to be the Rec. 779:11, 1954.)
684 • THE LIVER AND GALLBLADDER

a liver acinus and is defined as the tissue duct, are the axial structures of the smallest
supplied by a terminal branch of the portal portal canals. Small lateral branches given off
vein and of the hepatic artery and drained at short intervals from these terminal venules
by a terminal branch of the bile duct. The pass through fenestrations in the limiting
limits of the acinus are not defined by any plate of hepatic cells to become continuous
recognizable anatomical landmarks but ex¬ with the sinosoids of the neighboring liver
tend outward to the terminal branches of the lobules.
hepatic veins and to the imaginary outer The hepatic artery on entering the porta
limits of acini associated with neighboring branches into interlobar and then into intralob¬
portal canals. The parenchyma is continuous ular arteries. The great majority of the flow
from one acinus to the next, and indeed from these vessels is distributed to capillaries
from one classical lobule to the next. There¬ in the connective tissue stroma of the liver.
fore, if the supply and drainage of one unit Only a small volume continues into the ter¬
should fail, it would still be supplied and minal hepatic arterioles of the smallest portal
drained by others. This concept of liver struc¬ canals (Fig. 27-7). These give off collateral
ture, although still not universally accepted, branches to the hepatic sinusoids and nu¬
has proved useful in the understanding of merous small branches to a dense capillary
some aspects of liver physiology and in ac¬ network that surrounds the bile ductules.
counting for some manifestations of liver This peribiliary or periductal plexus was for¬
pathology, especially that following bile duct merly thought to drain into the portal ven¬
occlusion and that found in cirrhosis of the ules. Vascular casts studied by scanning elec¬
liver. tron microscopy have now clearly shown that
The classical lobule, the portal lobule, and the efferent vessels are given off to the hepatic
acinus should not be considered as conflicting sinusoids (Fig. 27-8). Thus, much of the
concepts of liver structure but as complemen¬ arterial blood reaches the sinusoids indirectly
tary ones. Because of the complexity of the via the peribiliary plexus. The extraordinar¬
function of the liver, it is sometimes useful ily rich vascularity of the small bile ducts
to think in terms of one and at other times suggests the possibility that some constituents
in terms of another. It is noteworthy that the of the bile may be reabsorbed in passage
traditional lobulation is not present in the through the intrahepatic bile ducts.
lower vertebrates nor in the mammalian em¬ The primary function of the hepatic cir¬
bryo. culation is carried out in the sinusoids. These
form an exceedingly elaborate three-dimen¬
sional plexus presenting an enormous sur¬
The Blood Supply
face for interchange of metabolites with the
Most organs receive their afferent blood hepatic parenchyma. Every liver cell in the
supply from one or more arteries. The liver radially arranged cellular plates of the lobule
is exceptional in that its principal afferent is exposed on at least one, and usually two,
blood supply is the portal vein carrying blood sides to blood circulating in the sinusoids.
from the intestinal tract and spleen. About Corrosion preparations of vascular casts
75 per cent of the blood to the liver is brought viewed with the scanning microscope illus¬
to it by the portal vein and only 25 per cent trate dramatically the richness of the micro¬
by the relatively small hepatic artery. The por¬ vasculature of the liver (Fig. 27—7). The blood
tal vein entering the porta of the liver leaves the lobule through a terminal radicle
branches into interlobar veins and these in turn of the hepatic vein, the central vein of the
into conducting veins 400 pm or more in classical lobule. Its wall is penetrated by in¬
diameter. Their walls are thinner and have numerable openings through which blood
less adventitial connective tissue than veins flows from the surrounding sinusoids into its
of comparable size elsewhere. The conduct¬ lumen (Fig. 27—9). The central veins drain
ing veins branch to give rise to interlobular into intercalated veins (.sublobular veins). Several
veins, which possess few longitudinal smooth of these unite to form collecting veins and
muscle cells in their walls. The terminal portal these in turn join to form hepatic veins, which
venules that arise from the interlobular veins pursue a course through the liver independ¬
are 280 pm or less in diameter and very thin ent of the portal venous system. Two or more
walled. These, accompanied by a terminal large hepatic veins enter the inferior vena
branch of the hepatic artery and a small bile cava.
 www: Slnusoidsr^

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m- •* _** J#
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J**W* •* i* %V V __ M

Figure 27-7. Scanning micrograph of a vascular cast of monkey liver illustrating the extraordinarily rich network of
hepatic sinusoids. Crossing the figure diagonally is a terminal branch of the hepatic artery and its associated vein.
(Micrograph courtesy of T. Murikami, from Johari, O., and I. Corvin, eds.: Scanning Electron Microscopy 1978. Part II.
Chicago, SEM Inc. AMF O’Hare, 1978.)

Figure 27-8. Scanning micrograph of a methacrylate cast of the blood vessels forming the periductal plexus, and its
communications with the sinusoids (at arrows). (Micrograph from Murikami, T. Arch. Histol. Jpn. 37:245, 1974.)
685
686 * THE liver and gallbladder

Figure 27-9. A, Scanning micrograph of a vascular cast of the central portion of a liver lobule, showing the confluence
of numerous sinusoids with the central vein. B, A comparable scanning micrograph of this region of the lobule cut
longitudinally through the central vein, showing the openings of sinusoids into its lumen. (A, Courtesy of T. Murikami.
B, From Mota, P. M., M. Muto, and T. Fujita. The Liver. Tokyo, Igaku, Shoin, 1978.)

Hepatic Sinusoids one as a typical endothelial cell and consid¬

ered the other to be a form of tissue macro¬
The hepatic sinusoids are larger than cap¬ phage called the cell of Kupffer. Other inves¬
illaries and are more irregular in shape, and tigators observed that, upon repeated
their lining cells are directly apposed to the injection of colloidal particulate matter into
epithelial cells of the parenchyma with essen¬ experimental animals, the phagocytic cells of
tially no intervening connective tissue. The the sinusoids became larger and more nu¬
lining of the sinusoids consists of a thin layer merous; they thought they could recognize
of cells that differ from typical capillary en¬ forms intermediate between endothelial cells
dothelium in two respects: (1) some of the and Kupffer cells. Therefore, it was thought
lining cells are actively phagocytic (Fig. 27— that the endothelial cells could transform into
10) and (2) the cell boundaries do not blacken phagocytic cells when the need arose and
with silver nitrate. This latter property led that the different appearance of the lining
some early investigators to suggest that the cells simply represented different functional
lining was a syncytium. Various cytological states of a single cell type. The application of
and experimental studies carried out in the electron microscopy and radiolabeling tech¬
era of light microscopy cast doubt upon this niques has now resolved the controversy in
interpretation, and subsequent studies with favor of distinct populations having different
the electron microscope have established be¬ origins and functions. In electron micro¬
yond doubt that the endothelium is made up graphs of liver fixed by perfusion, it is now
of individual cells. possible to identify three cell types associated
Whether the lining is composed of one or with the sinusoids—endothelial cells, Kupffer
two types of cells was long a subject of dis¬ cells, and perisinusoidal fat-storing cells.
pute. Those who distinguished two identified Endothelial Cells. These flattened cells
 THE LIVER AND GALLBLADDER 687

Figure 27-10. Low-magnification micrograph of rat liver, illustrating the intimate relationship of the cell plates to the
sinusoids. Kupffer cells in some of the sinusoids contain dense phagocytosed material. (Micrograph courtesy of E.
Wisse.)
688 • THE LIVER AND GALLBLADDER

form the greater part of the extremely thin be separated by intercellular openings 0.1 to
lining layer of the sinusoids. They are not 0.5 pm across. In addition, it can be seen in
signihcantly phagocytic to colloidal vital dyes, electron micrographs that the thin peripheral
although some pinocytotic activity is sug¬ portions of the endothelial cells are fenes¬
gested by electron micrographs in which trated, presenting a sievelike appearance
small plasmalemmal vesicles are seen associ¬ (Fig. 27-11). These openings are consider¬
ated with both the adluminal and the ablu- ably larger and more variable in size and
minal surfaces of the cell. shape than the pores in fenestrated capillar¬
Whether the sinusoidal lining was contin¬ ies. Thus, it appears that there are sizable
uous or discontinuous was a question long transcellular fenestrations as well as small
debated. Physiological observations on rates discontinuities between adjacent cells. It can
of clearance of substances from the blood be shown that in as short a time as 30 seconds
during its transit through the liver and ob¬ after injection of colloidal thorium dioxide
servations on the size of the molecules and into the portal vein, the dense particles can
colloidal particles that readily passed through be found on both sides of the endothelium
the wall of the sinusoid made it seem proba¬ and on the surface of the underlying hepatic
ble that there were openings in it that per¬ cells. Since the endothelium of the hepatic
mitted the blood plasma, but not the blood sinusoids also lacks a basal lamina in many
cells, to gain direct access to the surface of species, the particulate tracer evidently en¬
the hepatic cells. The electron microscope counters no barrier to passage through the
has provided visual confirmation of this inter¬ fenestrations.
pretation. In the common laboratory species Openings in the walls of the blood vessels
the endothelial cells have typical overlapping are rare in the circulatory system of verte¬
junctions in some places, but in others the brates, but it is now generally accepted that
attenuated margins of neighboring cells may the wall of the sinusoids is discontinuous in

-11’ E!ec,ron mic[°9raph of a sinusoid in rat liver fixed by vascular perfusion. The endothelium is extremely
wlsse E j uTrasfruSRes.^2!^ ]erlestrations ,ha< 9iva « a sievelike structure (see arrows). (Micrograph from
 THE LIVER AND GALLBLADDER • 689

most mammals, including the human. There

are, however, significant species differences
in the degree of endothelial discontinuity.
The hepatic sinusoids of sheep, goats, and
calves are reported to have a distinct basal
lamina and an endothelium with few fenes-
trae. In those species with a discontinuous
sinusoidal endothelium and no basal lamina,
there is no filtration barrier for macromole¬
cules or particles up to 0.5 fxm in diameter.
Chylomicra and very-low-density lipoprotein
particles can freely traverse inter- or trans-
cellular fenestrations.
Kupffer Cells. These stellate cells were
described by Kupffer in 1898 in liver stained
by a gold-chloride impregnation method.
They were depicted with their processes trav¬
ersing the sinusoids. Thus, they were thought
to lie within the sinusoid but fixed to the
endothelium. They frequently contain en¬
gulfed erythrocytes in various stages of dis¬
integration, and deposits of iron-containing
pigment. They actively phagocytize particu¬
late matter injected into the bloodstream (Fig.
27-10) and therefore are stained intensely
with such vital dyes as lithium carmine or
trypan blue. They also take up injected elec¬
tron-opaque particles of carbon or of thor¬
ium dioxide (Fig. 27—12). The Kupffer cells Figure 27-12. Liver of dog injected with India ink, showing
are therefore components of the diffuse uptake of carbon by the lining cells, x 675. (Courtesy of
A. J. Ladman.)
mononuclear phagocyte system. They may
retain for long periods phagocytosed mate¬
rial that cannot be digested by lysosomes.
Electron microscopic studies of Kupffer cells but have been reported in macrophages
cells in livers fixed by perfusion have done of other organs and in the Langerhans cells
much to clarify their cytological characteris¬ of the epidermis.
tics and their relations to other cells associ¬ The cytoplasm of Kupffer cells is richer in
ated with the sinusoids. They are usually organelles and more heterogeneous in ap¬
situated on the endothelium with processes pearance than that of endothelial cells. Clear
extending between the underlying endo¬ vacuoles of varying size and dense bodies
thelial cells. Their highly variable shape sug¬ presumed to be lysosomes are usually pres¬
gests that their form and relations to the ent. There is a juxtanuclear centrosome and
endothelium may change continually. They associated Golgi complex. Short cisternal pro¬
do not form desmosomes or other enduring files of granular endoplasmic reticulum are
specializations for attachment to the endo¬ scattered throughout the cytoplasm. These
thelial cells. The greater part of their irreg¬ are demonstrated with unusual clarity in
ular cell surface is exposed to the blood in preparations reacted for peroxidase activity
the lumen of the sinusoid. A thin glycocalyx (Fig. 27-13). Peroxidase is present in the
can be demonstrated. In addition to surface perinuclear cistern, in the lumen of the en¬
folds and slender villous projections, there doplasmic reticulum, and in the occasional
are peculiar sinuous invaginations of the annulate lamellae of Kupffer cells. This re¬
plasma membrane into the peripheral cyto¬ action serves clearly to distinguish them from
plasm. These have a central dense line endothelial cells, which have no peroxidase
between parallel membranes and a faint activity.
transverse striation, resulting in a highly It was formerly thought that increase in
characteristic appearance in thin sections that the numbers of Kupffer cells after experi¬
has led to their description as “vermiform mental stimulation of the reticuloendothelial
bodies.” These are never seen in endothelial system was due either to division of the
690 • THE LIVER AND GALLBLADDER

stimulated, the dividing Kupffer cells could

be shown to have originated from the bone
marrow of the donor. Similarly, when pairs
of unirradiated histocompatible mice were
maintained in parabiosis for several months,
karyotypic analysis of Kupffer cells revealed
limited numbers of partner-derived cells in
every instance. These experiments have led
to the conclusion that the Kupffer cells are
derived from a precursor in the bone mar¬
row, as are the free mononuclear phagocytes
of other organs. They are now considered to
be members of the mononuclear phagocyte
system.
Fat-Storing Cells. Cells that contain mul¬
tiple lipid droplets are found outside of the
sinuses, often occupying recesses between the
parenchymal cells but extending long proc¬
esses that are in contact with the endothe¬
lium. These cells have been described under
a variety of names: interstitial cells, lipocytes,
fat-storing cells, and stellate cells. They can be
stained with gold chloride (Figs. 27—14, 27—
15), and it is possible that some of the stellate
cells described by Kupffer were, in fact, fat-

Figure 27-13. Electron micrograph of a Kupffer cell,

showing a positive reaction for peroxidase in the nuclear
envelope and in cisternae of the endoplasmic reticulum.
This reaction serves to distinguish Kupffer cells from
endothelial cells. (Micrograph from Fahimi, D. J. Cell Biol.
47:247, 1970.)

preexisting cells of this type or to acquisition

of phagocytic capacity by endothelial cells
transforming into Kupffer cells. This latter
source is of course excluded since it is now
known that these are ontogenetically distinct
populations. The experimental stimulation of
phagocytic cells does result in a substantial
increase in the number of Kupffer cells in¬
corporating fT-thymidine. Thus, there seems
no doubt of their ability to increase by mito¬
sis. There is also compelling evidence that
Kupffer cells are replaced and are aug¬
mented by recruitment of monocytes from
the blood, which are transformed into Kupf¬
fer cells. In the relevant experiments, mice
were given sufficient whole body x-irradia-
tion to suppress division of their own cells.
They then received an intravenous injection
of bone marrow cells from another animal of Figure 27-14. Photomicrograph of normal rabbit liver
the same strain whose cells carried a chro¬ stained by Kupffer’s original gold impregnation method.
The perisinusoidal stellate or fat-storing cells are stained,
mosomal marker. When the reticuloendo¬
x 600. (Photograph by Wake, K. Am. J. Anat. 732:429,
thelial systems of the recipient mice were 1971.)
 THE LIVER AND GALLBLADDER • 691

Perisinusoidal Space (Space of Disse)

The relationship of the sinusoid lining to
the underlying liver cells has been settled by
the electron microscope. Formerly a contro¬
versy stemmed from the fact that in histolog¬
ical sections of human postmortem material,
a space, called the space of Disse, could be seen
between the sinusoid lining and the liver cells.
This was not apparent in biopsy material nor
in the usual sections of livers of laboratory
animals used in research. It was therefore
regarded by many histologists as a conse¬
quence of agonal or postmortem change in
the liver. In electron micrographs of well-
fixed material, the endothelium of the sinu¬
soids is not closely applied to a smooth pa¬
renchymal cell surface, as was previously
thought to be the case, but instead rests
lightly on the tips of a large number of
irregularly oriented microvilli on the surface
of the liver cell (Figs. 27—16, 27-17). There
is therefore a true perivascular space in the
normal liver into which the microvilli project.
The space of Disse described by pathologists
was evidently the result of an edematous
expansion of this space. The term has now
Figure 27-15. Fat-storing cells of rabbit liver after use of come to be applied freely to the narrow
Kupffer’s gold impregnation method, x 1800. (Photomi¬ perivascular space revealed by the electron
crographs from Wake, K. Am. J. Anat. 132:429, 1971.)
microscope in the normal liver.
Occasional unmyelinated nerve axons are
encountered in the space of Disse (Fig.
storing cells and not exclusively the phago¬ 27—18) and small bundles of collagen fibrils
cytic cells that now bear his name. forming the argyrophilic reticulum described
These stellate perisinusoidal cells have by light microscopists (Fig. 27—19), but the
some of the cytological characteristics of fi¬ space contains no true ground substance, and
broblasts. They tend to be more numerous plasma can apparently move freely through
in intermediate and peripheral portions of it. Although its content is plasma rather than
the hepatic lobule than they are in the central interstitial fluid, it must be considered an
zone. Their origin and functional significance interstitial space and not a lymphatic space,
are poorly understood. When exogenous vi¬ because it is not lined by lymphatic endothe¬
tamin A is administered it accumulates pref¬ lium. The space of Disse may, nevertheless,
erentially in the lipid droplets of the stellate be important in the formation of the abun¬
fat-storing cells. dant liver lymph.
Pit Cells. A fourth cell type associated with It is evident that direct access of the plasma
the sinusoids has recently been described. to the surface of the liver cell is a structural
The pit cell is also located in the perisinusoidal feature of great functional importance in the
space and attached to the endothelium. It active exchange of metabolites between the
possesses numerous short pseudopodia but liver and the bloodstream. The efficiency of
is nonphagocytic. Its most characteristic fea¬ this exchange is further promoted by the
ture is the presence in its cytoplasm of small increase in surface achieved by the microvilli.
dense granules resembling those of the en¬ From measurement of electron micrographs,
docrine cells of the gastrointestinal epithe¬ it is estimated that the length of the plasma
lium. Pit cells are present in small numbers membrane covering the microvilli and lining
in rat liver but have not yet been described the clefts between them is six times greater
in human liver. Their origin and function than the linear extent of cell surface meas¬
remain unknown. ured across the bases of the microvilli.
 Figure 27-16. Electron micrograph of plates of liver cells and intervening sinusoids. (Micrograph by E. Wisse.)

Figure 27 17. Electron micrograph of part of the surface of a rat liver cell bordering on a sinusoid Numerous irregularly
oriented microvilli project into a narrow space between the hepatic cell and the endothelium lining the sinusoid The
perivascular space is often called the space of Disse. Notice the small discontinuity in the lininq of the sinusoid (at
heavy arrow), x 18,000. (Courtesy of K. R. Porter and G. Millonig; labeling added.)
692
 THE LIVER AND GALLBLADDER • 693

Figure 27-18. In some species, nerve axons are found occasionally in the perisinusoidal space in apparent synaptic
contact with the hepatocytes. (Micrograph courtesy of E. Weihe and G. Metz.)

Figure 27-19. Photomicrograph of rat liver prepared by Pap’s silver method for demonstrating reticulum. A fine
meshwork of argyrophilic fibers is situated between the hepatic cells and the cells lining the sinusoids, x 1250.
694 • THE LIVER AND GALLBLADDER

Space of Disse

Lipoprotein

Agranular reticulum

Golgi complex
Granular
reticulum
canaliculus

Lysosome

Microbody

Figure 27-20. Drawing depicting the relationship of the liver cells to each other and to the sinusoids, and showing the
principal components of the hepatic cell as seen in electron micrographs. (Drawing by Sylvia Colard Keene.)

The Cytology of the Hepatic nucleus, but as many as 25 per cent are
Parenchymal Cells binucleate; 70 per cent or more of the nuclei
are tetraploid, and 1 to 2 per cent are octa-
The liver cells are polyhedral, with six or ploid. The nucleus is typically vesicular, with
more surfaces. The surfaces are of three a few scattered chromatin clumps and one or
sorts: those exposed to the perisinusoidal more prominent nucleoli. In electron micro¬
space; those exposed to the lumen of the bile graphs the liver cell nucleus has few features
canaliculus; and those in contact with adja¬ that distinguish it from the nuclei of other
cent liver cells (Fig. 27-20). The nuclei are cells. The chromatin is represented by ill-
large and round, with a smooth surface, but defined aggregations of filaments that appear
may vary in size from cell to cell. The varia¬ granular in section. Somewhat larger gran¬
tion in size has been shown to be an expres¬ ules 30 nm in diameter, called perichromatin
sion of polyploidy. Most cells have a single granules, are located near the masses of chro-
 THE LIVER AND GALLBLADDER • 695

matin. These are usually surrounded by a extensive smooth endoplasmic reticulum that
clear zone about 25 nm wide. These granules takes the form of a close-meshed plexus of
stain with uranyl acetate and indium and are branching and anastomosing tubules, some¬
therefore believed to contain nucleic acids. what variable in caliber (Fig. 27—23). Owing
The nucleoli consist of fine fibrils (6 nm) and to the thinness of the sections, however, the
dense granules (15 nm), and both of these continuity of the system is not always evident,
components are present in the anastomosing and it may appear as a congeries of separate
strands that constitute the nucleolonema. As profiles of irregular outline. Sites of conti¬
in other cell types, the nuclear envelope con¬ nuity between the rough- and smooth-sur¬
sists of two parallel membranes bounding a faced reticulum are frequently observed (Fig.
perinuclear cisterna. The conspicuous baso¬ 27-24). Small globules about 30 to 40 nm in
philic bodies seen in the cytoplasm with the diameter are often seen in the lumen of the
light microscope (Fig. 27-21) are found in smooth reticulum (Fig. 27-23). These rep¬
electron micrographs to be aggregations of resent the very-low-density serum lipoprotein
cisternae of an extensive rough endoplasmic (VLDL), which is synthesized in the liver and
reticulum (Fig. 27-22). The cisternae are released into the blood.
spaced somewhat farther apart and are less In histological sections the cytoplasm of
precisely parallel than the cisternae in the the liver cell presents an extremely variable
pancreas and other protein-secreting cell appearance, which reflects to some extent the
types. The cisternae are studded with nu¬ functional state of the cell. The principal
merous ribosomes, but the ends of their pro¬ source of variation is in the content of the
files are apt to be slightly expanded and free stored material—glycogen and fat. In the
of granules. In addition to the ribosomes preparation of histological sections, both fat
associated with the cytoplasmic membranes, and glycogen have been removed, but the
there are numerous polyribosomes free in presence of glycogen is indicated by irregular
the cytoplasmic matrix. empty spaces, and the presence of lipid is
The liver cell also contains a moderately represented by round vacuoles. By appropri-

Figure 27-21. Photomicrograph of rat liver stained with eosin and methylene blue. The deeply stained basophilic bodies
(at arrows) in the cytoplasm correspond to the aggregations of granular endoplasmic reticulum seen in electron
micrographs, x 1250.
696 • THE LIVER AND GALLBLADDER

Figure 27-22. Electron micrograph showing an area of granular endoplasmic reticulum from hamster liver, corresponding
to one of the basophilic bodies seen with the light microscope. The mitochondria and the granular reticulum are often
in close topographical relation to one another, x 34,000. (After Jones, A. L., and D. W. Fawcett. J. Histochem.
Cytochem. 74:215, 1966.)

ate methods of fixation, both fat and glyco¬ Mitochondria cannot be seen in the usual
gen can be preserved and stained. The con¬ histological preparation but can be revealed
tent of these materials in the liver may vary by special cytological techniques. They are
greatly with the diet or the time after feeding numerous and for the most part filamentous,
(Fig. 27-25). but they vary somewhat in size and shape in
When adequately preserved, glycogen ap¬ different parts of the lobule and in different
pears in electron micrographs of liver cells physiological conditions. The mitochondria
as dense aggregates or rosettes up to 0.1 pm are in no way unusual in their fine structure.
in diameter (alpha particles) composed of Lamellar or tubular cristae project into a
beta particles 20 to 30 nm in diameter. Gly¬ matrix of relatively low density. A number of
cogen is not uniformly distributed in the matrix granules are usually seen in each
cytoplasm but tends to be closely associated mitochondrial profile.
with the areas of smooth endoplasmic retic¬ The Golgi system of the cell consists of
ulum (Fig. 27-26). several parts, each situated near a bile can¬
Lipid occurs in the form of osmiophilic aliculus (Fig. 27—27). Each complex is made
droplets of varying size. These are few in up of three to five flat saccules or cisternae
number in the normal liver, but may be in close parallel array. The ends of the cis¬
dramatically increased after consumption of ternae are often dilated and contain numer¬
alcohol or other hepatotoxic substances. The ous moderately dense granules 30 to 60 nm
lipid droplets are not limited by a membrane. in diameter. These are identical to the low-
They may occur anywhere in the cytoplasm density lipoprotein particles observed in the
and have no special topographical relation to smooth reticulum. Associated with each of
any of the organelles other than mitochon¬ fthe Golgi complexes are several membrane-
dria, which may be closely applied to the limited dense bodies 0.2 to 0.5 pm in diam¬
surface of the droplet. eter. These peribiliary dense bodies contain
Figure 27-23. Electron micrograph of hepatocyte cytoplasm showing smooth-surfaced reticulum containing small,
spherical dense particles representing newly synthesized, very-low-density serum lipoprotein. Also present are two
microbodies or peroxisomes with eccentrically placed nucleoids. (Micrograph by R. Bolender.)
'Wfc ^ Si f*. V '-4
H? Oft fXA**.

.tr ' 81*^ **

Figure 27-24 Electron micrograph of a small area of hepatocyte cytoplasm including several mitochondria, cisternal
profiles of granular endoplasmic reticulum, and (at upper right) a close-meshed plexus of agranular reticulum. (Micrograph
by R. Bolender.) 0gy
Figure 27-25. Dietary differences in amount of stored glycogen are clearly illustrated by comparison of these
photomicrographs of rat liver. A, Liver of an animal fasted for 2 hours, containing 8.2 per cent glycogen. B, Liver of an
animal fasted for 21 hours, containing 0.9 per cent glycogen. (From Cardell, R., J. Larner, and M. B. Babcock: Anat.
Rec., 777:23, 1973.)

Figure 27-26. An area of hepatocyte cytoplasm containing a high concentration of glycogen in alpha granules. The
glycogen is always closely associated with profiles of smooth endoplasmic reticulum.
698
 THE LIVER AND GALLBLADDER • 699

histochemically demonstrable acid hydrolases functions. In the liver, despite its diverse
and therefore correspond to the lysosomes activities, the cells are all very similar in
isolated from homogenates of liver. appearance. All the parenchymal cells are
Scattered throughout the cytoplasm of the probably capable of carrying out all the func¬
hepatocyte are peroxisomes (Figs. 27-28, 27— tions of the liver. However, cytologists have
29). These are spherical bodies 0.2 to 0.8 pm long believed that the degree of their activity
in diameter enclosed by a membrane. In the under normal conditions depends primarily
laboratory rodents they contain a crystalline on their location within the lobule. The clas¬
nucleoid eccentrically placed in a moderately sical lobule can be divided into concentric
dense, finely granular matrix. The nucleoids zones on the basis of the cytological evidences
have been isolated and found to contain of activity of the cells. A zone of varying
uricase. This component is lacking in peroxi¬ width around the periphery of the lobule has
somes of the human. The positive staining of been designated the “zone of permanent
peroxisomes with the histochemical reaction function.” Next there is an intermediate
for peroxidase is attributed to the enzyme “zone of varying activity” and finally a narrow
catalase, which is present in the matrix. Per¬ zone around the central vein that is called
oxisomes are respiratory organelles that con¬ the “zone of permanent repose.” These zones
tain hydrogen peroxide generating oxidases correspond respectively to the portions of
and catalase. They are involved in plasmalo- the lobule where the liver cells are most
gen synthesis and oxidation of very-long- favorably situated, are intermediate, and are
chain fatty acids, but their significance in liver least favorably situated with respect to the
cell metabolism is still poorly understood. sequence in which oxygen and nutrients
reach them in the blood entering the sinu¬
soids from the terminal branches of the he¬
Zonation Within the Liver Lobule patic artery and portal vein at the periphery
of the lobule (at the center of the functional
In organs with multiple functions, it is unit of Rappaport). This zonation is quite
often possible to demonstrate cytological dif¬ striking in some species but less obvious in
ferences between cells performing different others.

Figure 27-27. Electron micrograph of the peribiliary region of adjacent hepatocytes, showing typical location of the
Golgi complex. Low-density lipoprotein (VLDL) is seen in distended cisternae of the Golgi and in the lumen of the
smooth endoplasmic reticulum (at arrows). (Micrograph courtesy of R. Bolender.)
700 • THE LIVER AND GALLBLADDER

staining so lightly that they are frequently

seen only with difficulty in mitochondrial
preparations. In the peripheral zone, how¬
ever, they are large, deeply staining spheres
or short rods that may crowd the cytoplasm.
In the intermediate zone, the rods become
progressively elongated until, as one ap¬
proaches the central zone, they are slender
filaments. The width of the intermediate
zone varies with the state of the diurnal tide
of alimentation. In other species, including
man, such changes are not demonstrable.
Under certain conditions, both pathologi¬
cal and physiological, fat may accumulate in
the liver. Usually it appears first in the cells
of the central zone as small spherical drop¬
lets; these become progressively larger by
coalescence as well as by further accumula¬
tion, until the cell may be distended by a
single large drop. In certain conditions, no¬
tably some sustained dietary deficiencies, fat
is deposited in the peripheral rather than in
the central zone. In both cases, fat disappears
when the condition responsible is corrected.
Position in the lobule may not be the only
determining factor in the relative activity of
liver cells. Application of the fluorescent an¬
tibody technique to localize the sites of pro¬
Figure 27-28. Photomicrograph of rat liver stained by a duction of plasma albumin has shown
histochemical reaction for demonstration of peroxidase marked differences among liver cells imme¬
activity. The distribution of reactive granules corresponds diately adjacent to one another, and the dis¬
to that of the microbodies or peroxisomes identifiable in
tribution of synthetically active cells in this
electron micrographs. (Micrograph from Fahimi, D. J. Cell
Biol. 43:275, 1969.) case appears to bear little relation to the
zonation within the lobule.

Typically, after the feeding of a large meal, Bile Canaliculi

glycogen is deposited first in the zone of
permanent function at the periphery of the Minute canals run between liver cells
lobule. During active digestion, glycogen fills throughout the parenchyma. As a rule, a
cells progressively farther into the interme¬ single canaliculus is found between each ad¬
diate zone until, in extreme cases, all but the jacent pair of cells. Thus, in a plate of liver
cells immediately adjacent to the central vein cells one cell thick, these bile canaliculi form
may be filled with it. With the conclusion of a network having hexagonal meshes with a
digestion, carbohydrate is returned to the single cell in each mesh. Because the laminae
blood, as needed, by removal of the glycogen, of parenchymal cells branch and anastomose,
beginning at the most centrally located de¬ the canaliculi form an extensive three-dimen¬
posits. If the fast is prolonged, glycogen may sional net with polyhedral meshes (Fig. 27—
ultimately disappear completely at the pe¬ 30). In amphibians, there are small branches
riphery. Thus, in an animal such as the that extend between cells from a core canal¬
mouse, which normally feeds at night, there iculus and end blindly. It is now generally
is a diurnal tide of glycogen within the lobule, agreed that there are no such blind branches
which may be spectacularly accentuated by in the mammalian liver. Instead the canaliculi
restricting the feeding time to one hour, with form a continuous network without interrup¬
the animal fasting the rest of the day. tion from lobule to lobule throughout the
Accompanying this tide is a corresponding parenchyma. The membrane lining the can¬
change in the mitochondria. Those in the aliculi is a site of adenosine triphosphatase
central zone are thin, elongated, and sparse, activity, and histochemical reactions for this
Figure 27-29. Microbodiesor peroxisomes of rodent liver are limited by a single membrane and contain a “nucleoid”
consisting of a paracrystalline array of tubular subunits. (Micrograph by R. Wood.) When stained for peroxidase activity,
the matrix of the microbodies reacts intensely but the nucleoid remains unstained. (From Fahimi, D. J. Cell Biol. 43:275,
1969.)

Figure 27-30. Scanning electron micrograph of rat liver fixed by perfusion and then broken open to reveal its internal
architecture. The micrograph affords a three-dimensional view of the plates of polygonal hepatic cells alternating with
sinusoids. Where the liver plates have been broken, the bile canaliculi can be identified on the contact surfaces of the
hepatic cells (at arrows). (Micrograph courtesy of M. Karnovsky.) 7Q1
702 • THE LIVER AND GALLBLADDER

Figure 27-32. Scanning micrograph of a cast of a branch

Figure 27-31. Photomicrograph of rat liver, showing the
of the biliary tree, showing the arborizing pattern of bile
branching pattern of bile canaliculi, which are demon¬
canaliculi. At the arrow is a canal of Herring communicat¬
strated here by their positive histochemical staining re¬
ing with the duct of a portal triad. (Micrograph from
action for adenosine triphosphatase, x 200. (Courtesy of
Murikami, T. Dig. Dis. Sci. 25:609, 1980.)
A. Novikoff.)

enzyme provide a useful method for selec¬ ject into its lumen. Along the margins of the
tively staining this system of minute intercel¬ canaliculus the membranes of the opposing
lular canals (Figs. 27-31, 27—32). cells come into close contact and form an
From observations with the light micro¬ occluding junction comparable with the zon¬
scope, it seemed reasonable to assume that ula occludens of other epithelia. These two
the bile canaliculi were distinct entities having bands of tight junction evidently seal the
walls of their own. Indeed, it was suggested commissures of the canaliculus and prevent
that their hexagonal network around the its contents from escaping into the intercel¬
hepatic cells contributed significantly to the lular cleft on either side. A narrow zone of
structural stability of the liver. This erro¬ cytoplasm immediately adjacent to the can¬
neous interpretation was abandoned when aliculus is free of organelles and has the finely
the electron microscope revealed that the fibrillar structure characteristic of a gelated
lumen of the bile canaliculus is merely an ectoplasmic layer.
expansion of the intercellular space and that In addition to the zonulae occludentes ad¬
its wall is simply a local specialization of the jacent to the canaliculus, a number of gap
surfaces of adjoining hepatic cells (Fig. 27- junctions are found on the boundaries be¬
27). Over most of their length, the apposed tween adjoining hepatic cells. These sites of
membranes of the two cells are relatively low electrical resistance permit communica¬
straight and separated by an intercellular tion between cells and provide for coordina¬
cleft about 15 nm wide. At the site of the bile tion of their physiological activities. These
canaliculus they diverge to form a canalicular junctional specializations are visible in elec¬
intercellular space 0.5 to 1 pm in diameter. tron micrographs of thin sections, but they
The portion of the cell membrane bordering are studied to best advantage in freeze-frac¬
on this space bears short microvilli that pro¬ ture preparations (Fig. 27-33).
 THE LIVER AND GALLBLADDER • 703

Figure 27-33. A, Electron micrograph of a portion of the boundary between two liver cells in thin section, showing the
cell membranes converging at the arrows and coming into close apposition to form a nexus or gap junction between
the arrows. (Micrograph courtesy of R. Wood.) B, Surface replica of an area of liver cell membrane freeze-fractured to
expose its P-face. In addition to the population of randomly distributed intramembranous particles, there are two gap
junctions exhibiting closely packed particles of uniform diameter. (Micrograph courtesy of A. Yee.)

The bile canaliculi vary in diameter, be¬ They have small diverticula that expand
coming somewhat distended with active se¬ against the adjacent parenchyma and are
cretion and more or less collapsed with de¬ applied tightly to it. The bile capillaries con¬
creasing activity. When distended, the tinue between the hepatic cells to empty into
microvilli are more widely scattered and ap¬ the lumen of the diverticula. The structure
pear to be shorter, and when the canaliculi of this transitional region of the duct system
are collapsed, the microvilli may pack the is well demonstrated if the cholangioles and
lumen so completely that it is virtually oc¬ bile canaliculi are distended, as they are fol¬
cluded. This may explain why the canaliculi lowing occlusion of the bile duct.
are hard to see with the light microscope.
The junction of the bile canaliculus with Connective Tissue Stroma
the bile duct system is not easily demon¬
strated. The fine terminal branches of the For an organ of its size, the liver has
bile duct leave the portal canal with the remarkably little stroma. A small amount of
terminal branch of the portal vein and pen¬ dense connective tissue forms a thin layer
etrate the parenchyma between two lobules (Glisson’s capsule) underlying the organ’s in¬
(i.e., in the core of a functional unit of Rap- vestment of peritoneal mesothelium. It ex¬
paport.) They are so small and have such tends into the organ in the portal canals
thin walls that they are recognized only with and follows them to their finest terminals en-
difficulty. These channels are quite different sheathing the branches of the portal vein,
in appearance from the smallest bile ducts hepatic artery, and bile duct. It also contains
and are called bile ductules or cholangioles. the network of lymphatics that drains lymph
704 • THE LIVER AND GALLBLADDER

from the liver. In these sites it is typical thirds of the liver can be removed and in a
connective tissue with dense collagenous fi¬ few days most of this tissue will be replaced.
bers and occasional fibroblasts. Within the Similarly, after administration of hepatotoxic
lobule, the only stromal structure is a network agents, notably the chlorinated hydrocar¬
of reticular fibers between the sinusoid lining bons, a substantial part of each lobule may
and the hepatic cell plates (Fig. 27-19). This be destroyed, and in this case too the lost
can be demonstrated by various techniques, tissue is rapidly replaced.
but especially well by some of the silver im¬ Regeneration after partial hepatectomy re¬
pregnation methods. At the periphery of the sults from cell division occurring throughout
lobule, where terminal branches of the portal the remaining liver mass. Therefore, the
veins enter the sinusoids, the collagenous original pattern of liver lobes is not restored.
fibers of the portal canals are continuous with Most of the research on regeneration has
the network of reticular fibers surrounding been done on the rat and the mouse, in
the sinusoids. This reticulum is the only sup¬ which the amount restored is usually as great
porting tissue of the liver parenchyma. It as the amount removed. In other animals,
contains no associated hbroblasts, the fibers the amount regenerated may be considerably
apparently being formed by the sinusoid lin¬ less, in inverse ratio to the size of the animal.
ing cells. Central necrosis .after a toxic dose of car¬
bon tetrachloride may involve as much as one
third to one half of each lobule and is re¬
Lymph Spaces
markably uniform throughout the liver. Only
The liver produces a large amount of the parenchymal cells are killed, leaving the
lymph. From one quarter to one half of the sinusoid linings intact, so that the circulation
total volume of the thoracic duct lymph through the lobule is maintained. The necro¬
comes from the liver, less from the intestine tic cells are removed by autolysis, while the
via the mesenteric lymphatics, and still less cells in the remaining part of the lobule
from the other organs of the posterior part divide rapidly by mitosis. The mass of normal
of the body. The hepatic lymph differs from liver tissue increases until in five or six days
the rest of the lymph in that it contains a the original lobular architecture of the liver
large amount of plasma protein, with the is completely restored. If the dose of carbon
ratio of albumin to globulin a little higher tetrachloride is repeated at regular intervals
than in the plasma. The network of lym¬ when regeneration is still in progress, thus
phatics follows the portal vein to its finest repeatedly producing new injury before the
terminal branches. Here it ends in the con¬ old has been repaired, fibrosis occurs, and if
nective tissue of the portal canals. No lym¬ it is continued long enough, cirrhosis of the
phatics have been demonstrated within the liver ensues.
liver lobules. It is believed that plasma trav¬ If the cellular injury is at the periphery of
ersing the discontinuities in the sinusoid lin¬ the lobule, as it is after bile duct occlusion or
ing and entering the space of Disse moves after treatment with other hepatotoxic
along toward the periphery of the lobule, agents, cell division occurs throughout the
bathing the microvilli of the hepatic cells as remaining tissue, but there is also considera¬
it goes. It then percolates into the extracel¬ ble mitotic activity in the epithelium of the
lular spaces around the interlobular twigs of bile ductules and smaller bile ducts, with a
the bile duct and the portal vein. It thus corresponding increase in their number. The
becomes the tissue fluid of portal canals, and ductules penetrate into the injured periph¬
the liver lymph is drained by the lymphatic eral part of the lobule, to reestablish the
capillaries that accompany ducts and blood pathway for bile drainage interrupted by the
vessels. death of the peripheral cells and dissolution
of their bile canaliculi. If the injury continues,
the increase in ductules and ducts may de¬
REGENERATION velop into a spectacular proliferation of the
bile ducts. If it does not continue, the normal
The liver parenchyma, compared with that architecture of the liver is rapidly restored,
of many other organs, is a fairly stable pop¬ with removal of the excess of the new ducts
ulation of cells that rarely need to be replaced and ductules. It is not clear whether the duct
in a normal adult. It is, however, capable of cells atrophy and disappear or transform into
spectacular regeneration. In the rat, two parenchymal cells. In repair after severe in-
 THE LIVER AND GALLBLADDER • 705

jury, the occurrence of cells of intermediate verted in the liver into glucose and then to
cytological appearance gives credence to the glycogen. As the need arises, glycogen is
latter possibility. broken down to glucose again in a process
The mechanism of initiation of mitosis in catalyzed by the enzyme phosphorylase. This
a normally quiescent tissue and of its cessa¬ enzyme usually occurs in an inactive form
tion when the lost tissue has been replaced but is specifically activated by the hormones
has been extensively investigated. Partial hep- epinephrine and glucagon, which act upon
atectomy with the ensuing regeneration has the liver and cause it to release glucose into
been a favored system in which to study this the blood.
and a variety of related problems. Many of the enzymes involved in glycogen-
esis and glycogenolysis are free in the cyto¬
plasmic matrix. These functions, therefore,
HISTOPHYSIOLOGY OF THE LIVER cannot be attributed to any particular cell
organelle. However, in electron micrographs,
The liver is a highly vascular organ receiv¬ the glycogen is usually localized in areas of
ing about 100 ml of blood from the portal cytoplasm rich in smooth endoplasmic retic¬
vein and 400 ml from the hepatic artery each ulum. The exact significance of this close
minute. In flowing through the sinusoids the topographical relationship is not yet clear,
blood is exposed to about 1.2 x 107 phago¬ but the enzyme glucose-6-phosphatase, which
cytic Kupffer cells per gram of liver. These is known to reside in these membranes, may
remove cellular debris and foreign particu¬ participate in some way in the release of
late matter. The liver therefore has an im¬ glucose to the blood.
portant blood-filtering function. The liver also plays a major role in the
The hepatic sinusoids normally present a metabolism and transport of lipids and in the
very low resistance to blood flow. However, maintenance of normal lipid levels in the
in heart failure pressure may increase in the circulating blood. The lipids in the blood
veins draining the liver and the sinusoids plasma are derived from ingested food, from
may then become distended with as much as mobilization of fat reserves in adipose tissue,
400 ml more blood than they normally con¬ or from synthesis from carbohydrate or pro¬
tain. Under these conditions the liver en¬ tein in the liver. The main vehicle for the
larges markedly. Conversely, after blood loss transport of lipids is the plasma lipoprotein,
from hemorrhage a substantial volume of and it is in the liver that the transformation
blood may be contributed by the liver to the of lipids into serum lipoprotein takes place.
general circulation to offset the blood loss. Small spherical particles 30 to 100 nm in
The liver thus serves as an important site of diameter are seen in electron micrographs of
blood storage. liver in agranular terminal expansions of the
In the disease cirrhosis, fibrosis greatly in¬ granular reticulum, in tubular elements of
creases the resistance to blood flow through the smooth reticulum, in exocytotic vesicles
the liver. Back pressure in the portal system at the cell surface, and in the space of Disse.
results in ascites—transudation and accumu¬ I hese particles contain a triglyceride lipid
lation of protein-rich fluid in the peritoneal core surrounded by a more water-soluble,
cavity. polar surface coat of protein, phospholipid,
Because of the remarkable range of its and cholesterol. In the isolated perfused
biochemical functions in intermediary metab¬ liver, the number of these particles is strik¬
olism and its strategic location in the circu¬ ingly increased when fatty acids are added to
lation, the liver is a vital organ for processing the perfusate. They represent very-low-density
nutrients absorbed from the gastrointestinal lipoproteins (VLDL) being formed in the liver
tract and for transforming them into mate¬ and released into the space of Disse. Triglyc¬
rials needed by the other specialized tissues erides are formed in the smooth reticulum
of the body. One of its most important func¬ from fatty acids, and these are combined
tions is maintenance of the normal blood there with protein synthesized in the granu¬
glucose concentration. Liver cells take up lar reticulum to form lipoprotein particles
glucose from the blood and by means of a (Figs. 27-23, 27-34). There is also good evi¬
series of enzymatic reactions polymerize it to dence that the reticulum, particularly the
form glycogen, the storage form of carbo¬ smooth form, plays an important part in the
hydrate. Simpler compounds, such as lactic synthesis of cholesterol in the liver. An ex¬
acid, glycerol, and pyruvic acid, can be con¬ perimentally induced increase in the abun-
706 • THE LIVER AND GALLBLADDER

Lipoprotein The hepatocytes have specific membrane re¬

ceptors for several of these proteins that are
taken up by receptor-mediated endocytosis,
moved through the cytoplasm by a vesicular
transport system, and secreted into the bile
canaliculi. Other plasma proteins, for which
there are no plasmalemmal receptors, are
taken up by nonselective fluid-phase endo¬
cytosis and are similarly transported. Using
radioactively labeled compounds, the trans-
cellular movements of the transport vesicles
can be traced autoradiographically. Direct and
indirect pathways are described (Fig. 27-35).
In the former, the endocytic vesicles form at
the sinusoidal surface, traverse the cell, and
fuse with the membrane of the bile canalic¬
ulus, discharging their ligands intact. In the
Figure 27-34. Diagram illustrating intracellular pathway indirect pathway, the vesicles fuse en route
of synthesis and release of very-low-density serum lipo¬ with small primary lysosomes that modify or
protein (VLDL). Fatty acids from the blood are esterified
in the smooth reticulum to form triglycerides. These are degrade the vesicle contents. The catabolic
combined with protein synthesized on ribosomes associ¬ products are then released into the bile.
ated with the rough reticulum. The particles are released This function of the hepatocytes is of spe¬
into the space of Disse and enter the blood in the cial interest in relation to the secretory im¬
sinusoids.
mune system of the intestine. As stated in
the foregoing chapter, immunoglobulin A
dance of smooth reticulum is accompanied synthesized by plasma cells in the lamina
by an enhanced capacity to synthesize choles¬ propria of the gut is complexed with a secre¬
terol from acetate. tory component in the intestinal epithelium
The liver is the site of synthesis of plasma and is released into the lumen to serve in the
proteins, and the rate of their production is immunological defense of the gut. However,
quite substantial. Studies on the isolated per¬ only a small fraction of the antibody pro¬
fused organ indicate that the liver of an adult duced in the lamina propria takes this direct
rat synthesizes in excess of 13 mg of albumin route to the lumen. The remainder is carried
per day. It is likely that the organelle princi¬ in the lymph to the thoracic duct and thence
pally involved is the granular endoplasmic into the general circulation. Much of the IgA
reticulum. A fine flocculent substance is in the plasma is destined to reach the intes¬
sometimes observed in the lumen of the cis- tinal lumen via the hepatobiliary pathway
ternae, and protein with the properties of described above. Immunoglobulin A is pres¬
albumin has been identified in liver micro- ent in the bile at five times its concentration
somes, but the entire secretory pathway of in the plasma. If the bile duct is surgically
plasma proteins has not been worked out. occluded in animal experiments, the level of
The liver also synthesizes many of the secretory IgA in the blood plasma rises mark¬
substances involved in blood clotting—fibrin¬ edly while its level in the gut falls to one
ogen, prothrombin, Factor III, and several tenth its normal concentration. Thus, biliary
others. The concentrations of these sub¬ IgA makes a very major contribution to the
stances in the blood can be used clinically as supply of antibodies in the intestinal lumen.
a measure of liver cell function. Liver disease The secretory component synthesized in the
may result in impaired synthesis of clotting hepatocytes is inserted as a transmembrane
factors and a tendency to bleed excessively protein in the plasmalemma at the sinusoidal
from minor injuries. surface of the cell, and its exposed portion
In addition to its many functions in nutri¬ serves as the receptor for plasma IgA that is
tional metabolism and bile secretion, the liver taken up by endocytosis and transported to
takes up and either catabolizes or secretes the bile.
some 20 proteins and glycoproteins that cir¬ The liver is responsible for the metabolism
culate in the blood. These include immuno¬ of a large variety of lipid-soluble drugs, in¬
globulins, albumin, transferrin, insulin, a2- cluding the barbiturates commonly used as
macroglobulin, and epidermal growth factor. sedatives. The enzymes responsible for the
 THE LIVER AND GALLBLADDER • 707

Figure 27-35. Schematic representation of the uptake, intracellular transport, and biliary secretion of four proteins and
glycoproteins of the plasma. Low-density lipoproteins (LDL) and epidermal growth factor (EGF) are taken up mainly by
fluid-phase endocytosis and are modified in transit by lysosomes. Insulin and immunoglobulin A take the direct path.
Taken up in vesicles by receptor-mediated endocytosis, they are released into the bile canaliculi unchanged. (Redrawn
and modified after Jones, A. L. K., R. H. Reston, and S. V. Burwen. In Popper, H., and F. Schaffner, eds.: Progress in
Liver Disease. Vol. 7. New York, Grune & Stratton, 1982.)

degradation of these compounds are local¬ ment bilirubin, which originates from the
ized mainly in the smooth-surfaced micro- breakdown of aged red blood cells being
some fraction of liver homogenates and eliminated from the circulation by Kupffer
hence reside in the smooth reticulum of the cells and phagocytes of the spleen. This sub¬
intact liver cell. Moreover, administration of stance, carried in the blood, is conjugated
such drugs induces a marked increase in the with glucuronide in the liver by enzymes in
smooth-surfaced membranes of the cyto¬ the endoplasmic reticulum. The conjugate,
plasm and this morphological change is ac¬ bilirubin glucuronide, is excreted into the
companied by a concomitant increase in the bile. In the intestine, it is reduced by the
drug-metabolizing enzymes. These changes action of bacteria into a group of compounds
do not represent toxic effects of the drug but collectively referred to as urobilinogens. While
rather an adaptive response of the liver cell, most of these are eliminated in the feces,
which enhances its efficiency in eliminating some are reabsorbed and reexcreted in the
the inducing drug. These morphological and bile. When bilirubin accumulates in the blood
biochemical changes thus appear to be the (hyperbilirubinemia), a person becomes jaun¬
basis of drug tolerance—the progressive loss of diced. This may result from (1) increased
effectiveness of a drug with continued use. production of bilirubin beyond the capacity
Certain of the steps in the metabolism of of the liver to excrete it—e.g., in hemolytic
steroid hormones in the liver also take place jaundice; (2) decreased uptake of bilirubin
in the endoplasmic reticulum and probably by the liver; (3) disturbance in conjugation
depend on some of the same hydroxylating of bilirubin in the liver—e.g., in jaundice of
enzymes that are involved in the metabolism the newborn; (4) interference with excretion
of exogenous drugs. due to obstruction of the bile duct system.
An important excretory function of the Determination of the relative amounts of
liver is the uptake and excretion of the pig¬ conjugated and unconjugated bilirubin in the
708 * THE liver and gallbladder

blood is therefore an important means of thrown into many folds and is said to secrete
evaluating liver function and of distinguish¬ an atypical variety of mucus. The scanty
ing among the several causes of jaundice in subepithelial connective tissue contains large
patients. numbers of elastic fibers and some lymphoid
A major function of hepatic cells is the cells. Many of these migrate through the
secretion of bile—a complex fluid consisting epithelium and pass into the lumen. Scat¬
of cholesterol, lecithin, fatty acids, electro¬ tered bundles of smooth muscles first appear
lytes, and bile salts. Bile salts, the most im¬ in the common bile duct. They run in the
portant component, are synthesized from longitudinal and oblique directions, and form
cholesterol by conversion to cholic and che- an incomplete layer around the wall of the
modeoxycholic acids, which then combine duct. As it nears the duodenum, the smooth
with glycine or taurine to form the corre¬ muscle layer of the ductus choledochus be¬
sponding conjugated acids. Their salts are comes more prominent, and at the duo¬
secreted in the bile in the amount of about denum its intramural portion functions as a
0.5 g/day. The cell organelles involved in the sort of sphincter, regulating the flow of bile.
biosynthetic pathway of bile acids are still
poorly understood. The bile salts have a
detergent or emulsifying action on ingested
fat. They form micelles with fatty acids and
THE GALLBLADDER
monoglycerides, facilitating their absorption
by the intestinal epithelium. In the absence The gallbladder is a pear-shaped, hollow
of bile secretion, much of the fat in the diet viscus occupying a shallow fossa on the infe¬
is not absorbed and passes out in the feces. rior surface of the liver. It consists of a
fundus, a body, and a neck that continues
into the cystic duct. Normally it measures
approximately 10 by 4 cm in the human, and
BILE DUCTS has a capacity of 40 to 70 ml. It shows marked
variations in shape and size and is frequently
The constituents of the bile are emptied the seat of pathological processes that change
into the bile canaliculi, which communicate its size and the thickness of its wall. The
with the interlobular bile ducts by the canals function of the gallbladder is to store, con¬
of Hering. The finest radicles of the bile centrate, and release into the duodenum the
ducts are 15 to 20 pm in diameter and have 600 to 1000 ml of bile secreted each day by
a small lumen surrounded by cuboidal epi¬ the liver.
thelial cells. They do not have a striated The gallbladder is covered over all but its
border. The cells show occasional mitoses. hepatic surface by a serosa continuous with
The interlobular bile ducts form a richly that of the liver. Its wall consists of a subse-
anastomosing network that closely surrounds rosal connective tissue layer overlying a layer
the branches of the portal vein. Closer to the of smooth muscle. Deep to this is the mucosa
porta, the lumen of the ducts of the second composed of the epithelium and its highly
order gradually becomes larger, while the vascular lamina propria. The mucosa is pli¬
epithelium becomes taller and has an accu¬ cated into numerous convoluted folds of
mulation of mitochondria at the base and varying height that demarcate narrow bays
another near the free border. These cells or clefts. In sections of the contracted gall¬
commonly contain fat droplets. Lymphocytes bladder these mucosal folds are tall and close
are frequently seen migrating through the together. When the organ is distended they
epithelium into the lumen. As the ducts be¬ are short and some distance apart. The
come larger, the surrounding layer of colla¬ changes in topography occurring in different
genous connective tissue becomes thicker and states of filling and contraction are most
contains many elastic fibers. At the transverse dramatically illustrated in surface views of
fossa of the liver, the main ducts from the the mucosa with the scanning electron micro¬
different lobes of the liver fuse to form the scope. In the contracted gallbladder each
hepatic duct, which, after receiving the cystic highly sinuous fold follows a course generally
duct, continues from the gallbladder to the parallel to that of neighboring folds (Fig. 27—
duodenum as the common bile duct (ductus 36). In the distended condition the folds are
choledochus). The epithelium of the extrahe- reduced to low ridges and it is apparent that
patic ducts is tall columnar. The mucosa is they branch and anastomose to form a net-
 THE LIVER AND GALLBLADDER • 709

Figure 27-36. Scanning micrograph of the contracted gallbladder. The mucosa is thrown up into highly convoluted
folds. A histological section through these has the appearance shown in the inset. Compare with Figure 27-37.
(Micrographs from Castellucci, M. J. Submicrosc. Cytol. 12:375, 1980.)

Figure 27-37. Scanning micrograph of the mucosal surface of a normally distended gailbladder. Relatively low mucosal
folds have the pattern of a network with polygonal meshes. A histological section of the gallbladder in this state has the
appearance shown in the inset. Compare with Figure 27-36. (Micrographs from Castellucci, M. J. Submicrosc. Cytol.
72:375, 1980.)
710 • THE LIVER AND GALLBLADDER

work outlining shallow polygonal recesses epithelium is 15 to 20 nm wide and is sealed

(Fig. 27-37). The loose organization of elastic near the lumen by a typical zonula occludens.
and reticular fibers of the lamina propria Toward the base, the intercellular space may
provides the flexibility to accommodate these be narrow or greatly widened. The width of
marked changes in mucosal topography. the intercellular clefts at the base depends on
The epithelium consists of a single layer of the functional state of the gallbladder epithe¬
tall columnar cells, with oval nuclei (Fig. 27— lium (see below).
38). The cytoplasm stains faintly with eosin. In the lamina propria and in the perimus-
An inconspicuous striated border is seen in cular layer near the neck of the gallbladder
histological sections, and with the electron there are simple tubuloalveolar glands. Their
microscope the apical surface of the colum¬ epithelium is cuboidal and clear, and the dark
nar cells is found to have numerous microvilli nuclei are compressed at the base of the cell.
(Fig. 27—39). These are somewhat shorter They thus stand out sharply against the
and less regular in their orientation than are darker, tall columnar epithelium of the gall¬
those of the striated border of the intestinal bladder. These glands are said to secrete
epithelium. At the tips of the microvilli the mucus.
membrane bears minute filiform appendages Outpocketings of the mucosa in this region
similar to those making up the glycocalyx on have sometimes beon confused with glands.
intestinal mucosa and on various other epi- These are lined by and are continuous with
thelia. the surface epithelium and extend through
The lateral cell boundaries are relatively the lamina propria and the muscular layer.
straight at the apical portion of the epithe¬ They are called Rokitansky-Aschoff sinuses and
lium, but from the level of the nucleus to the probably are indicators of a pathological
basal lamina there is a complex plication and change in the wall of the organ that permits
interdigitation of the cell surface. The inter¬ an evagination of the mucosa through en¬
cellular space in the upper portion of the larged meshes of the submucosal network of

sSj^ros^CytoL^ays^Igoo.)0* mUC0Sa °f rabbit 9allbladder' (Micrograph from Castellucci, M. J.

 THE LIVER AND GALLBLADDER • 711

Figure 27-39. Scanning electron micrograph of the luminal surface of gallbladder epithelium. The convex apical ends
of the cells are covered with short microvilli. (Micrograph from Mueller, J. C., A. L. Jones, and J. A. Long. Gastroenterology
63:856, 1972. © 1972 The Williams & Wilkins Co., Baltimore.)

smooth muscle. They are not found in the some of them connect with the bile ducts.
fetal gallbladder and apparently are acquired They never connect with the lumen of the
in adult life. gallbladder and are probably aberrant bile
The smooth muscle beneath the epithelium ducts laid down during the embryonic devel¬
is an irregular network of longitudinal, trans¬ opment of the biliary system. They have been
verse, and oblique fibers, with an associated called Luschka ducts, to distinguish them from
network of elastic fibers. The spaces between the epithelial outpouchings described above.
the bundles of muscle fibers are occupied by The cystic duct continues from the neck of
collagenous, reticular, and elastic fibers, and the gallbladder for a distance of 3 to 4 cm
occasional fibroblasts. and joins the hepatic duct to form the common
A fairly dense connective tissue layer ex¬ bile duct (ductus choledochus). Coursing down¬
ternal to the muscularis completely sur¬ ward behind the head of the pancreas, it
rounds the gallbladder and is in places con¬ approaches the pancreatic duct (ductus pancrea-
tinuous with the interlobular connective ticus). The two pass together through an
tissue of the liver. It contains many collagen¬ opening in the muscularis of the descending
ous and a few elastic fibers, scattered fibro¬ portion of the duodenum. In their oblique
blasts, macrophages and lymphoid cells, and course through the submucosa, the two ducts
small lobules of fat cells. The blood vessels, unite to form the hepatopancreatic ampulla (am¬
nerves, and lymphatics supplying the organ pulla of Vater), which opens into the duodenal
run in this layer and send branches into and lumen at the tip of a small papilla. In the
through the muscular layer to the mucosa. wall of the duodenum, the bile and pan¬
Not infrequently, particularly on the he¬ creatic ducts and ampulla are encircled by a
patic surface of the gallbladder and near its common annular band of smooth muscle
neck, peculiar ductlike structures may be called the sphincter of Oddi. This muscle com¬
seen. They can be traced for considerable plex is described as consisting of four parts:
distances in this connective tissue layer, and (1) a strong band of circular smooth muscle,
712 • THE LIVER AND GALLBLADDER

the sphincter choledochus, around the terminal continuously by the liver. The bile does not
portion of the bile duct; (2) a corresponding normally enter the intestine until a specific
sphincter pancreaticus around the pancreatic stimulus causes the gallbladder to contract.
duct; (3) longitudinal bundles of muscle fi¬ The stimulus is usually the presence of lipid
bers, the fasciculus longitudinalis, which span in the small intestine. Ingestion of fat auto¬
the space between the ducts; and (4) a mesh- matically causes discharge of the contents of
work of muscle fibers around the ampulla, the gallbladder. After a test meal of egg yolks
the sphincter ampullae. or cream, three fourths of its content are
The degree of development of these com¬ expelled within 40 minutes.
ponents of the sphincter of Oddi is subject When fat enters the small intestine, it
to great individual variation. Normally, con¬ causes release of a hormone, cholecystokinin,
traction of the sphincter choledochus stops from the mucosa. This is carried via the blood
the flow of bile. The longitudinal fasciculi to the gallbladder, inducing rhythmic con¬
shorten the intramural portion of the ducts tractions. In the peristalsis of the duodenum,
and probably facilitate flow of bile into the as waves of relaxation of its smooth muscle
duodenum. When the sphincter ampullae is pass by the ampulla of Vater, the tonic con¬
well developed, its contraction may have the traction of the sphincter of Oddi relaxes,
undesirable effect of causing reflux of bile permitting intermittent outflow of bile. Thus,
into the pancreatic duct, resulting in pan¬ the emptying of the gallbladder results from
creatitis. the combined action of cholecystokinin on
the musculature of the gallbladder and per¬
Blood Vessels, Lymphatics, and istalsis in the duodenum.
Nerves Of special clinical importance is the con¬
centrating function of the gallbladder. Its
The gallbladder is supplied with blood by mucosa reabsorbs water and ions from the
the cystic artery. The venous blood is col¬ bile. Experimental studies suggest the mech¬
lected by veins that empty primarily into anism. In gallbladders known to be trans-
small veins of the liver and only secondarily
into the cystic branch of the portal vein.
A prominent feature of the gallbladder is GALL BLADDER
LUMEN
its rich supply of lymphatic vessels, of which
there are two main plexuses, one in the
lamina propria and the other in the connec¬
tive tissue layer. The latter plexus receives
tributaries from the liver, thus affording a
pathway that accounts for hepatogenous cho¬
lecystitis. These plexuses are collected into
larger lymphatics, which pass through the
lymph node or nodes at the neck of the
gallbladder and then accompany the cystic
and common bile ducts. They pass through
several lymph nodes near the duodenum and
finally join the cisterna chyli. The nerves are
branches of the splanchnic sympathetic and
the vagus nerves. Study of the effects of
stimulation of these nerves has given rise to
contradictory reports by different investiga¬
tors. It is probable that both excitatory and
inhibitory fibers are contained in each of
them. Of greater clinical importance are the
sensory nerve endings, because overdisten¬
tion or spasms of the extrahepatic biliary tract
may inhibit respiration and set up reflex
disturbances in the gut.
Figure 27—40. Diagram illustrating the probable mecha¬
Histophysiology of the Gallbladder nism of concentration of the bile. Sodium is actively
pumped into the intercellular cleft below the occluding
The gallbladder serves as a site of concen¬ junction, creating a standing gradient that moves water
from the lumen to the blood vessels in the wall of the
tration and storage of bile, which is secreted gallbladder.
 THE LIVER AND GALLBLADDER • 713

A B
Figure 27-41. Photomicrograph of rabbit gallbladder epithelium. A, With a hyperosmotic solution in the lumen, the net
water flux is very low and the intercellular spaces at the base of the epithelium are relatively inconspicuous. B, In a
gallbladder actively transporting fluid in vitro, the intercellular spaces are greatly distended, x 1400. (From Kaye, G. J.
Cell Biol. 30:237, 1966.)

porting fluid, the intercellular spaces at the absorption of bile salts under such conditions
base of the epithelium are always distended is an important factor in the precipitation of
and the subepithelial capillaries are dilated. gallstones. After obstruction of the cystic duct
In experiments carried out in vitro, if either the bile may be resorbed in toto or replaced
sodium or calcium is omitted from the me¬ by a colorless fluid consisting largely of exu¬
dium, there is no fluid transport and the date and mucus. A small amount of mucus
intercellular spaces are narrow. If either ion is added to the bile as it passes down the
is replaced, fluid transport is restored and larger bile ducts, and mucus-secreting glands
the intercellular spaces again appear dis¬ are fairly numerous in the gallbladder neck.
tended. It is believed that during concentra¬ In some mammalian species, no gallbladder
tion of bile, active transport of solute (Na + ) is present. Its surgical removal in man is
across the lateral cell membrane increases the often followed by a marked dilatation of the
concentration of solute in the intercellular biliary passages.
space (Figs. 27-40, 27-41). Because of the
resulting osmotic gradient, water moves into
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Annu. Rev. Physiol. 39:323, 1977. barbital induced synthesis of microsomal drug me¬
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Harkness, R. D.: Regeneration of liver. Br. Med. Bull. W. N. Lotto: Subdivision of hexagonal liver lobules
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 THE LIVER AND GALLBLADDER • 715

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GALLBLADDER AND BILE DUCTS
Kaye, G. I., H. O. Wheeler, R. T. Whitlock, and N.
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Chapman, G. B., A. J. Chiardo, R. J. Coffey, and K. duct in the human fetus, with special reference to
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 -28
PANCREAS

The pancreas is the second largest gland portion is filled with highly refractile secre¬
associated with the alimentary tract. It con¬ tory granules. In histological sections stained
sists of an exocrine portion, which secretes daily with basic dyes, the base of the cells is in¬
about 1200 ml of digestive juice essential for tensely colored owing to the presence of a
the digestion of carbohydrates, fats, and pro¬ high concentration of ribonucleoprotein in
teins of the diet; and an endocrine portion this region (Fig, 28—3). There is a paler-
secreting hormones essential for the control staining supranuclear Golgi region that varies
of carbohydrate metabolism. in size in different 'phases of the secretory
The pancreas is a pinkish white organ lying cycle. The apical cytoplasm is crowded with
retroperitoneally at the level of the second secretory granules, usually called zymogen
and third lumbar vertebrae (Fig. 28-1). On granules because they contain the precursors
the right its head is adherent to the middle of the enzyme-rich pancreatic juice. They are
portion of the duodenum, and its body and most numerous in fasting and are relatively
tail extend transversely across the posterior few after the massive release of secretion
wall of the abdomen to the spleen. In the induced by a meal. After depletion of the
adult it measures from 20 to 25 cm in length zymogen granules, the Golgi complex en¬
and varies in weight from 65 to 160 g. It is larges as new secretory granules are being
covered by a thin layer of connective tissue, formed.
which does not, however, form a definite The pancreatic acinar cell has been a highly
fibrous capsule. It is finely lobulated, and the favored example of a protein-secreting cell
outlines of the larger lobules can be seen with and its ultrastructure and biochemistry have
the naked eye. probably been more intensively studied than
any other glandular cell. The participation
of the various cytoplasmic organelles in the
biosynthetic pathway and the mode of dis¬
THE EXOCRINE PANCREAS charge of the secretory product were de¬
scribed in some detail in Chapter 3 on Glands
Acinar Tissue and Secretion, and need only be reviewed
here.
The pancreas is a compound acinous gland The basal half of the cell is crowded with
organized in many small lobules that are parallel arrays of cisternae of rough endo¬
bound together by a loose connective tissue plasmic reticulum. The intervening cyto¬
stroma through which run the blood vessels, plasmic matrix is rich in free polyribosomes,
nerves, lymphatics, and interlobular ducts. and contains long mitochondria with well-
The acini of the exocrine pancreas are round developed cristae and numerous matrix
or elongate (Fig. 28-2). They consist of 40 to granules (Fig. 28—4). The endoplasmic retic¬
50 pyramidal cells in a single row around a ulum is the site of synthesis of the secretory
narrow lumen. The size of the lumen varies proteins and is therefore unusually extensive.
with the physiological state of the organ, Morphometric studies have shown that this
being small when the gland is at rest and organelle occupies some 20 per cent of the
becoming somewhat larger during active se¬ cell volume and presents a membrane surface
cretion. Between the acinar cells are short of about 800 pm2 per cell. The supranuclear
secretory canaliculi that open into the lumen Golgi complex consists of several curved
(Fig. 28-2). stacks of parallel cisternae and numerous
Examined in the living state with a dissect¬ small vesicles associated mainly with its con¬
ing microscope, the basal portion of the aci¬ vex cis-face. At the concave trans-face of the
nar cells is homogeneous, while the apical Golgi are one or more condensing vacuoles
716
 PANCREAS • 717

Figure 28-1, Drawing of the upper abdominal viscera with the stomach, transverse colon, and most of the liver cut
away to show the location and relationships of the pancreas. (Drawing by M. Brodel, from Trimble, I. R., J. W. Parsons,
and C. P. Sherman. Surg. Gynecol. Obstet. 73:711, 1941. By permission of Surgery, Gynecology & Obstetrics.)

with a homogeneous content of relatively low mogen “granules,” implying a semisolid con¬
density representing formative stages of new sistency, it is evident from electron micro¬
secretory granules. Occasional lipid droplets graphs of exocytosis that their content is fluid
and lysosomes may also be found in this at the time of its release, for it appears to
region. flow out through the opening formed by local
The apical pole of the cell is the site of fusion of its limiting membrane with the
accumulation and storage of the product and plasmalemma (Fig. 28-6). The secreted zy¬
between meals is packed with large, dense mogen is not in the form of granules but is
zymogen granules (Fig. 28—5). The narrow a moderately dense homogeneous material
free surface of the acinar cell usually has distributed throughout the lumen. Normally
sparse, irregularly oriented microvilli. the zymogen granules or droplets in the apex
In actively secreting glands, zymogen gran¬ of the cell remain discrete even though
ules may be found in the process of discharg¬ closely crowded together. In cells that are
ing their content into the lumen. Although it very actively secreting, however, one whose
is customary to call the stored products zy¬ membrane has become continuous with the
718 • PANCREAS

Capillary Acinar cells

Figure 28-2. Drawing of an acinus of the exocrine pancreas and its associated capillaries and nerves. Notice the
intercellular secretory canaliculi and the extension of the centroacinar cells into the acinus. (Reproduced from Krstic, R.
V. Die Gewebe des Menschen und Saugetiere. Berlin, Heidelberg, Springer-Verlag, 1978.)

mam
fH 1^™*' %*£?**■

ft* * 4
**

t 11-

' , .. -•... ■ .... .. :: .

> Ergastoplas/n ««*. ( J
' / , . * V v# \ _J
.LETK

complex ---7

ii'iiiSI:;';: • ■ ” ‘
MB
** v 44: \ rv
m
■ Hi I
j ■
^ M $ I
H

B ■■■
■■■ ■
% Jltl4‘?:’ f %
St -

Figure 28 3. Photomicrograph of human pancreas, showing an acinus and its centroacinar cells. The ergastoplasm
Golgi complex, and zymogen granules of the acinar cells are clearly identifiable. The fixation of the nuclei is less than
ideal, but adequate preservation of pancreas from postmortem material is difficult. (Courtesy of S Ito )
Figure 28-4. Electron micrograph of the basal region of a human pancreatic acinar cell, showing a portion of the
nucleus and the extensive development of cisternae of granular endoplasmic reticulum. (Micrograph courtesy of A. Like
and S. Ito.) x

Figure 28-5. A portion of two neighboring human pancreatic acinar cells. Below is the endoplasmic reticulum of the
paranuclear region of one cell, and above is the apical region of the adjacent cell filled with zymogen granules and
tubular elements of reticulum. (Micrograph courtesy of A. Like and S. Ito.) 7iQ
720 • PANCREAS

Figure 28-6. Electron micrograph of the lumen of an acinus and the apical portions of four acinar cells. Large, dense
zymogen droplets or granules are found in the cell apex. The limiting membrane of one of these has fused with the cell
membrane and its zymogen is being discharged into the lumen. The free surface of the acinar cells bears short microvilli.

plasma membrane may be joined by a second the bat and the dog, electron micrographs of
fusing with it and this one in turn by a third. stimulated glands reveal, in the lumen of the
In this way, a series of interconnected zy¬ reticulum, dense spherical bodies that resem¬
mogen granules may come to extend for ble small zymogen granules. In these species,
some distance downward into the apical cy¬ at least, concentration of the product evi¬
toplasm (Fig. 28-7). dently can occur in the reticulum as well as
The digestive enzymes of the pancreas are in the Golgi apparatus.
synthesized in the basal cytoplasm of the
acinar cells, where they accumulate in the
lumen of the endoplasmic reticulum. HISTOPHYSIOLOGY OF THE
Through it, they are channeled into the Golgi EXOCRINE PANCREAS
region, where they are segregated in vesicu¬
lar elements of Golgi complex origin and The exocrine secretory function of the
concentrated into typical zymogen granules. pancreas has a rhythmic cycle with a low
In most species it is only after the product basal rate of continuous secretion, periodi¬
has undergone this concentration that it is cally greatly increased by nervous and hor¬
sufficiently insoluble to resist extraction dur¬ monal stimulation associated with ingestion
ing specimen preparation. Consequently, zy¬ of food. Concurrently with the initiation of
mogen granules are visible in the Golgi re¬ gastric secretion, nerve impulses carried by
gion and apex of the cell, but their precursors the vagus nerve cause release of acetylcholine
in the endoplasmic reticulum are extracted, at the periphery of the pancreatic acini. This
and its lumen usually appears empty. Never¬ induces some release of enzymes into their
theless, the presence of the digestive enzymes lumen but no significant flow in the duct
within the reticulum has been established in system. Hormonal control of secretion ap¬
biochemical studies of the microsome frac¬ peals to be more important. The presence of
tion. In the guinea pig, and occasionally in
food in the gastric antrum and the passage
 PANCREAS • 721

coordinated action of secretin and cholecys¬

tokinin results in a copious secretion of en¬
zyme-rich pancreatic juice. Cholecystokinin
also activates contraction of the gallbladder,
adding bile to the duodenal contents.
Pancreatic juice contains enzymes for
digestion of the three major classes of nu¬
trients: carbohydrates, fats, and protein.
Starches and glycogen are broken down by
pancreatic amylase. Fat is hydrolyzed to glyc¬
erol and fatty acids by pancreatic lipase. The
proteolytic enzymes include trypsin, chymotryp-
sin, carboxypeptidase, ribonuclease, and deoxyri¬
bonuclease. These enzymes are released from
the acinar cells as inactive proenzymes or
zymogens and are activated by enzymes in
the lumen of the intestine. These powerful
proteases, if activated prematurely, would
digest the pancreas. Induced in the patholog¬
ical condition acute pancreatitis, this occurs
with rapid destruction of much of the gland.
To minimize this danger, the acini also syn¬
thesize trypsin inhibitor secreted with the en¬
zymes to prevent their premature activation
within the cells or the duct system. Under
some circumstances this intrinsic safeguard
is overwhelmed and acute pancreatitis rap¬
Figure 28-7. Electron micrograph of the apical portion of idly progresses, often with fatal outcome.
an acinar cell from a dog pancreas. A zymogen granule
or droplet opening onto the lumen may be joined to a
second and this to a third, so that zymogen may be
discharged through several intercommunicating mem¬
brane-limited vacuoles, x 24,500. (After Ichikawa, A. J. THE ENDOCRINE PANCREAS
Cell Biol. 24:369, 1965.)
Islets of Langerhans
of the acidic products of gastric digestion The endocrine function of the pancreas is
into the duodenum stimulates release of two segregated in small masses of endocrine cells
intestinal hormones, secretin and cholecystoki- forming the islets of Langerhans, which are
nin. scattered throughout the gland (Fig. 28—8).
Secretin is a polypeptide hormone of 27 They are not distinguishable with the naked
amino acid residues. Carried in the blood to eye, but if the gland is perfused with a dilute
the pancreas, it causes secretion of a large solution of neutral red, the islets are selec¬
volume of fluid containing a high concentra¬ tively stained and their number and distri¬
tion of bicarbonate. It does not stimulate the bution can then be studied. In the pancreas
acinar cells, and the secretion induced has of the adult human there are over a million
little or no enzymatic activity. This copious islets, but owing to their small size they con¬
alkaline juice serves to neutralize the acidic stitute only 1 to 2 per cent of the volume of
chyme entering the intestine from the stom¬ the gland. They are somewhat more numer¬
ach and creates the neutral or alkaline pH ous in the tail than in the body and head.
required for optimal activity of the pancreatic The islets are demarcated from the sur¬
enzymes. rounding acinar tissue by a thin layer of
Cholecystokinin is a polypeptide hormone reticular fibers, but there is very little reticu¬
of 33 amino acids secreted by the duodenum lum within the islet other than the delicate
and upper jejunum. Carried in the blood to fibrils associated with the capillaries. The
the pancreas, it causes secretion of large islets are often described as composed of
quantities of digestive enzy mes. Acting alone anastomosing cords or plates of epithelial
it does not significantly increase the volume cells, but this conveys a misleading impres¬
of outflow from the pancreatic ducts, but the sion of their organization. Reconstruction
722 • PANCREAS

Figure 28-8. Photomicrograph of an islet of Langerhans and surrounding acinar tissue in guinea pig pancreas.
Hematoxylin and eosin. x 500.

from serial sections reveals that each islet is D cells are selectively impregnated (Fig. 28—
a compact mass of epithelial cells permeated 102?), Fluorescein-conjugated antibodies to
by a labyrinthine network of capillaries. No the three hormones are now commonly used
cords or plates are evident in such recon¬ for definitive immunocytochemical identifi¬
structions. cation of the islet cells.
The islets contain three principal types of In humans the A cells tend to be located
cells, each secreting a different hormone: mainly at the periphery of the islet, but some
alpha cells (A2 cells) secreting glucagon; beta are scattered along the capillaries in its inte¬
cells (B cells) producing insulin', and delta cells rior. The B cells are the predominant cell
(D cells, Al cells) secreting somatostatin. In type distributed throughout the islet, making
histological sections prepared using the rou¬ up about 60 per cent of its mass. The D cells
tine hematoxylin and eosin method, these are least abundant. They are quite variable
cell types cannot be distinguished and the in shape, are often greatly elongated, and
islets simply appear as rounded masses of may occur anywhere in the islet. All the cell
cells staining less intensely than the sur¬ types are polarized with their secretory gran¬
rounding acinar tissue (Fig. 28-8). However, ules toward the capillaries.
techniques have been devised for selective Additional cells have been reported in
staining of each cell type. In islets stained some but not all species, and their physiolog¬
using the aldehyde-fuchsin trichrome ical significance is obscure. C cells found in
method, the secretory granules of the B cells the guinea pig pancreas are devoid of secre¬
are deeply colored (Fig. 28-9). With the Gn- tory granules and may be undifferentiated
melius silver impregnation technique, the A endocrine cells. E cells are described in opos¬
cells are blackened (Fig. 28-10A), and with sum islets, and F cells in the uncinate process
the Hellerstrom-Hellman silver method the of the canine pancreas and in several other
 PANCREAS • 723

Figure 28-9. Photomicrograph of an islet of Langerhans in the human pancreas stained with aldehyde-fuchsin, which
specifically colors secretory granules in the cytoplasm of insulin-producing B cells. The secretory material is often
polarized toward capillaries, x 500. (Micrograph from Hellerstrom, C. Acta Paediatr. Scand. Suppl. 270:7, 1977.)

Figure 28-10. A, Photomicrograph of a human pancreatic islet stained with the Grimelius silver technique for
demonstrating glucagon-producing A cells (A2 cells). The A cells are preferentially localized at the periphery of the islet
or along islet capillaries, x 250. B, Human pancreatic islet stained with the Hellerstrom-Hellman silver technique for
demonstration of somatostatin-producing D cells (A, cells). The cells are few in number and irregular in shape, x 400.
(Photomicrographs from Hellerstrom, C. Acta Paediatr. Scand. Suppl. 270:7, 1977.)
724 • PANCREAS

Figure 28-11. Electron micrograph of a juxtanuclear area of an alpha cell in a human islet of Langerhans. The alpha
granules have a very dense spherical core and a less dense outer region bounded by a membrane, x 24,000. (Courtesy
of A. Like.)

species. A cell type recently described and The ultrastructural characteristics of the
designated the Pp cell is believed to secrete principal cell types has been studied in con¬
pancreatic polypeptide. It occurs not only in the siderable detail. The A cells contain large
islets but also scattered among the acini of numbers of distinctive secretory granules,
the exocrine pancreas. which, after aldehyde fixation, have spherical
It has been assumed that the islets through¬ cores of high electron density and homoge¬
out the pancreas were similar in their content neous texture surrounded by a narrow zone
of endocrine cells. This has now been chal¬ of lower density (Fig. 28—11). After primary
lenged on the basis of immunofluorescent osmium fixation the outer zone is extracted,
staining of successive serial sections from the leaving a clear space between the dense core
rat pancreas. Marked differences are found and the limiting membrane of the granule.
in the number of cells containing glucagon The cytoplasm contains a few cisternae of
and pancreatic polypeptide in different re¬ rough endoplasmic reticulum and a juxta¬
gions of the gland. The pancreas develops nuclear Golgi region that may contain nas¬
from two primordia, and these receive their cent secretory granules. The mitochondria
blood supply from different arteries. In the are of varying length and orthodox internal
dorsal region supplied by the celiac artery, structure.
the islets are found to be glucagon rich and In the B cells the mitochondria are some¬
pancreatic polypeptide poor. In the ventral what larger and the Golgi complex more
region irrigated by the superior mesenteric prominent. The endoplasmic reticulum is not
artery, they are pancreatic polypeptide rich extensive. The ultrastructure of the B-cell
and glucagon poor. The question as to granules shows marked species differences.
whether this distinction of two types of islets Pale and dark categories of granules have
of Langerhans applies to other species has been described. In some species the granules
not been explored. are homogeneous and distinguishable from
 PANCREAS • 725

those of A cells only by their slightly lower produce large amounts of gastrin, resulting
density and larger size. In the human, cat, in hypersecretion of acid and severe gastric
and dog they have a very distinctive appear¬ ulcers.
ance, containing one or more dense crystals All the cells of the islets are polarized
in a matrix of low electron density (Fig. 28- toward the capillaries, which are of the fe¬
12). In the human the crystals are rectangular nestrated type as in other endocrine glands.
or polygonal in section, and at high magni¬
fication they have a very regular internal
periodic structure. The surrounding matrix HISTOPHYSIOLOGY OF THE
is often extracted in specimen preparation ENDOCRINE PANCREAS
and the crystals stand out in sharp contrast
against the clear background enclosed in a The hormones secreted by the three prin¬
loosely fitting membrane (Fig. 28-12). cipal cell types of the islets of Langerhans
D cells are unusually heterogeneous in the are all involved in controlling the level of
size, shape, and density of their granules. circulating glucose. When the blood glucose
Their identity as a separate cell type was falls below an optimal level, the A cells secrete
formerly a subject of dispute, some investi¬ glucagon, which raises blood glucose. When
gators considering them to be immature or the glucose level rises too high, the B cells
degenerating A cells. Their existence in release insulin, which lowers its level. The
many, if not all, mammalian species is no role of somatostatin produced by the D cells
longer in doubt. Their granules are larger is less completely worked out but it is capable
and considerably less dense than those of the of suppressing secretion of either insulin or
A cell (Fig. 28—13). The cytological features glucagon and is assumed to modulate the
that suggested that they were degenerative activity of the a and (3 cells to maintain
forms may have been attributable to their normal glucose levels.
inadequate preservation. Occasional cells Insulin is a polypeptide hormone com¬
bearing a superficial resemblance to D cells posed of 51 amino acids in two chains, des¬
are immunoreactive for gastrin and are called ignated A and B, linked by disulfide bonds.
G cells. Rare islet cell tumors of humans It is a very important hormone directly or

Figure 28-12. Electron micrograph of portions of two adjoining beta cells. The beta granules in man and several other
species are membrane-bounded spherical vesicles containing dense crystals of varying configuration. (Courtesy of A.
Like.)
726 • PANCREAS

Figure 28-13. Electron micrograph of a portion of a delta cell. The granules are homogeneous and tend to fill their
limiting membrane, but they vary considerably in density. It is not clear whether these cells represent a distinct type or
are altered alpha cells. (Courtesy of A. Like.)

indirectly affecting the function of nearly urea), which in turn causes dehydration and
every organ in the body. Its most general excessive thirst (polydipsia). Failure of insulin
effect is to facilitate the movement of glucose to affect the cells in the hypothalamus that
through cell membranes, especially those of control appetite leads to eating in excess
liver, muscle, and adipose tissue. Insulin (polyphagia). To generate the energy needed
binds to specific receptors in the cell mem¬ in the absence of glucose utilization, fat and
brane but the mechanism by which it aug¬ muscle protein are metabolized and body
ments glucose entry is still poorly under¬ weight is rapidly lost despite increased food
stood. Brain, muscle, and many other organs intake. Increased mobilization of fat results
are heavily dependent on glucose as an en¬ in accumulation of ketones in the blood
ergy source, and the facilitation of its entry plasma and their excretion in the urine (ke-
into cells is essential for normal metabolism. tonuria). The loss of sodium in excreting
Glucose is rapidly phosphorylated within the ketone salts alters the buffering capacity of
cells and in this form cannot diffuse out. the blood, which becomes excessively acid
Thus, insulin results in a trapping of glucose (acidosis). Without exogenous insulin, the di¬
in cells and a consequent lowering of its abetic is at risk of becoming comatose and
concentration in the circulating blood. In dying from dehydration and metabolic aci¬
addition to its effect on transport, insulin also dosis.
influences glucose utilization by enhancing Insulin is synthesized in the B cells in the
the activity of glucokinase and glycogen syn¬ form of proinsulin, a single polypeptide chain
thetase. In adipose tissue, insulin promotes of about 73 amino acid residues that under¬
accumulation of lipid by facilitating entry of goes post-translational processing involving
glucose and its conversion to fatty acids and removal of a 22-amino acid segment gener¬
triglycerides, and by inhibiting release of ating insulin consisting of two polypeptide
fatty acids from adipose cells to the blood. chains linked by two disulfide bonds. Until
In diabetes, glucose cannot enter cells and now, insulin used in the treatment of dia¬
be utilized owing to a deficiency in insulin betics has been extracted from the pancreas
production. The resulting excess of glucose of cattle and swine slaughtered for meat. This
in the blood (hyperglycemia) leads to excre¬ source may not long continue to meet the
tion of an abnormal volume of urine (poly¬ needs of the increasing number of diabetics
 PANCREAS • 727

in the population. Fortunately the gene for THE DUCT SYSTEM

human insulin has now been isolated and
introduced into E. coli bacteria, which then
synthesize human insulin. With this recom¬ The lumen of each acinus is continuous
binant DNA technology it will soon be pos¬ with the lumen of a small duct bounded by
sible to produce insulin in unlimited quantity. the centroacinar cells, so named because they
Histological examination of the pancreas are surrounded by and appear to extend into
of diabetic humans reveals hyalinization or the center of the acinus (Fig. 28-14). The
fibrosis of the islets of Langerhans with de¬ centroacinar cells are easily distinguished by
struction of a large portion of the B cells. their pale staining in histological sections and
Much less common than diabetes is the oc¬ by the very low density of their cytoplasm
currence of tumors of islet cells, either benign and the paucity of their organelles in electron
or metastasizing. The increased volume of B micrographs. Near the termination of the
cells results in hyperinsulinism and puts the duct system, part of the wall may be made
patient at risk of insulin shock. As the blood up of centroacinar cells and part by acinar
glucose level falls during massive release of cells (Fig. 28-14). The terminal portion of
insulin, the central nervous system becomes the duct system drains proximally into the
hyperactive; there is extreme agitation, intralobular or intercalated ducts. These are
tremor, and sweating, which may be followed lined by cells similar to the centroacinar cells
by convulsions and ultimately coma. A similar that form a low columnar epithelium. These
train of events occurs if a diabetic acciden tally ducts are tributaries of larger interlobular ducts
takes an overdose of insulin. Timely intra¬ (Fig. 28-15) lined by a low columnar epithe¬
venous administration of glucose will arrest lium in which goblet cells and occasional
progression of insulin shock and even restore argentaffin cells are interspersed. Small mu¬
comatose patients to consciousness. cous glands may bulge slightly from the duc-

Figure 28-14. Electron micrograph of a terminal segment of the duct system of the guinea pig pancreas showing the
lumen bounded on one side by acinar cells and on the other by centroacinar cells. (From Bolender, R. P. J. Cell Biol.
61:269, 1974.)
728 • PANCREAS

in the guinea pig, are said to be also present

in the human. Their epithelium is of a low,
irregularly cuboidal type. They show occa¬
sional mitoses. Occasional goblet cells may be
found. Some of the projections from these
tubules consist of islet cells, singly or in
groups, but the most striking feature of the
tubules is their connection, by one or more
short stalks, with large islets of Langerhans.
It has long been thought that these ductules
are composed of undifferentiated epithelium
from which new islets can arise after destruc¬
tion of the endocrine pancreas by disease or
injury. This interpretation is now less widely
accepted, for it has been shown that the islet
cells themselves can divide mitotically and
have considerable regenerative capacity.

BLOOD VESSELS,
LYMPHATICS, AND NERVES

The arterial supply of the pancreas is from

branches of the celiac and superior mesen¬
teric arteries. From the celiac it receives
Figure 28-15. Photomicrograph of a small interlobular branches through the pancreaticoduodenal
pancreatic duct. and splenic arteries; it also receives small
branches from the hepatic artery. The infe¬
rior pancreaticoduodenal artery is a branch
tal epithelium. The interlobular ducts join of the superior mesenteric. The vessels run
the main pancreatic ducts, of which there are in the interlobular connective tissue and give
two. The larger, or duct of Wirsung, begins in off fine branches that enter the lobules. Veins
the tail and runs through the substance of accompany the arteries throughout and lead
the gland, receiving throughout its course the blood either directly into the portal vein
numerous branches, so that it gradually in¬ or indirectly through the splenic vein.
creases in size as it nears the duodenum. In The lymphatic supply of the gland has not
the head of the pancreas, it runs parallel with been worked out in detail. The lymphatic
the ductus choledochus, with which it may drainage is principally into the celiac nodes
have a common opening in the ampulla of about the celiac artery.
Vater. The opening and closing of these ducts The nerve supply is mainly by unmyeli¬
are controlled by the sphincter of Oddi. The nated fibers arising from the celiac plexus.
accessory duct of Santorini is about 6 cm long. These fibers accompany the arteries into the
It is nearly always present and lies cranial to gland and end around the acini. There are
the duct of Wirsung. These larger ducts have also many sympathetic ganglion cells in the
around the epithelium a moderately thick interlobular connective tissues. The organ
layer of dense connective tissue containing also receives my elinated fibers from the vagus
some elastic fibers. nerves.
In addition to the system of ducts just In electron micrographs, axons are seen
described, the pancreas is said to contain a penetrating the basal lamina to end in inti¬
system of anastomosing small tubules that mate contact with the base of the acinar cells
arise from the large ducts and run in the (Fig. 28-16). These nerve terminals often
connective tissue surrounding them. These contain numerous synaptic vesicles. The
tubes have a diameter of 12 to 27 pm; they source of these nerves is not clear, but it is
are connected with the islets of Langerhans likely that they are the terminations of
and only occasionally with the acini. These branches from tbe vagus and may be involved
structures, although studied most extensively somehow in regulation of secretion.
 PANCREAS • 729

islets and acinar cells do not directly activate

the secretory mechanism, it is possible that
they may modulate the permeability of the
cell membranes or the sensitivity of the cells
to hormones.

REFERENCES

EXOCRINE PANCREAS
Caro, L. G., and G. E. Palade: Protein synthesis, storage
and discharge in the pancreatic exocrine cell: an
autoradiographic study. J. Cell Biol. 20:4, 1964.
Grossman, M. I.: Nervous and hormonal regulation of
pancreatic secretion. In de Reuch, A. V., and M. P.
Cameron, eds.: The Exocrine Pancreas. Ciba Foun¬
dation Symposium. Boston, Little, Brown 8c Co.,
1961.
Herzog, V., and M. G. Farquhar: Luminal membrane
retrieved after exocytosis reaches most Golgi cister-
nae. Proc. Natl. Acad. Sci. U.S.A. 74:5073, 1977.
Herzog, V., and H. Reggio: Pathways of endocytosis
from luminal plasma membrane in rat exocrine
pancreas. Eur. J. Cell Biol. 27:141, 1980.
Ichikawa, A.: Fine structural changes in response to
hormonal stimulation in the perfused canine pan¬
creas. J. Cell Biol. 24:369, 1965.
Jamieson, J. D., and G. E. Palade: Condensing vacuole
conversion and zymogen granule discharge in pan¬
Figure 28-16. Electron micrograph of a nerve axon creatic exocrine cells: metabolic studies. J. Cell Biol.
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cells from the bat. The axon contains synaptic vesicles Jamieson, J. D., and G. E. Palade: Production of secre¬
and lies within the basal lamina of the acini. tory proteins in animal cells. In Brinkley, R. B., and
K. R. Porter, eds.: International Cell Biology. New
York, Rockefeller University Press, 1976—1977, p.
The presence of unmyelinated nerves in 308.
the islets of Langerhans has also been re¬ Janawitz, H. D.: Pancreatic secretion of fluid and elec¬
trolytes. In: Code, C. F., and M. L Grossman, eds.:
ported, and some of these end on the endo¬ Handbook of Physiology. Sect. 6. Washington, DC,
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islet cells or in deep recesses in their bases Palade, G. E., P. Siekevitz, and L. G. Caro: Structure,
inside the basal lamina. Two types of endings chemistry, and function of the pancreatic exocrine
cell. In de Reuch, A. V. S., and M. P. Cameron,
are distinguishable. Those presumed to be
eds.: The Exocrine Pancreas. Ciba Foundation Sym¬
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Physiol. 39:373, 1977.
cles contain dense cores or granules of irreg¬
Sarles, H.: The exocrine pancreas. Int. Rev. Physiol.
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ENDOCRINE PANCREAS
largely on gastrointestinal hormones. There
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chemical and ultrastructural identification of the
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and cholecystokinin, and releases insulin in Baetens, D., F. Maisse-Lagae, A. Perrelet, and L. Oorci:
response to elevated blood sugar. The phys¬ Endocrine pancreas: three-dimensional reconstruc¬
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iological evidence concerning the role of
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730 • PANCREAS

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 .. . 29
RESPIRA TOR Y S YSTEM

All higher animals require oxygen to main¬ lamina that separates it from an underlying
tain their metabolic processes. The respira¬ connective tissue layer that contains mucous
tory system provides for the intake of oxygen glands (Fig. 29-1). The mucus from these
in the inspired air and the elimination of glands keeps the lining of the nasal cavity
carbon dioxide produced in cell metabolism moist. Beneath the epithelium on the lower
throughout the body. Oxygen is carried to, nasal conchae are rich venous plexuses that
and carbon dioxide from, the cells by the serve to warm the air as it passes through the
circulatory system. nose. The tissue containing these venous
The respiratory tract can be thought of as plexuses is capable of considerable engorge¬
having a conducting portion proximally, which ment, and in this respect bears a superficial
connects the exterior of the body with a resemblance to erectile tissue, but differs
respiratory portion, distally, where the ex¬ from it in the absence of septa containing
change of gases between the blood and air smooth muscle. This region is a common site
takes place. The conducting portion includes of “nose bleed.” Collections of lymphoid tis¬
the passages of the nose, the pharynx, the sue beneath the epithelium are a character¬
larynx, the trachea, and a branching system of istic feature of the mucous membrane of the
bronchi of progressively diminishing caliber. nose, especially near the nasopharynx.
The smallest branches, the bronchioles, are
continuous with the respiratory portion of The Olfactory Epithelium
the lungs. This consists of respiratory bron¬
chioles, alveolar ducts, and alveoli, which to¬ The receptors for the sense of smell are
gether make up the greater part of the vol¬ located in a specialized region of the nasal
ume of the lungs. epithelium that occupies the roof of the nasal
cavity and extends downward 8 to 10 fjim on
each side of the septum and onto the surface
of the upper nasal conchae. The olfactory
THE NOSE area is irregular in outline and has a total
surface area on both sides of about 500 sq
The nose is composed of bone, cartilage, mm.
muscle, and connective tissue. Its skin is pro¬ The olfactory epithelium is a tall, pseudostrat¬
vided with very fine hairs and unusually large ihed columnar epithelium about 60 pm thick
sebaceous glands. The integument continues (Fig. 29-2). It consists of three kinds of cells:
through the anterior nares into the vestibule supporting cells, basal cells, and olfactory cells.
of the nose. The stratified squamous epithe¬ The supporting cells were traditionally de¬
lium here bears large stiff hairs that project scribed as tall slender elements with an axial
into the airway and are believed to help in bundle of tonohbrils and a prominent “cutic-
excluding particles of dust from the inspired ular plate” immediately beneath the free sur¬
air. The remainder of the nasal cavity is lined face, inserting at either side into a prominent
with mucus-secreting, pseudostratihed cil¬ “terminal bar.” In electron micrographs the
iated epithelium. Dust particles trapped in cuticular plate is found to be a typical ter¬
the layer of mucus on the surface of the minal web associated on either side with the
epithelium are continually transported by cil¬ zonula adherens of well-developed junctional
iary action toward the pharynx, where they complexes that attach the supporting cells to
are disposed of by swallowing. the adjacent sensory cells. The free surface
The ciliated columnar epithelium contains of the cell bears numerous long slender mi¬
abundant goblet cells, and rests on a basal crovilli that project into the overlying blanket

731
732 • RESPIRATORY SYSTEM

Lamina propria Periosteum Bone

Opening
of duct
Blood
vessel

--Glands

Blood
vessel

Figure 29-1, Section of the mucosa of the osseous portion of the nose of a 22-year-old man. (After Sobotta.)

of mucus (Fig. 29—4). There is a small Golgi

complex in the apical cytoplasm, and pigment
granules that are responsible for the brown
color of the olfactory area. In some species
the supporting cells are secretory and contain
numerous mucigen granules in the apical
cytoplasm.

Figure 29-3. Photomicrograph of an Araldite-embedded

section of olfactory epithelium from a frog. The slender,
lighter-staining cells that extend to the surface of the
epithelium are the olfactory rods of the bipolar receptor
ceils. The darker cells making up the bulk of the upper
third of the pseudostratified epithelium are sustentacular
cells. At lower right is a portion of a gland of Bowman.
Figure 29-2. Photomicrograph of a celloidin-embedded Toluidine blue stain. (After Reese, T. J. Cell Biol. 25:209,
section of mammalian olfactory epithelium. Masson stain. 1965.)
 RESPIRATORY SYSTEM • 733

Between the bases of the supporting cells, The cytoplasm of the olfactory cell contains
the basal cells form a single layer of small a network of neurofibrils, which are espe¬
conical elements with dark nuclei and cially conspicuous around the nucleus. The
branching processes. cell may be slightly constricted at the level of
The olfactory cells, evenly distributed among its junctional complexes with the neighboring
the supporting cells, are bipolar nerve cells. supporting cells. Distal to this constriction,
Their round nuclei occupy a zone between the bulbous head of the olfactory cell den¬
the nuclei of the supporting cells and the drite projects above the general surface of
connective tissue. The apical portion of the the epithelium. This protruding portion is
cell is a modified dendrite and extends as a sometimes inappropriately called the olfactory
cylindrical process from the nucleus to the vesicle. Radiating from its surface are six to
surface of the epithelium (Fig. 29-4). The eight olfactory cilia originating from basal bod¬
basal portion of the cell tapers into a thin, ies embedded in a superficial ectoplasmic
smooth process about 1 pm thick. This is an layer of cytoplasm having the character of a
axon—one of the fibers of the olfactory terminal web. These olfactory cilia are for
nerve. It passes into the subepithelial connec¬ the most part nonmotile and extremely long.
tive tissue and there, with similar fibers, In the frog, in which they have been studied
forms small nerve bundles. These assemble in some detail with the electron microscope,
into about 20 macroscopically visible fila ol- they attain lengths of 150 to 200 pm. They
factona. have an atypical structure. The proximal seg¬
ment of the ciliary shaft is about 250 nm in
diameter and contains the usual 9 + 2 ar¬
rangement of longitudinal microtubules. A
few micrometers from their base there is an
abrupt narrowing of the shaft to about 150
nm. This slender portion of the shaft contin¬
ues to the tip of the cilium, constituting some
80 per cent of its overall length. In this
segment, the axoneme consists of 11 single
microtubules instead of the usual two singlets
and nine doublets. The slender distal seg¬
ments of the olfactory cilia course parallel to
the surface of the epithelium embedded
within a thick layer of mucus, but with their
tips near its surface. On the basis of the
anatomical and physiological evidence now
available, these specialized cilia appear to be
the component of the sense organ that is
excited by contact with odorous substances.
The unmyelinated fibers of the olfactory
nerve are enmeshed in a delicate connective
tissue rich in macrophages. The hla olfactoria
pass through openings of the cribriform plate
of the ethmoid bone and enter the olfactory
bulb of the brain, where the primary olfac¬
tory center is located. The olfactory mucosa
is also provided with myelinated nerve fibers
originating from the trigeminal nerve. After
losing their myelin sheaths, the fibers enter
the epithelium and end in fine arborizations
between the supporting cells. These endings
are receptors for stimuli other than odors.
The lamina propria of the olfactory mu¬
Figure 29-4. Diagrammatic representation of the essen¬ cosa is continuous with the dense connective
tial features of the olfactory epithelium based on electron tissue forming the periosteum of the cribri¬
microscopic studies. The height of the epithelium has
been foreshortened. The vertical lines in the rod or den¬
form plate. In it are numerous pigment cells
drite of the olfactory bipolar neuron represent microtu¬ and some lymphoid cells. The lamina propria
bules. contains a rich plexus of blood capillaries. In
734 • RESPIRATORY SYSTEM

its deeper layers it includes a plexus of large The paranasal sinuses are often a site of
veins and dense networks of lymphatic cap¬ painful inflammation, sinusitis, and occasion¬
illaries. The latter continue into large lym¬ ally require surgical drainage.
phatics, which course toward the lymph
nodes on either side of the head. If a colored
material is injected into the subarachnoid
spaces of the brain, it can penetrate into the
THE LARYNX
lymph capillaries of the olfactory region as
well as into the sheaths of the fila olfactoria. The larynx is an elongated structure of
This demonstrates a possible pathway for irregular shape, whose walls contain hyaline
infections to spread from the nasal mucosa and elastic cartilage, connective tissue,
to the meninges. striated muscles, and a mucosa with associ¬
The lamina propria in the olfactory area ated glands. It serves to connect the pharynx
also contains the branched, tubuloalveolar with the trachea. As a result of changes
olfactory glands of Bowman. The secretory por¬ resulting from the contraction of its muscles,
tions are oriented mainly parallel to the sur¬ it produces variations in the width of the
face, whereas the narrow ducts assume a opening between the vocal cords. The size of
perpendicular course and open onto the sur¬ this opening and the degree of muscular
face. Immediately under the epithelium the tension exerted upon the cords determine
duct is often considerably enlarged. The low the pitch of the sounds made by the passage
pyramidal cells of the secretory portion of of air through the larynx.
the glands are serous and contain obvious The framework of the larynx is made up
secretory granules. of several cartilages. Of these, the thyroid
Olfactory stimuli are of chemical nature. and cricoid cartilages and the epiglottis are
The secretion of the glands of Bowman keeps unpaired, whereas the arytenoid, corniculate,
the surface of the olfactory epithelium moist and cuneiform cartilages are paired. The
and furnishes the necessary solvent. As most thyroid and cricoid and the lower parts of
odoriferous substances are much more solu¬ the arytenoids are hyaline cartilage. The ex¬
ble in lipids than in water, and as the mem¬ trinsic muscles of the larynx support and con¬
branes and other constituents of olfactory nect it with surrounding muscles and liga¬
cells and their cilia contain lipids, odoriferous ments and their contraction raises it during
substances, even if present in extreme dilu¬ deglutition. The intrinsic muscles join together
tion, presumably become concentrated in the cartilages of the larynx. By their contrac¬
these structures. The continuous stream of tion they give different shapes to the laryn¬
the secretion of the olfactory glands, by re¬ geal cavity and thus play a role in phonation.
moving the remains of the stimulating sub¬ The anterior surface of the epiglottis, the
stances, keeps the receptors ready for new upper half of its posterior surface (the aryep-
stimuli. In this respect, the olfactory glands iglottic folds), and the vocal cords are all covered
doubtless have a function similar to that of with stratified squamous epithelium. In the
the glands associated with the taste buds. adult, ciliated epithelium usually begins at
the base of the epiglottis and extends down
the larynx, trachea, and bronchi.
PARANASAL SINUSES The cilia, which are 3.5 to 5 pm long, beat
toward the mouth, and thus move foreign
particles, bacteria, and mucus from the lungs
Connected with the nasal cavity, and form¬ toward the exterior of the body.
ing cavities in the respective bones, are the Goblet cells are scattered among the cylin¬
frontal, ethmoidal, sphenoidal, and maxillary
drical cells of the laryngeal epithelium in
sinuses—the accessory sinuses of the nose. These
varying numbers. The glands of the larynx
are lined with ciliated epithelium similar to
are of the tubuloacinous, mixed mucous va-
that of the nasal cavity but containing fewer
liety. The acini secrete mucus and may have
and smaller glands. The cilia beat so as to
seious crescents. A few taste buds are scat¬
move a blanket of mucus toward the nasal
tered on the undersurface of the epiglottis.
cavity. The mucosa of all the sinuses is thin
The true vocal cords contain the vocal or
and the lamina propria cannot be differen¬
inferior thyroarytenoid ligaments. Each of
tiated as a separate layer from the periosteum
these (one on each side of the midline) con¬
of the bones, to which is is tightly adherent.
sists of a band of elastic tissue bordered on
 RESPIRATORY SYSTEM • 735

its lateral side by the thyroarytenoid muscle nerves to the mucous membrane, and the
and covered medially by a thin mucous mem¬ inferior laryngeal nerve sends motor nerves
brane consisting of stratified squamous epi¬ to the muscles of the larynx.
thelium (Fig. 29—5). The anteroposterior di¬
mension of the space between the vocal cords
is said to be about 23 mm in men and 18 mm
in women. The shape of this opening be¬
CONDUCTING PORTION OF
tween the vocal cords undergoes great vari¬ THE RESPIRATORY TRACT
ations in the different phases of respiration
and in the production of various sounds in The airway or conducting portion of the res¬
talking and singing. Contraction of the thy¬ piratory tract is an arborescent system of
roarytenoid muscle approximates the aryte¬ tubules that begins with the trachea and con¬
noid and thyroid cartilages, and this relaxes tinues through 16 generations of dichoto¬
the vocal cords. mous branching to end in the terminal bron¬
The larynx is supplied by the upper, mid¬ chioles. This pattern of branching results in
dle, and lower laryngeal arteries, which in a progressive decrease in the diameter of the
turn arise from the superior and inferior channels, and a concurrent increase in their
thyroid arteries. The veins from the larynx number and in the total cross-sectional area
empty into the thyroid veins. The larynx of the system, as shown in Figure 29-6. It is
contains several rich plexuses of lymphatics, customary to refer to the branches distal to
which lead into the upper cervical lymph the trachea as bronchi, bronchioles, and terminal
nodes and to those about the trachea. The bronchioles, but it should be borne in mind
superior laryngeal nerve carries sensory that there are several generations of branch¬
ing within the first two categories, so that at
the level of terminal bronchioles the number
Edge of thyroid has reached about 65,000, each with a di¬
ca rti lage ameter of about 0.2 mm. The channels
throughout the bronchial tree serve simply
to conduct the inspired and expired air to
and from the respiratory portion of the lung
where the gaseous exchange with the blood
takes place.

The Trachea
False vocal cord
The trachea is a flexible tube about 11 cm
Duct of mixed glands long and 2 cm in diameter. It is lined by
ciliated, pseudostratihed columnar epithe¬
lium with an unusually thick basal lamina.
Many goblet cells are scattered throughout
the epithelium.
In electron micrographs, the ciliated cells
have a microvillous border through which
the cilia project into the lumen (Fig. 29-7).
Vocal muse
The apical cytoplasm contains a small Golgi
apparatus and numerous mitochondria. The
endoplasmic reticulum is not extensive and
Stratified
the cytoplasmic matrix contains only a mod¬
epitheliur
erate number of ribosomes. The goblet cells
Mixed mucc
are similar to those of the gastrointestinal
gland tract. Their expanded apical region is occu¬
pied by closely packed mucinogen granules
Ciliated ep
of low electron density and a somewhat com¬
pressed supranuclear Golgi apparatus. Their
narrow basal region is rich in cisternae of
Figure 29-5. Frontal section through the middle of the rough endoplasmic reticulum.
glottis of a 9-year-old boy. (After von Ebner.) Small pyramidal basal cells are intercalated
736 • RESPIRATORY SYSTEM

Generation Cross
of branching Name Diameter section Number
(cm) (cm2)

0 Trachea 2 3 1
1 Mainstem bronchi 1.3 1.35 2
0 2 Lobar bronchi 0.9 0.70 4 Figure 29-6. Table showing the
t- o 3 Segmental bronchi 0.7 0.38 8
numbers and average dimensions
4 Subsegmental bronchi 0.5 0.20 16
of the successive orders of dichot¬
§s
o11 12 Bronchiole 0.05 0.0021 4,096 omous branching of the bronchia!
o 13 Bronchiole 0.04 0.0012 8,192 tree and the respiratory portion of
the human lung inflated to three-
16 Terminal bronchiole 0.018 0.00024 65,536 fourths capacity. (Modified after
Weibel, E.R. Morphometry of the
Human Lung. New York, Aca¬
demic Press, 1963.)
>- 17 Respiratory bronchiole 1 0.015 0.00015 131,072
cc 18 Respiratory bronchiole 2
oz 0.012 0.00011 262,144
19 Respiratory bronchiole 3 0.011 0.00010 524,288
<8
£tE 20 Alveolar duct 1 0.010 0.00008 1,048,576
21 Alveolar duct 2 0.010 0.00008 2,097,152
in 22 Alveolar duct 3 0.010 0.00008 4,194,304
23 Alveoli 0.005 C.00002 8,388,608

between the bases of the columnar cells. The A less common cell type is the so-called
alignment of their nuclei below those of the brush cell—a slender columnar cell with a
ciliated columnar cells gives the epithelium microvillous luminal border and a well-de¬
its characteristic “pseudostratihed” appear¬ veloped smooth endoplasmic reticulum, but
ance. They have few organelles and are evi¬ no secretory granules. Small aggregations of
dently a reserve of relatively undifferentiated glycogen particles may be scattered through
cells capable of replacing other cellular ele¬ the cytoplasm. The function of the brush
ments of the epithelium. cells and their relationship to other cells of

Rgure 29-7 Scanning micrograph of the surface of tracheal epithelium from a horse The dome-shaoed anice<
LltisylfpAT) mlCr0V"" Can be am°n9 the ,U,,S of cilia ^e ciliated^epithellaf celte^^MkJroJt
 RESPIRATORY SYSTEM • 737

the epithelium remain to be clarified. They A characteristic feature of the trachea is its
have been variously interpreted as depleted supporting framework of 16 to 20 C-shaped
goblet cells or as intermediate stages in the hyaline cartilages that encircle it on its ventral
differentiation of basal cells to replace exfol¬ and lateral aspects (Fig. 29-8). These succes¬
iated columnar cells. The occasional obser¬ sive incomplete cartilaginous rings are sepa¬
vation of synapses with intraepithelial nerve rated by interspaces that are bridged by h-
fibers has led to the speculation that some broelastic connective tissue. Because of this
brush cells may function as sensory receptors, arrangement, the trachea has much more
but physiological validation of this suggestion pliability than it would have if enveloped in
is lacking. a continuous layer of cartilage. The tracheal
Sparsely distributed in the tracheobron¬ cartilages reinforce the wall and keep its
chial epithelium are cells resembling the ar¬ lumen open by resisting external forces that
gentaffin cells of the gastrointestinal tract. otherwise might constrict the airway. Outside
Viewed in electron micrographs, these cells of the cartilages, there is a layer of dense
contain numerous small, dense, secretory connective tissue containing many elastic fi¬
granules often concentrated near the basal bers.
lamina. There may be more than one cate¬ The posterior wall of the trachea adjacent
gory of such cells. Some do not reach the to the esophagus is devoid of cartilages (Fig.
lumen, and have the staining properties and 29-8). In their place is a thick band of bun¬
fluorescence characteristic of cells that store dles of smooth muscle, which, in the main,
catecholamines. Others are columnar and re¬ run transversely. At their ends, they are in¬
semble the peptide hormone—secreting cells serted among the dense elastic and collagen¬
of the enteroendocrine system. To date, the ous fiber bundles making up the layer of
small granule-containing cells of the respira¬ dense connective tissue peripheral to the tra¬
tory tract have received less investigative at¬ cheal cartilages. The smooth muscle is also
tention than those of the gut epithelium. joined to the mucosa by loose connective
The lamina propria of the tracheal epithe¬ tissue continuous with the lamina propria.
lium is a loose connective tissue, unusually A delicate network of lymphatics is found
rich in elastic fibers. It contains many sub¬ beneath the tracheal epithelium. These com¬
mucous glands consisting of tubular acini ra¬ municate with a much coarser plexus in the
diating from a duct that opens onto the
surface of the epithelium (Fig. 29—1). The
cells lining the duct are rich in mitochondria
but devoid of secretory granules. The elon¬
gate acini are lined, in their proximal portion,
by mucus-secreting cells, and their terminal
portion consists of serous cells. The granules
of the serous cells are discrete and electron
dense, while those of the mucous cells are
electron lucent and tend to be confluent. The
secretory product of both is glycoprotein, but
its composition is quite different in each.
The secretion of a blanket of mucus onto
the epithelium of the airway has an important
role in trapping inhaled dust and other par¬
ticulate matter, and in protecting the con¬
ducting and respiratory portions of the lungs
against potentially damaging toxic fumes.
The blanket of mucus is constantly moved by
ciliary action toward the pharynx, where it is
swallowed with the saliva. When the airway
is exposed to tobacco smoke or other irri¬
tants, the submucous glands increase in size,
and the goblet cells of the tracheal and bron¬
chial epithelium increase in number. The
composition of the glycoprotein secreted also
undergoes some modification. After removal Figure 29-8. Cross section through the trachea of a 9-
of the irritant, these changes are reversed in year-old boy. x 6. (Redrawn and modified from Kolliker-
a few months. von Ebner.)
738 • RESPIRATORY SYSTEM

tissue Cartilage
Figure 29-9. Cross section through a small bronchus of a man. x 30. (After Schaffer.)

submucosa, which drains into lymph nodes upper lobe, two in the middle lobe, and five
associated with the trachea along its entire in the lower lobe. In the left lung, there are
length. The blood supply comes mainly from five in the upper and live in the lower lobe.
the inferior thyroid artery. The nerves to the The segmental bronchi branch into subseg-
trachea arise from the recurrent branch of mental bronchi. The blood supply to the lung
the vagus and from the sympathetic chain. follows the same pattern of branching as the
The autonomic nerves of the trachea have bronchi. The parallel course of the bronchi
small ganglia from which fibers lead to the and blood vessels is of great importance to
smooth muscle in its posterior wall. Sensory the thoracic surgeon because it makes lobec¬
nerves are also associated with the mucosa tomy or segmental resection of part of a lobe
and are afferent pathways of the cough re¬ technically feasible.
flex. The structure of the primary bronchi is
very similar to that of the trachea up to the
The Bronchi point where they enter the lungs. Thereafter,
the cartilage rings disappear and are replaced
The trachea divides into two branches by cartilage plates of irregular outline, dis¬
called primary bronchi or main stem bronchi. tributed around the circumference of the
These enter the right and left lung at its tube (Fig. 29—9). Thus, the intrapulmonary
hilus. Coursing downward and outward, they bronchi are cylindrical and not flattened on
divide into lobar bronchi. The left lung has one side as are the trachea and extrapulmo-
upper and lower lobes; the right has upper, nary bronchi. Farther along the airway, the
middle, and lower lobes. Correspondingly, intramural cartilage plates become reduced
there are two lobar bronchi in the left and in size and number and disappear altogether
three in the right lung. The lobar bronchi in in subsegmental bronchi about 1 mm in di¬
turn divide into segmental bronchi to the sev¬ ameter.
eral bronchopulmonary segments in each As the cartilage in the wall of the bronchial
lung. There are three segments in the right tree diminishes, the smooth muscle becomes
 RESPIRATORY SYSTEM • 739

more prominent. It is organized in interlac¬ the level of the surrounding ciliated cells.
ing bundles, some of which have a prevailing Their most characteristic feature is the pres¬
circular or spiral course. In the larger intra- ence of numerous electron-dense secretory
pulmonary bronchi, the bundles are packed granules in their apical cytoplasm. The chem¬
closely enough to form a more or less contin¬ ical nature of the granule contents has not
uous layer. In sections, the bronchial mucosa been definitely established. It is not mucin, is
exhibits longitudinal folds, presumably owing resistant to lipid solvents, and seems to consist
to agonal contraction of the smooth muscle mainly of protein. In the rat, the granules
layer (Fig. 29-9). In more distal portions of have a crystalline substructure not evident in
the bronchial tree, the smooth muscle is more other species. The granules are discharged
attenuated and loosely organized. It is pres¬ by exocytosis. An extracellular material of
ent, however, throughout the conducting similar density is seen in electron micro¬
portion of the respiratory tract. graphs between the microvilli and around
The bronchial epithelium is ciliated col¬ the bases of the cilia. It is believed that the
umnar with goblet cells and associated sub¬ secretory product of the Clara cells forms a
mucosal glands. The latter diminish in num¬ surface coating, lining the bronchioles in
ber, and end at the level of the bronchioles. much the same way that mucus lines the
The height of the epithelium gradually de¬ larger bronchial tubes. A mucus lining would
creases along the tract, becoming cuboidal in not be suitable for such small passages be¬
the bronchioles and sparsely ciliated, low cu¬ cause of its stickiness. The walls of the bron¬
boidal in terminal bronchioles. The lamina chioles come into contact late in expiration
propria, which is set off from the epithelium and it is speculated that a nonsticky, protein¬
by a prominent basal lamina, is loose connec¬ aceous lining layer of low-surface tension
tive tissue rich in reticular and elastic fibers. may be required here to ensure their reopen¬
In addition to fibroblasts, it contains lympho¬ ing on inspiration. Thus, the secretion of the
cytes, mast cells, and occasional eosinophils. Clara cells may play a role analogous to that
of the surfactant lining the alveoli of the
Bronchioles lung.
Another ultrastructural feature of the
The bronchioles, representing the 12th to Clara cells in some species that has aroused
15th generations of branching of the bron¬ considerable speculation is an unusually ex¬
chial tree, are about 0.3 to 0.5 mm in diam¬ tensive smooth endoplasmic reticulum in the
eter. There are no glands or cartilaginous supranuclear cytoplasm. This organelle is
plates in their walls, and the muscularis does well developed in cells engaged in lipid me¬
not form a continuous circumferential layer, tabolism or in cholesterol and steroid secre¬
but is represented by discrete bundles of tion, whereas rough endoplasmic reticulum
varying orientation that join to form a loose is expected in cells synthesizing products rich
network with loose connective tissue filling in in protein. In the liver, the smooth reticulum
the meshes. The smooth muscle is innervated is rich in cytochrome P-450 and mixed-func¬
by parasympathetic nerve fibers. Although tion oxidases involved in the metabolism of
not a conspicuous feature of the bronchiolar drugs. It has been suggested that this orga¬
wall in sections, contraction of the smooth nelle in the Clara cells might metabolize tox¬
muscle very effectively constricts the lumen. ins in the inspired air, but to date this is
It is said to relax during inspiration and purely conjectural.
contract at the end of expiration. When con¬
traction is abnormally persistent, as in asthma
attacks, the constriction of the airway at this
level makes emptying of the lungs during RESPIRATORY PORTION OF
exhalation quite difficult. THE LUNGS
The epithelium lining the bronchioles lacks
goblet cells and consists mainly of ciliated
cells and Clara cells. Except for their reduc¬
Respiratory Bronchioles
tion in height, the ciliated cells are very The bifurcation of the terminal bron¬
similar in their ultrastructure to those more chioles gives rise to respiratory bronchioles,
proximally situated in the airway. The Clara short tubes 0.15 to 2.0 mm in diameter. In
cells are confined to the bronchiolar epithe¬ the human, there are three successive gen¬
lium. Their smooth-contoured, rounded ap¬ erations of respiratory bronchioles, constitut¬
ices or “domes” project into the lumen above ing the transition from the conducting to the
740 • RESPIRATORY SYSTEM

4-'C ;

_4~——- Ciliated epithelium

Respiratory bronchiole
Smooth muscle
j
f
'y»— —Cuboidal epithelium
» *0,
v- A
I |

T-J
$ r';

— Smooth muscle
o /
Alveolar ducts ^..- Cuboidal epithelium

Arteriole

I_Alveolar sacs

Vein

Smooth muscle
Figure 29-10. Drawing of a section through a respiratory bronchiole and two alveolar ducts of human lung. Note the
smooth muscle in the walls of the alveolar ducts. (Slightly modified from Baltisberger.)

respiratory portion of the lung. Their lining are so numerous and closely spaced that the
epithelium is cuboidal at this level, becoming limits of the duct are discernible in sections
low cuboidal and nonciliated in the subse¬ only by the alignment of the free edges of
quent branches. They differ from other thin connective tissue septa between the ad¬
bronchioles in that their walls are interrupted jacent alveoli (Fig. 29—10). The thickened
at intervals by thin saccular outpocketings, adluminal ends of these interalveolar septa
alveoli. Gas exchange can take place in these are covered by a few bronchiolar epithelial
thin-walled appendages, hence the name cells overlying delicate strands of smooth
“respiratory” bronchioles. The number of muscle in the septal connective tissue.
alveoli increases with each branching so that After two or three branchings, each alveo¬
much of the bronchiolar wall is replaced by lar duct ends in a minute space. This is simply
openings into alveoli. the terminal portion of the alveolar duct with
no distinguishing histological features. Al¬
though it is often called the atrium, it is
Alveolar Ducts debatable whether it deserves a separate des¬
ignation (Figs. 29—11, 29—12). It is merely a
The respiratory bronchioles are continu¬ common lumen or vestibule into which clus¬
ous with the alveolar ducts. Here the alveoli ters of four or more alveoli open.
 RESPIRATORY SYSTEM • 741

Figure 29-11. Schematic representation of the respiratory unit of the lung: respiratory bronchiole, alveolar ducts,
alveolar sacs, and alveoli. The atria indicated by the circles are spaces bounded on one side by the termination of the
alveolar duct and on the other by the openings of the alveolar sacs. (Slightly modified after Sorokin, S. In Greep, R. O.,
ed.: Histology. 2nd ed. New York, McGraw-Hill Book Co., 1966.)

Alveoli to the Type II alveolar cells by occluding

junctions.
Physiologically the most important com¬ The Type II alveolar cell is rounded and
ponents of the lung are the thin-walled, ter¬ its margins are partially overlain by the
minal saccular compartments, the pulmonary neighboring squamous cells of the epithe¬
alveoli (Figs. 29—13, 29—14). Their number in lium. The free surface has a limited number
the human lung is estimated to be 300 mil¬ of short microvilli, especially at the periphery
lion, presenting a surface of about 143 sq m, (Fig. 29—16). The rounded basal surface of
which is available for gaseous exchange be¬ the cell bulges into the connective tissue of
tween the inspired air and the blood. the alveolar septa. These cells are most com¬
The alveolar epithelium consists of two cell monly found in niches in the alveolar wall
types, the Type I alveolar cell (Type I pneu- along the junctions of the alveolar septa.
monocyte) and the Type II alveolar cell (also Although the thinly spread Type I alveolar
called Type II pneumonocyte, granular cells are quite inconspicuous in sections of
pneumonocyte, and great alveolar cell). The the alveoli, their cytoplasmic volume is twice
Type I alveolar cells are highly specialized that of the thicker Type II cells and their
squamous cells covering a large area, but they surface area exposed to the lumen is 50 times
are less than 0.2 pm in thickness except greater. Both cell types rest on a thin basal
where they are locally thickened to accom¬ lamina.
modate the nucleus (Fig. 29-15). The free The Type II alveolar cell is a secretory cell
surface of these cells is usually smooth con¬ that is the source of the thin layer of surface
toured. They are attached to each other and active phospholipid, pulmonary surfactant, that
742 • RESPIRATORY SYSTEM

Pulmonary artery

Mucosa of the bronchus

Small bronchus

Nerve

Bronchial artery
Fibrous layer

Smooth muscle '

Cartilage plate

Gland of the mucosa "

Window in muscular layer to show ..

elastic network

Network of elastic fibers

Network of smooth muscles

Bronchiole (without cartilage) —1

Pulmonary vein Respiratory bronchiole

Bronchia! veins

Section of
adjacent
lobule

Opening of alveolar sacs

(atria) into alveolar duct
Interlobular pigment Capillary net¬
work in alveolar walls
3 layers of the pleura (with elastic networks)

Figure 29—12. Portion of a pulmonary lobule from the lung of a young man. Free reconstruction by Vierling, somewhat
foreshortened. Mucosa and glands, green; cartilage, light blue; muscles and bronchial artery, orange; elastic fibers,
blue-black; pulmonary artery, red; pulmonary and bronchial veins, dark blue. (After Braus.)
Figure 29-13. Scanning micrograph of lung from a gazelle, prepared by instillation of the fixative to prevent collapse of
the alveoli and terminal segments of the bronchial tree. (Micrograph courtesy of P. Gehr.)

Figure 29-14. Scanning micrograph of a cut surface of human lung showing the thin alveolar sept& and the interior of
numerous alveoli, x 68. (Micrograph courtesy of P. Gehr.)

743
Figure 29-15. High-magnification scanning micrograph of the interior of an alveolus from bovine lung. The extremely
thin Type I alveolar cells conform to the rounded contours of the underlying capillaries. The boundaries of adjacent
polygonal cells appear as linear ridges (at arrows). (Micrograph courtesy of P. Gehr.)

Figure 29 16. Scanning micrograph of the interior of human pulmonary alveolus, showing a Type II alveolar cell with
its short microvilli located mainly at the periphery of its bulging free surface, x 7600. (Micrograph^courtesy of P Gehr)
744
 RESPIRATORY SYSTEM • 745

coats the epithelial lining of the alveoli. In mately spreads to form a monomolecular film
addition to the usual cytoplasmic organelles, over the alveolar surface (Fig. 29—19).
they have conspicuous dense secretory gran¬ The interstitium of the alveolar septa is the
ules usually located in the supranuclear re¬ tissue between the two layers of pulmonary
gion (Fig. 29—17). These secretory granules epithelium lining adjacent alveoli. It includes
contain the precursors of pulmonary surfac¬ a very close-meshed network of capillaries,
tant and have a characteristic lamellar sub¬ pericytes, septal cells (interstitial fibroblasts),
structure in electron micrographs. Their con¬ mast cells, monocytes, and occasional lym¬
tent is very rich in phospholipid, the phocytes (Fig. 29-20). The most numerous
phospholipid: protein ratio varying from 3:1 of the cellular elements of the interstitium
to 6:1. In freeze-fracture preparations, the are the septal cells. They are considered by
lamellae within the granules are straight and some to be a form of fibroblast and to func¬
stacked with a very regular periodicity. It is tion in maintenance and repair of the con¬
reported, however, that when rapidly frozen nective tissue of the lung. Strips of peripheral
with liquid helium without prior aldehyde lung tissue contract in vitro in response to
fixation, no lamellar structure is seen. The hypoxia. Actin and myosin have been found
formation of the lamellae may therefore be in the septal cells by immunocytochemical
induced during fixation and may not accu¬ methods, and it is speculated that contraction
rately reflect their structure in the living state. of these cells may decrease alveolar size and
Nonetheless the lamellar substructure of the decrease capillary perfusion under certain
granules in routine preparations is a useful physiological conditions. There is also some
identifying feature of Type II alveolar cells. experimental evidence that they participate
The granules are discharged by exocytosis in resynthesis of elastin after enzymatic in¬
(Fig. 29-18). The recently released surfactant duction of emphysema.
often forms patterns resembling myelin In sections, capillaries occupy most of the
forms of hydrated phospholipid, but it ulti¬ thickness of the alveolar septa, and where

Figure 29-17. Electron micrograph of a Type II alveolar cell from fetal rat lung, illustrating the lamellar bodies.
(Micrograph courtesy of M. C. Williams.)
746 • RESPIRATORY SYSTEM

Figure 29-18. Micrograph of a lamellar body being ex¬

truded from an alveolar cell in a 21-day fetal rat lung. Figure 29-19. After release from the Type II epithelial
(Micrograph courtesy of M. C. Williams.) cell, the phospholipid of the lamellar bodies forms complex
myelin figures in the alveolar lumen, as shown in this
micrograph from fetal rat lung near term. The surfactant
ultimately spreads over the lining of the alveoli in a
monomolecular film. (Micrograph courtesy of M. C. Wil¬
the pulmonary epithelium overlies the capil¬ liams. Reproduced from The Journal of Cell Biology
laries, the interstitial space is reduced to an 72:260, 1977 by copyright permission of The Rockefeller
University Press.)
exceedingly thin layer between the basal lam¬
ina of the epithelium and that of the capillary
endothelium. The barrier to diffusion be¬
tween the alveolar air and the blood thus spaces between the capillaries of the alveolar
consists of (1) a layer of fluid and surfactant, wall (Fig. 29—22). From one to six may be
(2) the attenuated epithelium, (3) its basal found and larger numbers have been re¬
lamina, (4) a thin interstitial space, (5) the ported. Their significance is not entirely clear
basal lamina of the capillary, and (6) the but it is thought that they may provide for a
capillary endothelium. The total thickness of collateral air circulation that would tend to
these six components may be as little as 0.2 prevent alveolar collapse (atelectasis) when pe¬
pun but averages about 0.5 pm (Fig. 29-21). ripheral branches of the bronchial tree be¬
Between the neighboring capillaries, the come obstructed. They may also be disadvan¬
interstitium is somewhat thicker, and it is in tageous in that they provide pathways for the
these areas that the septal cells reside to¬ spread of bacteria from alveolus to alveolus
gether with a delicate network of elastic fi¬ in pneumonia.
bers. In the early phase of various forms of The interstitial tissue near the mouths of
pulmonary pathology, there is a leakage of the alveoli is rich in elastic fibers and contains
fluid from the capillaries, resulting in inter¬ a wreath of wavy bundles of collagen fibers.
stitial edema and accumulation of fluid in the These fibers are continuous with those encir¬
alveoli. cling the mouths of neighboring alveoli and
Small openings, alveolar pores (pores of thus tend to give some substance to the wall
Kohn), traverse the thin wall between adja¬ of the alveolar duct. The wavy collagen fibers
cent alveoli. These minute apertures are only probably straighten out as the alveoli expand
7 to 9 pm in diameter and are located in the during inspiration. The extensive network of
 Alveolus

Figure 29-20. Micrograph of equine lung, showing erythrocyte-filled capillaries in the thin septa between alveoli,
x 640. (Micrograph courtesy of P. Gehr.)

Alveolus

Type I alveolar Basal Capillary endothelial

cell

Figure 29-21. A, Micrograph of the septum between two alveoli of rabbit lung. Capillaries covered by Type I alveolar
cells bulge into the lumen, exposing a large surface to the inspired air. B, Higher magnification of an area such as that
enclosed in the rectangle (*) in A, showing the layers making up the diffusion barrier for gaseous metabolites.
(Micrographs courtesy of P. Gehr.) 7-7
748 • RESPIRATORY SYSTEM

Pore of Kohn

Capillary

Surfactant
Figure 29-22. Micrograph of a section of an alveolar septum from dog lung. Occasional small openings, called the
pores of Kohn, traverse the thin wall between adjacent alveoli. The opening here is partially obscured by an accumulation
of surfactant. (Micrograph courtesy of P. Gehr.)

septal reticular fibers is continuous with the In cigarette smokers, the cytoplasm of
collagen bundles that encircle the mouths of these cells is crowded with large, irregularly
the alveolar sacs. These latter in turn merge shaped masses of pigment representing un-
with collagen fibers in the walls of the bron¬ digestible residues of phagocytized material
chioles and small arteries and veins. (Fig. 29-24). These heterogeneous inclusions
Alveolar Macrophages. The pulmonary al¬ have electron-dense and electron-lucent
veolar macrophage is the principal mononu¬ areas, and contain lipid, myelin figures, and
clear phagocyte of the lungs. It is not a part occasionally crystals of unknown provenance.
of the alveolar wall, but a free cell resting on In persons with heart disease attended by
its surface. In this location, it is directly ex¬ pulmonary congestion, the alveolar macro¬
posed to inhaled dust, environmental toxins, phages contain many vacuoles filled with he¬
and bacteria and serves as the primary de¬ mosiderin resulting from phagocytosis of ex-
fense against such particulate matter. travasated erythrocytes and degradation of
Alveolar macrophages vary in size from 15 their hemoglobin.
to 40 pm in diameter and have an irregularly The origin of alveolar macrophages was a
shaped nucleus with a prominent nucleolus. subject of disagreement, some workers be¬
The cytoplasm contains numerous vacuoles lieving that they arose by exfoliation of alveo¬
and may appear “foamy” with the light mi¬ lar lining cells, others insisting that they were
croscope owing to its high degree of vacuoli¬ derived from fixed macrophages present in
zation. In electron micrographs, the cyto¬ the alveolar septa. This controversy now
plasmic organelles include a prominent Golgi seems to have been resolved by use of DNA-
apparatus and a few mitochondria. The en¬ labeling techniques and chromosomal mark¬
doplasmic reticulum is not extensive. Free ers for identifying cells and tracing their
ribosomes and (3 particles of glycogen are origins. It is now widely accepted that pul¬
present in moderate numbers in the cyto¬ monary macrophages are part of the general
plasmic matrix. The most striking ultrastruc- mononuclear phagocyte system of the body
tural feature of the cell is the large number (see Chapter 5). They originate from stem
and heterogeneity of its membrane-bounded cells in the bone marrow and are transported
inclusions (Fig. 29—23). Many of these are in the blood as monocytes that enter the
primary lysosomes, 0.5 pm or less in diameter interstitium of the lung, where they undergo
and round or oblong in shape. These contain certain maturational changes. The mature
a large number of hydrolytic enzymes. The cells then migrate to the surface of the alveoli.
specific activities of acid phosphatase, (3- Most of the proliferative activity giving rise
glucuronidase, and lysozyme are reported to to pulmonary macrophages takes place in the
be substantially higher in alveolar macro¬ bone marrow, but the maturing monocytes
phages than they are in comparable phago¬ in the interstitium of the lung also have a
cytes from the peritoneal cavity. limited capacity for division.
Figure 29-23. Electron micrograph of a human alveolar macrophage from a nonsmoker. Numerous small lysosomes
are present but relatively few heterophagic vacuoles are seen. (Micrograph courtesy of S. Pratt and A. J. Ladman.)

Figure 29-24. Electron micrograph of an alveolar macrophage from an 18-year-old cigarette smoker. The cytoplasm is
crowded with pigment masses representing undigestible residues of material phagocytized from the alveoli. (Micrograph
courtesy of S. Pratt and A. J. Ladman.)
749
750 • RESPIRATORY SYSTEM

In common with other phagocytes, the pleura are believed to be branches of the
alveolar macrophages have receptors on their vagus and the sympathetic nerves supplying
surface for IgG and the C3b component of the bronchi.
complement. Their ability to ingest bacteria
is enhanced in the presence of specific anti¬
body. Intracellular destruction of bacteria is
INNERVATION OF THE LUNGS
carried out by their phagolysosomal system
supplemented by the generation of super¬
oxide ion. The lung is innervated by parasympathetic
The alveolar macrophages form the first nerves via the vagus nerve and by sympa¬
line of defense of the lungs and are remark¬ thetic nerves arising from the second to
ably efficient. Although large numbers of fourth thoracic sympathetic ganglia. These
infectious organisms are continually carried nerves form plexuses around the hilus of the
into the lungs in the inspired air, the alveolar lung and give rise to intrapulmonary nerves
surface is usually sterile. Alveolar macro¬ that accompany the ramifications of the bron¬
phages greatly outnumber all other cell types chial tree and its associated blood vessels.
in the lung and are constantly being elimi¬ Parasympathetic and sympathetic nerves to
nated and replaced. They migrate or are the lung contain both afferent and efferent
passively transported from the alveolar sur¬ fibers. The bronchoconstrictor fibers are
face to the bronchioles, and then are carried from the vagus, and the bronchodilator fibers
by ciliary action through the upper airways are sympathetic.
to the pharynx, where they mingle with sali¬ Both sensory and motor nerves have been
vary secretions and are swallowed. The mech¬ identified extending as far peripherally as
anism of their directed movement to the the terminal bronchioles. Light microscopic
bronchioles is not entirely clear, but the con¬ observations based on methylene blue and
tinuous production of surfactant and the silver stains left some doubt as to whether
transudation of fluid from the capillaries may the innervation of the lung extended beyond
form a moving surface film for transport of the respiratory bronchioles. Electron micro¬
the macrophages. The number cleared from scopic studies have now shown nerve endings
the lungs each day is astronomical. On the in the alveolar ducts and alveolar walls of
basis of counts of mononuclear cells in the laboratory rodents and they probably also
respiratory fluid of cats with tracheal can¬ occur in humans. These endings are of two
nula, it has been estimated that the clearance types. One located in the interstitium, and
rate is about 2 X 106 alveolar macrophages containing many small mitochondria, is con¬
per hour. Comparable measurements for hu¬ sidered to be sensory. It may possibly corre¬
mans would no doubt give much higher num¬ spond to the juxtacapillary (J-type) receptor
bers. postulated by physiologists and believed to
be stimulated by an increase in pulmonary
capillary pressure. The second type of ending
is of less common occurrence and contains
THE PLEURA many dense-cored vesicles. It is found in close
association with Type II pneumonocytes and
The cavities containing the lungs are lined it is speculated that it may be the morpho¬
by a serous membrane, the pleura, which logical basis for the reported effect of the
consists of a thin layer of collagenous tissue nervous system on the secretion of pulmo¬
containing some fibroblasts and macrophages nary surfactant.
and several prominent layers of elastic fibers.
It is covered by a layer of mesothelial cells
like those of the peritoneum. The layer lining Neuroepithelial Bodies
the wall of the thoracic cavity is called the Physiologists have long postulated the ex¬
parietal pleura; that reflected over the surface istence of chemoreceptors in the lung sensi¬
of the lungs is the visceral pleura. A prominent tive to the composition of the inhaled air. A
feature of the pleura is the great number of morphological basis for this function has only
blood capillaries and lymphatic vessels dis¬ recently been identified. Isolated groups of
tributed in it. The few nerves of the parietal specialized cells are found throughout the
pleura are branches of the phrenic and in¬ epithelium lining the airways. These cells are
tercostal nerves. The nerves to the visceral selectively stained by silver salts (argentaffin
 RESPIRATORY SYSTEM • 751

reaction) and emit fluorescence at the wave¬ neuroendocrine cells, as in other chemore-
length characteristic of serotonin (5-hydrox- ceptors such as the carotid body. The re¬
ytryptamine). These aggregations of cells sponse of the neuroepithelial bodies may
have been called neuroepithelial bodies. They therefore be modulated by the central ner¬
occur in the epithelium of bronchi, and dis- vous system.
tally to the terminal bronchioles, and seem
to be preferentially located at or near sites of
bifurcation. The groups of cells forming
these bodies have the general shape of a BLOOD SUPPLY
truncated cone, 15 pm high with a rounded
base 30 to 45 pm wide. The apex projects The lungs receive most of their blood from
slightly above the surface of the surrounding the pulmonary arteries. These are of large
bronchiolar epithelium. caliber and of elastic type. The branches of
In histological sections, the neuroepithelial these arteries accompany the bronchi and
bodies consist of two cell types—modified their branches as far as the respiratory bron¬
Clara cells that serve as supporting elements chioles. The arterial paths in the lung, how¬
and 10 to 15 neurosecretory cells containing ever, are subject to considerable variation.
small granules in their basal cytoplasm. In From the respiratory bronchioles they divide,
electron micrographs, the apical pole of the and a branch passes to each alveolar duct
secretory cells extends to the lumen, and and is distributed in a capillary network over
bears numerous slender microvilli and occa¬ all the alveoli that communicate with this
sionally a solitary cilium. The nucleus is large duct. The venules arise from the capillaries
and indented, with the clumps of chromatin of the pleura and from the capillaries of the
situated mainly at the periphery. The Golgi alveolar septa and portions of the alveolar
complex is supranuclear, and most mitochon¬ ducts. They run in the intersegmental con¬
dria are in the apical cytoplasm. Granular nective tissue, independently of the arteries,
endoplasmic reticulum is moderately exten¬ and join to form the pulmonary veins. In
sive in the apical and paranuclear regions. passing through the lung, the pulmonary
Free ribosomes are abundant in the cyto¬ artery is usually above and behind its accom¬
plasmic matrix and there are a few granules panying bronchial tube, whereas the vein is
of glycogen. below and in front of it. The bronchial arter¬
The most characteristic feature of the cells ies and veins are much smaller than the
is the presence of small, dense-cored vesicles pulmonary vessels. These arteries arise from
or granules throughout the cytoplasm, but the aorta or the intercostal arteries and follow
most concentrated near the base. Their flu¬ the bronchi. They are distributed to the walls
orescence, positive chromaffin reaction, and of the bronchi, their glands, and the interlob¬
argentaffin staining leave little doubt that ular connective tissue beneath the pleura.
these granules contain serotonin. There is Most of the blood carried by the bronchial
suggestive evidence that these cells may also arteries is brought back by the pulmonary
synthesize or store other biologically active veins. In the alveoli that arise from the res¬
peptides, some of which may cause contrac¬ piratory bronchioles, there are capillary an¬
tion of bronchiolar smooth muscle. T he gran¬ astomoses between the terminations of the
ules are released by exocytosis in response to pulmonary and the bronchial arteries.
hypoxia and hypercapnia. It is probably sig¬
nificant that the capillaries associated with
the neuroepithelial bodies are fenestrated,
while those elsewhere in the lamina propria
LYMPHATICS
of the bronchial tree are not.
It is believed that the neuroepithelial bod¬ There are two main divisions of the lym¬
ies are directly responsive to diminished ox¬ phatics of the lungs. One set is in the pleura
ygen content in the inspired air, releasing and the other in the pulmonary tissue. They
serotonin and possibly other mediators into communicate infrequently. Both drain into
the blood, thereby regulating alveolar capil¬ the lymph nodes at the hilus of the lung. I he
lary circulation and affecting the ventilation: lymphatics of the pleura form a dense net¬
perfusion ratio. The bodies appear to have a work with large and small polygonal meshes.
special innervation with unmyelinated affer¬ The large meshes are formed by large vessels
ent and efferent fibers ending among the and demarcate the lobules; the small mesh-
752 • RESPIRATORY SYSTEM

work is formed of smaller vessels that mark area. Air flowing over it is warmed and mois¬
out the anatomical units. There are many tened before it reaches the trachea and lungs.
valves in these lymphatics that control the The importance of the partial saturation of
flow of lymph so that it passes toward the the air with water vapor during its passage
hilus and not into the pulmonary tissue. through the nose is evident in patients with
These pleural lymphatics join to form several a tracheotomy who are obliged to take air
main trunks, which drain into the lymph directly into the trachea. Under these condi¬
nodes at the hilus. tions, excessive drying of the mucosa may
The pulmonary lymphatics may be divided lead to encrustation, interference with clear¬
into several groups, which include those of ance of the upper airway, and ultimately to
the bronchi, of the pulmonary artery, and of serious infection. Similarly, if one nostril of
the pulmonary vein. The lymphatics accom¬ a dog is surgically closed, the increased ven¬
panying the bronchi extend peripherally as tilation of the other side of the nasal cavity
far as the alveolar ducts, and their end exceeds its humidifying capacity, and the
branches join the lymphatic radicles of the drying effect may result in transformation of
plexuses about the pulmonary artery and the ciliated columnar epithelium into strati¬
vein. There are no lymphatic vessels beyond fied squamous epithelium—a phenomenon
the alveolar ducts. The pulmonary artery is called squamous metaplasia.
accompanied by two or three main lymphatic The filtration of the inspired air during its
trunks. The lymphatics associated with the passage through the nose is also important.
pulmonary vein begin with its radicles in the Targe particles are excluded by the coarse
alveolar ducts and in the pleura. All the hairs in the external nares, while the smaller
lymphatics of the pulmonary tissue drain ones tend to be entrapped in the layer of
toward the hilar nodes. Efferent trunks from mucus in the lining epithelium, which is con¬
the hilar nodes anastomose to form the right tinually moved by ciliary action toward the
lymphatic duct, which is the principal chan¬ pharynx, where it is swallowed.
nel of lymph drainage from both the right The clearing of the respiratory passages by
and left lungs. There are no valves in the coordinated beating of cilia on the lining
intrapulmonic lymphatics except in a few epithelium also takes place in the paranasal
vessels, in the interlobular connective tissue sinuses and throughout the trachea and
near the pleura, which accompany the bronchi. The secretion of goblet cells and
branches of the pulmonary veins. These lym¬ submucosal glands forms a coherent sheet of
phatic vessels connect the pulmonary and mucus covering the surface of the mucosa.
pleural lymphatic plexuses. As their valves Cilia constantly beating at about 14 cycles a
point only toward the pleura, they provide a second beneath it move this blanket of mucus
mechanism whereby lymph can flow from at a speed of as much as 1 cm per minute,
the pulmonary tissue into the pleural lym¬ carrying adherent dust particles, bacteria, cel¬
phatics if the normal flow of lymph from the lular debris, and adsorbed chemical pollu¬
pulmonary tissue toward the hilus is inter¬ tants toward the pharynx, where they are
rupted. disposed of by swallowing. The physiological
As mentioned, the mucosa of the bronchi importance of ciliary activity is clearly dem¬
is infiltrated with lymphocytes and often con¬ onstrated in those rare individuals who have
tains germinal centers. There are other ac¬ the inherited condition known as Kartagener’s
cumulations of lymphatic tissue in the adven¬ disease or immotile cilia syndrome. Their genetic
titia of the pulmonary arteries and veins. defect results in failure to synthesize dynein,
the ATPase-containing protein that forms
the “arms” on the microtubules of ciliary and
flagellar axonemes (see Chapter 2). Since
HISTOPHYSIOLOGY OF THE dynein is essential for conversion of the
RESPIRATORY TRACT chemical energy in ATP to the mechanical
work of microtubule sliding, males with this
deficit have immotile spermatozoa and are
The functions of the nasal passages in infertile, and individuals of both sexes suffer
warming, humidification, and filtration of the from chronic sinusitis, chronic bronchitis,
inspired air are often not fully appreciated. and bronchiectasis owing to the absence of
The mucous membrane lining the nasophar¬ motile cilia to clear their respiratory passages.
ynx and covering the turbinates is richly
Another essential device for removal of
vascularized and presents a large surface
particulate matter and irritant chemicals
 RESPIRATORY SYSTEM • 753

from the airway is the cough reflex. This de¬ itates expansion. Deficiency of surfactant se¬
pends on the presence of sensory nerve end¬ cretion occurs occasionally in the newborn,
ings in the lining epithelium. Afferent im¬ resulting in respiratory distress syndrome (hyaline
pulses pass from these to the medulla in the membrane disease), a condition that often re¬
central nervous system, triggering an auto¬ sults in the death of the infant because of
matic sequence of events that involves deep inadequate pulmonary ventilation.
inspiration; closure of the epiglottis and vocal Destruction of elastic fibers and diminished
cords; and forceful contraction of abdominal elastic recoil are prominent features of the
and internal intercostal muscles that raises pathophysiology of emphysema, a condition
pressure on the air entrapped in the lungs. characterized by destructive changes in the
The epiglottis and larynx are then suddenly alveolar walls, resulting in great enlargement
opened and the air under pressure bursts of the air spaces distal to the terminal bron¬
out, attaining velocities as high as 100 mph. chioles and progressive inefficiency of gas¬
The rush of air carries with it the irritant eous exchange. A number of environmental
foreign matter, removing it from the trachea factors, including cigarette smoking and air
and bronchi. The sneeze reflex involves a similar pollution, may contribute to the development
train of events that results in clearance of the of the disease. However, it may occur in
nasal passages. certain susceptible individuals in the absence
The primary function of the lungs is, of of these factors. Susceptible persons lack nor¬
course, to provide for assimilation of oxygen mal blood levels of ay-antitrypsin, a versatile
from the air and removal of carbon dioxide inhibitor of various proteases, including elas-
from the body. In inspiration, contraction of tase. Such individuals may be unable to con¬
the diaphragm lowers intra-alveolar pres¬ trol the effects of hydrolytic enzymes released
sure, drawing air into the lungs. The network from the lysosomes of macrophages and
of blood capillaries in the walls of the alveoli granulocytes that accumulate in chronic pul¬
is separated from the air by a very thin moist monary infections, and enzymatic destruction
sheet of alveolar epithelium that permits of the connective tissue framework of the
rapid diffusion of oxygen into, and carbon alveolar wall then follows.
dioxide out of, the blood. The exchange of The lungs also have important nonrespi-
gases takes place by passive diffusion, but the ratory functions. Since the entire output of
liberation of carbon dioxide from carbonic the heart passes through the lungs, the en¬
acid is accelerated about 5000-fold by the dothelium of the pulmonary vessels is favor¬
enzyme carbonic anhydrase, which resides in ably situated to metabolize or transform var¬
the red blood cells. In addition to gas ex¬ ious substances that circulate in the blood.
change, there is a loss of approximately 800 Although the endothelium of these vessels is
ml of water a day in the expired air. Ether, morphologically identical to that of capillaries
other volatile anesthetics, and alcohol in the elsewhere in the body, it possesses distinctive
blood may also be eliminated via the lungs. enzymatic properties. It contains monoamine
Expiration results from relaxation and el¬ oxidase, which enables it to break down
evation of the diaphragm, diminishing the serotonin that is released into the blood by
size of the thorax. A significant component other organs. Enzymes that transform the
of expiration is attributable, however, to the vasoactive substance angiotensin I to angio¬
inherent elastic properties of the lung. The tensin II, and enzymes that inactivate brady-
parenchyma of the lung is rich in delicate kinin, also have been demonstrated in pul¬
extracellular elastic fibers that stretch during monary endothelium. These and other bio¬
inspiration and recoil during expiration. In logically active substances are metabolized
addition, intermolecular forces within the almost completely in a single passage through
thin him of fluid on the alveolar lining main¬ the vascular bed of the lungs.
tain a surface tension that tends to reduce The lung can also be considered to have
the surface area of the alveoli. Surface ten¬ an endocrine function. In response to certain
sion is responsible for about two thirds of the stimuli, it releases into the blood prostaglan¬
recoil of the lungs, and elastic fibers account dins, histamine, slow-reactive substance of
for the rest. The resistance to alveolar expan¬ anaphylaxis, and other mediators. Thus, the
sion imposed by surface tension would be lung is not merely concerned with exchange
very much greater were it not for the secre¬ of gaseous metabolites, but also plays a sig¬
tion of surfactant by the Type II epithelial nificant role in control of the blood levels of
cells, which lowers surface tension and facil¬ many biologically active substances.
754 • RESPIRATORY SYSTEM

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Swindle, P. F.: The architecture of the blood vascular Thurlbeck, W. M.: Structure of the lungs. Int. Rev.
networks in the erectile and secretory lining of the Physiol. 74:1, 1977.
nasal passages. Ann. Otol. Rhinol. Laryngol. Weibel, E. R. Morphological basis of alveolar-capillary
744:913, 1935. gas exchange. Physiol. Rev. 55:419, 1973.
TRACHEA, BRONCHI, AND BRONCHIOLES Weibel, E. R., P. Gehr, D. Haies, and J. Gil: The cell
population of the normal lung. In Bouhuys, A., ed.:
Boyd, M. R.: Evidence for the Clara cell as a site of Lung Cells in Disease. Elsevier/North Holland
cytochrome P450-dependent mixed function oxi¬ Biomedical Press, 1976.
dase activity in lung. Nature (Lond.) 269:713, 1977. Williams, M. C.: Conversion of lamellar body mem¬
Foliguet, B., and J. L. Cordonnier: Pulmonary neuro¬ branes to tubular myelin in alveoli of fetal rat lung.
epithelial bodies. Bull. Europ. Physiopathol. Respir. J. Cell Biol. 72:260, 1977.
77:113,1981.
Hoyt, R. F., Jr., S. P. Sorokin, and H. Feldman: Number, PULMONARY MACROPHAGES AND MAST
subtypes, and distribution of small granule neu¬ CELLS
roendocrine cells in the infracardiac lobe of hamster Brain, J. D., J. J. Godleski, and S. P. Sorokin. Quantifi¬
lung. Exp. Lung Res. 3:273, 1983.
cation, origin and fate of pulmonary macrophages.
Hung, K. S., M. S. Hertwick, J. D. Hardy, and C. L.
In Brain, J. D., D. F. Proctor, and L. Reid, eds.:
Loosli: Ultrastructure of nerves and associated cells
Respiratory Disease Mechanisms. New York, Marcel
in bronchiolar epithelium of the mouse lung. J. Dekker, 1977, p. 849.
Ultrastruct. Res. 45:426, 1973.
Hocking, W. G., and D. W. Golde: The pulmonary-
Jeffery, P. K., and L. Reid: New observations of rat
alveolar macrophage. N. Engl. I. Med. 597:580,
airway epithelium: a quantitative and electron mi¬ 639, 1979.
croscopic study. J. Anat. 729:295, 1975.
Sorokin, G.: Phagocytes in the lungs: incidence, general
Lauweryns, J. M., and M. Cokelaere: Hypoxia-sensitive
behavior, and phytogeny. In Brain, J. D., D. F. Proc¬
neuro-epithelial bodies. Intrapulmonary secretory
tor, and L. Reid, eds.: Respiratory Disease Mech¬
neuro-receptors modulated by the CNS. Z. Zell-
anisms. New York, Marcel Dekker, 1977, p. 711.
forsch. 745:521, 1973.
Wasseiman, G. I.: The lung mast cell: its physiology and
Lauweryns, J. M., M. Cokelaere, P. Theunynck, and M.
potential relevance to defense of the lung. Environ.
Delearsnyder: Neuroepithelial bodies in mammalian
Health Perspect. 55:153, 1980.
 30
THE URINARY SYSTEM

The urinary system consists of the kidneys, When the cut surface of the hemisected
ureters, urinary bladder, and urethra. The kidney is viewed with the naked eye, a darker
system functions to clear the blood of the reddish brown cortex is readily distinguishable
waste products of metabolism and to regulate from a lighter medulla. The medulla is made
the concentration of many constituents of the up of 5 to 11 conical subdivisions called renal
body fluids. In addition to their excretory pyramids, each having its base toward the
function, the kidneys have an endocrine cortex and its apex or papilla projecting into
function—producing and releasing into the the lumen of a minor calyx. The lateral
bloodstream a humoral agent that affects boundaries of each pyramid are defined by
blood formation (erythropoietin) and an¬ inward extensions of the darker cortical tis¬
other that influences blood pressure (renin). sue forming the renal columns (of Bertin). A
In the male, the urethra not only conveys the renal pyramid together with the cortical tis¬
urine to the outside but also serves the re¬ sue overlying its base and covering its sides
productive system as the pathway for the constitutes a renal lobe. Each lobe of the hu¬
discharge of semen. man kidney corresponds to the entire uni-
pyramidal kidney of common laboratory ro¬
dents. During embryonic development, each
lobe arises in association with a different
KIDNEYS minor calyx, and during fetal life the several
lobes are recognizable as distinct convexities
on the surface of the organ, but later in
The human kidneys are paired organs sit¬ development they fuse into a continuous
uated retroperitoneally on the posterior wall smooth-contoured cortex.
of the abdominal cavity on either side of the The gray substance of each pyramid is
vertebral column. They are roughly bean¬ radially striated with brownish lines that con¬
shaped, 10 to 12 cm in length, 5 to 6 cm in verge toward the apex of the papilla. These
width, and 3 to 4 cm in thickness. A concavity, striations are a reflection of the orientation
the hilus, is found on the medial border. The of the straight portions of the microscopic
large excretory duct, the ureter, emerges from uriniferous tubules and of the blood vessels
the hilus and courses downward to the uri¬ that course parallel to them. The tip of each
nary bladder, which is situated in the pelvis papilla, called the area cribrosa, is perforated
directly behind the pubis. The kidney is by about 25 small openings, where the ter¬
closely invested by a thin but strong capsule minal segments of the uriniferous tubules
of dense collagenous fibers. The parenchyma open into a minor calyx.
of the kidney surrounds a large cavity, the The myriad renal tubules that constitute
renal sinus, that extends inward from the hilus the parenchyma of the kidney are specialized
and contains the renal pelvis. The remainder along their length for different functions,
of the sinus around the renal pelvis is occu¬ and each of the specialized segments tends
pied by loose connective tissue and adipose to be located at a particular level. This con¬
tissue, through which the blood vessels and sistency in distribution of corresponding seg¬
nerves pass into the renal tissue. ments is reflected in grossly distinguishable
The renal pelvis is a funnel-shaped expan¬ zones in the medulla, which differ slightly in
sion of the upper end of the ureter, which color or pattern. There is an inner and outer
sends into the substance of the kidney two or zone of the medulla, and the outer zone is
three sizable outpocketings called major ca¬ sometimes further subdivided into a darker
lyces. These in turn have a number of smaller and thicker inner band or stripe and a lighter
branches called minor calyces (Fig. 30—1). and thinner outer band.
755
756 • THE URINARY SYSTEM

Arcuate vessel

Renal column (cortical)

Pyramid

Cortical substance

Incised minor calyx Figure 30-1. Human kidney, seen

Hilus from behind, after removal of part of
Renal artery the organ. Three fifths natural size.
Medullary substance
(After Braus.)
Renal vein .
Major calyx
Renal pelvis
Ureter Minor calyx

Cortical substance

From the bases of the medullary pyramids, urine, and serves as the excretory duct con¬
thin, radially directed striations extend into veying the urine to the renal pelvis. These
the cortical substance. These bear some re¬ two components arise in the embryo from
semblance to the striations in the pyramid separate primordia that become connected
but do not extend through the entire thick¬ secondarily. This is in contrast to the devel-
ness of the cortex. These markings are called
the medullary rays (of Ferrein) and represent
continuations of bundles of tubules from the Capsule
pyramid into the cortex (Figs. 30-2, 30-3).
Each medullary ray and its immediately as¬
sociated cortical tissue are referred to as a
renal lobule, although they are not separated
from one another by connective tissue septa,
as is often the case in lobules of other glands. Glomeru lus

The structural basis of the patterns that are

visible on the cut surface of the normal kid¬
ney will be better understood after the de¬
Medullary
scription of the uriniferous tubules. It is
ray
sufficient here to note that recognition of
these gross markings is not only an aid to
understanding of the complex microscopic
organization of this organ, but is also of
practical value to the pathologist to the extent
that loss or distortion of the normal pattern
is associated with particular disease entities.

Arcuate
URINIFEROUS TUBULES artery

The tubules composing the kidney have Outer zone

two principal portions. The first portion, the of medulla

nephron, corresponding to the secretory ele¬

ments of other glands, is concerned with the
formation of urine; the second portion, the
collecting tubule, carries out a final concentra¬ Figure 30-2. Section of kidney of Macacus rhesus.
Fixation by vascular perfusion—hence the empty blood
tion of urinary solutes to form a hypertonic vessels. Photomicrograph (slightly retouched), x 13.
 THE URINARY SYSTEM • 757

Juxtamedullary
glomeruli

Inner
cortex <

\ Juxtamedullary
cortex

Outer
medulla <
Medullary rays
and vasa recta

V.

Inner
medulla

Figure 30-3. Section of dog kidney, showing medullary rays and vasa recta extending from juxtamedullary glomeruli to
the border of the inner medulla. Mallory stain, x 30. (After Thorburn, W., et al. Circulation Res. 73:290, 1963.)

opment of other glands, in which the ducts which is deeply indented by a globular tuft
and secretory portions arise from a single of capillaries, the glomerulus (Fig. 30—4). This
primordium that branches dichotomously mass of capillaries and its surrounding chal¬
and becomes secretory in the distal part of ice-shaped epithelial capsule together consti¬
its arborescent pattern. tute the renal corpuscle. It has a vascular pole
where the afferent and efferent vessels enter
The Nephron and leave the glomerulus, and a urinary pole
where the slitlike cavity within the capsule of
The nephron is the tubular functional unit Bowman is continuous with the lumen of the
of this organ. There is estimated to be about next segment of the nephron, the proximal
two million in each kidney, and the output tubule (Fig. 30—5). This segment consists of a
of urine represents a summation of the func¬ convoluted and straight portion. The latter
tions of this very large number of units. is followed by a thin segment, and this in turn
Along the length of the nephron are several by the straight and convoluted portions of
morphologically distinct segments, each hav¬ the distal tubule. The convoluted portion of
ing a characteristic configuration and occu¬ the proximal tubule (proximal convoluted tu¬
pying a definite position in the cortex or bule) and the convoluted portion of the distal
medulla. Each segment is lined with a specific tubule (distal convoluted tubule) are both lo¬
type of epithelium specialized for a particular cated in the cortex close to the renal corpus¬
role in the formation of urine. cle. The portion of nephron between the two
At the proximal end of each is a thin- convoluted segments (namely, the straight
walled expansion called Bowman’s capsule, portion of the proximal tubule, the thin seg-
758 • THE URINARY SYSTEM

Figure 30-4. Scanning micrographs of plastic casts of two glomeruli. Left, viewed at the vascular pole; right, from the
capsular side. Observe in both that the caliber of the efferent arteriole is smaller than that of the afferent arteriole. This
no doubt ensures an adequate filtration pressure in the capillaries. (Micrographs from Takizawa, J., et al. Lab. Invest.
40:519, 1979.)

ment, and the straight portion of the distal from the radially oriented straight collecting
tubule) forms a loop, the loop of Henle, ex¬ tubules. The latter are continuous with the
tending from the cortex for a variable dis¬ papillary ducts, which deliver urine to the
tance into the medulla. The radially oriented minor calyces.
descending and ascending limbs of the loop The tortuous convoluted tubules of neigh¬
run parallel to each other and are connected boring nephrons in the cortex intermingle so
by a sharp bend (Fig. 30—5). These several extensively that the identity and shape of the
segments are represented in the same se¬ individual units cannot be ascertained from
quence in all nephrons, but the length of the study of histological sections. What is known
loop and the proportions of the segments of of their three-dimensional configuration has
which it is formed vary with the position of been established by their reconstruction from
the glomerulus in the cortex. Nephrons serial sections or by maceration of the tissue
whose glomeruli are in the outer or subcap- and teasing out the individual nephrons by
sular portion of the cortex have short loops time-consuming micromanipulation. It has
of Henle with abbreviated thin segments, and also been possible by micropuncture to pen¬
these loops extend only a very short distance etrate individual glomerular capsules and to
into the outer zone of the medulla. Nephrons observe directly the progress of injected con¬
whose glomeruli are situated in the deep or trast media through the lumen of a single
juxtamedullary region of the cortex form a nephron.
loop with long descending and ascending The renal corpuscles have evolved as effi¬
limbs and an extensive thin segment, which cient ultrafiltering devices for clearing the
penetrates deep into the inner zone of the blood of wastes. About 1200 ml of blood
medulla. Nephrons with glomeruli midway flows through the kidneys per minute and
in the cortex have intermediate characteris¬ about one fifth of the plasma volume is fil¬
tics. In addition, there are significant differ¬ tered off in the renal corpuscles. Thus, about
ences in the vascular supply to these three 120 ml of a fluid, called the glomerular filtrate,
categories of nephrons. These will be de¬ enters the renal tubules each minute. As this
scribed later. The distal convoluted tubules fluid passes through the various segments of
are joined to the collecting duct system by a the nephron, its composition is modified by
short connecting segment, often referred to secretion of certain substances into it and
as the arched collecting tubule to distinguish it reabsorption of water and other constituents
 THE URINARY SYSTEM • 759

Cortical from it. The final product, urine, is drained

Nephron
through the collecting ducts into the renal
pelvis.
Renal Corpuscle. The capsule of Bowman
around the tuft of glomerular capillaries is a
double-walled cup composed of squamous
epithelium. Although not strictly true, it is
conceptually useful to consider that in the
embryonic development of the renal corpus¬
cle, the glomerulus is pushed into, and deeply
indents, a blind terminal expansion of the
uriniferous tubule like a fist thrust into a
balloon. From this mode of development it
would follow that there is a visceral layer of
epithelium (the glomerular epithelium) applied
to the capillaries, as well as a parietal layer
(the capsular epithelium). Between these is a
narrow chalice-shaped cavity, the capsular
space (Bowman’s space). At the vascular pole
of the renal corpuscle, the visceral layer is
reflected off the afferent and efferent glo¬
merular vessels to become continuous with
the epithelium of the parietal layer. At the
urinary pole, the squamous capsular epithe¬
lium is continuous with the cuboidal epithe¬
lium in the neck of the proximal convoluted
tubule (Fig. 30—6).

Figure 30-5. Highly schematic drawing of a cortical Figure 30-6. Highly schematic representation of the renal
nephron and a juxtamedullary nephron, comparing the corpuscle. The parietal layer of Bowman’s capsule is
renal corpuscles, proximal convoluted tubules, straight depicted considerably thicker than it actually is, and the
descending portions of proximal tubule, thin segments, visceral layer overlying the capillaries of the glomerulus
straight ascending portions of the distal tubule, distal is greatly simplified, with only the major processes of the
convoluted tubules and collecting tubules. The cortical podocytes depicted. Although earlier described as a clus¬
nephrons have a short loop of Henle with a very short ter of simple loops, the capillaries are now believed to
thin segment, whereas these structures are long and branch and anastomose to form a network. (Redrawn and
extend deep into the medulla in juxtamedullary nephrons. modified after Bargmann.)
760 • THE URINARY SYSTEM

cells of the visceral layer become so exten¬

sively modified that, in the adult, they bear
little resemblance to any other epithelial cells.
These cells, called podocytes, are closely ap¬
plied to the capillaries and are basically stel¬
late, with several radiating primary processes
that embrace the vessels in a manner remi¬
niscent of the pericytes of other capillaries
(Fig. 30—7). The primary processes give rise
to very numerous secondary processes, also
Nwf J*m 1 iSilM f j 111; called foot processes or pedicels. These interdi-
gitate with corresponding elements of neigh¬
* Mi
boring podocytes to create an extraordinarily
~ ■mm" elaborate system of intercellular clefts, called
Figure 30-7. Highly schematic representation of the in- filtration slits (Figs. 30—8, 30—9).
terdigitating pattern of secondary processes (foot proc¬ In thin sections examined with the electron
esses or pedicels) of the podocytes on the outer surface microscope, the cell bodies of the podocytes
of a glomerular capillary loop. This arrangement provides
are rarely found in extensive contact with the
a very large area of slender filtration slits or slit pores
between adjacent processes. (Redrawn and modified after basal lamina. Instead, they stand off 1 or 2
Gordon. In Ham, A. W.: Histology. 5th ed. Philadelphia, pm and are attached to it via their primary
J. B. Lippincott Co., 1965.) and secondary processes, which ramify over
the surface of the basal lamina. The cell body
and major processes of one podocyte may
In the development of the renal corpuscle, arch over undermining primary processes of
the parietal layer remains a typical squamous neighboring podocytes (Fig. 30—9). As a con¬
epithelium of flat polygonal cells, but the sequence of these relationships, the greater

Rgure 30-8. Scanning electron micrograph of a small area of the glomerulus, showing the visceral epithelium 'consist™
of podocytes with their interdigitating processes forming an elaborate filigree around the cylindrical capillaries (Micrograph
courtesy of F. Spinelli, Ciba-Geigy Ltd. Research Laboratories, Basle, Switzerland.) ^ ^
 THE URINARY SYSTEM • 761

Figure 30-9. Scanning micrograph of two adjacent capillary loops of rat glomerulus, illustrating the remarkable
complexity of the primary and secondary processes of the podocytes, whose ramification and inierdigitation leaves an
enormous area of filtration slits through which the glomerular filtrate passes into the capsular space. (Micrograph
courtesy of F. Spinelli, Ciba-Geigy Ltd. Research Laboratories, Basle, Switzerland.)

part of the capsular surface of the glomerular barrier requires the higher resolution avail¬
capillaries is carpeted by interdigitating foot able in transmission electron micrographs.
processes, thus providing a maximal area of The podocytes have nuclei of complex form,
slit pores for filtration. By laborious study of often deeply infolded. Their cytoplasm con¬
thin sections with the transmission electron tains a well-developed Golgi complex, cister¬
microscope and imaginative reconstruction, nal profiles of granular endoplasmic reticu¬
students of renal cytology have ventured to lum and abundant free ribosomes.
depict the three-dimensional configuration of Cytoplasmic filaments and microtubules are
the podocytes, as shown in Figures 30—6 and plentiful, both in the cell body and in the
30—7. With the scanning elecron microscope, primary and secondary processes. The foot
it is now possible to obtain three dimensional processes are aligned upon the outer surface
images of the cellular topography of this of a thick basal lamina, which they share with
remarkably specialized epithelium. Its actual the endothelium of the underlying glomer¬
complexity (Figs. 30-8, 30—9) far exceeds the ular capillary (Fig. 30—12). Adjacent proc¬
most daring earlier interpretations of its esses are not in contact but are separated by
structure based on conventional light and filtration slits about 25 nm wide. The plasma
electron microscopy. membrane of the foot processes has a prom¬
Demonstration of the cytology of the po¬ inent surface coat, or glycocalyx, that stains
docytes and the finer details of the filtration intensely with ruthenium red and osmium
762 • THE URINARY SYSTEM

hanced. Fine filaments appear to connect the

densities to the membranes of the epithelium
and endothelium, respectively. Similar dis¬
crete binding sites for cationic molecules are
seen between other epithelia and endothelia
and their basal lamina. It is speculated that
these sites may play a role in attachment of
the cells to the meshwork of macromolecules
of Type IV collagen and proteoglycans that
make up the basal lamina. A major
glycosaminoglycan of the glomerulus is hep¬
aran sulfate, which contributes to the strongly
anionic properties of its basal laminae, to¬
gether with the carboxyl groups of collage¬
nous and noncollagenous glycoproteins.
The endothelium of glomerular capillaries
is quite thin and is perforated by pores or
fenestrae 70 to 90 nm in diameter. In other
fenestrated endothelia, the pores have a thin-
pore diaphragm. This is not present in glo¬
merular endothelium. The abundance and
distribution of endothelial pores are best seen
in the extended en face views provided by the
freeze-fracture method of specimen prepa¬
ration (Fig. 30—14). The thicker portions of
Figure 30-10. Scanning micrograph showing the relation¬ the endothelial cells containing the nucleus
ship of primary processes and small secondary processes are usually on the side of the capillary away
of the podocytes, called pedicels. Notice that the cell body from the capsular space.
of the podocyte may be elevated from the capillary sur¬ The intercapillary spaces of the glomerulus
face, and processes of a neighboring podocyte may
extend under it (at arrow) to interdigitate with its own
that radiate from its hilus are occupied by
pedicels. (Micrograph courtesy of F. Spinelli, Ciba-Geigy the mesangium, consisting of mesangial cells
Ltd. Research Laboratories, Basle, Switzerland.) and an extracellular matrix resembling the
material of basal laminae. The mesangial cells
are stellate in form and have a number of
cytological characeristics in common with the
tetroxide owing to its sialic acid content. In pericytes of other capillaries, but these cells
the living state, the 1- to 2.5-nm filamentous are phagocytic. It is believed that they are
molecules forming the surface coat of adja¬ involved in maintenance of the basal lamina
cent foot processes largely fill the filtration of the glomerular capillaries, removing and
slits. At the outer edge of the basal lamina, disposing of residues of filtration. They prob¬
the slits are bridged by a thin slit diaphragm 4 ably also participate in its turnover by re¬
to 6 nm in thickness (Fig. 30-13) extending moval of the older deep portions of the basal
between the membranes of adjacent foot lamina as it is renewed at the epithelial sur¬
processes. face.
The basal lamina is 0.1 to 0.15 pm in The ultrahltration of blood plasma in the
thickness and consists of three zones of dif¬ renal glomeruli is of fundamental impor¬
fering electron density: the lamina rara ex¬ tance in the control of the extracellular fluid
terna, adjacent to the epithelium; the lamina volume, plasma volume, cardiac output, and
rara interna, adjacent to the capillary endo¬ systemic blood pressure. The structural com¬
thelium; and a denser central zone, the lam¬ ponents of the filtration barrier between blood
ina densa. In routine electron micrographs, flowing through the glomerular capillaries
the basal lamina shows little resolvable sub¬ and the capsular space consist of (1) the
structure, but when it is stained with ru¬ fenestrated endothelium, (2) the basal lam¬
thenium red or exposed to cationized ferri¬ ina, and (3) the filtration slits between the
tin, a pattern of regularly spaced densities is interdigitating foot processes of the podo¬
observed in the laminae externa and interna, cytes (Fig. 30—13). Which of these compo¬
and the density of the lamina densa is en¬ nents is the primary filter serving to retain
 THE URINARY SYSTEM • 763

Figure 30-11. A thin plastic section of a renal corpuscle from a rat kidney fixed by perfusion and showing the open
lumens of the capillaries. The irregularity of their outer surface is due to the sections of podocyte processes on their
exterior, x 700. (Courtesy of A. Aoki.)

plasma proteins in the circulation has been a ing of filtration residues that accumulate
subject of debate. Experiments using elec¬ against it.
tron-dense particulate tracers of differing The Proximal Tubule. At the urinary pole
molecular weight led some investigators to of the renal corpuscle, the squamous parietal
conclude that the basal lamina was only a epithelium of Bowman’s capsule is continu¬
course prefilter and that the filtration slits ous with the cuboidal epithelium of the prox¬
were the most significant barrier. Others re¬ imal tubule (Fig. 30—6). This segment of the
garded the basal lamina as the critical barrier. nephron is about 14 mm long and 60 pm in
Additional experimental evidence using par¬ diameter. The proximal convoluted tubules
ticulate tracers has provided strong support make up the bulk of the renal cortex. Each
for the basal lamina as the main barrier to is composed of a convoluted portion (pars
passage of molecules in the same size range convoluta) and a straight portion (pars recta).
as plasma proteins (32,000 to 125,000 M.W.). In addition to many small loops, the convo¬
The filtration depends not only on the size luted portion usually forms a large loop di¬
and shape of the molecules retained but on rected toward the kidney capsule. The recur¬
their charge, with cationic molecules binding rent limb of this loop returns to the vicinity
firmly to anionic binding sites in the basal of the renal corpuscle and then courses to¬
lamina. The prevailing view of the functions ward the nearest medullary ray, where it
of the structural components of the glomer¬ straightens out to become the pars recta,
ulus is that the basal lamina is the main filter; running inward toward the medulla.
the endothelial fenestrae act only as a coarse The epithelium of the proximal convoluted
sieve holding back the formed elements of tubule consists of a single layer of cells with
the blood and controlling access of its ma- a conspicuous brush border on their luminal
cromolecular constituents to the filter; and surface. In kidney tissue fixed by routine
the mesangium serves to unclog and recon¬ methods, the lumen of the proximal tubule
dition the filter by phagocytosing and dispos¬ is often occluded by the apposition of the
764 • THE URINARY SYSTEM

L ; * * * '* ,f . ^W ’ V
. x-x..rx;:- «#xx ipj,, ,;#i
IJJ 'X' ' ,: - ^ I £:
i H vt*' * IMA rn- ::«»-%
. i. 'Ah r . ^ ■
✓* < p
Pedicels m
X HH ■i
■■i
m « jMb
n wRmB|
’
l

% - •:
H
^m
. .

■ ;vlP^iBwjyg.ol
-

Endothelial
cell *. .
I
p i
*

Glomerular
|gp
capillar^ IS
If
i!
lumetl x !' ■ 1 j&

j|. ^ llS
*■'■

it wM'-
fifvt %
y Jra&T «M& :

A ^kX
■I
kl* % U
Fenest rat ionsc^
*" « :s “ \ /*
Basal \4 A ’ \
lamina
Podocyte 3KX/
processes rM®aL: > •:.l-
V " i'+ ^<* ^ *t|
I W* ~**Gbv w

:..: ;:::

J Podocyte r

Figure 30 12. Transmission electron micrograph of a transverse section of a glomerular capillary, showino its basal
lamina interposed between the fenestrated capillary endothelium on the inside and the slit pores between pedicels of
the visceral epithelium on the outside. An area comparable with that in the rectangle is shown at higher magnification
in Figure 30-13. (Micrograph from Tyson, G., and R. Bulger. Anat. Rec 772'669 1972)
 Basement: lamina

Capillary lumen

Figure 30-13. Electron micrograph of a portion of the wall of a glomerular capillary, showing pores in the extremely
attenuated endothelium. On the outer surface of the basal (basement) lamina are the foot processes of the podocytes,
with the narrow filtration slits between them, x 70,000. (Courtesy of D. Friend.)

Figure 30-14. Freeze-cleaving preparation of rat kidney glomerulus. At right, the membrane of the endothelium of a
capillary has been cleaved, showing the very uniform size and regular distribution of the endothelial pores. At upper
left, the membranes of a number of pedicels on the outer aspect of the capillary have been cleaved. (Micrograph
courtesy of D. Goodenough.)
765
766 * THE URINARY SYSTEM

WMBwmm

.n
HI M

■ mm? * "*■'■
mmrni
M ij;: •gf; -■'“• ’«■' %

iiip
.1
”•< „ Proximal
3g|gji
•. ^ ■'
-r -> >.
Wgftt
SSsk-i
✓• ■ „* Distal M

■w A#i
g ■'•-•'••': »
it- ?#Jr
*■«• t -%■ -7\J *
*■4
•# <*' •£.#! 1 ‘JR# ,• | m
^ wW H ? **
<Rff&3if!>!f&&'i>k> f
tmr
*§. * I
*# I pi
. m
IT iaf ** ' ■ ■ * ^0Wkftj
gi*
m Hi Proximo Ǥ& ft;
«

'*‘ </ *
stc ■
••« T

&]|Sgp!
W#i:* £~i fp?i *
mmm
Proximal
. jSsh ■: i
Jlfjl::;|;#. v;#1;,
Figure 30-15. Low-magnification electron micrograph from the cortex of rat kidney fixed by vascular perfusion. The
field includes five proximal convoluted tubules in transverse section, showing their open lumen and prominent brush
border. Two distinct segments of the proximal convolution can be recognized. The first portion has a deeper brush
border (section at lower right). The second portion has a thinner brush border and prominent dense bodies in the
cytoplasm (other three labeled sections), x 2000. (After Maunsbach, A. J. Ultrastruct. Res. 75:252, 1966.)

brush borders of the surrounding cells. It without special staining of the mitochondria.
was formerly thought that this was the nor¬ The lateral limits of the cells are rarely re¬
mal condition, and that the glomerular fil¬ solved with the light microscope because their
trate might percolate through the interstices sides are elaborately fluted and deeply inter-
among the microvilli of the brush borders. It digitated with complementary ridges and
is now known that this constriction of the grooves on the neighboring cells. In favora¬
tubules and obliteration of the lumen is an ble preparations, affording surface views of
artifact. If the fixative is dripped onto the the epithelium, some of the major interdigi-
surface of the living kidney, or if the organ tations of the cell surfaces can be seen, but
is perfused under conditions that involve no the true complexity of the shape of these
agonal fall of blood pressure, all the proximal cells can only be appreciated by careful study
tubules will have a wide open lumen (Figs. of electron micrographs. There are columnar
30-15, 30-16). Each proximal tubule cell lateral ridges that extend the full height of
contains a single spherical nucleus in an eo¬ the cell. An even greater number of slender
sinophilic cytoplasm. In cytological prepara¬ lateral processes near the cell base extend
tions, the Golgi apparatus forms a crown
under adjacent cells (Fig. 30—17). The result¬
around the upper pole of the nucleus, and
ing compartmentation of the base of the
long rodlike mitochondria in the basal half
epithelium seen in micrographs of thin sec¬
of the cell tend to be oriented parallel to the
tions was originally attributed to a simple
cell axis. In well-preserved tissues, this ori¬
infolding of the basal plasma membrane.
entation of the mitochondria may result in a
However, most of the membrane-bounded
faint vertical striation of the cell base, even
basal compartments seen in micrographs are
 THE URINARY SYSTEM • 767

Figure 30-16. Electron micrograph of a sector of the wall of a proximal tubule of rat kidney, x 6000. (Courtesy of R.
Bulger.)

Figure 30-17. Drawing of the shapes and interrelations of the cells of the proximal convoluted tubule. As in fluted
columns, some of the interdigitated lateral processes extend the full height of the cell; others are confined to the base
and extend beneath adjacent cells. (After Bulger, R. Am. J. Anat. 116:237, 1965.)
768 • THE URINARY SYSTEM

cytoplasm are numerous tubular invagina¬

tions called apical canaliculi (Fig. 30—18). The
membrane lining them has a conspicuous
surface coat and short spiny projections into
the cytoplasm as on the membrane of coated
vesicles in other cell types. Vacuoles are also
found in the apical cytoplasm, often in close
relationship to the ends of the apical canalic¬
uli. Some of these have a content of appre¬
ciable density and stain with histochemical
reactions for acid phosphatase, indicating
that they are secondary lysosomes. It is well
established that albumen and other small
proteins that are not retained by the glomer¬
ular filter are absorbed by the proximal con¬
voluted tubule. The apical canaliculi and as¬
sociated vacuoles are involved in endocytotic
uptake of protein and its transport to lyso¬
somes for intracellular digestion. When up¬
take of protein in the proximal convoluted
tubules is experimentally suppressed by
administration of certain positively charged
dibasic amino acids (lysine, arginine), there
is a marked increase in the protein content
of the urine.
A very rapid degradation of small peptides
to amino acids has also been demonstrated
Figure 30-18. Electron micrograph of a portion of the in the proximal tubule. This is not attribut¬
brush border of the proximal convoluted tubule a few
seconds after intravenous injection of myoglobin. The able to endocytosis and lysosomal hydrolysis,
myoglobin has passed into the glomerular filtrate and but probably occurs at the luminal surface
appears between the microvilli and in the apical canaliculi. through the action of peptidases in the mem¬
It is also present in vacuoles in the apical cytoplasm. brane of the brush border. This mechanism
(Micrograph courtesy of W. Anderson.)
is thought to be important in (1) conservation
of amino acids, (2) inactivation of toxic pep¬
tides, and (3) regulation of circulating levels
not open at any point to the cytoplasm of the of small peptide hormones.
overlying cell. It is clear, therefore, that many In addition to uptake of proteins and
of them are, in fact, sections of undermining amino acids, the proximal tubule reabsorbs
basal processes of neighboring cells. In ad¬ nearly all the glucose in the glomerular fil¬
dition to the basal processes, there are smaller trate and certain essential vitamins, while
lateral processes that are confined to the allowing waste products and substances of no
juxtaluminal region of the epithelium. nutritional value to be excreted in the urine.
The elaborate mterdigitation of the lateral Although the entire length of the proximal
and basal portions of the cells greatly ampli¬ tubule seems to have much the same struc¬
fies the area of the cell surface exposed to a ture in ordinary histological preparations and
labyrinthine system of intercellular clefts. has customarily been divided, on the basis of
This region of the cell membrane is rich in macroscopic criteria, into the convoluted por¬
Na+ + K+ - ATPase and is the site of active tion and the straight portion, these designa¬
pumping of sodium out of the cell to create tions do not adequately express the structural
the electrochemical gradient responsible for and functional heterogeneity found along the
movement of water and solute from the tu¬ length of this portion of the tubule. Three
bule lumen to the peritubular capillaries. uitrastructurally distinct segments (S1? S2, and
At the cell apex, the thousands of microvilli S3) are identifiable in the proximal tubule of
forming the brush border are long, regularly all the mammals studied to date, and their
oriented and closely packed, increasing the distribution does not consistently conform to
surface area exposed to lumen more than the division into pars convoluta and pars
20-fold. Arising from the clefts between the recta. Cells in Sj are taller than in the other
microvilli and extending downward into the two segments, and the basal and lateral inter-
 THE URINARY SYSTEM • 769

digitation of adjacent cells is extensive. The way to very sparse, irregularly oriented, short
apical canaliculi and endocytosis vesicles are microvilli on the luminal surface of the thin
more conspicuous and the number of mito¬ segment. The nuclei cause the central por¬
chondria and compartmentation of the base tions of the cells to bulge into the lumen.
of the epithelium are greater than in suc¬ Owing to the small caliber of the tubule, its
ceeding segments. In S2, the cell height is thin wall, and the bulging of the perikaryon
lower, mitochondrial length and number are into the lumen, the thin segment of Henle’s
lower, and interdigitation with adjacent cells loop in cross section bears a superficial resem¬
is less. The cells of S3 are cuboidal and have blance to a capillary or venule (Fig. 30-20).
only a few small basal interdigitations, and The mammalian kidney usually contains
the mitochondria are fewer and randomly short-looped nephrons and long-looped
oriented. All three segments have a promi¬ nephrons (Figs. 30—5, 30—21). The short loop
nent brush border, but the microvilli are of Henle has a descending thin limb that is
noticeably longer in S3. continuous at the flexure with the ascending
The Thin Segment. The loop of Henle con¬ straight portion of the distal tubule, but has
sists of the straight portion of the proximal no ascending thin limb. The long loop con¬
tubule, the thin segment, and the ascending sists of a descending thin limb and an ascend¬
straight portion of the distal tubule. In the ing thin limb. The thin limbs of the loop are
outer zone of the medulla, the descending lined by epithelium that exhibits marked re¬
portion of the proximal tubule abruptly nar¬ gional differentiations in its ultrastructure.
rows from a width of about 60 pm to con¬ Four structurally distinct segments are rec¬
tinue as the thin segment, about 15 pm in ognizable (Fig. 30-21). Type I, characteristic
diameter (Fig. 30-19). The epithelium of the abbreviated thin limb of the short
changes from cuboidal to squamous, with a loop, is a very simple flat epithelium with no
height of only 0.5 to 2 pm. There is a sudden interdigitation of neighboring cells. Its zon-
termination of the brush border, which gives ulae occludentes consist of two to four anas-

Figure 30-19. Electron micrograph of the abrupt junction of the straight portion of the proximal tubule with the thin limb
of the loop of Henle. Slightly oblique section through the junction. The brush border stops suddenly and the epithelium
becomes very thin, x 4200. (After Osvaldo and Latta. J. Ultrastruct. Res. 75:144, 1966.)
770 • THE URINARY SYSTEM

Descending
thin limb

Ascending
thick limb

Capillary

Figure 30-20. Electron micrograph of the inner portion of the outer zone of the medulla in rat kidnev. Descendino thin
?hpdpaSH'th'C^hlimHbS °f ^ loop of ^enle are seen above> and capillaries of the vascular bundle below. Note that
the endothelium in the descending arterial limb of the capillary loops is continuous while that of the ascendina venous
mb is fenestrated. Interstitial cells are indicated by arrows. (Micrograph from Boh^S* O ^ 47 329
1 y 14. j ’
 THE URINARY SYSTEM • 771

SHORT long tional strands in freeze-fracture prepara¬

LOOP LOOP tions.
There is a gradual simplification of the
epithelium as the loop descends to the inner
portion of the medullary pyramid. There the
Type III epithelium is again composed of
flat, noninterdigitating cells with several
strands in their occluding junctions. In the
ascending thin limb, Type IV epithelium is
thin and again has elaborately interdigitated
cell processes and very few microvilli. The
shallow junctions consist of only one or two
strands.
The segmental differentiation of the thin
region of the loop of Henle is as described
above in the common laboratory rodents, but
differs significantly in lagomorphs and feli-
dae. It has been studied in less detail in the
human, but comparable differences are
found in the height of the epithelium and
the degree of interdigitation of the cells in
the descending and ascending portions of
the long loops.
In rats and certain other species whose
kidneys have a single pyramid, the corre¬
sponding segments of the loops tend to be in
register. As mentioned earlier, this regularity
of arrangement is reflected in a zonation
within the medulla that is detectable on na¬
ked eye inspection of the cut surface. An
outer and inner zone of the medulla can be
m recognized, and an inner and outer band or
stripe are distinguishable within the outer
Figure 30-21. Schematic representation of the ultrastruc- zone of the medulla. In these species, the
tural features of the epithelium of the thin limb of the loop boundary between the outer and inner band
of Henle in its several segments. Nuclear region of the
cells is not drawn. (Modified after Kriz, W., et al. In of the outer medulla is at the junctions of
Maunbach, A. L., et al., eds.: Functional Ultrastructure of the straight portion of the proximal tubules
the Kidney. London, Academic Press, 1980.) with the descending thin limbs of Henle’s
loops. The transition between the inner and
outer medulla is at the junctions of the as¬
tomosing junctional strands. In the outer cending thin limbs with the ascending
portion of the descending thin limb of long straight portion of the distal tubule. The
loops, the Type II epithelium is somewhat inner medulla therefore contains collecting
higher and has numerous microvilli on its tubules, thin limbs of the loop of Henle, and
luminal surface. In micrographs of thin sec¬ blood vessels (Figs. 30-22B, 30—23).
tions, typical cellular units containing a nu¬ In the human, a similar zonation of the
cleus make up only a small portion of the medulla is detectable, though less easily, be¬
wall. Most of the wall is composed of small cause the loops of Henle are of different
membrane-bounded profiles 1 to 3 pm lengths, depending on the position of their
across, separated from one another by inter- renal corpuscle in the cortex and the length
membranous clefts extending from the basal of their thin segment (Fig. 30—5). The short
lamina to the lumen. These short anucleate loops are associated with the renal corpuscles
units of the epithelium are sections through located nearer the surface of the kidney.
deeply interdigitating processes of adjacent Loops of this type are about seven times as
cells. Some contain one or two mitochondria; numerous as the long ones. Their bend,
others are largely devoid of organelles. They which is distal to the thin segment, is located
are attached to one another by juxtaluminal in the outer part of the medulla and is
zonulae occludentes having one or two junc¬ formed by the thick ascending limb. The thin
772 • THE URINARY SYSTEM

Figure 30-22. A, Photomicrograph of a transverse section through the outer medulla, which is composed of thin
segments (TL) ascending straight portions of distal tubules (DT), and collecting tubules (CT). B, Transverse section
through the inner medulla, composed of thin limb (TL) of the loop of Henle and collecting tubules (CT).

segment is in the descending limb of the loop imal tubule and is composed of three parts:
and may be very short or in some instances the straight portion (pars recta); the portion
may even be absent. In the latter event, the adjacent to the renal corpuscle, containing
straight descending portion of the proximal the macula densa (pars maculata); and the
tubule continues directly into the thick as¬ convoluted portion (pars convoluta). The
cending limb. In the longer loops, which are straight portion begins in the inner band of
associated with the deeper-lying juxtamedul- the outer zone of the medulla and constitutes
lary renal corpuscles, the bend is formed by the ascending thick limb of the loop of Henle.
the thin limb. These long loops may extend The transition from the thin segment to the
nearly to the apex of the papilla. In this ascending limb of the distal tubule is abrupt.
event, the length of the thin limb may be 10 The epithelium is cuboidal, but the lumen is
mm or even more. generally wider than that of the proximal
The junction of the outer and inner zones tubule. A typical brush border is lacking.
of the medulla is marked by the transition of Some cells have numerous microvilli, while
the thin limb to the ascending thick limb of others have a smooth luminal surface except
the long loops of Henle. The boundary be¬ near the cell margins, where a few microvilli
tween the outer and inner bands of the outer are found. These smooth-surfaced cells pre¬
medulla in the human kidney is somewhat dominate in the medullary segment of the
obscured by the prevalence in this region of distal tubule. The apical vesicles and canalic-
short loops having the junctions between uli characteristic of the proximal tubule are
their successive segments at different levels. absent in the distal tubule. The base of the
The Distal Tubule. The distal tubule is
epithelium is elaborately compartmentalized
shorter and somewhat thinner than the prox¬ by infoldings of the basal membrane that
 THE URINARY SYSTEM 773

Capillary

Collecting
tubule

Henle

Capillary

Collecting
tubule

Henle

Figure 30-23. Electron micrograph of a transverse section through the inner zone of the medulla of rat kidney. Collecting
tubules, loops of Henle, and capillaries are seen in cross section surrounded by an interstitium with a homogeneous
extracellular matrix of low electron density. (Micrograph from Bohman, S.-O. J. Ultrastruct. Res. 47:329, 1974.)
774 * THE URINARY SYSTEM

Figure 30-24. Electron micrograph of a portion of the base of a distal convoluted tubule of guinea pig kidney, illustrating
the small and large basal compartments. The latter contain long mitochondria oriented perpendicular to the cell base.
Notice that the peritubular capillary is of the fenestrated type with several of its pores indicated by arrows. (Courtesy of
A. Ichikawa.)

make deep incursions into the cell (Fig. The thick ascending limb of Henle’s loop
30-24). In addition, there are undermining or straight portion of the distal tubule enters
basal processes of neighboring cells compa¬ the cortical tissue, returns to the renal cor¬
rable with those described for the proximal puscle of the same nephron, and attaches to
tubule. Long mitochondria are lodged in its vascular pole, particularly to the afferent
these basal compartments, and their orien¬ arteriole. The side of the tubule in contact
tation parallel to the axis of the cell results with the afferent arteriole forms an elliptical
in the prominent striation of the basal cyto¬ disc of taller cells measuring 40 by 70 pm in
plasm observed with the light microscope. the human kidney. This area, called the mac¬
The mitochondria have many cristae and ula densa has been reported to have some
numerous matrix granules. The Golgi comp¬ function in the hemodynamics of the kidney,
lex is small and forms a crown around the but its precise role has not been defined.
upper pole of the nucleus. There are a few From here the straight portion continues as
cisternal profiles of granular endoplasmic re¬ the convoluted portion of the distal tubule.
ticulum and a moderate number of free ri¬ This portion of the tubule has many short
bosomes. A pair of centrioles is located in the loops and irregular contortions. It usually
apical cytoplasm, and from one of these a courses toward the surface above the corre¬
single flagellum projects into the lumen. sponding renal corpuscle.
The junctional complexes between cells are
shallow and permeable to lanthanum. This
Collecting Ducts
finding, together with the relatively low
transepithelial electrical resistance, suggests The connections of the nephrons with the
that there is a functional paracellular path¬ collecting tubules are located in the cortex of
way for solute and water movement across the kidney along medullary rays. The distal
the epithelium in addition to the high-resis¬ tubules are continuous with arched collecting
tance transcellular pathway. tubules, which are tributaries of straight col-
 THE URINARY SYSTEM • 775

lecting tubules located in the medullary rays. renal tubules and the blood and lymph ves¬
The collecting tubules pass inward in the sels. In the cortex, it constitutes about 7 per
medullary ray, through the outer zone of the cent of the tissue volume, while the vascula¬
medulla. When they reach the inner zone, ture occupies an additional 6 per cent. The
they join at acute angles with other, similar relative volume of the interstitium increases
tubules. There are about seven such conver¬ considerably toward the inner region of the
gences in the medulla near the pelvis, and medulla and in the papilla. The interstitial
they result in the formation of large, straight tissue of the kidney attracted little investiga¬
tubules called papillary ducts (of Bellini). tive interest until it was discovered that the
These have a lumen 100 to 200 pm in di¬ concentration of solutes in the intertubular
ameter and open on the area cribrosa at the spaces of the medulla has an important role
apex of each papilla. in concentration of the urine by reabsorption
The system of the intrarenal excretory of water.
ducts has an epithelium quite different from Two types of interstitial cells have been
that of the various parts of the nephron. In described in the cortex, one bearing a super¬
the smallest collecting tubules, the cells are ficial resemblance to fibroblasts and the other
cuboidal and very distinctly outlined; they a lymphocyte-like cell. The fibroblast-like
contain a darkly staining round nucleus, and cells are of irregular shape, with long taper¬
most of the cells have a clear pale cytoplasm. ing processes often in contact with processes
There are a few mitochondria and, near the of neighboring cells of the same type. The
surface, a pair of centrioles with a central endoplasmic reticulum is well developed and
single flagellum. its cisternae may be somewhat distended by
In addition to the principal or light cells, a flocculent material. The cortical cytoplasm
which have a relatively smooth convex apical is rich in actin filaments and microtubules.
surface, there are small numbers of interca¬ Occasional lipid droplets are present in the
lated or dark cells with more abundant organ¬ cytoplasm. The lymphocyte-like cells have a
elles, a denser cytoplasm, and an apical sur¬ nucleus with abundant heterochromatin, and
face bearing longer microvilli. The dark cells their sparse cytoplasm is rich in free ribo¬
are rarely found in the inner medullary seg¬ somes. The function of cortical interstitial
ment. The principal cells gradually increase cells is not firmly established, but it seems
in height from low cuboidal in the outer part likely that the fibroblast-like cells produce
of the medulla to cuboidal, and finally to low collagen and glycosaminoglycans of the ex¬
columnar in the papillary ducts. They are tracellular matrix of the interstitium. The
always arranged in a single layer, with all the small round cells may be blood-derived lym¬
nuclei at one level and with the free surfaces phocytes or early cells of the monocyte-phag¬
bulging slightly into the lumen of the tubule. ocyte series.
The cytoplasm keeps its pale appearance. In the renal medulla, three types of inter¬
The centrioles remain at the bulging free stitial cells have been described. Type I is a
surface. In the area cribrosa, the simple col¬ highly pleomorphic cell containing multiple
umnar epithelium of the ducts continues small lipid droplets. Type II is lymphocyte¬
onto the surface of the papilla. like, and Type III is a pericyte associated
It was formerly believed that the collecting with the descending vasa recta. The medul¬
ducts were merely inert conduits conveying lary interstitial cells, Type I, have been most
fluid from the distal tubule to the renal pelvis. thoroughly studied owing to the suggestion
This is now known to be untrue. The cortical that they may have an endocrine function.
collecting ducts respond to vasopressin by They are distributed at regular intervals be¬
swelling and dilatation of the lateral intercel¬ tween the parallel tubules and vessels (Fig.
lular clefts, and the medullary collecting 30—25). They are variable in form and their
ducts play a major role in the countercurrent processes are often in close apposition to the
mechanism for urine concentration in the basal lamina of thin limbs of the loop of
mammalian kidney. Henle and to the capillaries (Fig. 30-20).
Gap junctions occur at sites of contact be¬
tween processes of neighboring Type I cells.
The endoplasmic reticulum resembles that of
THE RENAL INTERSTITIUM the comparable cells in the cortex and is
generally more extensively developed in cells
The interstitium is usually defined as the of the outer zone of the medulla. The mito¬
space outside both the basal laminae of the chondria and Golgi apparatus are unremark-
776 • THE URINARY SYSTEM

Figure 30-25. Photomicrograph of the renal medulla showing the location (at arrows) of the interstitial cells.
(Photomicrograph from Bohman, S.-O. Cell Tissue Res. 789:1, 1978.)

able. A few lysosomes are present, as well as din-producing cells. This hypothesis has now
peculiar inclusions called “cylindrical bodies.” been abandoned for lack of direct evidence.
These consist of bundles of 5 to 50 parallel The most attractive possibility is that they are
cylinders — 0.15 |Jim in diameter and up to endocrine cells involved in regulation of
11 pm in length. These are believed to be blood pressure. The belief that the medullary
derivatives of the endoplasmic reticulum. interstitial cells may secrete a hormone rests
Their function is unknown. mainly upon experiments in which medullary
A characteristic feature of these cells are tissue was transplanted subcutaneously in an¬
lipid droplets about 0.5 pm in diameter and imals with high blood pressure. The trans¬
generally exhibiting little osmiophilia (Fig. plants developed into nodules consisting of
30-26). The number of lipid droplets in¬ cells identified as proliferating interstitial
creases in salt depletion and water loading, cells, and the blood pressure fell. Upon re¬
and decreases in water deprivation and in moval of the nodule, the pressure rose again
hypertension. The isolated lipid droplets to hypertensive levels. These observations
have been found to consist of triglycerides strongly suggest secretion by interstitial cells
unusually rich in long-chain polyunsaturated of a hormone with antihypertensive activity.
fatty acids. There is no evidence that the The postulated hormone has not been iso¬
droplets are released from the cells, but they lated.
may contain precursors of some secretory
product.
A number of functions have been sug¬ JUXTAGLOMERULAR COMPLEX
gested for the Type I interstitial cell, but with
little supporting evidence. The renal medulla In addition to their function in excretion,
is known to be unusually rich in prostaglan¬ the kidneys have a role in the regulation of
din, and prostaglandin synthesis can be stim¬ blood pressure. There is a clear association
ulated by antidiuretic hormone. These obser¬ between certain types of kidney disease and
vations led to the suggestion that the Type I high blood pressure (hypertension). The kid¬
interstitial cells were specialized prostaglan¬ ney produces and releases into the blood a
 THE URINARY SYSTEM • 777

Figure 30-26. A type I interstitial cell in the renal papilla in close relationship to a capillary and a loop of Henle. The
cytoplasm contains numerous small lipid droplets. (Micrograph from Bohman, S.-O. J. Ultrastruct. Res. 38:225, 1972.)

substance called renin. This has no vasomotor ent arterioles at the vascular pole of the
effect itself but is an enzyme that acts upon glomerulus. The interrelations of the gran¬
a plasma globulin, angiotensinogen, to split off ular juxtaglomerular cells, the macula densa,
a decapeptide, angiotensin I. A converting and the extraglomerular mesangial cells are
enzyme in the blood plasma then acts upon poorly understood. They are believed to have
this to split off two more amino acids, con¬ related functions, however, and together they
verting it to angiotensin II—the most potent constitute the juxtaglomerular apparatus or com¬
vasoconstrictor known. Renin is synthesized plex. The juxtaglomerular cells are described
in the juxtaglomerular region of the ne¬ as “myoepithelioid” because they appear to
phron, where the ascending straight portion be highly modified smooth muscle cells. They
of the distal tubule returns to the renal cor¬ have a slightly basophilic cytoplasm and their
puscle and comes into intimate relation with specific granules are most clearly demon¬
its vascular pole (Fig. 30-27). strated by the Bowie stain, the PAS reaction,
Among the smooth muscle cells in the wall or the fluorochrome dye thioflavine T. In
of the afferent arteriole just proximal to its electron micrographs they have a moderately
entrance into the glomerulus are cells that abundant granular endoplasmic reticulum
contain conspicuous cytoplasmic granules. and a well-developed Golgi complex. The
These granular juxtaglomerular cells are in granules appear to arise in the cisternae of
contact with the intima of the arteriole on the Golgi complex, as in other glandular cells.
the one side, and on the other side they are When first formed the granules are of vari¬
intimately related to the base of the epithelial able shape, and have a crystalline internal
cells making up the macula densa in the wall structure with a periodicity of 5 to 10 nm.
of the distal tubule (Fig. 30-28). Also asso¬ Coalescence of these elements gives rise to
ciated with the granular cells are a few non- mature granules, which are irregularly
granular ones and a group of pale-staining shaped conglomerates that may retain evi¬
extraglomerular mesangial cells (also called dences of crystalline order but more often
lacis cells, polkissen, or polar cushion) located appear homogeneous.
in the angle between the afferent and effer¬ The secretory nature of these granules was
778 • THE URINARY SYSTEM

Figure 30-27. Photomicrograph of a thin plastic section of rat kidney, showing a glomerulus and clusters of
juxtaglomerular cells in the wall of its afferent arteriole. (Micrograph courtesy of R. Bolender.)

established in experimental studies that dem¬ fluorescence technique using antibody to

onstrated changes in granule content of the highly purified antigen localizes renin exclu¬
juxtaglomerular cells secondary to renal is¬ sively in the granules of myoepithelial cells
chemia or to experimental alteration of salt in the juxtaglomerular apparatus, and not in
intake. These studies led to the hypothesis the macula densa.
that these cells are the site of production of The kidney is apparently not the only site
renin. Support for this thesis has come from of renin production. In the mouse and other
the finding that the solubility characteristics species that exhibit sexual dimorphism of the
of renin and of the granules of the juxta¬ submaxillary gland, renins that cross-react
glomerular cells are similar, and that there is with that of the kidney are localized by im¬
a direct correlation between the level of renin munofluorescence in certain segments of the
determined by bioassay and the degree of gland.
granulation of the juxtaglomerular cells. Mi¬ The bases of the cells of the macula densa
crodissection methods have localized the are in very close relation to the juxtaglomer¬
renin to the immediate vicinity of the renal ular cells, and the basal lamina between them
corpuscle, and application of the immuno¬ is exceedingly thin. This close topographical
 THE URINARY SYSTEM • 779

the zona glomerulosa of the adrenal cortex

to release aldosterone; this acts in turn upon
the collecting ducts of the kidney to induce
sodium and water retention, which tends to
correct the reduction of plasma and intersti¬
tial tissue fluid volume.

BLOOD SUPPLY

Because the kidneys serve to clear the

blood of accumulated waste products of me¬
tabolism, they have a very large blood flow,
averaging about 1200 ml/min through both
kidneys. A knowledge of the blood supply of
the kidney is essential to an understanding
of its function.
The renal artery enters the hilus of the
kidney and divides into two main sets of
branches directed toward the dorsal and ven¬
tral aspects of the organ. In the adipose tissue
surrounding the pelvis, these branches in
turn divide into smaller interlobar arteries that
enter the substance of the kidney and course
peripherally in the renal columns between
Figure 30-28. Photomicrograph of two renal corpuscles the pyramids or lobes of the kidney. At the
from macaque kidney, showing (at arrows) two typical level of the base of the medullary pyramids,
examples of the macula densa, an area of the wall of the the interlobar arteries arch over to run par¬
distal tubule where the cells are thicker and the nuclei
are crowded together and superimposed, x 175.
allel to the surface of the organ as the arcuate
arteries at the corticomedullary junction.
Small interlobular arteries, given off from the
arcuate arteries at regular intervals, course
relationship has been interpreted as suggest¬ radially toward the kidney surface (Fig.
ing some interchange of substances between 30-29).
the macula densa and the juxtaglomerular The interlobular arteries running radially
cells. Consistent with such a relationship is in the cortex give off numerous afferent arte¬
the report that the polarity of the Golgi rioles to the glomeruli. The blood is carried
complex in the cells of the macula densa is from the glomeruli via efferent arterioles. The
toward the juxtaglomerular cells, whereas in efferent vessels of glomeruli situated in the
the remainder of the circumference of the outer part of the cortex are of small diameter
distal tubule, it is toward the lumen. and break up to form the cortical intertubu¬
The finding that the cells of the macula lar capillary network. The efferent vessels of
densa show changes in their histochemically the more deeply situated juxtamedullary glo¬
demonstrable enzymatic activities when the meruli are of larger caliber and pass down¬
rate of secretion of the juxtaglomerular cells ward into the medulla, breaking up into
is altered provides further indication that the bundles of thin-walled vessels somewhat
two structures are functionally related. The larger than ordinary capillaries, called vasa
juxtaglomerular complex also has an impor¬ recta (Fig. 30-30). The efferent vessels of the
tant function in regulation of tissue hydration juxtamedullary glomeruli and the vasa recta
and blood volume. Any condition that re¬ both contribute branches to an intertubular
duces blood or extracellular fluid volume capillary network in the medulla.
seems to be sensed by the afferent arteriole The vasa recta form hairpin loops at var¬
acting as a baroreceptor or by the macula ious levels in the medulla, turning back to¬
densa as a sensor of sodium concentration. ward the cortex and running close to and
The juxtaglomerular cells are stimulated to parallel with the vessels from which they
synthesize and release renin. The resulting recur. The descending vessels penetrate the
angiotensin II in the blood directly stimulates outer medulla to different depths before
780 • THE URINARY SYSTEM

Capsular A.

Figure 30-29. Schematic representation of the finer arterial branching in dog kidney. (After Kugelgen, A., and K. J.
Otto. In Zwanglose Abhandlungen aus dem Gebiet der normalen und pathologischen Anatomie. Vol. 5. Stuttgart, Georg
Thieme, 1959.)

turning back. The descending and ascending responding number of interlobular arteries.
limbs of these loops form a countercurrent The blood in these flows inward to the ar¬
system of vessels referred to as a vascular cuate veins and thence to the interlobar veins,
bundle or rete mirabile. As more vessels turn which Anally become confluent in the hilus
back, the vascular bundles taper down as they to form the renal vein.
approach the inner medulla. The descending The hemodynamics of the renal circulation
vessels forming the arterial limbs of the vas¬ are such that the flows to various zones of
cular bundle or rete are slightly smaller than the kidney are very different. Measurements
the recurrent vessels that constitute the ve¬ of blood flow distribution in the unanesthe¬
nous limbs (Fig. 30—31). The fine structure) tized dog give values of 472 ml/100 g/min in
of the vessel walls also differs, the arterial the cortex, 132 ml/100 g/min in the outer
component having a continuous endothe¬ medulla, and 17 ml/100 g/min in the inner
lium, whereas the venous component has a medulla. Although the cortical flow is nor¬
thin fenestrated endothelium. The proximity mally very rapid, strong stimulation of sym¬
of the vessels in the vascular bundles and the pathetic nerves may diminish it almost to
large surface they present to one another zero. Under various stressful circumstances,
facilitate rapid movement of diffusible sub¬ the cortex of the kidney becomes pale, and
stances between the ascending and descend¬ red blood may appear in the renal vein.
ing limbs of the loops. The vasa recta thus Evidently, under these conditions, the renal
serve as efficient countercurrent exchangers cortex is relatively ischemic and the bulk of
for diffusible substances. the blood that would normally pass through
1 he capillaries of the outermost layers of the cortical glomeruli for filtration is by¬
the cortex are drained toward the surface by passed through the juxtamedullary glomeruli
radially arranged branches, the superficial cor¬ and the vasa recta into the interlobular veins,
tical veins, which join veins of characteristic and thence to the renal vein.
configuration on the surface of the kidney,
called stellate veins. This outer mantle of ve¬
nous channels is drained by a relatively small LYMPHATICS
number of interlobular veins into the arcuate
veins that accompany the arteries of the same The distribution of lymphatics in the kid¬
name. The capillaries in the deeper part of ney has been a subject of controversy. A
the cortex empty into radially oriented deep number of investigators have contended that
cortical veins, of which there are some 400 per intrarenal lymphatics are found only in as¬
square centimeter running parallel to a cor¬ sociation with the larger blood vessels and
 THE URINARY SYSTEM • 781

Cortical
Nephron
Collecting
Tubu e

Figure 30-31. Photomicrograph of a rete mirabile from

dog kidney. Notice that the vessels are of two morpho¬
logical types: the descending arterial limbs are capillaries
with a round cross section and walls of appreciable
thickness; the ascending venous limbs are larger, are
more irregular in outline, and have exceedingly thin walls.
The latter are filled here with gray precipitate of plasma,
while the former appear empty or contain a few erythro¬
cytes. x 250.

extending peripherally only as far as the

interlobular vasculature. Others have re¬
ported that the initial lymphatic capillaries
are tributaries of interlobular lymphatics and
are found among the tubular elements mak¬
ing up the parenchyma of the renal lobules.
The latter interpretation has now been
strongly supported by combined light and
electron microscopic studies employing im¬
proved methods for identification and delin¬
eation of lymphatics. In addition to the gen¬
erally accepted association of lymphatics with
Figure 30-30. Schematic drawing of the blood supply the larger arterial vessels, lymphatic capillar¬
associated with cortical and juxtamedullary nephrons. In
ies are present within the parenchyma of the
the latter, the efferent arteriole runs downward into the
medulla, where it gives rise to a bundle of vasa recta. renal cortex. Morphometric analysis has
These, together with their recurrent venules, form long shown that, in the dog, as many as one third
bundles of parallel vessels called retia mirabilia. Here the of the cortical lymphatics are intralobular.
arterial and venous elements have been separated to These drain mainly to interlobular lymphat¬
illustrate their continuity in long loops. In life, the arterial
ics and thence to lymphatic plexuses in the
and venous limbs of the loops intermingle, as shown in
Figure 30-31. hilus of the kidney. Some in the outer cortex
782 • THE URINARY SYSTEM

drain to a capsular lymphatic plexus. The The blood circulates through the glomer¬
volume of lymph in the cortex is estimated ular capillaries with a hydrostatic pressure of
to be about 1 per cent of the volume of blood about 70 mm Hg. This tends to press the
in peritubular capillaries. fluid constituents of the blood through the
pores and intercellular spaces of the endo¬
thelium, across the basal lamina, through the
NERVES filtration slits between foot processes of the
podocytes, and into Bowman’s capsule. The
Macroscopic dissection shows that the sym¬ hydrostatic pressure in the capillaries is op¬
pathetic celiac plexus sends many nerve fibers posed by an average colloid osmotic pressure of
into the kidney. Their distribution inside the about 32 mm Hg and a capsular pressure of
organ has not been worked out satisfactorily. about 20 mm Hg. The net filtration pressure is
It is relatively easy to follow nonmyelinated thus about 18 mm Hg. With some 1300 ml
and myelinated fibers along the course of the of blood flowing through the glomeruli of
larger blood vessels. They provide the adven¬ both kidneys each minute, approximately 125
titia with sensory nerve endings and the mus¬ ml of glomerular filtrate is produced. Anal¬
cular coat with motor endings. Along with ysis of fluid aspirated from Bowman’s capsule
the afferent arterioles, nerve fibers may reach by micropuncture has established that it is an
the renal corpusles, and some of them seem ultrafiltrate of blood plasma with nearly the
to end on their surfaces. A nerve supply of same composition as the interstitial fluid. It
the uriniferous tubules, however, has not contains small molecules such as phosphates,
been convincingly demonstrated. Some in¬ creatine, uric acid, and urea, and small
vestigators describe plexuses of fine nerve amounts of albumin, but is free of larger
fibers that surround and seem to penetrate protein molecules and substances combined
the basal lamina. On its inner surface they with them. Molecules of molecular weight
are said to form another plexus, from which 100,000 and larger are arrested in the basal
terminal branches arise to end between the lamina. However, the capillary wall not only
epithelial cells. There is a good possibility is a size-selective barrier in the filtration of
that the silver stains on which these descrip¬ macromolecules, but also exhibits selectivity
tions are based were impregnating reticulum based on charge. Anionic molecules are more
and not axons. The finer innervation of the limited in their passage across the glomerular
kidneys has not been systematically studied filter than are neutral molecules of the same
at the electron microscope level, where the size. The filter thus serves to retain albumin
fine nerve fibers can be identified with and other anionic proteins in the circulation.
greater certainty. The negatively charged sites responsible for
this selectivity are located on the surface of
the capillary endothelium; on the surface
HISTOPHYSIOLOGY OF THE KIDNEYS coat of the podocyte foot processes; and
within the basal lamina of the glomerulus
In forming urine, the kidneys do not pro¬ (Fig. 30—32). The polyanion of the podocyte
duce new material in significant amounts, but surface coat is a 140,000-M.W. sialoprotein,
eliminate water and some of the waste prod¬ which has been isolated and called podoca-
ucts of metabolism that are carried in solution lyxin. The basal lamina contains Type IV
in the blood. In addition to their excretory collagen, which possesses acidic amino acids
function, in which they dispose of waste and with free carboxyl groups and a heteropoly¬
foreign substances, the kidneys have equally saccharide region that includes sialic acid. In
important conservative functions, by which they addition to these the basal lamina contains a
retain the amounts of water, electrolytes, and proteoglycan, rich in heparan sulfate. This
other substances needed by the body, while highly charged anionic macromolecule oc¬
eliminating excesses of these substances. curs in a more or less regular lattice in the
They therefore play an important role in the laminae interna and externa (Fig. 30-32).
maintenance of the organism. The kidney Of the 125 ml of filtrate formed per minute
carries out its functions by a combination of in the glomeruli, 124 ml are reabsorbed as
filtration, passive diffusion, active secretion, the fluid passes through the various segments
and selective absorption. The form, topo¬ of the nephrons and the collecting ducts,
graphical relations, and microscopic organi¬ leaving a volume of only 1 ml to be excreted
zation of its components represent structural as urine. This small remainder is not simply
adaptations favoring these processes. derived by absorption of water; its contents
 THE URINARY SYSTEM • 783

Podocyte Filtration slits Polyanionic

fenestrae lamina heparan sulfate

Figure 30-32. Diagrammatic representation of the glomerular filtration barrier. The anionic sites reside in the surface
coat of the endothelium and of the podocyte processes, and in proteoglycan macromolecules, rich in heparan sulfate,
that form a network of angular particles in the lamina rara externa and interna of the basal lamina. (From Farquhar, M.
G. Reproduced from The Journal of Cell Biology 81:137, 1979 by copyright permission of The Rockefeller University
Press.)

are modified in its passage along the tubules on the fluid aspirated. The change in the
by (1) diffusion of some substances back into composition of the filtrate as it passes along
the blood, (2) absorption of some by osmotic the nephron can thus be studied directly.
work, and (3) excretion of others into the Similarly, fluid of known composition can be
lumen. perfused through a segment of tubule be¬
The transport of certain substances into tween two pipettes, and the substances added
and out of the nephron can be fairly precisely to or subtracted from it can be determined.
measured in healthy humans and is the basis By these and other methods, it has been
of a variety of clinical measurements of kid¬ shown that 85 per cent or more of the sodium
ney function. Inulin is a nonmetabolized car¬ chloride and water of the glomerular filtrate
bohydrate that, when injected intravenously, is reabsorbed in the proximal tubule. In this
rapidly appears in the glomerular filtrate but process, the cells actively transport sodium
is not secreted or absorbed by the tubules. It from the lumen, and the water and chloride
can be used, therefore, as a means of meas¬ passively follow it to maintain osmotic equi¬
uring the amount of plasma filtered by all librium.
the glomeruli of both kidneys. This calcula¬ The remarkable functional efficiency of
tion is based on the concentrations of inulin the proximal convoluted tubule is attribut¬
in the urine and in the plasma during the able in large part to the configuration of its
experiment. The volume of plasma contain¬ cells, which are specialized to maximize the
ing the same amount of inulin as that found surface area of the physiologically important
in the urine is the amount of plasma that has regions of their plasma membrane. In the
been “cleared” of this substance by filtration current interpretation of proximal tubule
during the period of the test. The inulin function, water and solutes may pass from
clearance furnishes a standard from which it the tubule lumen via a paracellular route
is possible to estimate what proportions of through the juxtaluminal intercellular junc¬
other substances are reabsorbed or excreted tions, or via a transcellular route that involves
by cells in various parts of the tubule. flux across the luminal surface and then
Much of our knowledge of the functions across the lateral membranes into the inter¬
of renal tubules has been derived from stud¬ cellular clefts below the junctions. The pas¬
ies on amphibian species, in which it is pos¬ sive movement of water along this path is
sible to puncture the glomerular capsule and due to an osmotic gradient generated by
the tubules at different levels in living ani¬ active transport of solute across the lateral
mals and carry out microchemical analyses cell membranes. Stereological morphometric
784 • THE URINARY SYSTEM

studies of proximal convoluted tubule have body, are not completely reabsorbed but are
shown that the microvilli of the brush border allowed to remain in the urine and are elim¬
increase the apical surface 36 times, and the inated from the body. While some 99 per
amplification of the lateral surfaces by proc¬ cent of the water of the glomerular filtrate is
esses interdigitating with adjacent cells gives conserved, only 40 per cent of the urea and
them an area equal to that of the brush none of the creatinine is reabsorbed.
border, and both are 20 times greater than In addition to its capacity for active reab¬
the area of the cell base. In the straight sorption, the proximal tubule has the capacity
portion of the proximal tubule, the luminal to secrete creatinine, para-aminohippuric
surface is amplified only 15 times by its less acid, the organic iodine compound Diodrast,
well-developed brush border, and the lu¬ and sulfonic dyes such as phenol red. The
minal and lateral surfaces are only 10 times secretory capacity of the proximal tubule
greater than the cell base. These configura¬ does not have the physiological importance
tional differences in the cells of the two in man that it does in some lower animals,
segments of the proximal tubule are reflected particularly those fish that have aglomerular
in quantitative differences in resorption of kidneys. However, the substances that are
water from the glomerular filtrate. secreted are useful in the clinical evaluation
Normally, all the glucose in the filtrate is of kidney function and renal blood flow.
also reabsorbed in the proximal convoluted When introduced into the blood in moderate
tubule and it is calculated that nearly half a concentrations, Diodrast and para-aminohip¬
pound of glucose and more than three puric acid are entirely removed during a
pounds of sodium chloride are recovered per single passage of blood through the kidneys.
day from the glomerular filtrate of man. If Since it is impossible to remove by filtration
the level of glucose in the blood is raised all the substances dissolved in the blood and
experimentally above a certain level, the glu¬ have any fluid plasma left, the complete re¬
cose is not completely absorbed and appears moval of a substance in one passage of blood
in the urine. This tubular maximum for re ab¬ through the kidney must occur in part by
sorption of glucose (glucose Tm) is a useful filtration and in part by excretory work.
index of the reabsorptive capacity of the Knowledge of the concentration of such a
kidney tubules. substance in the blood and the amount found
Other metabolically important substances in the urine produced in a given time makes
that are reabsorbed in the proximal tubule it possible to calculate the blood flow through
are amino acids, protein, acetoacetic acid, and the kidney. The blood flow is equal to the
ascorbic acid. On leaving the proximal tu¬ plasma flow plus the cell volume found by
bules, the fluid contains essentially none of hematocrit. From the values for Diodrast
these substances. The absorption of proteins clearance and for inulin clearance, one can
in the proximal tubule has been followed determine the fraction of renal plasma flow
morphologically by intravenous administra¬ that is filtered by the glomeruli. If one then
tion of peroxidase and its subsequent detec¬ raises the concentration of Diodrast in the
tion by a histochemical method. Similar stud¬ blood, a point is reached at which the kidney
ies have been carried out by administration fails to remove all the material from the
of ferritin and 125I-labeled albumin by micro¬ blood. The maximal concentration that is
puncture, followed by direct or autoradio¬ completely cleared is taken as a measure of
graphic visualization of the tracer substance. the excretory capacity of the tubule (Tm, or tubular
The results of these studies are in close agree¬ maximum). If the values are known for renal
ment, all showing uptake in the apical inva¬ plasma flow and inulin clearance, the meas¬
ginations and apical vacuoles of the proximal urement of other substances (such as urea,
tubule within a very few minutes. In 30 to uric acid, and phosphate and bicarbonate
60 minutes, the label is localized in dense buffers) in the blood and urine can be related
granules containing acid phosphatase, inter¬ to the activities of the total number of neph¬
preted as secondary lysosomes. It is assumed rons with regard to these substances. In this
that absorbed albumin is degraded by the way it has been determined which substances
lysosomes and is not returned to the blood¬ are secreted, which are reabsorbed, and
stream. which diffuse passively from the glomerular
Thus, useful substances are conserved by filtrate. The localizations of these specific
reabsorption. On the other hand, the end events in various portions of the tubule are
products of metabolism, urea, uric acid, and less well known.
creatinine, which are of little or no use to the The loop of Henle is essential for the
 THE URINARY SYSTEM • 785

production of hypertonic urine. Only those tenance of this gradient depends on the ar¬
birds and mammals that have a thin segment rangement of the vascular supply and loop
in the loop excrete urine that is hypertonic of Henle, and on the permeability properties
to the blood plasma. The length of this seg¬ of its various segments (Fig. 30-33, 30—34).
ment, and of the renal papilla where it is The thin descending limb is highly permea¬
located, are correlated with the degree to ble to water but not to NaCl. The thin as¬
which the species can concentrate the glo¬ cending limb is impermeable to water but
merular filtrate. Mammals such as beaver permeable to salt. The thick ascending limb,
living in an aqueous environment have little which continues into the cortex as the distal
need to conserve water, and the osmolarity tubule, is impermeable to diffusion of salt
of their urine is only about twice that of and water but has considerable capacity for
plasma. Animals living in the desert, on the active transport of salt from the glomerular
other hand, have a long renal papilla and filtrate to the interstitial fluid. This active salt
very long loops of Henle, and can concen¬ transport creates hypertonicity in the outer
trate urine to over 20 times the osmolarity of medulla and inner cortex. Water diffuses
plasma. The maximal concentration achieved from the descending limb of the loop, which
by humans is about fivefold. is relatively impermeable to salt and urea.
Fluid in the outer renal cortex is approxi¬ Diffusion of water into the interstitium cre¬
mately isosmotic with plasma, but from the ates an increased intratubular concentration
corticomedullary junction to the tip of the of solute, mainly salt. After the fluid rounds
papilla, there is a continuous increase in the bend of the loop, the permeability of the
osmolarity of the interstitial fluid. The main¬ thin ascending limb allows salt to diffuse out

DISTAL TUBULE

Figure 30-33. Countercurrent multiplier mechanism. Both the thin ascending limb in the inner medulla and the thick
ascending limb in the outer medulla, as well as the first part of the distal tubule, are impermeable to water, as indicated
by the thickened lining. In the thick ascending limb, active chloride reabsorption, accompanied by passive sodium
movement (1), renders the tubule fluid dilute and the outer medullary interstitium hyperosmotic. In the last part of the
distal tubule and in the collecting tubule in the cortex and outer medulla, water is reabsorbed down its osmotic gradient
(2), increasing the concentration of urea that remains behind. In the inner medulla both water and urea are reabsorbed
from the collecting duct (3). Some urea reenters the loop of Henle (not shown). This medullary recycling of urea, in
addition to trapping of urea by countercurrent exchange in the vasa recta (not shown), causes urea to accumulate in
large quantities in the medullary interstitium (indicated by the larger type), where it osmotically extracts water from the
descending limb (4) and thereby concentrates sodium chloride in descending-limb fluid. When the fluid rich in sodium
chloride enters the sodium chloride-permeable (but water-impermeable) thin ascending limb, sodium chloride moves
passively down its concentration gradient (5), rendering the tubule fluid relatively hypo-osmotic to the surrounding
interstitium. (From Jamieson, R. L., and R. H. Maffly. N. Engl. J. Med. 295:1059, 1976.)
786 • THE URINARY SYSTEM

The thick ascending limb, distal tubule,

and cortical and outer medullary portions of
the collecting duct can be thought of as a
second loop. These segments are permeable
to water but impermeable to salt and urea.
In this loop, tubular fluid diluted by active
transport of salt in the thick ascending limb
and distal tubule continues to lose some
water, and the concentration of urea in the
collecting duct increases. In the inner me¬
dulla, the collecting duct is permeable to
water and urea. Urea leaves the tubule fluid
down its concentration gradient, contributing
to the high osmolarity of the interstitial fluid,
and water diffuses out of the collecting duct
to achieve the final concentration of the ur¬
ine.
Maintenance of the osmotic gradient that
drives this passive countercurrent multiplier
thus involves two solutes: (1) salt, which dif¬
fuses from the thin ascending limb and is
actively transported in the thick limb; and (2)
urea, which diffuses from the collecting duct
in its passage through the inner medulla.
The classical countercurrent multiplier
mechanism advanced by physiologists in the
1960s to account for concentration of the
urine assumed active sodium transport by
the entire ascending limb of the loop of
Figure 30-34. Countercurrent exchange by the vasa Henle, and assigned no significant role to
recta. The medullary circulation, unlike the ioops of Henle,
consists of a network of channels with main thoroughfares urea in maintenance of the gradient. Electron
(the vasa recta) and branch connections. Pr denotes micrographs of the thin ascending limb re¬
plasma protein. The size of type indicates the relative vealed a very simple structure incompatible
concentrations of each solute with respect to its location with active transport. Subsequent develop¬
in the medulla but not necessarily with respect to other
solutes. The progressive rise in the concentration of
ment of a technique for perfusion of isolated
sodium chloride and urea in the medullary interstitium is segments of tubules in vitro showed that the
due to the loop of Henle and collecting tubule. Since the descending and ascending thin limbs were
capillaries are permeable to sodium chloride and urea, quite different in their permeability proper¬
these solutes enter descending vasa recta and leave
ties, and neither had a mechanism for active
ascending vasa recta. This transcapillary exchange helps
“trap” the solutes in the medulla. Conversely, water leaves sodium transport. The new passive countercur¬
the descending vasa recta, causing the plasma protein rent multiplier mechanism described above is
concentration to increase. In the ascending vasa recta, now consistent with these observations and
the sum of oncotic (that due to plasma protein) and
with the ultrastructure of these segments of
osmotic (that due to nonprotein solutes) pressures results
in capillary fluid uptake. Thus, water reabsorbed from the the nephron.
collecting tubule and Henle’s descending limb is removed The water permeability of collecting ducts
from the medulla and returned to the general circulation. is controlled by antidiuretic hormone (ADH)
Vasa recta function in a dual capacity, trapping solute from the posterior lobe of the hypophysis.
and removing water, to preserve the hyperosmolality of
the renal medulla. (From Jamieson, R. L., and R. H. In the presence of this hormone, fluid within
Maffly. N. Engl. J. Med. 295:1059, 1976.) the duct can come to osmotic equilibrium
with the interstitial fluid surrounding the
duct. In the absence of the hormone, the
duct is relatively impermeable to water, and
of the tubule down its own concentration the urine passing through the medulla re¬
gradient, resulting in dilution of the ascend¬ mains dilute in spite of the concentration
ing-limb fluid. The thin ascending limb thus gradient in the surrounding interstitium.
passively contributes to maintenance of high The efficiency of reabsorption of sodium
interstitial osmolarity in the inner medulla. ions is also under hormonal control. Aldo-
 THE URINARY SYSTEM • 787

sterone secreted by the zona glomerulosa of mucosa, and the lamina propria of the mu¬
the adrenal cortex acts specifically upon the cosa blends with the smooth muscle coat,
renal tubules to increase their rate of sodium which in turn is covered by an adventitial
transport to the interstitium. In the absence layer of connective tissue.
of aldosterone, there is a serious loss of All the excretory passages of the urinary
sodium via the urine. When the hormone is tract are lined with transitional epithelium.
present in normal amount, some 1200 g of In the calyces, it is two or three cells thick,
sodium are reabsorbed each day and only a and in the ureter, four or five. When the
few hundred milligrams escape in the urine. wall of the bladder is contracted, the epithe¬
lium is six to eight cells thick and its super¬
ficial cells are rounded or even club-shaped
(Fig. 30-35). When the bladder is distended,
PASSAGES FOR THE the epithelium is thin and the cells are greatly
EXCRETION OF URINE flattened and stretched.
Electron micrographs of transitional epi¬
The excretory passages convey urine from thelium reveal fine structural features pecul¬
the parenchyma of the kidney to the outside. iar to this tissue. The free surface of the cells
Their walls are provided with a well-devel¬ at the lumen has a characteristic scalloped
oped coat of smooth muscle. Its contractions appearance. Segments of membrane of vary¬
move the urine forward. ing length are quite straight and seemingly
The calyces, the pelvis, the ureter, and the stiff (Fig. 30-36). Neighboring straight seg¬
bladder all have a similar structure, but the ments may be so oriented as to produce
thickness of the wall gradually increases from angular surface contours not seen on other
the upper to the lower part of the urinary cells. There is a superficial ectoplasmic layer
tract. The inner surface is lined with a mu¬ of cytoplasm rich in fine filaments, and bun¬
cous membrane. There is no distinct sub¬ dles of filaments course through the deeper

Capillary
Epithelium

Capiilary

Lamina
propria

Vein

Artery
Vein (cut
tangen¬
tially )

Muscle ——

Figure 30-35. Section of wall of human urinary bladder in contracted condition; capillaries penetrate the epithelium,
x 150. (After A. A. Maximow.)
788 • THE URINARY SYSTEM

Figure 30-36. Electron micrograph of portions of two transitional epithelial cells from the bladder. Notice the flattened
elliptical vesicles in the cytoplasm of the upper cell and the peculiar angular appearance of the luminal surface. This
apparently results from insertion of relatively stiff segments of membrane into the surface when the lenticular vesicles
fuse with the plasma membrane. (Micrograph from Hicks, M., and B. Ketterer, J. Cell Biol. 45:542, 1970.)

cytoplasm as well. Flattened, elliptical, or len¬ to be a hexamer composed of twelve smaller

ticular vesicles are present in the superficial subunits arranged in a stellate configuration
cytoplasm, and these are bounded by thick (Fig. 30-37). The significance of this lattice
membranes of the same character as that on structure in relation to the function of the
the luminal surface. Vesicles of this kind are bladder epithelium is by no means clear. It
peculiar to the cells of transitional epithe¬ is known, however, that in man the tonicity
lium. It is speculated that they may be formed of the bladder urine may be two to four times
within the cell and may be added to the higher than that of the plasma in the capil¬
surface membrane, providing for its replace¬ laries of the lamina propria. If the transi¬
ment or for its rapid expansion in distention tional epithelium were to act as a semiperme-
of the bladder. able membrane, water would pass from blood
The luminal plasma membrane of the su¬ to urine, and the latter would become di¬
perficial cells of bladder epithelium has a luted. This does not occur, and it seems
unique ultrastructure and unusual physiolog¬ evident therefore that the epithelium pos¬
ical properties. It is thicker (12 nm) than sesses an effective barrier preventing water
most cell membranes and asymmetrical in loss. I he barrier function is diminished or
sections, with the outer dense line of the unit lost if the thick surface membrane is chemi¬
membrane significantly thicker than the in¬ cally altered or mechanically damaged. The
ner dense line. When this membrane is iso¬ barrier is believed to reside in occluding
lated and examined by negative staining and junctions between the superficial cells that
optical diffraction, it is found to have a highly close the intercellular spaces, and in the spe¬
ordered substructure consisting of hexago- cial properties of their thick luminal mem¬
nally arranged subunits. Each subunit seems brane.
 THE URINARY SYSTEM • 789

looser layer of connective tissue is especially

abundant so that in the contracted condition
of the organ, the mucous membrane forms
numerous thick folds.
The muscular coat of the urinary passages,
in contrast to that of the intestine, does not
form clearly defined separable layers. In¬
stead, it occurs as loose anastomosing strands
of smooth muscle separated by abundant
collagenous connective tissue. In general, the
muscular coat consists of an inner longitudi¬
nal and an outer circular layer, but their
limits are ill defined. Beginning in the lower
third of the ureter, a third external longitu¬
dinal layer is added. This is especially prom¬
inent in the bladder.
In the small calyces capping the papillae of
the pyramids, the strands of the inner lon¬
gitudinal muscle layer end at the attachment
of the calyx to the papilla. The outer circular
strands reach higher up and form a muscular
ring around the papilla. The calyces show
periodic contractions. This muscular activity
is believed to assist in moving the urine out
Figure 30-37. Electron micrograph of a negatively stained
of the papillary ducts into the calyces. The
portion of the cell membrane at the free surface of muscular coat of the ureter also performs
transitional epithelium. This membrane has a unique slow peristaltic movements. The waves of
substructure (see inset) consisting of hexagonally ar¬ contraction proceed from the pelvis toward
ranged subunits. This structure may be related to the
the bladder.
unusual permeability properties of the bladder epithelium.
(Micrograph from Hicks, M., and B. Ketterer. J. Cell Biol. Because the ureters pierce the wall of the
45:542, 1970.) bladder obliquely, their openings are usually
closed by the pressure of the contents of the
bladder and are open only when the urine is
No true glands are present in the calyces, forced through them. A fold of the mucous
the pelvis, or the ureter, but glands may be membrane of the bladder extending over the
simulated here by small, solid nests of epithe¬ ureteral orifice and acting as a valve usually
lial cells within the thickness of the epithelial prevents the backflow of the urine. In the
sheet. In the urinary bladder, however, and intramural part of the ureters, the circular
in the vicinity of the internal urethral orifice, muscular strands of their wall disappear, and
small invaginations of the epithelium into the the connective tissue of the mucous mem¬
subjacent connective tissue can be found. brane contains longitudinal muscular strands
They contain numerous clear, mucus-secret¬ whose contraction opens the lumen of the
ing cells and are similar to the glands of ureter.
Littre in the urethra. The muscular coat of the bladder is very
There is a thin basal lamina between the strong. Its thick strands of smooth muscle
epithelium and the lamina propria. The con¬ cells form three layers, which intermingle at
nective tissue of the latter forms thin folds their margins and cannot be separated from
that may penetrate deep into the epithelium. one another. The outer longitudinal layer is
The connective tissue underlying the mucosa developed best on the dorsal and ventral
is abundant and contains elastic fiber net¬ surfaces of the viscus, while in other places
works and sometimes small lymphatic nod¬ its strands may be wide apart. The middle
ules. Its deeper layers have a loose arrange¬ circular or spiral layer is the thickest of all.
ment. The mucous membrane in the empty The inner layer in the body of the bladder
ureter, therefore, is thrown into several lon¬ consists of relatively sparse longitudinal or
gitudinal folds, which in cross section give a oblique strands. In the region of the trigone,
festooned appearance to the margin of the thin dense bundles of smooth muscle form a
lumen (Fig. 30—38). In the bladder, the deep, circular mass around the internal opening of
790 • THE URINARY SYSTEM

Fat tissue

Adventitial
connective tissue

mina propria
Internal longitudinal
muscle bundles
Circular
muscle bundles
Adventitial
connective tissue

Vein
External longitudinal
Artery
muscle bundles

Artery

Figure 30-38. Cross section of markedly contracted human ureter, x 30. (After Schaffer.)

the urethra, forming the internal sphincter of A sympathetic nerve plexus in the adven¬
the bladder. titial coat of the bladder, the plexus vesicolis,
is formed in part by the pelvic nerves, which
Blood Vessels, Lymphatics, and originate from the sacral nerves, and in part
Nerves by the branches of the hypogastric plexus.
The vesical plexus sends numerous nerves
The blood vessels of the excretory passages into the muscular coat. A continuation of the
penetrate first through the muscular coat and nerve plexus, but seemingly without nerve
provide it with capillaries. They then form a cells, is found in the connective tissue of the
plexus in the deeper layers of the mucous mucosa. Flere the sensory nerve endings are
membrane. From here, small arteries pass located. Many fibers penetrate into the epi¬
toward the surface and form a rich capillary thelium between the cells, forming varicose
plexus immediately under the epithelium. free endings.
The deeper layers of the mucosa and the
muscularis in the renal pelvis and the ureters
also contain a well-developed network of
URETHRA
lymph capillaries. In the bladder, lymph ves¬
sels are said to be present only in the mus¬
cularis.
Male Urethra
Nerve plexuses, small ganglia, and scat¬ The male urethra has a length of 18 to 20
tered nerve cells can be found in the adven¬ cm. Three parts can be distinguished. The
titial and muscular coats of the ureter. Most short proximal segment surrounded by the
of the fibers supply the muscle, but some prostate is the pars prostatica (prostatic urethra).
fibers apparently of efferent nature have Here the posterior wall of the urethra forms
been traced into the mucosa and the epithe¬ an elevation, the colliculus seminalis (verumon-
lium. tanum). On its surface in the midline is the
 THE URINARY SYSTEM • 791

Encapsulated sensory nerve ending

Artery

Vein

Lacuna Vein Intraepithelial group

of glandular cells

Figure 30-39. Section of cavernous part of male human urethra, x 165. (After A. A. Maximow.)

opening of the utriculus prostaticus, the rudi¬

mentary homologue of the uterus in the
male. Located to the right and to the left of
this are the two slitlike openings of the ductus
ejaculatorius (ejaculatory duct) and the numer¬
ous openings of the ducts of the prostate
gland. The second, very short segment of the
urethra (18 mm long), the pars membranacea
(membranous urethra), extends from the lower
pole of the prostate to the bulb of the corpus
spongiosum of the penis. The third portion,
pars spongiosa (penile urethra), which is about
15 cm long, passes longitudinally through
the corpus spongiosum of the penis.
The prostatic part is lined with the same
transitional type of epithelium as the bladder.
The pars membranacea and the pars spon¬
giosa are lined with a stratified or pseudo-
stratihed columnar epithelium (Figs. 30—39,
30-40). Patches of stratified squamous epi¬
thelium are common in the pars spongiosa.
In the terminal enlarged part of the canal,
the fossa navicularis, stratified squamous epi¬
thelium occurs as a rule. In the surface epi¬
thelium, occasional mucous goblet cells may
be found. Intraepithelial cysts containing a
colloid-like substance are common.
The lamina propria of the mucosa is a Figure 30-40. Photomicrograph of stratified columnar
loose connective tissue with abundant elastic epithelium of human urethra.
792 * THE URINARY SYSTEM

Stratified Epithelium with Outpocketings of clear mucous cells Blood vessel

columnar clear cells
epitheiiu

Figure 30-41. Section of urethral gland (gland of Littre) from cavernous part of male human urethra, x 165. (After A.
A. Maximow.)

networks. No separate submucous layer can lium is transformed into compact intraepi¬
be distinguished. This connective tissue con¬ thelial nests of clear cells, which have the
tains numerous scattered bundles of smooth staining reactions of mucus. In old age some
muscle, mainly oriented longitudinally. In of the recesses of the urethral mucosa may
the outer layers, however, circular bundles contain concretions similar to those of the
are also present. The lamina propria has no prostate.
distinct papillae extending into the epithe¬
lium; these appear only in the fossa navicu-
Female Urethra
laris. The membranous portion of the ure¬
thra is surrounded by a mass of striated The female urethra is 25 to 30 mm long.
muscle, a part of the urogenital diaphragm. Its mucosa forms longitudinal folds and is
The surface of the mucous membrane of lined with stratified squamous epithelium. In
the urethra shows many recesses, the lacunae many cases, however, pseudostratihed col¬
of Morgagni. These outpocketings continue umnar epithelium can be found. Numerous
into deeper, branching tubules, the glands of invaginations are formed by the epithelium
Littre (Fig. 30—41). The larger ones among (Fig. 30—42). The outpocketings in their wall
them are found especially on the dorsal sur¬ are lined in many places with clear mucous
face of the pars spongiosa of the urethra. cells, as in the glands of Littre of the male
They run obliquely in the lamina propria urethra. The glands may accumulate colloid
and are directed with their blind end toward material in their cavities or may even contain
the root of the penis. They sometimes pene¬ concretions. The lamina propria, devoid of
trate far into the corpus spongiosum. The papillae, is a loose connective tissue with
glands of Littre are lined with the same abundant elastic fibers. It is provided with a
epithelium as the surface of the mucous highly developed system of venous plexuses
membrane, but in many places this epithe¬ and has, therefore, a character resembling
 THE URINARY SYSTEM • 793

Figure 30-42. Cross section through female human urethra. The darker portions of the lamina propria are smooth
muscle bundles, x 10. (After von Ebner.)

the corpus spongiosum of the male. The Elema, J. D., J. R. Hoyer, and R. L. Vernier: The
glomerular mesangium: uptake of intravenously in- <
mucous membrane with its veins is sur¬ jected colloidal carbon. Kidney Int. 9:35, 1976.
rounded by a thick mass of smooth muscles; Farquhar, M. G.: The primary glomerular filtration
the inner layers of the latter have a longitu¬ barrier—basement membrane or epithelial slits?
dinal, the outer layers a circular, arrange¬ Kidney Int. 5:197, 1975.
Farquhar, M. G., and Y. S. Kanwar: Characterization of
ment. Distally, the smooth muscles are
anionic sites in the glomerular basement membranes
strengthened by a sphincter of striated mus¬ of normal and nephrotic rats. In Leaf, A., et al.,
cle. eds.: Penile Pathophysiology. New York, Raven
Press, 1980, pp. 57-74.
Farquhar, M. G., and G. E. Palade: Functional evidence
for the existence of a third cell type in the renal
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RENAL CORPUSCLE Latta, H. W., W. H. Johnston, and T. M. Stanley:
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Ericsson, J. L. E., A. Bergstrand, G. Andres, H. Bucht, P. Brown: Endocrine-type antihypertensive function

and G. Cinotti: Morphology of the renal tubular of renomedullary interstitial cells. Kidney Int. 5:271,
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Microbiol. Scand. 65:361, 1965. Schifferli, J., A. Grandcamp, and F. Chatelanat: Ultra¬
Kokko, J. P., and F. C. Rector: Countercurrent multi¬ structure of the interstitial cells of the rat renal
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Oswaldo, L., and H. Latta: The thin limb of the loop of Tobian, L.: Relationship of juxtaglomerular apparatus
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Pricam, C., F. Humbert, A. Perrelet, and L. Orci: A
BLOOD VESSELS
freeze etch study of the tight junctions of the rat
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renal ultrastructure. I. Proximal tubule of healthy
individuals. Lab. Invest. 75:1357, 1966. LYMPHATICS
Tisher, C. C., R. E. Bulger, B. F. Trump: Human renal Albertine, K. H., and C. C. O’Morchoe: Distribution and
ultrastructure. III. The distal tubule in healthy density of the canine renal cortical lymphatic system.
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border and lateral cell walls in the rabbit proximal L. P. Henry. Renal lymphatics: the internal distri¬
nephron. Kidney Int. 5:343, 1975. bution. Nephron 5:454, 1968.
Welling, L. W., and D. J. Welling: Shape of epithelial Pierce, E. C.: Renal lymphatics. Anat. Rec. 99:315, 1944.
cells and intercellular channels in the rabbit proxi¬ Rojo-Ortega J. M., E. Yeghiayan, and J. Genest: Lym¬
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INTERSTITIUM Invest. 29:336, 1973.

Bohman, S. O.: The interstitial cells of the renal medulla. HIS TOPHYSIOLOGY
Thesis, University of Aarhus, Denmark, 1979. Beeuwkes, R.: Renal countercurrent mechanisms or how
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 THE URINARY SYSTEM • 795

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URINARY BLADDER epithelium of the urinary bladder of rat. Lab. Invest.
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water. J. Cell Biol. 28:21, 1966.
 31
MALE REPRODUCTIVE
SYSTEM

The male reproductive system includes the On the inner aspect of the tunica albugi¬
gonads, two testes and their duct system (the nea, dense connective tissue gives way to a
ductuli efferentes, ductuli epididymides, ductus def¬ looser layer provided with numerous blood
erens, and ejaculatory ducts) together with as¬ vessels, the tunica vasculosa testis. A loose con-
s

sociated accessory glands—the seminal vesicles, nective tissue of similar character extends
prostate, and bulbourethral glands (Cowper’s inward from this layer to fill all of the inter¬
glands) (Figs. 31—1, 31-2). stices among the seminiferous tubules. It con¬
tains fibroblasts, macrophages, mast cells, and
perivascular mesenchymal cells. In addition,
there are clusters of epithelioid interstitial cells,
TESTIS also called Leydig cells. These constitute the
endocrine tissue of the testis.
The testis is a compound tubular gland Each testis is suspended in the scrotum at
enclosed in a thick fibrous capsule, the tunica the end of a long vascular pedicle, the sper¬
albuginea. On the posterior aspect of the matic cord, which consists of the excretory
organ, a thickening of the connective tissue duct of the testis, the ductus deferens, and the
capsule projects into the gland as the medias¬ blood vessels and nerves supplying the testis
tinum testis. Thin fibrous septa, called the on that side. The epididymis, an organ closely
septula testis, extend radially from the medias¬ applied to the posterior surface of the testis,
tinum to the tunica albuginea, dividing the is made up of the convoluted proximal part
organ into about 250 pyramidal compart¬ of the excretory duct system (Figs. 31-2,
ments, the lobuli testis. The septula may be 31—3). Each testis and epididymis is sur¬
incomplete toward the periphery, so that the rounded on its anterior and lateral surfaces
lobules intercommunicate, but where their by a cleftlike serous cavity that arises late in
apices converge upon the mediastinum they embryonic development as a detached por¬
are more completely separated. tion of the peritoneal cavity.
Each lobule is composed of one to four The testes develop early in embryonic life
highly convoluted seminiferous tubules. These in the dorsal wall of the abdominal cavity
are 150 to 250 pm in diameter, 30 to 70 cm and later descend into the scrotum, each
long, and extremely tortuous (Fig. 31-3). carrying with it an outpocketing of the peri¬
The seminiferous tubules constitute the ex¬ toneum, the tunica vaginalis propria testis,
ocrine portion of the testis, which is in es¬ which forms the serous cavity around the
sence a cytogenous gland whose holocrine testis. It consists of an outer parietal and an
secretory product is whole cells, the sperma¬ inner visceral layer that is closely applied to
tozoa. The tubules are usually highly convo¬ the tunica albuginea of the testis on its ante¬
luted loops, but they may also branch or end rior and lateral surfaces. On the posterior
blindly. At the apex of each lobule its semi¬ aspect of the testis, where the blood vessels
niferous tubules pass abruptly into the tubuli and nerves enter the organ, the visceral layer
recti, the first segment of the system of excre¬ is reflected from its surface and is continuous
tory ducts. They in turn are confluent with with the parietal layer. After removal of the
the rete testis, a plexiform system of epithe¬ parietal layer, the visceral coat covering the
lium-lined spaces in the connective tissue of testis appears as a glistening, smooth surface
the mediastinum. covered with mesothelium, which is the rem-
796
 MALE REPRODUCTIVE SYSTEM • 797

URINARY URETER
PERITONEUM BLADDER

SEMINAL
PUB/S- VESICLE

AMPULLA

RECTUM
Figure 31-1. Diagrammatic represen¬
CA VERNOUS
tation of the male genital system. The BODIES
midline structures are shown in sagittal PROSTATE
section; bilateral structures, such as
testis, epididymis, vas deferens, and
seminal vesicle, are depicted intact. EJACULATORY
DUCT
(After C. D. Turner.)
URETHRA—VfflfS
PRO STATIC UTRICLE

COWPER'S GLAND

GLANS
PENIS

SCROTUM EPIDIDYMIS

TUNICA VAGINALIS TESTIS

nant of the coelomic epithelium that covered Ductus

deferens
the primordium of the gonad in the embryo.
The tunica vaginalis enables the testis, which Ductuli
is sensitive to pressure, to glide freely in its
envelopes.

Caput
epididymidis

Tunica
albuginea

Rete testis

Septula
testis

Seminiferous
tubules

Tubuli recti

Corpus
epididymidis

Cauda
epididymidis
Figure 31-3. Cutaway diagram of the architecture of the
testis and excurrent duct system. The septula divide the
organ into a number of compartments occupied by highly
Figure 31-2. Photomicrograph of human testis, epidid¬ convoluted seminiferous tubules. One has been unraveled
ymis, and ductus deferens. The epididymis has been and drawn out to show its length and the fact that it is a
dissected free and drawn away from the testis to reveal loop terminating in the rete testis. (Drawing modified from
more clearly the coni vasculosi. (Photograph courtesy of Hamilton, W. J. Textbook of Human Anatomy. London,
A. F. Holstein.) Macmillan & Co., 1957.)
798 • MALE REPRODUCTIVE SYSTEM

Sertoli Cells. The three-dimensional con¬

SEMINIFEROUS TUBULES
figuration of the Sertoli cell is extraordinarily
complex, but it can be thought of as basically
The Lamina Propria or Boundary
columnar, resting upon the basal lamina and
Tissue extending upward through the full thickness
The seminiferous tubules are enclosed by of the epithelium to its free surface. From
one or more layers of adventitial cells derived the columnar portion of these cells an elab¬
from primitive connective tissue elements of orate system of thin processes radiate lat¬
the interstitium. The organization of this erally to surround the spermatogenic cells
boundary tissue varies from species to spe¬ and occupy all of the interstices among them.
cies. In the common laboratory rodents, The earliest of the germ cells, the spermato¬
there is a single layer of flattened polygonal gonia, also rest upon the basal lamina, while
cells that meet edge to edge to form a contin¬ the more advanced stages of the germ cell
uous epithelioid sheet surrounding the tu¬ line are found at successively higher levels in
bule. In their ultrastructure, these cells have the epithelium (Figs. 31—7, 31—8). The pro¬
the cytological characteristics of smooth mus¬ liferative activity in the epithelium is confined
cle and are contractile. They cannot be called to the spermatogonia and spermatocytes near
true smooth muscle cells because of their the base. The continual formation of new
atypical shape and epithelioid organization, generations of cells in this region displaces
and therefore are referred to as myoid cells or the more advanced cells to higher levels until,
peritubular contractile cells. They are believed as mature sperm, they come to border di¬
to be responsible for the rhythmic shallow rectly upon the lumen. To understand sper¬
contractions that can be observed in the sem¬ matogenesis, it is important to bear in mind
iniferous tubules of these species. The con¬ that the seminiferous epithelium consists of
tractions seem to be intrinsically generated, (1) a fixed population of nonproliferating
since no nerves have been observed in or supporting cells and (2) a proliferating and
near this layer. differentiating population of germ cells that
In larger species, such as ram, boar, and move slowly upward along the sides of the
bull, there are multiple layers of adventitial Sertoli cells to the free surface. This dynamic
cells. In these, only the innermost layer is relationship of the cells makes the lining of
muscle-like, the next layer has some of the the seminiferous tubules unique among epi-
characteristics of smooth muscle, and outer thelia.
layers have the appearance of fibroblasts. In Owing to the elaborate shape of the Sertoli
monkey and man there are also multiple cells and the limitations of resolution of the
layers of cells, but these are not epithelioid light microscope, their outlines cannot be
and the cells do not resemble smooth muscle seen distinctly. Earlier, this gave rise to the
as much as in other species. Contractility of widespread belief that they constituted a syn¬
seminiferous tubules lias not been observed cytium, but this interpretation is now known
in primates. In many cases of human male to be erroneous. In sections parallel to the
infertility, the boundary tissue becomes basal lamina of the epithelium, the bases of
greatly thickened. the Sertoli cells can be seen with the light
microscope as distinctly outlined polygonal
The Seminiferous Epithelium areas. Electron micrographs clearly show
pairs of apposed membranes at the boundary
In the adult, the seminiferous tubules are between adjacent Sertoli cells and between
lined by a complex stratified epithelium com¬ the latter and the germ cells. Therefore, the
posed of two major categories of cells: sup¬ spermatogenic cells are not embedded in a
porting cells and spermatogenic cells. The sup¬ “Sertolian syncytium,” but instead occupy
porting elements are of a single kind, the deep recesses of conforming shape in the
Sertoli cells, whereas the spermatogenic cells lateral and apical surfaces of the Sertoli cells.
include several morphologically distinguish¬ 4 he elaborate shape of these cells (Fig. 31—9)
able types: spermatogonia, primary spermatocytes, is probably attributable to their close coapta¬
secondary spermatocytes, spermatids, and sperma¬ tion to the highly irregular and changing
tozoa (Figs. 31—4 to 31—8). These germ cells contours of the differentiating germ cells that
are not ontogenetically distinct cell types but they surround.
are successive stages in a continuous process The nucleus of the Sertoli cell is generally
of differentiation of the male germ cells. ovoid in outline but may have one or more
 MALE REPRODUCTIVE SYSTEM • 799

Figure 31-4. Photomicrograph of several seminiferous tubules and the interstitial tissue of a typical mammalian testis.
Notice that the plane of section transects neighboring tubules in different stages of the spermatogenic cycle. Tubules A
and B contain nearly mature sperm about to be released; in C and D, elongating spermatids are the most advanced
germ cells; in E, F, and G, spermatogenesis has proceeded only to the round spermatid stage.
800 • MALE REPRODUCTIVE SYSTEM

Blood vessel

Fibroblast
Inter¬ Spermato¬
stitial gonium
cell Sertoli cell
Sper¬ 'rimary
mato¬ spermatocyie
gonium
Sertoli
cell
Sertoli
cell

Primary
spermat¬
ocyte in Sperm
mitosis
Spermatid
Sertoli cells '

with bunches <

Basement
of maturing membrane
sperm
Interstitial cell

Blood vessel
Interstitial cell

Secondary spermatocyte
Figure 31-5. Section of human testis (obtained at operation). The transected tubules show various stages of
spermatogenesis, x 170. (After A. A. Maximow.)

deep infoldings of its surface. It is about 9 lipid droplets and occasional lipofuscin pig¬
by 12 pm in average size, with a relatively ment granules are found near the cell base.
homogeneous nucleoplasm, except for a The granular endoplasmic reticulum is
large and highly characteristic nucleolus, con¬ sparse but the agranular reticulum is well
sisting of a round or oval central body developed, especially near the cell base. It
flanked by two rounded basophilic masses. usually occurs in the form of a network of
In electron micrographs, the tripartite struc¬ smooth-surfaced tubules, but in some species
ture of this complex is confirmed. The cen¬ it may form concentric systems of membranes
tral element consists of a typical nucleolo- around lipid droplets. The presence of a well-
nema organized around a homogeneous developed smooth endoplasmic reticulum at
central area of relatively low density. The two the cell base has been interpreted by some as
adjacent darker masses of finely granular evidence for secretion or modification of ste¬
material appear to be nucleolus-associated roid hormones, but such a function has yet
chromatin. to be clearly established for the Sertoli cell.
The cytoplasm contains numerous slender In certain stages of the spermatogenic cycle,
elongated mitochondria often oriented par¬ close aggregations of smooth reticulum are
allel to the long axis of the cell. Numerous found in the cytoplasm immediately adjacent
 MALE REPRODUCTIVE SYSTEM • 801

to the developing acrosomal cap of each lysosomes, and to large irregular conglom¬
associated spermatid (Fig. 31—9). The signif¬ erates of lipochrome pigment with a very
icance of this striking localization remains heterogeneous content of globular and gran¬
unexplained, but may be an expression of ular components of varying density. Al¬
the “nurse cell” function of Sertoli cells. though the lysosomal digestive system of
The Golgi complex is extensive but shows these cells is well developed, the amount of
no morphological indication of involvement accumulated pigment is not large when one
in secretory activity. The slender mitochon¬ considers that they are responsible for deg¬
dria have the usual foliate internal membrane radation and disposal of the residual cyto¬
structure and are remarkable only for their plasm of generation after generation of sper¬
length. The lysosomal system of the cell ex¬ matids.
hibits a diversity of components, ranging The filamentous component of the cyto¬
from membrane-limited, spherical primary plasmic matrix of Sertoli cells is more abun¬
lysosomes to pleomorphic, dense, secondary dant than in many other cell types. A feltwork

Primary spermatocyte
Spermatogonium with crystalloid Basement membrane

Spermato¬
gonium
Crystalloid in Crystalloid in
Sertoli cell Sertoli cell

Sertoli cell

Sertoli cell

Fibril in
Sertoli cell

Primary
spermatocyte

Primary
.. spermatocyte
in first mei¬
Primary otic division
spermatocyte
in first meiotic
division

Sperm Spermatid
Figure 31-6. Section of same testis as in Figure 31-5; seminiferous epithelium with primary spermatocytes in first
meiotic division. Iron-hematoxylin stain, x 750. (After A. A. Maximow.)
802 • MALE REPRODUCTIVE SYSTEM

Abnormal spermatid Primary spermatocytes

I ^--'

Spermatid ill 1

Sertoli cell
Crystal

Spermato¬
gonium with
crystal

Spermato¬
Diuiding gonium
secondary
spermatocyte
Secondary
spermatocyte

permato-
gonium with
crystal
Spermatid

Basement
membrane

DiuidIng~secondary Secondary spermatocyte

spermatocytes
Figure 31-7. Section of same testis as in Figure 31-5; seminiferous epithelium with mitoses of secondary spermatocytes
in second meiotic division. The loosening of the spermatocytes and spermatids from their normal attachment to the
Sertoli cells is an artifact of specimen preparation, x 750. (After A. A. Maximow.)

of 8- to 10-nm filaments excludes organelles talloid. They are often rather poorly ordered
and inclusions and results in a thin clear zone and there may be irregular defects in the
surrounding the nucleus. Filaments in lower interior of these crystalloids, occupied by
concentrations are randomly dispersed in the cytoplasmic matrix. Their chemical nature
cytoplasm. Occasionally they associate lat¬ and physiological significance are unknown.
erally to form fascicles parallel to the long The Sertoli cells provide mechanical sup¬
axis of the cell. At certain stages of the port and protection for the developing germ
spermatogenic cycle, microtubules are also cells, and they participate in their nutrition.
abundant in the supranuclear columnar por¬ They also seem to play an active role in the
tion of the Sertoli cell. They are very uni¬ release of the mature spermatozoa. The Ser¬
formly spaced and oriented parallel to the toli cells are never observed in division in the
cell axis. The escalation of the germ cells in mature testis. They are resistant to heat,
the epithelium and the release of spermato¬ ionizing radiation, and various toxic agents
zoa probably depend upon active changes in that destroy the more sensitive spermato¬
shape of the supporting cells. The filaments genic cells.
and microtubules of the Sertoli cells are very
probably the agents of these cytoplasmic
movements. THE SPERMATOZOON
Inclusions peculiar to the human Sertoli
cell are the crystalloids of Charcot-Bottcher (Fig. It will facilitate the student’s understanding
31—8). These are slender, fusiform structures of the complex cytological changes that take
10 to 25 pm long, often visible with the light place in the germ cells of the seminiferous
microscope. In electron micrographs, they epithelium if the structure of the end prod¬
consist of dense straight filaments 15 nm in uct—the spermatozoon—is described first.
diameter. These subunits are generally par¬ The mature spermatozoon is an actively
allel or converge toward the end of the crys¬ motile, free-swimming cell consisting of a
 MALE REPRODUCTIVE SYSTEM • 803

Fibrils in Sertoli cell Sertoli cell

*

Crystalloid
in Sertoli
cell

Sperm Interstitial
cell

Spermato¬
gonium

Cell of
lamellaied
connective
tissue
Maturing
sperm

Spermatid

Basement
membrane

Primary spermatocyte

Figure 31-8. Section of same testis as in Figure 31-5; seminiferous epithelium with clusters of maturing sperm,
connected with Sertoli cells. Iron-hematoxylin stain, x 750. (After A. A. Maximow.)

head, which contains a nucleus endowed with greater part of its bulk consists of the nucleus,
all the genetic traits a father can transmit to whose chromatin has become greatly con¬
his offspring, and a tail or flagellum, which densed to diminish its volume for greater
provides the motility that assists in transport mobility and to protect its genome from dam¬
of the sperm to the site of fertilization and age in transit to the egg. The anterior two
ensures that it is appropriately oriented for thirds of the nucleus is covered by the acro¬
penetration of the coatings of the ovum (Figs. somal cap—an organelle containing enzymes
31-10 to 31-12). that have an important role in sperm pene¬
The human sperm head is ovoid in outline tration during fertilization (Figs. 31-11,
in frontal view and pyriform when seen on 31-13). The mammalian sperm head varies
edge, being thicker near the base and taper¬ greatly in size and shape from species to
ing toward the tip. The head is 4 to 5 pm in species.
length and 2.5 to 3.5 pm in width. The The sperm tail is about 55 pm long and
804 • MALE REPRODUCTIVE SYSTEM

Figure 31-9. Drawing illustrating the ultrastructure of the Sertoli cell and its relationship to the germ cells. The
spermatocytes and early spermatids occupy niches in the sides of the columnar supporting cell, while late spermatids
reside in deep recesses in its apex. (After Fawcett, D. W., In Male Fertility and Sterility. Serono Symposium, 1973.)
 MALE REPRODUCTIVE SYSTEM • 805

acrosomal membrane. The two membranes

run parallel throughout most of their course
and enclose a narrow cavity occupied by a
homogeneous amorphous material—the en¬
zyme-rich acrosomal contents. In the human
spermatozoon, the acrosome is relatively
small and does not extend anteriorly much
beyond the leading edge of the nucleus. In
many other mammalian species, however,
there is a conspicuous thickening of the cap
that extends well beyond the nucleus and
may exhibit a species-specific shape. It is
useful to designate this region as the apical
segment of the acrosomal cap. The main por-

- Acrosomal Cap

Head

POSTACROSOMAL REGION

Neck

- Middle Piece

Figure 31-10. Photomicrograph of chinchilla sperm

stained by the Feulgen reaction and counterstained with
light green. The crescentic apical segment of the acro-
some can be seen at the leading edge of the sperm head,
x 3000.

varies in thickness from about 1 pm near the

base to 0.1 pm near its tip. It presents four
segments along its length recognizable with
► Principal Piece
the light microscope by slight differences in
Tail
thickness and in the nature of their sheaths.
From proximal to distal, these regions are
the neck, the middle piece, the principal piece,
and the end piece (Fig. 31 — 11). There are
significant differences in the internal struc¬
ture of these segments. These cannot be
clearly resolved in fresh preparations but
require special cytological techniques, or elec¬
tron microscopy, for their demonstration.
The description of spermatozoon structure
that follows is based largely on electron mi¬
croscopic studies.
Sperm Head. The acrosome is a caplike, - End Piece

membrane-limited organelle that closely con¬

forms to the contours of the tapering anterior
portion of the sperm nucleus. The inner
acrosomal membrane, which is adherent to Figure 31-11. Drawing of a mammalian spermatozoon
the nuclear envelope, is continuous at the as seen with the light microscope, presenting the terms
posterior margin of the cap with the outer used in describing its various regions.
806 • MALE REPRODUCTIVE SYSTEM

Figure 31-12. Scanning micrograph of rabbit sperm migrating over the surface of the uterine endometrium. (Micrograph
courtesy of P. Motta. )

tion of the acrosome is then called the prin¬ adhering cells of the corona radiata and will
cipal segment. In addition, in all mammalian digest the zona pellucida from recently ovu¬
species, there is a specialized caudal region lated eggs. Sperm in the vicinity of ova in the
where there is an abrupt narrowing of the oviduct undergo a sequence of structural
cap and a slight condensation of its contents. changes called the acrosome reaction: the outer
With special staining techniques, this region membrane of the acrosome fuses at multiple
was visible to light microscopists as a band points with the overlying plasma membrane
around the middle of the head and was of the sperm head to create numerous open¬
therefore called the equatorial segment (Fig. ings through which the enzyme-rich contents
31-13). The functional significance of this of the acrosome are liberated in a process
specialization has yet to be discovered. not unlike the release of secretory products
The acrosome of mammalian spermatozoa from a glandular cell. The release of the
stains with the periodic acid—Schiff reaction enzymes is believed to facilitate sperm entry.
and hence contains appreciable amounts of The nucleus of the mature mammalian
carbohydrate. In addition, it is known to spermatozoon is usually dense and homoge¬
contain several enzymes of lysosomal nature. neous in electron micrographs (Figs. 31—13,
Among those identified to date are hyaluron- 31—14). Human spermatozoa are subject to
idase, neuraminidase, acid phosphatase, considerably more variation in size, in shape,
P-A-acetylglucosamidase, and aryl suifatase. and in texture of their chromatin than are
There is also a proteolytic enzyme called those of other species. In ejaculates of fertile
acrosin, remarkably similar to trypsin in its men, the chromatin in a certain proportion
substrate specificity, pH optimum, and range of the sperm nuclei is not homogeneous but
of inhibitors. The precise role of these en¬ is a dense conglomerate of closely packed
zymes in fertilization is still unclear, but ex¬ coarse granular subunits. In these, the proc¬
tracts of sperm acrosomes will disperse the ess of chromatin condensation during sper-
 MALE REPRODUCTIVE SYSTEM • 807

Caudal to the acrosome, a specialized dense

Apical
segment
layer is found between the plasma membrane
and the nuclear envelope (Fig. 31-14) that
Cell exhibits a characteristic pattern of fine struc¬
membrane
ture that differs from species to species. It
seems more closely adherent to the inner
aspect of the cell membrane than to the
Outer
acrosomal
underlying nucleus. This component corre¬
membrane sponds to the “postnuclear cap” of classical
light microscopy. Flowever, inasmuch as it is
not continuous over the posterior surface of
Acrosomal m the nucleus but is simply a broad band encir¬
Principal
contents —
segment cling the postacrosomal region, the term
postacrosomal cap is no longer appropriate.
Although the chemical nature and functional
significance of this layer are not known, it is
Inner
thought to be an important structure, for it
acrosomal
membrane is specifically in this region that the mem¬
brane of the sperm first fuses with that of
the egg during fertilization.
The plasma membrane is firmly adherent
to the nuclear envelope along a line, called
the posterior ring, which encircles the sperm
head at the caudal edge of the postacrosomal
dense layer (Fig. 31—14). Behind this line or
Equatorial
segment
groove the membrane diverges somewhat
from the underlying structures of the nu¬
cleus. The nuclear envelope, which is gener¬
ally closely applied to the condensed chro¬
Figure 31-13. Electron micrograph of a monkey sper¬ matin, diverges from it caudal to this line of
matozoon, illustrating the relationship of the acrosomal membrane adherence and forms a fold of
cap to the condensed nucleus and to the cell membrane. variable size extending back into the neck
region. Pores and annuli that are absent from
the nuclear envelope over the condensed
miogenesis does not seem to have progressed nucleus are abundant in this fold or scroll
to completion. Another peculiarity of human that extends back into the neck. This portion
sperm nuclei is the frequent occurrence of of the envelope is interpreted as a redun¬
irregularly shaped clear spaces of varying size dancy resulting from the diminution in vol¬
in the chromatin. These are commonly re¬ ume of the nucleus associated with nuclear
ferred to as nuclear vacuoles, although they condensation during spermiogenesis. The
are not membrane bounded. They seem to nuclear envelope over the caudal surface of
result from randomly occurring defects in the head is again devoid of pores, and its
condensation of the chromatin, and there is membranes are in close apposition. The nu¬
no evidence at present that they interfere clear envelope lines the shallow implantation
with fertilizing capacity. Despite the absence fossa where the tail attaches to the head. In
of resolvable order in the fine structure of this area there are regular periodic densities
the nucleoplasm, there are indications that bridging the 10-nm interspace between the
the chromosomes retain their identity and leaves of the nuclear envelope. These may
have a consistent arrangement within the help strengthen this region of attachment of
nucleus. For example, quinacrine mustards tail to head.
have been found to stain the Y chromosome The Neck. Immediately behind the head
of man more or less selectively. When this is a connecting piece that has a dense capitulum
method is applied to human sperm, a yellow conforming in shape to the implantation
fluorescent spot is located in approximately fossa to which it attaches (Fig. 31—27). Ex¬
the same position in the heads of all of the tending backward from this capitulum are
male determining sperm. nine segmented columns 1 to 1.5 pm long
808 • MALE REPRODUCTIVE SYSTEM

Mitochondrial
sheath

^ \ m

droplet s t ) 4;
Figure 31-14. Electron micrograph of portions of two spermatozoa of the dormouse (Glis glis). In the postacrosomal
region, the membrane is reinforced by a dense lamina. This specialization may be important for fertilization, for it is this
region that first fuses with the egg membrane. The asterisk indicates the site of the posterior ring, a circumferential iine
of fusion of the plasmalemma and the outer and inner membranes of the nuclear envelope. (Micrograph by D. Phillips,
from Fawcett, D. W. Biol. Reprod. 2:Suppi. 2, 1970.)

that are continuous at their caudai ends with man sperm, a sizable mass of residual cyto¬
nine outer dense fibers of the sperm flagellum. plasm may surround the neck, but in most
In the interior of the connecting piece im¬ mammals this cytoplasmic droplet is either ab¬
mediately subjacent to the articular surface sent from mature sperm or located at the
of the capitulum is a transversely oriented caudal end of the middle piece.
proximal centriole (Fig. 31-27). The triplet The Middle Piece. In the core of the sperm
microtubules of the centriole wall are embed¬ flagellum is the axoneme consisting of two
ded to varying degrees in the dense material central singlet microtubules surrounded by
composing the articular surface and seg¬ nine evenly spaced doublets, an arrangement
mented columns of the connecting piece. A commonly described by the formula (9 + 2).
distal centriole oriented in the axis of the This microtubule complex extends without
sperm flagellum is usually absent from the significant change in structure throughout
mature spermatozoon, but vestiges of its nine the length of the tail from the neck to near
triplets may be associated with the inner the tip of the end piece. The sperm flagellum
aspect of the segmented columns. The cen¬ differs from other flagella in that the axo¬
tral pair of microtubules of the flagellar ax- neme is surrounded by nine outer dense fibers
oneme may extend anteriorly in the interior (9 + 9 + 2). Each dense fiber is continuous
of the connecting piece as far as the proximal anteriorly with one of the segmented col¬
centriole. umns of the connecting piece, and each
One or two longitudinally oriented mito¬ courses longitudinally in close relation to one
chondria may be found in the neck region of the nine doublets of the axoneme. The
outside the connecting piece, and these may axoneme is considered to be the motor com¬
have processes that extend between the seg¬ ponent, and the outer dense fibers to be
mented columns into its interior. In the hu¬ resilient stiffening structures of the tail.
 MALE REPRODUCTIVE SYSTEM • 809

Three regions of the sperm tail are defined die piece is basically similar in all mammalian
by the nature and extent of the sheaths that spermatozoa*, but the length of the mitochon¬
surround the 9 + 9 + 2 core complex (Fig. drial helix varies from about 15 gyres in
31-20). The middle piece is characterized by a primates to over 300 in some rodents. In the
sheath of circumferentially oriented mito¬ human, the middle piece is from 5 to 7 (xm
chondria arranged end to end in a tight helix long and somewhat more than 1 pm thick.
(Figs. 31-14, 31-15, 31—20). These mito¬ Immediately caudal to the last turn of the
chondria generate the energy for sperm mo¬ mitochondrial sheath is the annulus, a ring of
tility. The structural organization of the mid- dense material to which the flagellar mem¬
brane is firmly adherent (Fig. 31—16). The
annulus and its attachment to the membrane
are presumed to prevent caudal displacement
of the mitochondria during the tail move¬
ments.
The Principal Piece. The principal piece
of the spermatozoon is about 45 pm long
and about 0.5 pm thick at the base, gradually
tapering toward the end piece. It has a highly
characteristic fibrous sheath (Figs. 31—17 to 31 —
20). Studied by electron microscopy, the lat¬
ter is seen to consist of continuous dorsal and
ventral longitudinal columns connected by
regularly spaced circumferential ribs that ex¬
tend halfway around the sheath and are
continuous at their ends with the longitudinal

Retro-
annular
recess
Annulus

Figure 31-16. Electron micrograph of the junction of the

midpiece and principal piece of a rodent spermatozoon.
Figure 31-15. Electron micrograph of the midpiece of a This is the site of the annulus, a dense ring fused to the
spermatozoon in longitudinal section, showing the close plasma membrane. In some species, the cell membrane
apposition of the mitochondrial sheath to the outer dense forms a deep groove or recess immediately behind the
fibers of the flagellum. annulus. In others, the membrane is relatively smooth.
810 • MALE REPRODUCTIVE SYSTEM

of the sperm tail. The principal plane of

bending appears to be perpendicular to the
dorsoventral axis of the tail, and the more
rapid “power stroke” observed in the proxi¬
mal portion of the tail is assumed to be
toward the side having four outer dense
fibers, two of which (numbers 5 and 6) are
especially large. The details of the mecha¬
nism of sperm tail movement have yet to be
worked out. It is now believed that the mi¬
crotubules of the axoneme produce bending
by a sliding mechanism comparable with that
of skeletal muscle (See Cilia, Chapter 2.)
The End Piece. The fibrous sheath ends
abruptly 5 to 7 pm from the tip of the
flagellum. The terminal portion, distal to this
point, consisting of the axoneme covered
only by the flagellar membrane, is called the
end piece (Fig. 31-20). Its structure is essen¬
tially identical to that of a simple flagellum
or cilium. The manner in which the axoneme
ends varies somewhat from species to species.
In some, the doublets terminate at different
levels in the tapering tip, but in primates, the
nine doublets dissociate into 18 single micro¬
tubules. Thus, including the central pair,
cross sections through the terminal half mi¬
crometer may show 20 closely spaced single
microtubules.
Figure 31-17. Longitudinal sections of two adjacent
sperm tails. One has been cut tangentially to the fibrous
sheath and shows some of its ribs in surface view; the
other is a midline section and therefore shows the circum¬ SPERMATOGENESIS
ferential ribs of the sheath in cross section.
Spermatogenesis comprises the entire se¬
quence of events by which spermatogonia are
columns (Fig. 31—18). In cross sections of the transformed into spermatozoa. For conven¬
principal piece, it is apparent that outer ience of description it may be divided into
dense fibers 3 and 8 terminate a short dis¬ three principal phases. In the first, called
tance beyond the annulus (Fig. 31—18). Distal spermatocy to genesis, the most primitive sper¬
to their termination, the tapering inner edges matogonia (Type A) proliferate by mitotic
of the dorsal and ventral columns of the division to replace themselves and to give rise
sheath extend into the position of these fibers to several successive generations of sperma¬
and are attached to a short flange projecting togonia, each somewhat more differentiated
radially from doublets 3 and 8 of the axo- than the preceding one. Division of the last
neme (Fig. 31—18). A plane through the generation of spermatogonia (Type B) yields
longitudinal columns of the fibrous sheath preleptotene spermatocytes. In the second
coincides approximately with the plane phase, meiosis, the spermatocytes undergo two
through the centers of the central pair of maturation divisions, which reduce the chro¬
microtubules of the axoneme. This plane mosome number by half and produce a clus¬
divides the cross section of the tail asymmet¬ ter of spermatids. In the third phase, called
rically (Fig. 31-18). On one side is a minor spermiogenesis, the spermatids undergo a re¬
compartment containing three outer dense fi¬ markable series of cytological transforma¬
bers, and on the other is a major compartment tions leading to the formation of spermato¬
containing four. The asymmetry in the dis¬ zoa.
tribution of these fibers in the cross section For spermatogenesis to continue without
is believed to be reflected in the movements exhausting the supply of stem cells, the sper-
 MALE REPRODUCTIVE SYSTEM • 811

Dorsal
longitudinal
column

Rib of the Rib of the

fibrous fibrous
sheath sheath

Minor Major
compartment compartment
x

Ventral
longitudinal
column

Figure 31-18. Transverse section through the principal piece of a hamster spermatozoon. Outer fibers 3 and 8 have
terminated and their place is filled by inward extensions of the dorsal and ventral longitudinal columns of the fibrous
sheath. The cross section is asymmetrical, with a major compartment containing four dense fibers and a minor
compartment containing three.

matogonia must perpetuate themselves and ical nucleus containing chromatin granules
also produce generation after generation of of varying size, many of which are distributed
spermatocytes. In human testicular tissue along the nuclear envelope. The single nu¬
preserved in Zenker-formol fixative, at least cleolus is centrally located and often has
two types of spermatogonia (A and B) can be granules of chromatin associated with it. The
distinguished with little difficulty. The Type cytoplasm is not significantly different from
A spermatogonium has a spherical or ellipsoid that of the Type A spermatogonium.
nucleus with very fine chromatin granules The Type A spermatogonium undergoes
and one or two irregularly shaped nucleoli a series of divisions that give rise to other
attached to the inner aspect of the nuclear Type A spermatogonia. Of these progeny,
envelope. The cytoplasm is homogeneous certain ones serve as stem cells for future
and pale-staining. In some spermatogonia of cycles of spermatogonial renewal and sper-
this type, the nucleoplasm is dark, and a matocytogenesis. Others proceed to differ¬
large, pale-staining nuclear vacuole is pres¬ entiate through recognizable intermediates
ent. These cells in the human and monkey into Type B spermatogonia. The division of
are designated as dark Type A spermato¬ Type B spermatogonia then gives rise to
gonia, to distinguish them from the others primary spermatocytes.
with paler nucleoplasm and no nuclear vac¬ In somatic cells, chromosomes are present
uole. The Type B spermatogonium.i has a spher¬ in pairs. These cells are conventionally de-
812 * MALE REPRODUCTIVE SYSTEM

fertilization the spermatozoon and ovum

each contribute a haploid set of chromosomes
to the zygote, reestablishing the diploid chro¬
mosome number. The special type of nuclear
division that results in formation of the hap¬
loid gametes is called meiosis (See also Chapter
1). In spermatogenesis it occurs in the sper¬
matocytes.
The primary spermatocytes at first resemble
in size and cytological characteristics the sper¬
matogonia from which they arise, but as they
move away from the basal lamina of the
germinal epithelium, they accumulate more
cytoplasm and become distinctly larger. Al¬
most immediately after their formation, the
spermatocytes enter prophase of the first
maturation division. Their chromatin be¬
comes reorganized into thin threadlike chro¬
mosomes characteristic of the leptotene stage
of meiosis. The homologous chromosomes,
which have duplicated themselves during the
preceding interphase, undergo intimate pair¬
ing during the zygotene stage through for¬
mation of synaptonemal complexes. Because of
the greater thickness and deeper staining of
the paired chromosomes at this stage, they
show up more clearly than those of the lep¬
totene stage. When the pairing of the chro¬
mosomes to form bivalents or tetrads is com¬
plete, they continue the process of coiling
and shortening to form the much coarser
and more obvious chromosomal strands typ¬
ical of the pachytene stage. At this stage, the
duplicated chromosomes can be identified as
dyads or sister chromatids held together at their
centromeres. Each pachytene element consists
of four chromatids. It is also at this period
that crossing over occurs, in which correspond¬
ing regions of the chromatids of the paired
chromosomes are exchanged. During the en¬
suing diplotene stage the chromosomes com¬
Figure 31-19. Cross sections at successive intervals
along the length of the Chinese hamster sperm tail,
plete their process of shortening and the
illustrating the reduction in diameter of the outer dense synaptonemal complexes disappear.
fibers and the tapering of the tail as a whole. A is a These stages of meiotic prophase are ex¬
section at the level of the midpiece. B-F, Successive tremely prolonged, extending over about 22
levels in the principal piece. G and H are through the end
days. For this reason, a great many sperma¬
piece. (Micrographs courtesy of D. Phillips.)
tocytes in different stages of prophase can be
seen in cross sections of seminiferous tubules.
At the end of prophase, the nuclear mem¬
scribed as diploid in chromosome number, brane disappears. The tetrads, or bivalents,
whiie the gametes that contain only one chro¬ arrange themselves at the equatorial plate in
mosome of each pair are haploid. The reduc¬ metaphase I. At anaphase I the centromeres of
tion in chromosome number of the gametes each homologous pair move to opposite poles
to half the somatic number is part of an of the spermatocyte taking both chromatids
orderly process that maintains a constant (dyads) along with them. This is in contrast
number of chromosomes for each species. At to mitosis, during which the duplicated chro-
 MALE REPRODUCTIVE SYSTEM • 813

mosomes line up on the equatorial plate and quickly completed, resulting in the formation
the centromeres divide, sending copies of of secondary spermatocytes carrying only
each chromosome to the opposite poles. The half the number of chromosomes originally
chromosomes that separate in meiosis are present. Since the chromosomes have already
also unique in that they may differ from both been duplicated, the secondary spermato¬
maternal and paternal chromosomes because cytes remain in interphase only briefly and
of exchanges that have taken place during are therefore encountered only infrequently
crossing over. Anaphase I and telophase I are in sections of seminiferous tubules. The sec-

Connecting

th
Ac

En

Figure 31-20. Generalized diagram of the structure of a mammalian spermatozoon as revealed by electron microscopy.
814 * MALE REPRODUCTIVE SYSTEM

Figure 31-21. Electron micrograph showing two guinea pig spermatids and the intercellular bridges by which they are
joined to each other and to two other spermatids of the same cluster. The small arrows indicate the local thickening of
the cell membrane encircling the bridges. The large arrows passing through the bridges indicate the sites of continuity
of the cytoplasm from cell to cell, x 9000.

ondary spermatocytes quickly complete the jugation. Therefore of any two secondary
second phase of the meiotic division with a spermatocytes resulting from the first matu¬
brief prophase II followed by metaphase II with ration division, one will contain 22 autosomes
the chromosomes aligned on the equatorial and an X chromosome, while the other will
plate. Anaphase II differs from anaphase I in contain 22 autosomes and a Y chromosome.
that the centromeres divide as in mitosis, Since all the eggs produced by the female
permitting the sister chromatids to move to are the same, containing 22 autosomes plus
opposite poles. Upon completion of meiosis X, those sperm developing from spermato¬
at telophase II, spermatids are formed that cytes containing X will be female determining,
have a haploid set of chromosomes. because fertilization will result in a zygote
In the human spermatogonium there are containing 44 + XX (female), whereas sperm
46 chromosomes, consisting of 22 pairs of developing from secondary spermatocytes
autosomes and one pair of sex chromosomes (XX containing a Y chromosome will be male de¬
or XY). The different pairs of autosomes termining, for the zygote will contain 44 +
vary in size and in the location of the kine- XY (male).
tochore, but the two members of any given Electron microscopic studies have shown
pair of autosomes are morphologically iden¬ that the division of all male germ cells, except
tical. The sex chromosomes in the female the most undifferentiated spermatogonia,
(XX) are also identical, but those of the male differ from somatic cell divisions in another
(XY) differ markedly in size. At the end of important respect. Following division of the
the first maturation division in spermatogen¬ nucleus (karyokinesis), division of the cell body
esis, each bivalent, including the XY pair, (cytokinesis) is incomplete, and the daughter
separates into its two constituent chromo¬ cells remain connected by protoplasmic
somes along the line of their previous con¬ bridges at the site where the constricting
 MALE REPRODUCTIVE SYSTEM • 815

cleavage furrow encounters the spindle rem¬ cytokinesis is incomplete, resulting in groups
nant. Such spindle bridges occur as transient of conjoined spermatogonia and larger syn¬
structures in mitosis of somatic cells, but in cytial clusters of primary spermatocytes.
the seminiferous epithelium they remain, These produce double the number of inter¬
after dispersal of the spindle, as sizable com¬ connected secondary spermatocytes. Their
munications between the daughter cells (Fig. division in turn produces very large numbers
31-21). They persist to a late stage in the of conjoined spermatids (Fig. 31-22). The
differentiation of the spermatids into sper¬ progeny of a single spermatogonium thus
matozoa. It has been the traditional view that form a cluster of germ cells that remain in
each spermatogonium divided and the protoplasmic continuity throughout their dif¬
daughter cells each developed into a primary ferentiation. This arrangement is probably
spermatocyte; this ultimately divided into two responsible for the synchrony of develop¬
secondaries, and these in turn divided to ment of large numbers of germ cells in any
form four individual spermatids. This inter¬ one area of the seminiferous tubule. Individ¬
pretation is now known to be incorrect. In ual sperm are separated from the syncytia at
all but the earliest spermatogonial division, the moment of their release from the epithe-

A. SPERMATOGONIA

A2 SPERMATOGONIA

A3 SPERMATOGONIA

A4 SPERMATOGONIA

In. SPERMATOGONIA

Figure 31-22. Diagram illustrating

the clonal nature of the male germ
cells. Only the most primitive sper¬
matogonia, dividing to replace the
stem cell population, complete cy¬
toplasmic division and give rise to
separate daughter cells. Once com¬
mitted to differentiation, the daugh¬
ter cells of all subsequent sper¬
matogonial divisions and the two
meiotic divisions remain connected
by intercellular bridges. Individual
sperm are ultimately separated
from syncytial chains of residual
bodies still connected by bridges.
The numbers of interconnected
cells are considerably larger than
depicted in this figure.

RESIDUAL
BODIES
816 • MALE REPRODUCTIVE SYSTEM

Acrosomal
vesicle
Granule

Endoplasmic
reticulum

Figure 31-23. Electron micrographs of the juxtanuclear region of macaque spermatids, illustrating successive stages
in formation of the acrosome. A, The initial event is the appearance of an acrosome vesicle and granule at the trans¬
face of the Golgi complex. B, The vesicle and granule are enlarged by further contributions from the Golgi complex and
become closely adherent to the nuclear envelope. C, The acrosomal vesicle then spreads over the anterior hemisphere
of the nucleus to form the acrosomal cap. The substance of the granule subsequently spreads laterally to occupy the
entire interior of the acrosomal cap. (Micrographs courtesy of M. Dym.)
 MALE REPRODUCTIVE SYSTEM • 817

lium. Partial failures of this process may ultimately to cover its entire anterior hemi¬
account for the frequency of abnormal dou¬ sphere as a membranous head cap. The ac¬
ble spermatozoa in the ejaculate. rosomal granule meanwhile remains localized
at the pole of the nucleus.
Spermiogenesis In the third or acrosomal phase of spermi¬
ogenesis, there is redistribution of the acro¬
The term spermiogenesis describes the se¬ somal substance, a condensation of the nu¬
quence of developmental events by which cleoplasm, and an elongation of the
spermatids are transformed into spermato¬ spermatid (Fig. 31-24). The bulk of the ac-
zoa. Each of the relatively small spherical or rosome remains localized at the anterior pole
polygonal spermatids resulting from division of the nucleus, but during this phase of
of the secondary spermatocytes has a nucleus spermiogenesis its substance gradually
5 to 6 pm in diameter with pale-staining spreads in a thin layer into the fold of mem¬
finely granular chromatin. A small Golgi ap¬ brane composing the head cap until the ac-
paratus can be seen in the juxtanuclear cy¬ rosome and head cap are coextensive and
toplasm. The first sign of differentiation of constitute the acrosomal cap (often simply
a specific component of the spermatozoon is called the acrosome). In its definitive form it
the appearance of one or more small gran¬ is a caplike structure, limited by a membrane
ules within the Golgi apparatus (Fig. and containing a substance rich in carbohy¬
31-23A). In some species these are first ob¬ drate and hydrolytic enzymes. It varies in size
served in the spermatocytes; in others they and shape in different species but is present
are not seen until the spermatid stage. These on the sperm of all mammals. The spermatid
proacrosomal granules are rich in carbohydrate nucleus becomes elongated and flattened
and are most clearly demonstrated in speci¬ during this period. Its uniformly dispersed,
mens stained for light microscopy by the finely granular nucleoplasm becomes trans¬
periodic acid—Schiff reaction. In electron mi¬ formed into thin strands or filaments that
crographs, each is found to be enclosed subsequently shorten and thicken into coarse
within a membrane-limited vesicle associated dense granules.
with the Golgi apparatus. Although the gen¬ During the fourth or maturation phase of
eral features of spermiogenesis can be fol¬ spermiogenesis, there is little further change
lowed with the light microscope, the finer in the simple acrosome of primate sperm,
details described here can be visualized only but in other species it continues to undergo
with the electron microscope. As develop¬ further alterations and gradually takes on
ment progresses, the several separate gran¬ the shape characteristic of the species.
ules in the Golgi region coalesce into a single The dense granules in the condensing nu¬
large globule, the acrosomal granule, contained cleus become coarser, increasing in size at
within a membrane-bounded acrosomal vesicle the expense of the intervening spaces until
or vacuole. This becomes adherent to the they finally coalesce and the nucleus is trans¬
outer aspect of the nuclear envelope (Fig. formed into a homogeneous dense mass de¬
31—235). The point of its adherence marks void of visible substructure. By the time this
the future anterior tip of the sperm nucleus. condition has been reached, the nucleus has
The Golgi apparatus remains closely associ¬ attained the flattened pyriform shape char¬
ated with the surface of the acrosomal vesicle, acteristic of the human sperm head. Defects
and it continues to form smaller vesicles that in the condensation of the nucleoplasm often
coalesce with the membrane of the acrosomal leave one or more clear areas of variable
vesicle, contributing their contents to its en¬ shape and size, recognized with the light
largement. microscope as nuclear “vacuoles.” These are
It is convenient to divide spermiogenesis large and of frequent occurrence in the hu¬
into four phases. That period from the ap¬ man sperm but are small and relatively un¬
pearance of the proacrosomal granules to the common in the spermatozoa of other species.
development of a hemispherical acrosomal While the early stages of acrosome forma¬
granule fixed to the nuclear envelope is re¬ tion are in progress at the anterior pole of
ferred to as the Golgi phase. In the second or the nucleus, the centrioles migrate to the
cap phase, the limiting membrane of the ac¬ opposite end of the spermatid. There the
rosomal vesicle increases its area of adher¬ distal centriole becomes oriented perpendic¬
ence to the nuclear envelope, forming a thin ular to the cell surface and gives rise to a
fold that spreads over the pole of the nucleus, slender flagellum that grows out into the
818 • MALE REPRODUCTIVE SYSTEM

Figure 31-24. Electron micrographs illustrating successive stages in condensation of the chromatin and shaping of the
nucleus of the spermatid late in spermiogenesis. (Micrographs courtesy of M. Dym.)

narrow extracellular cleft between the sper¬ The flagellum at this time consists only of
matid and the surrounding Sertoli cell (Fig. the axial filament complex or axoneme, with
31—25). As the nucleus begins to elongate two central fibrils and nine peripheral doub¬
and condense, the pair of centrioles and the lets. The latter are continuous with the wall
base of the flagellum recede from the surface of the distal centriole. The centriole is encir¬
and take up a position at the caudal pole of cled by a ring of moderately dense filamen¬
the nucleus (Fig. 31—26). At about the same tous or granular material. This annular struc¬
time, cytoplasmic microtubules arise and be¬ ture was called the “ring centriole” by classical
come laterally associated to form a roughly cytologists in the belief that it arose by une¬
cylindrical structure, called the manchette, qual division of one of the centrioles. Elec¬
which extends caudally from a ringlike spe¬ tron microscopic studies provide no evidence
cialization of the cell membrane located at in support of this interpretation. Instead, the
the posterior margin of the acrosomal cap ring appears to be a derivative of the chro-
(Fig. 31—28). Concurrently with the appear¬ matoid body (Fig. 31— 27A). This loose ring is
ance of the manchette, there is a marked intimately associated with another small
elongation of the spermatid, so that the bulk dense ring that arises as a local specialization
of the cytoplasm is displaced well behind the on the inner aspect of the plasma membrane,
caudal pole of the nucleus, where it sur¬ where the latter is reflected from the cell
rounds the proximal part of the flagellum body onto the flagellum.
(Fig. 31-28C,D,E). In the further differentiation of the tail,
 MALE REPRODUCTIVE SYSTEM • 819

Proximal
centriole
Connecting

a•-■
' piece

FT'S? Distal..
jBsaa, centriole

Centriolar
adjunct
A cr>

Figure 31-25. The earliest events in formation of the tail

consist of migration of the centrioles to the cell surface in
the postnuclear region, and polymerization of microtubule
protein on the template provided by the distal centriole. A Connecting
typical 9 + 2 axoneme is formed and the simple flagellum
> Proximal fV Piece
elongates by accretion of microtubule subunits to its distal centriole
end.

Remnant
f of distal
* centriole
Redundant j
nuclear ‘ft

envelope 1

Outer
dense
fiber
/
Mitocbondrfal
sheath ■
Axoneme

Figure 31-27. A, After establishing contact with the

implantation fossa, the proximal or juxtanuclear centriole
forms, at its distal end, the centriolar extension or adjunct.
Figure 31-26. The proximal member of the pair of cen¬ The segmented connecting piece begins to form around
trioles comes to occupy a shallow groove, the implantation the distal centriole. 8, Later in development, the centriolar
fossa, in the caudal pole of the nucleus. The anlage of adjunct disappears and the distal centriole disintegrates,
the future annulus then appears as a ringlike density leaving the proximal centriole in a vault or niche in the
adjacent to the membrane at its site of reflection onto the connecting piece that joins the nucleus to the outer dense
flagellum. fibers of the tail.
820 • MALE reproductive system

Acrosome
Manchette
Acrosomal
cap

Nuclear
ring

Nucleus

Mitochondrial
sheath

Flagellar
canal Annulus

Intercellular Fibrous
bridge sheath

Figure 31-28. Diagram of successive stages in guinea pig spermatid differentiation, including nuclear condensation,
appearance of the manchette, elongation of the cell, appearance of the fibrous sheath, caudal migration of the annulus,
and formation of the mitochondrial sheath. (From Fawcett, D. W., W. A. Anderson, and D. M. Phillips. Dev. Biol. 26:220,
1971.)

nine longitudinally oriented segmented col¬ oriented ribs or hoops are deposited around
umns arise around the centrioles. These are the tail distal to the annulus to form the
joined to each other proximally and to the fibrous sheath of the principal piece.
base of the nucleus to constitute the connect¬
ing piece. Distally the nine structural ele¬
Spermiation
ments forming the connecting piece are
joined to nine longitudinal dense fibers that With the completion of differentiation of
develop just peripheral to the doublets of the the tail, the spermatozoon is separated from
axoneme. The distal centriole and the large the excess cytoplasm, which remains in the
ring that earlier encircled it gradually disap¬ epithelium (Fig. 31—29) as a membrane-lim¬
pear as the connecting piece and outer fibers ited anucleate mass called the residual body (of
of the tail develop. The smaller dense ring Regnaud), consisting of fine granules, lipid
fixed to the flagellar membrane persists, and droplets, and degenerating excess organelles
in the further elongation of the tail, it is not used in formation of the spermatozoon.
carried distally several micrometers. As it
moves back, the manchette disappears and
The Cycle of the Seminiferous
mitochondria gather around the segment of
Epithelium
flagellum between the annulus and the nu¬
cleus and become disposed helically around Spermatogenesis has been most thoroughly
it to complete the differentiation of the mid¬ studied in the common laboratory rodents,
dle piece (Fig. 31-28C,D,E). where it displays a degree of order and
While these developmental events are in regularity that facilitates a systematic analysis
progress, a succession of circumferentially of the process. As stated earlier, several steps
 MALE REPRODUCTIVE SYSTEM • 821

Figure 31-29. Diagrammatic representation of successive stages in sperm release. The axial components of the sperm
are gradually extruded while the syncytial mass of spermatid cell bodies is retained in the epithelium. The attenuated
stalk connecting the sperm to the residual cytoplasm finally gives way, freeing the spermatozoon. (From Fawcett, D. W.
In Segal, S. J., et al., eds.: The Regulation of Mammalian Reproduction. Springfield, IL, Charles C Thomas, 1973.)

of germ cell development are found at dif¬ tion into a spermatozoon, passes through all
ferent levels in the germinal epithelium, with the cell types encountered by starting at the
the stem cells found at the base and the more bottom of column I in this figure and reading
differentiated cells located at successively from left to right along the horizontal rows
higher levels (Fig. 31-30). The development from the bottom to the top row.
of any one generation of germ cells goes on Spermatids at different phases of differ¬
concurrently with the development of earlier entiation are always associated with sperma¬
and later generations at other levels in the tocytes and spermatogonia at their specific
epithelium. The cells in different phases of phases of development. The particular asso¬
development are not randomly distributed ciation of cells found at any point along the
within the epithelium but occur in a number length of a seminiferous tubule changes with
of well-defined and easily recognized combi¬ time, passing successively through all 12
nations or associations. The number of dis¬ stages and then repeating the sequence. The
tinguishable cell associations varies with the cycle of the seminiferous epithelium is defined as
species. In the guinea pig, for example, 12 the series of changes occurring in a given
such cellular associations or stages of sperma¬ area of the epithelium between two successive
togenesis are identified. These are illustrated appearances of the same cellular association.
in the 12 vertical columns of Figure 31—31 The duration of the cycle has not been de¬
and are designated by the Roman numerals termined for the guinea pig, but it is about
at the bottom of each column. In any histo¬ 12 days in the rat, and a spermatogonium
logical section of guinea pig testis, the cross takes about four cycles or 48 days to complete
sections of neighboring seminiferous tubules its differentiation and be released as a mature
will vary in their appearance because of the spermatozoon.
different cell associations they contain (com¬ The various cell associations also occur in
pare Figs. 31—32 and 31—33). If enough a numerically orderly sequence along the
tubules are examined, all 12 cell associations length of the seminiferous tubule. Thus, in¬
or stages will be found, corresponding to the stead of considering the changes at a given
vertical columns in Figure 31—31. In studying point in the tubule, one can look at it from a
this figure, it should be realized that a sper¬ different point of view, namely, as a series of
matogonium, in the course of its differentia¬ successive cell associations found along the
822 • MALE REPRODUCTIVE SYSTEM

Figure 31-30. Photomicrograph of a guinea pig seminiferous tubule and portions of adjacent tubules. Notice that the
epithelium in these four tubules exhibits different associations of cell types, representing different stages of spermato¬
genesis.

1
i Jf . * 1

| J*
w. Wt si si Si S j s*
13 13 14 14 14 15 15 i •i . |

V vv
j

fii m$ P pm / ■L J
« » t | * j V *
*
■ rp' : *. *.
f "I
Vi
u V
i 2 3 4 5 6 7 8 9 10 11 12

* § <| %
# *» « ."T*
W rssc*’ Ip* V;/
p p P p P p P p P p Di n
J*
■*; 4 A %
C y vS # if #
in in in B B R R L L z Z P

/t: ® ' ■i ' * 4

*
6 &
•prT'^x'
I'Ll; ^ ■a* '
£Y; V: •*> «5S> Ci o W *
A A A A A A A A A AP A A

I n ffl EZ Y ffl ffll an IX X XI xn

Figure 31-31. In these vertical columns, all 12 different cell associations or stages found in guinea pig seminiferous
tubules have been assembled in their correct temporal sequence. Reading from lower left to right and from bottom to
top, one follows the morphogenetic events from spermatogonium to release of spermatozoa. The time from the
appearance of any one of these cell associations at a given point along the tubule until the reappearance of the same
cell association is defined as one cycle of the seminiferous epithelium. (From Clermont, Y. Fertil. Steril. 6:563, 1960.)
Figure 31-32. Photomicrograph of Stage I of guinea pig spermatogenesis, which includes type A spermatogonia (A),
intermediate spermatogonia (in), pachytene spermatocytes (P), spermatids at the beginning of acrosome formation (1),
and a more advanced generation of elongated spermatids with flattened, condensed nuclei (13). (Inset from Clermont,
Y. Fertil, Steril. 6:563, 1960.)

Figure 31-33. Photomicrograph of Stage VII of guinea pig spermatogenesis, characterized by the presence of
spermatogonia (A); large pachytene spermatocytes (P); spermatids in the cap phase of acrosome formation (6); nearly
mature spermatids projecting into the lumen (15); and residual bodies with dense cytoplasm forming a layer adjacent to
the lumen. ooo
824 • MALE REPRODUCTIVE SYSTEM

length of the same tubule. The wave of the dents, the recognizable stages or cell associa¬
seminiferous epithelium is then the distance be¬ tions in man occupy small wedge-shaped
tween two successive identical cell associa¬ areas in the tubular epithelium (Fig. 31-34).
tions. That portion of the length of a wave The human germinal epithelium is there¬
occupied by one cell association is referred fore a mosaic of irregularly shaped areas
to as a segment. made up of the six different cell associations.
The sequence of pictures along a wave is Three or more stages of the cycle may be
similar to the sequence of events taking place seen in a single cross section of a tubule. The
in one given area during a cycle of the situation is further complicated by the fact
seminiferous epithelium. In the rat there are that the cells at the borders of these areas
said to be about 12 waves along the length may intermingle to give atypical or hetero¬
of each tubule. The length of each segment geneous associations of cells. The six typical
in the wave corresponds roughly to the rela¬ associations are depicted in Figure 31-35.
tive duration of that particular cell association The duration of the cycle of the human
or stage of the cycle. seminiferous epithelium has been deter¬
In contrast to the very regular ordering of mined by autoradiographic analyses of testic¬
germinal elements in rodents, the appear¬ ular biopsies from volunteers. Within one
ance of the seminiferous epithelium in his¬ hour of local injection of tritiated thymidine,
tological sections of the human testis at hrst the label was found in nuclei of preleptotene
suggests a haphazard arrangement of its cell spermatocytes in Stage III, but not in the
types. Because of this apparent disorder, it pachytene spermatocytes of that stage nor in
was formerly believed that no synchronicity any other cells more advanced in their de¬
of germ cell development comparable with velopment. With the passage of time these
that found in rodents existed in man, and labeled cells would be expected to pass
that no “cycle of the seminiferous epithe¬ through leptotene, zygotene, and early pach¬
lium” could be defined. ytene stages of meiotic prophase and reap¬
This has now been found to be erroneous. pear at the end of the cycle in a Stage III
Six well-defined stages can be recognized, cell association as midpachytene spermato¬
but instead of each occupying the entire cross cytes. Serial biopsies revealed that the mid¬
section of the seminiferous tubule, as in ro- pachytene spermatocytes of Stage III hrst
showed that label 16 days after the initial
injection of thymidine. It was thus established
that the duration of one cycle is 16 days. As
expected, labeled spermatids in Stage III
were found at 32 days (two cycles). Assuming
one cycle for the cells to develop from sper¬
matogonia to preleptotene spermatocytes,
and one to advance from spermatids to re¬
lease of spermatozoa, the total duration of
spermatogenesis in man is estimated to be
four consecutive cycles or 64 days.

Degenerative and Regenerative

Phenomena
In seasonally breeding mammals, active
spermatogenesis, beginning at puberty, is dis¬
continued and reinitiated periodically for the
rest of the life of the animal. Each time, it
continues only during the period of rut, at
the end of which most of the spermatogenic
cells are eliminated by degeneration or mat¬
Figure 31-34. In the human, the stages of spermatogen¬ uration depletion. Concomitantly, the semi¬
esis do not occupy the whole circumference of a tubule
as in other species. In this photomicrograph, for example, niferous tubules shrink and gradually come
four different associations of cells are found in the same to contain only Sertoli cells and some sper¬
cross section. (From Clermont, Y. Am. J. Anat. 112:50, matogonia. In this condition, they resemble
1963.) the tubules of a prepubertal testis. At the
 MALE REPRODUCTIVE SYSTEM • 825

STAGE III STAGE IV

Figure 31-35. Diagram of the six recognizable cell associations or stages of the cycle of the human seminiferous
epithelium. Ser, Sertoli cell; Ad and Ap, dark and pale type A spermatogonia; B, type B spermatogonia; R, resting
primary spermatocyte; L, leptotene spermatocyte; Z, zygotene spermatocyte; P, pachytene spermatocyte; Di, diplotene
spermatocyte; Sptc-lm, primary spermatocyte in division; Sptc-ll, secondary spermatocyte in interphase; Sa, Sb, Sc,
Sd, spermatids in various stages of differentiation; RB, residual bodies of Regnaud. (From Clermont, Y. Am. J. Anat.
112:35, 1963.)
826 • MALE REPRODUCTIVE SYSTEM

beginning of a new period of sexual activity, rest of the body. Testes that fail to descend
spermatogonia multiply and rapidly regen¬ into the scrotum during development and
erate the various generations of spermato- remain in the abdomen never produce ma¬
genic cells. In the lower vertebrates, these ture sperm. They show atrophic tubules con¬
seasonal changes of the testis are even more taining only Sertoli cells and scattered sper¬
prominent. matogonia. Failure of descent of the testis is
In man and other mammals that are not called cryptorchidism. In experimentally pro¬
seasonal breeders, spermatogenesis is contin¬ duced cryptorchidism, the testis soon de¬
uous. Nevertheless, in an active human testis creases in size and comes to contain only
the tubules contain scattered degenerating Sertoli cells and spermatogonia. The seminif¬
spermatogenic cells in the seminiferous epi¬ erous tubules also atrophy in experimental
thelium. This is not pathological unless it animals fed a diet lacking vitamin E; they
exceeds certain limits. The degenerating cells also undergo regression in vitamin A defi¬
are seen in segments of the tubule in which ciency.
the seminiferous epithelium is active and In all such cases, the Sertoli cells are more
normal spermatogenesis is in full progress. resistant than spermatogenic cells. Some
The significance of the normal degeneration spermatogonia frequently remain. Thus, un¬
of a certain number of germ cells is not der favorable conditions, when the noxious
known. factor is removed or the vitamin deficiency
Abnormal spermatogenic cells can often be corrected, a more or less complete regener¬
found. In spermatogonia and spermatocytes, ation of the seminiferous epithelium may
giant forms, as well as cells with two nuclei, take place. In mammals with a short life span,
are not uncommon. Multinucleated giant spermatogenesis continues undiminished un¬
spermatids likewise are not infrequent. These til death. Although spermatogenesis contin¬
abnormalities appear to be a consequence of ues far into senility in man, the seminiferous
the peculiar mode of cytokinesis in the divid¬ tubules do undergo gradual involution with
ing germinal cells, which normally leaves advancing age. A testis of a man older than
them connected by intercellular bridges. Fail¬ 35 usually shows scattered atrophic tubules;
ure of initial constriction between the daugh¬ in the remainder of the organ, however,
ter cells or a subsequent opening up of the spermatogenesis may continue without visible
bridges between two or more cells may lead alterations. In very old men, all the tubules
to the formation of multinucleated sperma¬ may be depleted of spermatogenic cells.
tocytes or spermatids. Spermatids with two
nuclei may continue to develop; thus, mon¬
strous sperm with two tails or with one tail INTERSTITIAL TISSUE
and two heads, may arise. These abnormal
sperm are carried with the normal mature The endocrine component of the testis, the
sperm into the epididymis, where some de¬ Eeydig cells, are located in the angular inter¬
generate, but others persist and are also stices between the convoluted seminiferous
found in ejaculated semen. tubules. The blood vessels form peritubular
The germ cells of the seminiferous epithe¬ plexuses in the intertubular spaces. The or¬
lium are sensitive to noxious agents. In path¬ ganization of the interstitial tissue varies con¬
ological conditions of general (infectious dis¬ siderably from species to species. In the hu¬
eases, alcoholism, dietary deficiencies) or man the extravascular interstitial tissue is a
local (inflammation) character, degenerative loose connective tissue exceptionally rich in
changes, especially the formation of multi¬ extracellular fluid (Fig. 31-36). In addition
nucleated giant cells by the coalescing sper¬ to small clusters of Leydig cells, there are
matids, may become prominent. Exposure of infrequent fibroblasts, a few macrophages,
the testis to a sufficient dose of x-rays causes occasional mast cells, and some relatively un¬
an extensive degeneration of spermatogenic differentiated cells of mesenchymal origin
cells and may result in sterility. These cells that are capable of developing into Leydig
are also sensitive to high temperature. Even cells in response to gonadotropic stimulation.
the normal internal temperature of the body In other species, especially in the pig, horse,
is incompatible with spermatogenesis. In the and opossum (Fig. 31—37), Leydig cells are
majority of adult mammals, the testes are very abundant, occupying most of the inter¬
therefore lodged in a scrotum, which has a tubular space.
temperature a few degrees lower than the The interstitial cells of Leydig occur in
Figure 31-36. Photomicrograph of the interstitium between seminiferous tubules of human testis, showing several
clusters of Leydig cells. Most of the extravascular space is occupied by a protein-rich interstitial fluid, which appears a
uniform gray in this figure. There is little collagen in the interstitial tissue of the normal human testis. (Photomicrograph
courtesy of W. Neaves.)

Figure 31-37. Photomicrograph of opposum testis for comparison with the human testis in Figure 31-36. Here, Leydig
cells occupy nearly all the interstitial space.
827
828 • MALE REPRODUCTIVE SYSTEM

tend to be tubular instead of lamellar, but

this is not true in all mammalian species. The
cytoplasm is acidophilic in routine prepara¬
tions and may contain a number of vacuoles
where lipid droplets have been extracted
(Fig. 31-38). In common with other steroid-
secreting endocrine cells, the most striking
ultrastructural feature of the interstitial cell
is its extensive smooth endoplasmic reticulum
(Figs. 31—39, 31—40). Cisternal profiles of the
granular reticulum are also present, but the
bulk of the cytoplasm is filled with a branch¬
ing and anastomosing system of smooth-sur¬
faced tubules. T hese membranes contain the
enzymes necessary for several of the steps in
the biosynthesis of androgenic steroids. Per¬
oxisomes and lysosomes are found in abun¬
dance, but their function in the economy of
these cells is not understood. Golden-brown
deposits of lipochrome pigment occur in the
Leydig cells in men of all ages, but they
become increasingly prominent with advanc¬
ing age (Fig. 31-39).
A feature peculiar to the human Leydig
cell is the presence of conspicuous cyto¬
plasmic crystals 3 pm or more in thickness
and up to 20 pm in length. These crystals of
Reinke are highly variable in size and shape;
Figure 31-38. Photomicrograph of a group of Leydig cells they may be rounded or pointed at the ends
in human testis. These cells often contain numerous lipid (Fig. 31—41). They have little affinity for the
inclusions, which are extracted in specimen preparation common histological stains and appear nearly
and appear as clear vacuoles. (Photomicrograph courtesy colorless in routine preparations. They can,
of W. Neaves.)
nevertheless, be recognized in negative image
in ordinary preparations and, if desired, they
clusters of varying size, sometimes closely can be stained by azocarmine. They are iso¬
associated with blood vessels. They are usu¬ tropic in polarized light and have the solu¬
ally irregularly polyhedral, 14 to 20 pm bility properties of protein. In electron mi¬
across, where they are closely packed, but at crographs they present a highly ordered
the periphery of the clusters or where occur¬ structure that differs in its pattern, depend¬
ring individually they may be elongated or ing on the plane of section. The crystal con¬
spindle shaped. The large spherical nucleus sists of filamentous molecules about 5 mm
contains a small amount of peripherally dis¬ thick. The crystals occur in the testes of most
posed heterochromatin and one or two men from puberty to senility, but their abun¬
prominent nucleoli. Binucleate cells are com¬ dance is subject to considerable variation.
mon. Adjacent to the nucleus is a large clear They are found in no other mammalian
area that is found in electron micrographs to species and their significance is unknown.
be occupied by a well-developed Golgi ap¬
paratus. Although the Golgi complex is
prominent and responds to gonadotropic BLOOD VESSELS AND LYMPHATICS
stmulation by enlargement, the role of this OF THE TESTIS
organelle in the biosynthetic and secretory
processes of this cell type is not known. There The blood supply of the human testis is
is no visual evidence of accumulation of a from a branch of the abdominal aorta called
product in secretory granules in the Golgi the internal spermatic or testicular artery. It
region, and at present we have little under¬ divides either before reaching the testis or
standing of the mechanism of release of ste¬ °n its surface, giving rise to several main
roid hormones by these cells. Mitochondria branches that penetrate the organ. These in
are abundant and quite variable in size and turn give rise to centripetal branches that course
shape. In electron micrographs, their cristae toward the rete testis. Major branches of
Figure 31-39. Electron micrograph of a portion of a human Leydig ceil. As in other steroid-secreting cells, its most
striking ultrastructural feature is the very extensive smooth endoplasmic reticulum. The dense bodies at the top of the
figure are deposits of lipofuscin pigment that accumulate with age. (Micrograph courtesy of W. Neaves.)

Figure 31-40. Interstitial cells in some species contain abundant droplets of lipid, whereas others are virtually devoid
of lipid. The electron micrograph of opossum interstitial cell shown here is free of lipid. The cytoplasm is occupied by a
very extensive agranular endoplasmic reticulum, x 10,000. (After Christensen, A., and D. W. Fawcett. J. Biophys.
Biochem. Cytol. 9:653, 1961.)
829
830 • MALE REPRODUCTIVE SYSTEM

Figure 31-41. Electron micrograph of Leydig cell cytoplasm from human testis showing lipochrome pigment deposits
vesicular elements of smooth reticulum, and several crystals of Reinke. (Micrograph from Naqano T and I Ohtsuki
J. Cell Biol. 57:148, 1971.)
 MALE REPRODUCTIVE SYSTEM • 831

these vessels run in the opposite direction as The corresponding vein on the left joins the
centrifugal branches. The branching of the left renal vein. This evidentlv results in some
m J

centripetal and centrifugal arteries gives rise slight impairment of venous return, so that
to many intertubular arterioles located in the the left pampiniform plexus is often more
angular columns of interstitial tissue between distended than the right, and the left testis is
the seminiferous tubules. The intertubular generally lower. The veins of the left pam¬
capillaries derived from these form networks piniform plexus are sometimes varicose, a
in the interstitial tissue. The capillaries in condition described as varicocele.
neighboring interstitial columns are con¬ In many animal species and possibly in
nected by ladder-like, circumferentially ori¬ man, the arrangement of blood vessels in the
ented peritubular capillaries (Fig. 31—42). spermatic cord (the internal spermatic artery
Postcapillary venules join to form collecting surrounded by the pampiniform plexus) con¬
venules, and the confluence of these in turn stitutes a countercurrent heat exchange sys¬
forms veins that course toward the tunica tem that allows the arterial blood to lose heat
albuginea (centrifugal veins) or toward the rete to the cooler venous blood in the pampini¬
testis (centripetal veins). The former drain into form plexus. This precooling of the arterial
veins of the tunica; the latter, running in the blood helps maintain the temperature of the
septula testis, converge upon a venous plexus testis a few degrees below deep body temper¬
associated with the rete testis. Upon reaching ature. This lower temperature is essential for
the surface, these veins join those of the continued spermatogenesis.
tunica in forming the pampiniform plexus of The intertubular areas of the human testis
veins in the spermatic cord. The right pam¬ also contain thin-walled lymphatic vessels that
piniform plexus drains directly into the in¬ drain into larger lymphatics in the septula
ferior vena cava via the internal spermatic vein. testis and tunica albuginea, and thence up-

Figure 31-42. Scanning micrograph of a resm cast of the microvasculature surrounding several seminiferous tubules
that have been dissolved out of the preparation. Intertubular vessels coursing parallel to the tubules are interconnected
by closely spaced peritubular capillaries to form a circumferentially oriented ladder-like pattern. (Micrograph from Suzuki,
F. Am. J. Anat. 763:309, 1982.)
832 • MALE REPRODUCTIVE SYSTEM

ward in lymph vessels of the cord to para¬ formation and activation of protein kinases.
aortic lymph nodes and nodes associated with Cholesterol esterase liberates free cholesterol
the renal blood vessels. from cholesterol esters in lipid droplets of
The lymphatics show considerable varia¬ the Leydig cells. Mitochondrial enzymes
tion in pattern from species to species. In the cleave off the side chain of cholesterol to
common laboratory rodents, they form very form pregnenolone and enzymes in the
extensive peritublar sinusoids. The aggrega¬ smooth endoplasmic reticulum, then carry
tions of Leydig cells in these species are out the several biosynthetic steps in transfor¬
centrally located and closely associated with mation of this molecule to testosterone.
the walls of the blood vessels. They are sur¬ Although the Leydig cells constitute only a
rounded by sinusoidal lymphatics. The ste¬ small percentage of the total volume of the
roid-secreting cells are thus interposed be¬ testis, their steroidogenic potential is impres¬
tween the blood vascular elements on the one sive. In the rat, where they make up only 1
side and the lymphatic sinusoids on the other, per cent of the testis volume, there is esti¬
and release their hormone into both. In mated to be 22 million Leydig cells per gram
larger species—ram, bull, and man—the lym¬ of testis. The smooth endoplasmic reticulum,
phatics are not sinusoidal but form thin- which contains the steroidogenic enzymes,
walled vessels more or less centrally located has an estimated surface area of 10,500 pm2,
in the intertubular areas. The Leydig cells and the mitochondria number about 600 per
are not intimately related to either the blood cell. From this morphometric data it is cal¬
vessels or the lymphatics, but evidently they culated that an average Leydig cell can pro¬
release androgen into the abundant extracel¬ duce about 10,000 molecules of testosterone
lular fluid of the interstitial tissue, whence it per second. Testosterone appears to be pro¬
diffuses to the tubules for its local effect on duced as needed, and not stored in secretory
spermatogenesis, and to the vessels for its granules for release on demand as in many
effect on distant target organs. other glandular cells. Detailed stereological
analysis of the human testis has not been
carried out, but it is likely that Leydig cells
occupy less than 3 per cent of the total
HISTOPHYSIOLOGY OF THE volume of the organ.
TESTIS The hypophyseal control of Leydig cell
function has proved to be more complex than
Endocrine Function was formerly thought. The release of LH
from the pituitary gonadotropes is now
The endocrine function of the testis resides known to be a discontinuous process, occur¬
mainly in the interstitial cells of Leydig. ring mainly during the night in pulses at
These synthesize and release the male sex intervals of about 90 minutes. Moreover, hor¬
hormone testosterone, which is required in mones other than LH have been found to
high local concentration to sustain sperma¬ influence testosterone production. Prolactin
togenesis in the seminiferous tubules. In ad¬ binds to specific receptors on Leydig cells,
dition to its local action, testosterone circulat¬ affecting the number of their LH receptors
ing in the blood is essential for maintenance and impairing their capacity to store choles¬
of function in the accessory glands of the terol esters as precursors of testosterone. LH-
male reproductive tract—the seminal vesicles, releasing hormone (LHRH) not only affects
prostate, and bulbourethral glands. Testos¬ pituitary gonadotropes but is reported to act
terone is also responsible for development directly upon Leydig cells, affecting their LH
and maintenance of the male secondary sex binding and their steroidogenic responsive¬
characteristics—male pattern of pubic hair; ness.
growth of beard; low-pitched voice; and mus¬ There is increasing evidence that the sem¬
cular body-build. iniferous tubules also exert a local regulatory
Production of testosterone by the Leydig effect on the ultrastructure and function of
cells depends primarily on the hypophyseal the Leydig cells. An LHRH-like peptide,
luteinizing hormone (LH), formerly referred to thought to be produced by the Sertoli cells,
as interstitial cell stimulating hormone (ICSH). is inhibitory to Leydig cells. Local damage to
The action of LH on the Leydig cells involves seminiferous tubules appears to remove this
binding to specific receptors in their plasma inhibition and results in hypertrophy of the
membrane. This, in turn, induces cyclic AMP neighboring Leydig cells.
 MALE REPRODUCTIVE SYSTEM • 833

Exocrine Function cells in the seminiferous epithelium, inhibin

is believed to be continuously released and
The spermatozoa can be thought of as a to act upon the hypophysis to suppress FSH
secretory product of the seminiferous tu¬ production. Depletion of germ cells results
bules. The numbers of sperm produced are in an increase in circulating levels of FSH.
astronomical. Improved methods for esti¬ The FSH-inhibiting substance, inhibin, has
mating daily sperm production in the human yet to be completely characterized and its
yield a mean value of 94.6 x 106 per testis physiological importance remains a subject
or 5.6 x 10b per gram of testicular tissue. of controversy.
Although this seems a very large number, it The secretory function of the seminiferous
is low compared with other species. Daily epithelium thus includes the synthesis and
sperm production per gram of testis in the release of androgen-binding protein; an
rat is at least 20 x 106; in the rabbit, 25 x LHRH-like peptide; and possibly inhibin. It
106; and in the boar, 23 x 106. Taking into also elaborates a considerable volume of fluid
account the weight of the testis in the boar, that serves as a vehicle for transport of the
the daily production of the two testes would spermatozoa to the rete testis and epididymis.
amount to about 16.2 x 109 spermatozoa. This fluid is rich in potassium and other ions,
In the human, a normal ejaculate is 2 to 5 in glutamate, and in inositol. These constit¬
ml in volume and contains 40 to 100 million uents are probably important for mainte¬
sperm per milliliter. This represents only a nance of the spermatozoa in their transit
fraction of the number stored in the epidi¬ through the excretory duct system.
dymis. Men with counts below 20 million per
milliliter are usually infertile. The production
Blood-Testis Permeability Barrier
of such large numbers of spermatozoa is less
surprising if one considers that the two testes It has been known for several decades that
contain 800 to 1200 seminiferous tubules, vital dyes and many other substances intro¬
each 30 to 70 cm long, which, in the aggre¬ duced into the blood stream readily leave the
gate, add up to one third of a mile of contin¬ vessels and enter the extracellular spaces of
uously proliferating seminiferous epithelium. most of the tissues and organs except the
Spermatogenesis depends on testosterone brain. This vital organ is protected by a blood-
produced by the Leydig cells in response to brain barrier that resides in the endothelial
stimulation by LH and on follicle stimulating junctions of the brain capillaries. More re¬
hormone (FSH), which acts upon the semi¬ cently a blood-testis barrier has been described.
niferous tubules, binding specifically to the The permeability barrier in this case is not
Sertoli cells. FSH is believed to be necessary in the walls of the blood vessels, which are in
for completion of spermatogenesis, but its fact unusually permeable in the interstitial
mode of action remains controversial. It is tissue. Instead, the exclusion of dyes and
known that FSH increases Sertoli cell synthe¬ other large molecules from the seminiferous
sis of an androgen-binding protein that is re¬ tubules is due to the presence of special
quired to maintain a high concentration of junctional complexes between adjacent Ser¬
testosterone within the seminiferous epithe¬ toli cells near the base of the seminiferous
lium. It may also promote synthesis of other epithelium. These consist of many parallel
products involved in the nurse-cell function lines of fusion of the apposed membranes,
of the Sertoli cells. Androgen-binding pro¬ which effectively prevent entry of substances
tein is secreted into the lumen of the tubules into the system of intercellular clefts in the
and transports testosterone downstream to upper parts of the epithelium. These occlud¬
maintain normal function of the epithelium ing junctions are situated at the interface
lining the ductuli efferentes and epididymis. between overarching Sertoli cell processes
The release of LH by the hypophysis is just above the spermatogonia (Fig. 31-43).
regulated by a negative feedback mechanism. Thus, they divide the epithelium into a basal
Elevated levels of circulating testosterone compartment, containing the spermatogonia
suppress the release of LH, and abnormally and preleptotene spermatocytes and an ad-
low levels of testosterone result in increased luminal compartment, containing the more ad¬
LH release. The feedback regulation of FSH vanced stages of germ cell differentiation.
is less well understood. It is thought to in¬ Substances in the extracellular spaces of the
volve a nonsteroidal factor called' inhibin, interstitium have relatively unimpeded access
probably produced by the Sertoli cells. In the to the basal compartment but are barred
presence of a normal complement of germ from deeper penetration into the epithelium
834 • MALE REPRODUCTIVE SYSTEM

Figure 31-43. Drawing illustrating how the occluding junction (at arrows) between overarching processes of adjacent
Sertoli cells divides the epithelium into basal and adluminal compartments. Substances diffusing from the interstitium
have direct access to the spermatogonia in the basal compartment, but are excluded from the adluminal compartment
by membrane fusion in the junction complexes. These then constitute the main structural basis of the blood-testis
permeability barrier. (From Fawcett, D. W. In Handbook of Physiology. Vol. 3. Baltimore, Williams and Wilkins Co.,
1975.)

by the Sertoli cell junctional specializations. tion. Moreover, the postmeiotic germ cells
At the appropriate stage of the spermato- are genetically different from the parent
genic cycle, spermatocytes must move from cells. The blood-testis barrier no doubt serves
the basal compartment into the adluminal to prevent foreign protein from these cells
compartment. This is accomplished without from reaching the blood and inducing the
disruption of the permeability barrier. Un¬ formation of antibodies, which would result
dermining processes are extended from the in autoimmune infertility.
Sertoli cells to join beneath the spermatocytes
and form new occluding junctions. The
preexisting Sertoli junctions above the sper¬
matocytes then dissociate, permitting upward EXCRETORY DUCTS OF THE
movement of this cohort of conjoined germ TESTIS
cells.
The full significance of this arrangement
The Tubuli Recti and Rete Testis
is still being studied, but it is clear that a
barrier near the base of the epithelium ena¬ The seminiferous tubules occupy lobules
bles the Sertoli cells to maintain in the or compartments within the testis that are
adluminal compartment a microenvironment demarcated by connective tissue septula ex¬
especially favorable for germ cell differentia¬ tending inward from the tunica albuginea.
 MALE REPRODUCTIVE SYSTEM • 835

The several tubules in each lobule form con¬ are lysosomes. There is no evidence that the
voluted loops, the ends of which converge epithelium of the ductuli efferentes is secre¬
toward a posteriorly situated region of highly tory. The nonciliated cells have short micro¬
vascular connective tissue described as the villi and canalicular invaginations of the free
mediastinum of the testis. Within its substance is surface that are pathways of endocytosis. In
a labyrinthine plexus of epithelium-lined animals injected with vital dyes, the cells
channels called the rete testis (Fig. 31-3). accumulate dye inclusions, as a result of ab¬
Near the ends of the seminiferous tubules, sorption from the lumen. The tall ciliated
the germ cells disappear from the epithelium, cells usually have a conical form, with the
leaving a short terminal segment lined by broad end toward the lumen. The cilia on
Sertoli cells only. An abrupt narrowing then the free surface beat toward the epididymis
occurs where the seminiferous tubule is con¬ and move the sperm in this direction.
tinuous with the tubulus rectus, a short, narrow Outside the basal lamina of the epithelium
channel connecting the seminiferous tubule is a thin layer of circularly arranged smooth
to the rete testis. The tubuli recti and rete muscle cells. In the distal portion of the
testis are lined by a simple cuboidal epithe¬ ductuli forming the coni vasculosi, the mus¬
lium. The cells are relatively simple in their cular layer becomes more prominent.
fine structure and do not give the appearance
of being highly active. Their free surface
Ductus Epididymidis
bears a sparse covering of microvilli.
Many—perhaps all—of the lining cells have The convoluted tubules of the coni vascu¬
a single flagellum projecting into the lumen. losi gradually fuse with one another to form
This is presumed to be motile, although it is the single highly coiled ductus epididymidis. It
not obvious what function it could serve other forms a compact organ less than 7.6 cm long,
than some degree of agitation of the fluid but if the duct were uncoiled and straight¬
contents of the rete. ened out, it would be over 6 m long. The
In the guinea pig, the cells of the tubuli epididymis is the site of accumulation and
recti and the proximal portion of the rete storage of spermatozoa. In addition, there is
testis store a remarkable amount of glycogen, evidence from work on experimental animals
which displaces the nucleus toward the lumen that the spermatozoa are not physiologically
and all other organelles toward the cell pe¬ mature when they leave the testis but grad¬
riphery. This has not been reported in other ually acquire the ability to fertilize and the
species. capacity for normal motility as they slowly
move through the long epididymal duct. The
Ductuli Efferentes epithelium lining this organ may play an
important role in creating a fluid environ¬
From the posterosuperior aspect of the ment favorable for continued maturation of
testis, 12 or more ductuli efferentes arise from the spermatozoa. The epididymis is custom¬
the rete and emerge on the surface of the arily subdivided into three regions for de¬
testis. Through numerous spiral windings scriptive convenience: the caput (head), corpus
and convolutions they form five to ten conical (body), and cauda (tail) (Fig. 31-3). At the
bodies about 10 mm in length called the coni distal end of the cauda, the duct gradually
vasculosi. These have their bases toward the straightens out and continues as the ductus
head of the epididymis and their apices to¬ deferens.
ward the mediastinum testis (Figs. 31—2, The epididymis is lined with a pseudostrat-
31—3). They are held together by connective ified columnar epithelium in which at least
tissue and constitute part of the head of the two cell types are distinguishable. The prin¬
epididymis. cipal cells in the initial segment of the caput
The ductuli efferentes have a characteristic are very tall but gradually become lower in
epithelium. The lumen has a festooned out¬ successive segments and are low columnar or
line because it is lined by alternating groups cuboidal in the cauda epididymidis. The free
of tall ciliated and lower nonciliated cells. surface of each principal cell bears a tuft of
The shorter cells may form small, cuplike very long, nonmotile stereocilia (Figs. 31-44,
excavations in the thickness of the epithe¬ 31—45). These have no basal bodies and in
lium. The nonciliated cells may contain gran¬ electron micrographs appear to be enormous
ules. These were formerly interpreted as se¬ microvilli. They have a bundle of fine fila¬
cretory material, but ultrastructural and ments in their core that extends downward
histochemical studies now indicate that they for some distance into the apical cytoplasm.
836 • MALE reproductive system

Figure 31-44. Section of ductus epididymidis from an adult man. Spermatozoa are seen in the lumen, x 180.

The cell surface between stereocilia is irreg¬ tions of the epididymal epithelium. Over 90
ular in contour, exhibiting numerous invag¬ per cent of the fluid leaving the testis is
inations suggestive of active pinocytosis. absorbed in the ductuli efferentes and ductus
Large numbers of coated vesicles and large epididymidis. If horseradish peroxidase or
multivesicular bodies are present in the apical an electron-opaque particulate marker is in¬
cytoplasm. These structural elements are be¬ jected into the rete testis, it can be shown to
lieved to participate in the absorptive func- be taken into vacuoles and ultimately into
multivesicular bodies of the principal cells of
the epididymal epithelium. Lysosomes are
also found in these cells, and in certain seg¬
ments of the epididymis of some species they
are so large and numerous as to be easily
visualized with the light microscope. They
were interpreted as secretory granules by a
number of early investigators, but this view
has now been abandoned.
The principal cells have a number of cy-
tological characteristics typical of actively syn¬
thesizing secretory cells. Their basal cyto¬
plasm is filled with cisternae of granular
endoplasmic reticulum, and the apical por¬
tion of the cell contains many profiles of
smooth or sparsely granulated reticulum that
are distended with a homogeneous content
Figure 31-45. Photomicrograph of human epididymal of low density. The supranuclear Golgi com¬
epithelium, showing the characteristic row of basal cells plex is remarkably large, but is not obviously
and the stereocilia on the free surface. involved in segregation of a secretory prod-
 MALE REPRODUCTIVE SYSTEM • 837

uct. There are no secretory granules and no with pale cytoplasm and dark heterochro-
unambiguous indication of exocytosis. The matic nuclei. These have been termed “halo
functional significance of the high degree of cells” by some authors, but recent electron
differentiation of the principal cells still microscopic studies identify them as intra¬
eludes us. The epididymal epithelium is epithelial lymphocytes.
known to produce glycerophosphorylcholine, External to the epithelium of the ductus
and a sperm-coating glycoprotein. epididymidis is its smooth musculature,
Cells of the second type, the basal cells, are which exhibits a gradual proximodistal in¬
small round or pyramidal elements lodged crease in thickness. In the caput, the contrac¬
between the bases of the columnar cells (Fig. tile cells are very slender, and the bundles
31—45). Their cytoplasm has little affinity for they form are, for the most part, oriented
stains and is of low density in electron micro¬ circumferentially. In the corpus, sparse
graphs. The organelles are few and relatively strands of longitudinally and obliquely ori¬
simple in their structure. Lipid droplets are ented cells form an incomplete outer layer.
common in the basal cells of some species. At the transition from the corpus to the
The basal cell surface may interdigitate ex¬ cauda, typical large smooth muscle cells are
tensively with the neighboring principal cells. added to the smaller contractile cells charac¬
The function of the basal cells is even more teristic of more proximal portions of the
obscure than that of the principal cells. epididymis (Fig. 31-46). These progressively
Scattered among the columnar cells at var¬ increase in number. In the distal portion of
ious levels in the epithelium are small cells the cauda, the two-layered muscle coat is

Figure 31-46. A, Drawing showing circumferentially oriented thin contractile cells underlying the epithelium of the initial
portion of the ductus epididymidis. B, Corresponding illustration of the transitional zone between the corpus and the
cauda. Slender contractile cells are found immediately subjacent to the epithelium, and several layers of typical large
smooth muscle cells are found peripheral to these. (From Baumgarten, H., A. Holstein, and E. Rosengren. Z. Zellforsch.
720:37, 1971.)
838 * MALE reproductive system

transformed into a three-layered coat, which Ductus Deferens

continues into the ductus deferens.
The regional differences in the cytology of On passing into the ductus deferens, the
the musculature are associated with differ¬ excretory pathway acquires a larger lumen
ences in the motility of the ductus epididy- and a thicker wall. The epithelium and the
midis. In the caput and upper corpus, where lamina propria mucosae form longitudinal
slender muscle cells predominate, the duct folds, which result in the highly irregular
undergoes spontaneous rhythmic peristaltic outline of the lumen seen in cross section
contractions that serve to transport the sper¬ (Figs. 31-49, 31-50). The pseudostratified
matozoa slowly along the tract. These con¬ columnar epithelium is lower than in the
tractions are independent of nervous stimu¬ epididymis, and the cells usually have ster¬
lation and continue when the duct is excised eocilia. The connective tissue of the mucous
and maintained in vitro. Contractions of this membrane contains extensive elastic net¬
character are much reduced in the cauda, works. The highly developed muscular coat
the principal site of sperm storage. The forms a layer 1 mm thick. It consists of inner
larger smooth muscle fibers that predominate and outer longitudinal layers, and a powerful
in this region evidently require adrenergic intermediate layer of circular muscle. Out¬
sympathetic nervous stimulation. The sym¬ side the muscle there is an adventitial coat of
pathetic nerve net increases in density along connective tissue. The firm duct is easily
the cauda and reaches a maximum in the palpable through the thin skin of the scro¬
intra-abdominal portion of the ductus def¬ tum.
erens, which is principally involved in the The spermatic cord consists of the ductus
powerful contractions that expel sperm dur¬ deferens and its accompanying spermatic ar¬
ing the ejaculatory reflex. tery, pampiniform plexus of veins, and
The epididymis is a highly vascular organ nerves of the spermatic plexus. The cord is
with the highest volume flow associated with enclosed by the cremaster muscle, a discontin¬
the metabolically active initial segment of the uous layer of loose, mainly longitudinal
caput (Figs. 31-47, 31-48). strands of striated muscle. This layer extends

Figure 31 47. Surface view of the vascular architecture of the initial segment of the mouse epididymis. Scanning
micrograph of a resin cast. (Micrograph from Suzuki, F. Am. J. Anat. 763:309, 1982.)
 MALE REPRODUCTIVE SYSTEM • 839

Figure 31-48. Transverse section through the initial segment of mouse epididymis, showing the extraordinarily rich
periductal capillary network. (Micrograph from Suzuki, F. Am. J. Anat. 163:309, 1982.)

Epithelium
Lamina propria
Figure 31-49. Cross section of Internal longitu¬
human ductus deferens, x 30. dinal muscle
(After Schaffer.) Intermediate cir¬
cular muscle
External longitu¬
dinal muscle

Nerve

Nerve

Artery
840 • MALE reproductive system

downward to invest the testes and serves to

raise them in response to cold, fear, and
other stimuli.
The ductus deferens, after crossing the
ureter in the abdominal cavity, forms a fusi¬
form enlargement, the ampulla. At the distal
end of the ampulla it receives the duct of a
large gland, the seminal vesicle. Then, as the
short (19-mm) straight ejaculatory duct, it
pierces the body of the prostate gland, at the
base of the urinary bladder. It opens by a
small slit into the prostatic urethra, on a
thickening of its posterior wall, called the
colliculus seminalis or verumontanum (Figs.
31-51, 31—53). The openings of the ejacula¬
tory ducts are located on either side of a
blind invagination on the summit of the col¬
liculus, the utriculus masculinus. This vestigial
organ represents in the male the homologue
of the uterus.
In the ampulla of the ductus deferens, the
mucosa is thrown into numerous thin, irreg¬
ularly branching folds, which in many places
fuse to give the appearance in sections of a
netlike system of partitions with angular
meshes. The epithelium shows evidence of
Figure 31-50. Histological section of the human ductus
deferens, showing its irregular lumen, pseudostratified secretion. From the excavations between the
epithelium, lamina propria, and surrounding bundles of folds, numerous tortuous branched outpock-
longitudinal smooth muscle. (Micrograph courtesy of A. etings reach far into the surrounding mus¬
Hoffer.) cular layer and are lined with a single layer

U rethra

Utriculus of prostate Stratified epithelium

Accessory prostatic gland

Lacuna in cross
Prostatic duct
section
Stratified columnar

Stratified columnar v. epithelium

epithelium
A) Ejaculatory
Ejaculatory duct ■
' '5Q* duct
-OAft .
Lymphoid tissue

Figure 31 51. Cross section through colliculus seminalis of a young man. The urethra has been incised above. The
utriculus of the prostate has prostatic ducts emptying into it. x 10. (After von Ebner, from Schaffer.)
 MALE REPRODUCTIVE SYSTEM • 841

Figure 31-52. Photomicrograph of monkey seminal vesicle.

Prostatic Colliculus
ducts seminalis Surface toward the pubic bone

Cross-striated
muscle fibers

Glands

Pars prostatica
/ \ urethrae

Utriculus
prostaticus
Ductus ejaculatorii

Openings of the
ductus prostatici
Dilated prostatic
duct

Capsule

Figure 31-53. Cross section of human prostate, x 4. (After Braus.)

842 • MALE REPRODUCTIVE SYSTEM

of columnar, clear cells of glandular nature superficial cells the centrioles are located just
containing secretion granules. The muscula¬ beneath the surface and give rise to a central
ture is much less regularly arranged here flagellum. The cells contain numerous gran¬
than in the other parts of the ductus defer¬ ules and clumps of yellow lipochrome pig¬
ens. ment that first appears at puberty. Similar
pigment may also be found in the smooth
muscles and in the connective tissue of the
Ejaculatory Ducts
seminal vesicles. The epithelial cells contain
The epithelium lining the ejaculatory ducts secretion granules. The secretion of the sem¬
is a simple or pseudostratifred columnar ep¬ inal vesicles is a slightly yellowish, viscid liq¬
ithelium, probably endowed with glandular uid. In sections it appears as coagulated,
functions. Its cells contain a large quantity of netlike, deeply staining masses in the lumen.
yellow pigment granules. Near the openings After castration, the epithelium atrophies,
of the duct, the epithelium often assumes the but it can be restored to normal by injections
structure of “transitional” epithelium. The of testosterone.
mucosa of the ducts forms many thin folds The muscular wall of the seminal vesicles
reaching far into the lumen; its connective is provided with a plexus of nerve fibers and
tissue is provided with abundant elastic net¬ contains small sympathetic ganglia.
works. The dorsomedial walls of the ducts
contain a series of glandular outpocketings,
which may be accessory seminal vesicles. The Prostate Gland
ducts proper are surrounded only by con¬
The prostate is the largest of the accessory
nective tissue.
glands of the male reproductive tract. Its
secretion, together with that of the seminal
vesicles, serves as a diluent and vehicle for
ACCESSORY GLANDS OF THE transport of sperm from the male to the
MALE REPRODUCTIVE TRACT female. The prostate is about the size of a
horse chestnut and surrounds the urethra at
its origin from the urinary bladder. It is a
Seminal Vesicles
conglomerate of 30 to 50 small, compound
The seminal vesicles are elongated organs tubuloalveolar or tubulosaccular glands,
with numerous lateral outpocketings from an from which 16 to 32 excretory ducts open
irregularly branching lumen. They arise as independently into the urethra on the right
evaginations of the ductus deferens and are and left sides of the colliculus seminalis (Fig.
basically similar to it in structure. The wall 31—53). The form of the glands is irregular.
consists of an external connective tissue layer Large cavities, sometimes cystic, alternate
rich in elastic fibers, a middle layer of smooth with narrow branching tubules. The blind
muscle thinner than in the ductus deferens, ends of the secreting portions are sometimes
and an epithelium resting on a thin layer of narrower than the excretory ducts. In many
loose connective tissue. The mucosa forms places, branching papillae and folds with a
an intricate system of thin, primary folds, thin core of connective tissue project far into
which branch into secondary and tertiary the lumen. In sections they may appear as
folds. These project far into the lumen and isolated, epithelium-lined islands in the cavi¬
anastomose frequently. In this way, numer¬ ties. The basal lamina is indistinct, and the
ous cavities in different sizes are formed, glandular epithelium rests on a layer of con¬
separated by thin branching partitions (Fig. nective tissue with dense elastic networks and
31-52). All these cavities open into the larger numerous blood capillaries. In the larger
central cavity, but in sections many of them alveolar cavities the epithelium may be low
may seem to be isolated. cuboidal or even squamous, but in most
The epithelium shows great individual var¬ places it is simple or pseudostratihed colum¬
iations, which probably depend on age and nar. The cytoplasm of the cells contains nu¬
physiological conditions. As a rule, it is pseu- merous secretory granules. The epithelial
dostratihed and consists of rounded basal cells become smaller and lose their secretion
cells lodged between larger cuboidal or low granules after castration. Injections of testos¬
columnar cells. All basal cells have a supra¬ terone restore the cells quickly to their nor¬
nuclear pair of centrioles, whereas in the mal appearance and activity.
 MALE REPRODUCTIVE SYSTEM • 843

Dense connective tissue forms a capsule The prostate is abundantly provided with
around the organ and extends into it to plexuses of nonmyelinated nerve fibers con¬
constitute its abundant stroma. Thick septa nected with small sympathetic ganglia. Sen¬
of collagenous fibers and smooth muscle ra¬ sory nerve endings of various kinds (end
diate from the colliculus seminalis toward the bulbs, genital corpuscles, and so on) are scat¬
capsule partitioning the glandular tissue. tered in the interstitial connective tissue. Free
Around the urethra, smooth muscle forms a nerve endings have been described in the
thick ring—the internal sphincter of the blad¬ epithelium.
der. The utriculus prostaticus, situated deep in
The secretion of the prostate is a thin, the interior of the prostate gland and open¬
opalescent liquid with a slightly acid (pH 6.5) ing on the colliculus seminalis, may be a
reaction. It has a rather low protein content minor accessory gland of the male sexual
but contains diastase, beta glucuronidase, sev¬ apparatus. It is a blind vesicle of considerable
eral proteolytic enzymes, and a potent fibrin- size lined with an epithelium with many folds
olysin. It is the main source of the citric acid and glandlike invaginations. The epithelium
and acid phosphatase of the semen. In sec¬ is similar to that of the prostate. Sometimes,
tions, the secretion in the glandular cavities patches of ciliated columnar epithelium can
appears granular. It contains occasional des¬ also be found.
quamated cells and spherical or ellipsoid con¬ The prostate is of great medical interest
centrically lamellated bodies—the prostatic because benign nodular hyperplasia of the prostate
concretions (Fig. 31-54). These are believed to is the most common tumor in the aging male.
originate through condensation of the secre¬ The disease begins at about 45 years of age,
tions. They may become calcified, and may and by the time the age of 80 is reached, 80
exceed 1 mm in diameter. The smaller con¬ per cent of the male population is affected
cretions pass into the semen and can be with varying degrees of obstruction of the
found in the ejaculate. The larger ones are bladder neck and urinary retention. Cancer
unable to pass out through the ducts and of the prostate is also the most common
may remain in the gland lodged in cysts. malignant tumor in the male.
Their number increases with age. Unfortunately, the physiology of the gland

A B
Figure 31-54. Photomicrographs of human prostate. A, The character of the epithelium. B, Typical concretions.
Hematoxylin and eosin.
844 • MALE REPRODUCTIVE SYSTEM

is not well understood, and even its mor¬ smooth muscles. The latter may penetrate
phology has been a subject of controversy with the connective tissue into the interior of
and terminological confusion. On the basis the lobules.
of embryological studies, the prostate has The structure of the epithelium in the
traditionally been subdivided into middle, secreting portions and in the ducts is subject
lateral, and posterior lobes, all pyramidal in to great functional variations. In the enlarged
form and situated respectively anterior, lat¬ alveoli the cells are usually flattened; in the
eral, and posterior to the axis determined by other glandular spaces they are cuboidal or
the course of the ejaculatory ducts through columnar, with the nuclei at the base (Fig.
the gland. These lobes merge at their bound¬ 31-55). The cytoplasm contains small mucoid
aries as development progresses, and in the secretion droplets and spindle-shaped inclu¬
adult they have no reality. There is now a sions staining with acid dyes. It has been
tendency to abandon these regional desig¬ suggested that they leave the cell body as
nations. Some authors do, however, distin¬ such and then dissolve and mix with the
guish central and peripheral zones on the basis mucin. The excretory ducts are lined with a
of histological criteria. In the central zone, pseudostratified epithelium resembling that
the stroma is denser, the branching of the of the urethra and may contain large patches
duct system is more elaborate, the epithelium of secreting cells. They are also provided
is more exuberant, and the sacculations are with small accessory glandular outpocketings
larger, with prominent intraluminal parti¬ having the structure of the glands of Littre
tions. In the peripheral zone, the stroma is in the urethra.
looser, the duct system simpler, and the sac¬ After fixation, the secretion appears in the
culations smaller and less partitioned. This lumen of the glandular spaces and ducts as
histological heterogeneity may well be re¬ a precipitate that stains brightly with eosin.
flected in functional differences. In life the secretion is a clear, viscid, mucus¬
For the interpretation of prostatic disease, like lubricant, which can be drawn out into
it is probably more important to realize that long thin threads. Unlike true mucus, it does
in addition to the glandular tissue of the not form a precipitate with acetic acid. In the
prostate proper, there are smaller glands that
originate as diverticula of the urethra above
the verumontanum. These extend radially
into the periurethral connective tissue and
the smooth muscle of the surrounding
sphincter. Although surrounded by the pros¬
tate and formerly considered a part of it,
these urethral glands are ontogenetically and
functionally distinct. It is now widely ac¬
cepted that these are the site of origin of
benign “prostatic” hyperplasia (BPH). On the
other hand, the prostate proper is the site of
predilection for the development of cancer.

Bulbourethral Glands
The bulbourethral glands (Cowper’s glands),
each the size of a pea, are of the compound
tubuloalveolar variety. In some respects they
resemble mucous glands. Their ducts enter
the membranous urethra or the posterior
portion of the cavernous urethra. The ducts
as well as the secreting portions are of irreg¬
ular size and form, and in many places they
show cystlike enlargements. The terminal
portions end blindly. The connective tissue
partitions between the glandular lobules
Figure 31-55. Part of the lobule of the bulbourethral
measure 1 to 3 mm across and contain elastic
gland of a 23-year-old man. Zenker, x 120. (Slightly
fibers and thick strands of striated and modified from Stieve.)
 MALE REPRODUCTIVE SYSTEM • 845

boar, the secretion is extremely viscous and groove occupied by the corpus cavernosum
rubbery, and plays an important role in the urethrae (Fig. 31-56). The latter is traversed
gelation of the seminal plasma that takes throughout its length by the penile urethra
place soon after ejaculation in this species. and ends with an acorn-shaped enlargement,
the glans penis, bearing on its posterior aspect
a pair of concavities that cap the conical ends
of the two corpora cavernosa penis.
THE PENIS The erectile tissue of the corpora cavernosa
is a vast, spongelike system of irregular vas¬
The penis is formed of three cylindrical cular spaces fed by the afferent arteries and
bodies of cavernous or erectile tissue, the two drained by the efferent veins. In the flaccid
corpora cavernosa penis and the unpaired corpus condition of the organ, the cavernous spaces
cavernosum urethrae (corpus spongiosum). Aris¬ contain little blood and appear as collapsed
ing from the ascending rami of the pubis on irregular clefts. In erection they become large
either side, the corpora cavernosa converge cavities engorged with blood under pressure.
and join at the pubic angle. From there they This increased inflow of blood and relative
run distally side by side to their conical distal restriction of outflow causes the enlargement
ends, forming the dorsal two thirds of the and rigidity of the erect penis.
shaft of the penis. On the upper surface of The three cavernous bodies making up the
the penis, along the line of their junction, is shaft of the penis are surrounded by a thick,
a shallow longitudinal groove occupied by resistant, fibrous capsule, the tunica albuginea.
the dorsal artery and vein. On their lower In the flaccid state the tunica albuginea is 2
surface the corpora cavernosa form a deep mm in thickness; it is reduced in erection to

Figure 31-56. Cross section of a penis of a 21-year-old man The septum in the corpus cavernosum penis is incomplete,
because the section is from the distal part of the organ. The penis was fixed by injection of formalin into the corpus
cavernosum. (Slightly modified from Stieve.)
846 • MALE REPRODUCTIVE SYSTEM

about 0.5 mm. Its bundles of collagenous The tunica albuginea of the corpus caver-
fibers are arranged mainly longitudinally in nosum urethrae is much thinner than that of
its outer layer and circularly in its inner the corpora cavernosa penis and contains
portion. The collagen fiber bundles have a circularly arranged smooth muscle fibers in
wavy course in the flaccid state, but straighten its inner layer. It also is provided with abun¬
out during erection to accommodate the con¬ dant elastic networks. The blood lacunae
siderable increase in volume of the corpora here, unlike those of the corpora cavernosa,
cavernosa. Elastic fibers are relatively sparse. are everywhere the same in size. The trabec¬
The extensibility of the tunica is attributable ulae between them contain more numerous
very largely to the arrangement of the colla¬ elastic fibers, whereas smooth muscle fibers
gen bundles and their ability to lengthen to are relatively scarce. The cavernous spaces
a fixed limit by changing from sinuous to occupying the axis of the corpus cavernosum
straight. By analogy, an elastic bandage is urethrae gradually pass into the venous
made up of inelastic cotton fibers, but can be plexus of the urethral mucosa.
stretched greatlv and can then return to its The glans penis consists of dense connective
initial length because of the pattern in which tissue containing a plexus of large anasto¬
they are woven. The few elastic fibers in the mosing veins, with circular and longitudinal
tunica albuginea may play some role in the smooth muscles in their thick walls. The
recoil of the collagen to its resting pattern. longitudinal muscle strands often bulge into
The tunica albuginea also forms a fibrous the lumen of the veins.
partition between the two corpora cavernosa. The skin covering the penis is thin and is
In the distal half of the shaft, this septum is provided with an abundant subcutaneous
fenestrated so that the cavernous spaces of layer containing smooth muscle, but is devoid
the two corpora communicate. Dense fibrous of adipose tissue. The skin on the distal part
trabeculae extend inward from the tunica, of the shaft of the penis is devoid of hair and
branching and anastomosing to form a com¬ has only sweat glands in limited numbers.
plex framework around the cavernous The glans is covered by an encircling fold of
spaces. This trabecular meshwork is also con¬ skin, the prepuce. Its inner surface, adjacent
tinuous with a dense connective tissue sheath to the glans, is moist and has the character
that surrounds the deep artery, which is of a mucous membrane. The dermis of the
centrally located in each corpus. Bundles of glans penis is fused with the deeper connec¬
smooth muscle fibers run longitudinally in tive tissue of the glans. In this region there
the fibrous sheath of the central artery and are peculiar sebaceous glands (glands of Ty¬
along the trabecular meshwork throughout son), which are not associated with hairs.
the corpora, and insert upon the inner aspect They show great individual variations in
of the tunica albuginea. The abundance of number and distribution.
smooth muscle in the organ has not been
fully appreciated, but recent studies indicate
Mechanics of Erection
that it is the predominant tissue of the par¬
enchyma of the corpora cavernosa. Erection of the penis is a vascular phenom¬
The central artery gives off helicine enon controlled by complex neural pathways
branches that course through the fibrous activated by both psychic and tactile stimuli.
trabeculae, terminating in end arteries that Reflex erection results from afferent sensory
open into the cavernous spaces or sinuses. input carried via the pudendal nerve to the
These are lined by endothelium and are spinal cord where vasomotor impulses are
irregularly shaped clefts 1 mm or so across generated in parasympathetic fibers of the
in the flaccid state, but which fill with blood nervi erigentes. These produce vasodilatation
and expand to several times that diameter of the arteries of the penis, filling the cavern¬
when the corpora are engorged. The cavern¬ ous spaces in the corpora and causing erec¬
ous spaces are largest in the central region tion. In addition to this simple reflex arc,
of the corpora and gradually diminish in size there is a thoracolumbar pathway that can be
toward the cavernous venules on the inner activated by psychic stimuli including mem¬
aspect of the tunica albuginea. These in turn ory and imagination, as well as visual and
are continuous with larger veins that pene¬ tactile input to the cerebral cortex. Higher
trate the tunica albuginea to join circumflex neural centers appear to control vasodilator
veins that are tributaries of the deep dorsal fibers originating in thoracic sympathetic
vein of the penis. ganglia, d hese can cause erection independ-
 MALE REPRODUCTIVE SYSTEM • 847

ently of reflex stimulation. An adrenergic pressure. It is argued that since the helicine
nerve supply to the corpora from sacrococ¬ arteries open mainly into the larger axial
cygeal sympathetic ganglia is believed to be cavernous spaces, the expansion of these
involved in the vasoconstriction that results spaces compresses the smaller peripheral
in detumescence. spaces and the thin-walled veins underlying
The hemodynamics of erection continues the relatively unyielding tunica adventitia. In
to be a subject of debate. It is agreed that this way, the outflow of blood is throttled
erection involves rapid filling of the corpora down as the blood accumulates in the corpora
following vasodilatation of the deep arteries under increasing pressure. Contraction of
and their helicine branches, and accom¬ the intrinsic smooth muscle is thought to
panied by relaxation of the intrinsic smooth contribute to the high intracavernosal pres¬
muscle of the parenchyma. Dilatation of the sure and rigidity of the erect penis. In further
arteries results in an initially greater rate of support of the concept of restriction of out¬
inflow than outflow. A majority of investiga¬ flow is the observation that the walls of the
tors believe that this is sufficient to explain circumflex veins are unusually muscular. In
erection, and experimental evidence, ob¬ addition, these veins consistently exhibit
tained mainly from dogs, seems to support unique specializations of their intima called
this view. Others believe that there must also polsters (Fig. 31-57). These are local accu¬
be constriction of venous outflow. Noninva- mulations of fibroblasts and smooth muscle
sive measurement of intracavernosal pres¬ cells beneath the endothelium that form con¬
sure in man during erection indicates that it spicuous longitudinal thickenings or ridges
is several times higher than systolic blood that can be followed through hundreds of

Figure 31-57. Photomicrographs of specializations of the veins of the penis believed to be involved in the hemodynamics
of erection. A, Cross section through a circumflex vein at the proximal end of the corpora cavernosa of a 68-year-old
man, showing (at arrows) thick muscular polsters. x 350. B, Sections through circumflex veins at the distal end of the
corpora cavernosa. The polsters are indicated by arrows, x 40. (Photomicrographs from Goldstein, A. M. B., et al.
Urology 20:259, 1982.)
848 • MALE REPRODUCTIVE SYSTEM

serial sections. These are believed to have a SEMEN

role in constricting the lumen and retarding
venous outflow during erection.
As the sperm pass along the excretory
Superficially similar intimal thickenings
ducts, the secretions of the ducts and acces¬
have been reported in the arteries of the
sory glands are added to them. The final
human penis, but these are regarded as
product is the semen. The sperm in the sem¬
prearteriosclerotic changes. These so-called
iniferous tubules are nonmotile. They are
arterial polsters are found only occasionally
slowly forwarded into the tubuli recti and the
and are probably pathological, whereas ve¬
rete testis by the fluid secreted into the tu¬
nous polsters are a constant finding and are
bules. The seminiferous tubules may also
more likely to have physiological significance.
actively move the quiescent sperm by execut¬
ing peristaltic movements. Tfie flattened cells
Lymphatics investing the seminiferous tubules have been
shown to have many of the fine structural
Dense, superficial networks of lymphatic characteristics of smooth muscle cells. In
capillaries are found in the skin of the pre¬ some species, slow contractile movements of
puce and shaft of the penis. They form a the excised seminiferous tubules have been
dorsal superficial lymph vessel, which runs observed. In the ductuli efferentes, the epi¬
toward the medial inguinal lymph nodes. thelium with its cilia beating toward the epi¬
Deep nets of lymphatic capillaries collect the didymis contributes to the transport of the
lymph from the glans; they form a plexus on sperm.
each side of the frenulum and continue into The long, winding duct of the epididymis
a dorsal subfascial lymph vessel. is slowly traversed by the sperm propelled by
peristaltic contractions of smooth muscle in
the duct wall. They are kept here, especially
Nerves in the tail, for a long time, sometimes for
several weeks. In some species, the sperm
The nerves of the penis come both from acrosome undergoes continuing morpholog¬
the sacral plexus, via the pudendal nerve, ical differentiation during the passage
and from tfie pelvic sympathetic system. The through the epididymis, and the fertilizing
former supply the striated muscles of the capacity of the spermatozoa is known to in¬
penis (such as the bulbocavernosus) and also crease progressively. As previously noted, the
furnish the sensory nerve endings in the skin ductus epididymidis has an outer layer of
and the mucosa of the urethra. Among these smooth muscle that is responsible for
sensory endings, free nerve endings can be rhythmic peristaltic movements that move
demonstrated in the epithelium of the glans, the sperm along the duct. During ejaculation,
the prepuce, and the urethra. In addition, contraction of the ductus deferens is of pri¬
there are free nerve endings in the subepi- mary importance in expulsion of stored
thelial connective tissue of the skin and the sperm.
urethra. Finally, numerous encapsulated cor¬ The epididymis is the site of storage of
puscles of various types are present: corpuscles spermatozoa. The sperm do not accumulate
of Meissner in the papillae of the skin of the to any great extent in the ductus deferens.
prepuce and the glans; genital corpuscles in This part of the excretory system, with its
the deeper layers of the stratum papillare of heavy muscular coat, is adapted for their
the dermis of the glans and in the mucosa of speedy transportation during sexual activity.
the urethra; and corpuscles of Vater-Pacini, The function of the seminal vesicles is
which occur along tfie dorsal vein in the primarily glandular. Their thick secretion
subcutaneous fascia, in the deeper connective contributes substantially to the volume of the
tissue of the glans, and under the albuginea ejaculate. It is rich in fructose, which is the
in the corpora cavernosa. The sympathetic principal sugar of the semen and provides
nervous plexuses are connected with the the carbohydrate substrate utilized as an en¬
smooth muscles of the vessels and form ex¬ ergy source by motile spermatozoa of the
tensive, nonmyelinated networks among the ejaculate. The secretion contains small
smooth muscles of the trabeculae in the cor¬ amounts of yellowish pigment, mainly flavins,
pora cavernosa. which give the semen a strong fluorescence
 MALE REPRODUCTIVE SYSTEM • 849

in ultraviolet light—a property of some med¬ abundant secretion of the seminal vesicles is
icolegal importance in the detection of semen coagulated in the vagina by an enzyme con¬
stains. tained in the prostatic juice, and thus a solid
In the process of ejaculation, the muscular plug is formed in the vagina that temporarily
tissue of the prostate also contracts and dis¬ occludes its lumen and prevents the escape
charges its abundant liquid secretion. The of the semen.
semen, entering the urethra and mixing with
the secretion of the glands of Cowper and
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Huckins, C.: The spermatogonial stem cell population cytochemistry of the ductus epididymis in stallions,
in adult rats. I. Their morphology, proliferation rams, and bulls. Acta Morphol. Neerl. Scand. 7:337,
and maturation. Anat. Rec. 769:533, 1971. 1958.
Setchell, B. P.: The functional significance of the blood- Orgebin-Crist, M. C.: Studies on the function of the
testis barrier. J. Androl. 7:3, 1980. epididymis. Biol. Reprod. (Suppl.) 7:155, 1969.

INTERSTITIAL TISSUE ACCESSORY GLANDS OF MALE

Christensen, A. K.: Leydig cells. In Hamilton, D. W.,
REPRODUCTION
and R. O. Greep, eds.: Handbook of Physiology: Brandes, D., D. Kirchheim, and W. W. Scott: Ultrastruc¬
Endocrinology. Vol. 5, Sect. 7, Male Reproductive ture of the human prostate; normal and neoplastic.
System. Washington, DC, American Physiological Lab. Invest. 75:1541, 1964.
Society, 1975. Franks, L. M.: Benign nodular hyperplasia of the pros¬
Christensen, A. K.: The fine structure of the testicular tate: a review. Ann. R. Coll. Surg. Engl. 74:92, 1954.
interstitial cells of the guinea pig. J. Cell Biol. Flarbitz, T. B., and O. A. Haugen: Histology of the
26:911, 1965. prostate in elderly men. Acta Pathol. Microbiol.
Crabo, B.: Fine structure of the interstitial cells of the Scand. [Sect. A] 80:756, 1972.
rabbit. Zeitsch. Zellforsch. 67:587, 1963. McNeal, J. E.: The prostate and prostatic urethra—a
Fawcett, D. W., W. B. Neaves, and M. N. Flores: Com¬ morphologic synthesis. J. Urol. 797:1008, 1973.
parative observations on intertubular lymphatics
and the organization of interstitial tissue of the PENIS
mammalian testis. Biol. Reprod. 9:500, 1973. Conti, G.: L’Erection du penis humain et ses bases
Johnson, L, and W. B. Neaves: Age related changes in morphologicovasculaires. Acta Anat. 74:217, 1952.
the Leydig cell population, seminiferous tubules, Goldstein, A. M., J. P. Meehan, R. Zakhary, P. A.
and sperm production in stallions. Biol. Reprod. Buckley, and F. A. Rogers: New observations on the
24:703, 1981. microarchitecture of the corpora cavernosa in man
Kaler, L. W., and W. B. Neaves: Attrition of the human and their possible relationship to the mechanism of
Leydig cell population with advancing age. Anat. erection. Urology 26:259, 1982.
Rec. 792:513, 1978. Meehan, J. P., and A. M. B. Goldstein: High pressure
Mori, H., and A. K. Christensen: Morphometric analysis within the corpus cavernosum in man during erec¬
of Leydig cells in the normal rat testis. J. Cell Biol. tion. Its probable mechanism. Urology 27:385,
84:340, 1980. 1983.
BLOOD VESSELS AND LYMPHATICS Shirai, M., and N. Ishii: Hemodynamics of erection in
man. Arch. Androl. 6:27, 1981.
Lawcett, D. W., P. M. Heidger, and L. V. Leak: Lymph Weiss, H.: The physiology of human penile erection.
vascular system of the interstitial tissue of the testis Ann. Intern. Med. 76:793, 1972.
as revealed by electron microscopy. J. Reprod. Fer¬
til. 79:109, 1969.
 32
FEMALE REPRODUCTIVE
SYSTEM

The female reproductive system includes tive life of the woman, the ovaries and the
the internal organs—ovaries, oviducts, uterus, endometrium of the uterus undergo a regu¬
and vagina; and the external genitalia—mons larly repeated cycle of hormonally controlled
pubis, labia majora, labia minora, and clitoris histological changes about every 28 days. Ces¬
(Fig. 32—1). The size and microscopic struc¬ sation of menses, or menopause, occurs at a
ture of these organs change greatly with age. mean age of 51 years as a result of depletion
They are relatively small and functionally of ovarian follicles and consequent reduction
quiescent until puberty, which involves grad¬ in estrogen secretion.
ual breast development and other changes of
body form; the appearance of axillary and
pubic hair; and growth and differentiation
of the internal reproductive organs. These OVARY
changes culminate in menarche, the first men¬
strual flow, which occurs at about 13 years of The human ovaries are. slightly flattened
age. Thereafter, throughout the reproduc¬ paired organs, each measuring 2.5 to 5 cm

Figure 32-1. Drawing of a sagittal

section of the female pelvis, showing
the reproductive organs and their re¬
lationships to bladder, urethra, and
rectum. (From Anson, B. J. Atlas of
Human Anatomy. 2nd ed. Philadel¬
phia, W. B. Saunders Co., 1963.)

851
852 • FEMALE REPRODUCTIVE SYSTEM

in length, 1.5 to 3 cm in^width, and 0.6 to The ovary is covered by a continuous sheet
1.5 cm in thickness. One of the edges, the of squamous or cuboidal epithelium, which
hilus, is attached by the mesovarium to the was named the germinal epithelium in the mis¬
broad ligament, which extends from the uterus taken belief that the primordial oocytes orig¬
laterally to the wall of the pelvic cavity. inated from it (Fig. 32-3). The term persists
The ovary has a thick peripheral zone, or although the evidence now overwhelmingly
cortex, which surrounds the medulla. Embed¬ favors the extragonadal origin of the primor¬
ded in the connective tissue of the cortex are dial germ cells. Beneath the germinal epithe¬
follicles containing the female sex cells, oocytes. lium is a layer of dense connective tissue, the
The follicles are present in a wide range of tunica albuginea (Fig. 32—4).
sizes representing various stages of their de¬
velopment (Fig. 32—2). When a follicle Ovarian Follicles
reaches maturity it ruptures at the surface of
the ovary to release the ovum, which then Embedded in the stroma of the cortex deep
enters the open end of the neighboring ovi¬ to the tunica albuginea are the follicles. The
duct. The boundary between the ovarian younger the woman, the more numerous
cortex and medulla is poorly defined. The they are. In a normal young adult, over
medulla consists mainly of loose connective 400,000 have been counted in serial sections
tissue and a mass of contorted blood vessels of both ovaries. Of this large number, fewer
that are large in proportion to the size of the than 500 ova are released by the process of
ovary. ovulation during a woman’s reproductive life

Small
Corpus follicle
luteum
Adipose
Large tissue
follicle

Atretic
follicle

follicles

Figure 32-2. Photomicrograph of a transected ovary of Macacus rhesus. (Courtesy of H. Mizoguchi.)

Figure 32-3. A, Cells of the germinal epithelium of the ovary as seen with the transmission electron microscope in a
thin section. The surface bears irregularly oriented microvilli. B, Scanning electron micrograph of the germinal epithelium
of the ovary. The bulging free surfaces have many more microvilli than one would infer from study of thin sections of
this epithelium. The germinal epithelium resembles the peritoneal mesothelium covering other abdominal and pelvic
organs. (Micrographs courtesy of E. Anderson.)

Primordial Tunica

Figure 32-4. Photomicrograph of cat ovary, showing numerous primordial follicles and two secondary follicles with a
well-developed antrum, x 100.
853
854 • FEMALE REPRODUCTIVE SYSTEM

During each menstrual cycle, from five to 15 Primary Follicles. Development of follicles
follicles begin to grow and develop, but only in the ovary occurs continuously from in¬
one of these is destined to proceed to ovula¬ fancy to the menopause, uninterrupted by
tion. The others become arrested at various anovulatory cycles or pregnancies. The tran¬
stages of development, and degenerate in the sition from a quiescent primordial follicle to
process called follicular atresia. The number a developing primary follicle involves cyto-
of follicles decreases progressively through¬ logical changes in the oocyte, the follicular
out life, and at menopause follicles are hard cells, and the adjacent stromal cells. As the
to find, although a few may persist into old oocyte enlarges, the follicular cells become
cuboidal or low columnar and by mitotic
age- •

The vast majority of follicles are primordial

,
proliferation give rise to a stratified epithe¬
or unilaminar follicles. They are found mainly lium of granulosa cells, thus transforming the
in the periphery of the cortex, immediately unilaminar primordial follicle to a multilam-
beneath the tunica albuginea (Figs. 32-4, inar primary follicle (Figs. 32—7, 32—9). The
32-5). Each consists of a large round oocyte stratified epithelium rests on a thick basal
surrounded by a single layer of flattened lamina, the membrana limitans externa, which
follicular cells. Owing to the large size of the separates the granulosa from the surround¬
oocyte, there may be several follicular cells ing stromal cells that have become concen¬
around its circumference in sections. Primor¬ trated around the follicle and have differen¬
dial follicles may contain more than one oo¬ tiated to form the theca folliculi (Figs. 32—9,
cyte, but such polyovular follicles are quite 32-10).
uncommon. In the concurrent growth of the oocyte,
Primordial Follicles (Unilaminar Folli¬ there is a noticeable change in the distribu¬
cles). The oocyte of the primordial follicle is tion of its organelles. The single large juxta¬
enveloped by a single layer of flattened fol¬ nuclear Golgi gives rise to multiple complexes
licular cells. It has a large, eccentrically placed widely dispersed in the ooplasm. The rough
vesicular nucleus with a conspicuous nucleo¬ endoplasmic reticulum becomes more exten¬
lus. In favorable thin sections the meiotic sive, and the number of free polyribosomes
chromosomes can be seen (Fig. 32—6). Asso¬ increases. Mitochondria proliferate and dis¬
ciated with the juxtanuclear cell center is a perse throughout the ooplasm. Small vesicles
well-developed Golgi apparatus surrounded and multivesicular bodies increase in num¬
by numerous small mitochondria. The sur¬ ber. Between the oocyte and the surrounding
faces of the oocyte and the enveloping follic¬ granulosa cells, a space develops into which
ular cells at this stage are smooth and in close irregularly shaped microvilli project from the
apposition. oolemma and from the neighboring granu¬
In electron micrographs the prominent losa cells (Figs. 32—8, 32-11). An amorphous
juxtanuclear Golgi complex consists of short material accumulates in this space around
parallel arrays of cisternae and large num¬ the microvilli and gradually condenses to
bers of small vesicles. Similar vesicular pro¬ form the zona pellucida, a highly refractile
files are distributed in smaller numbers glycoprotein layer that stains intensely with
throughout the ooplasm, and it is believed the periodic acid—Schiff reaction. The micro¬
that these may originate in the Golgi appa¬ villi of the oocyte and the slender processes
ratus. Annulate lamellae are often found of the neighboring granulosa cells are in
adjacent to the nucleus or free in the neigh¬ contact within the substance of the zona pel¬
boring ooplasm. The spherical or short lucida throughout the subsequent develop¬
plump mitochondria tend to congregate in ment of the follicle. The zona pellucida,
the vicinity of the cell center. Later, when which may attain a thickness of 5 pm in the
the oocyte begins to grow, they become dis¬ mature follicle, is usually considered to be a
persed throughout the ooplasm (Fig. 32-8). product of the granulosa cells, but there is
The endoplasmic reticulum in the early some evidence that the oocyte may contribute
stages of oocyte development takes the form to its formation.
of vesicles or slightly elongated profiles bear¬ As the primary follicle develops, the mito¬
ing a few ribosomes. Longer cisternal profiles chondria, rough endoplasmic reticulum, and
subsequently arise in limited numbers and free ribosomes of the granulosa cells increase
finally become associated in parallel arrays. in abundance and the Golgi complex be¬
Multivesicular bodies are a common compo¬ comes more prominent. Lipid droplets are
nent of the ooplasm but are more numerous common in the cytoplasm in some species.
in later stages of development. Large gap junctions are found in areas of
 FEMALE REPRODUCTIVE SYSTEM • 855

Figure 32-5. Photomicrograph of a number of primordial follicles in the cortex of monkey ovary. The chromosomes of
the dictyate stage of meiosis are visible in the oocyte nuclei. The oocytes are surrounded by a single layer of flattened
follicular cells.

Nucleus of ovum
Cytoplasm of ovum
Nucleolus of ovum Perinuclear mitochondria

Follicular cell with mitochondri Basal lamina

Interstitial
connective tissue

Blood vessel

Figure 32-6. Primary unilaminar follicle in an early stage of its growth. Drawing from a preparation of human ovary by
C. M. Bensley. (After A. A. Maximow.)
856 FEMALE REPRODUCTIVE SYSTEM

Figure 32-7. Bilaminar primary follicle from monkey

ovary.

*><• •2%?

^* 7." , •■ sfc- $\ :»#A>V:-«d

Figure 32-8. Electron micrograph of an early primary follicle. The cytoplasm of the oocvte contains numerous
mitochondria. A space has appeared between the oocyte and the follicular cells, and amorphous materialTs accumulatha
in it to form the zona pellucida. (Micrograph courtesy of P Motta ) dmorpnous material is accumulating
 FEMALE REPRODUCTIVE SYSTEM • 857

Basal lamina Mitosis of follicular cell

Blood vessel
with erythrocytes

^ Developing ovu m

— Nucleus of ovum

L2-Zona pellucida

Follicular epithelium

Theca folliculi Call-Exner body

Figure 32-9. Drawing of a growing follicle from human ovary. The follicular epithelium is now several layers in thickness
and the oocyte is considerably enlarged. (After A. A. Maximow.)

contact between adjacent granulosa cells, pro¬ externa do not undergo further cytological
viding low-resistance pathways for their elec¬ differentiation and continue to resemble fi¬
trical and metabolic coupling. broblasts. The boundaries between the theca
As the follicles increase in size, they grad¬ interna and theca externa and between the
ually move deeper into the cortex (Fig. 32—2). theca externa and the surrounding stroma
Concurrently with the proliferation of the are indistinct.
granulosa cell layer, the sheath of stromal Secondary Follicles (Antral Follicles). In
cells forming the theca folliculi becomes more the course of the continuing proliferation of
prominent, and a wedge-shaped thickening the follicular cells, the enlarging follicle be¬
extends from the follicle toward the surface comes oval in shape and the oocyte eccentric
of the ovary, forming the theca cone. The in position. When the follicle reaches a di¬
theca subsequently differentiates into a ameter of about 0.2 mm and has six to 12
highly vascular inner layer of secretory cells, layers of cells, irregular spaces filled with
the theca interna, and an outer layer, the theca clear fluid appear among the granulosa cells.
externa, composed mainly of connective tis¬ This fluid, called liquor folliculi, increases in
sue. Numerous small vessels penetrate the amount as the follicle enlarges and the irreg¬
theca externa to supply a rich capillary plexus ular spaces among the granulosa cells become
in the theca interna, but the granulosa cell confluent to form a single crescentic cavity,
layer remains avascular throughout the the antrum. Thenceforth the follicle is de¬
growth of the follicle. scribed as a secondary follicle, or antral follicle
The cells of the theca interna are initially (Fig. 32—13). By the time the formation of
hbroblast-like, fusiform cells, but they sub¬ the antrum begins, the oocyte has usually
sequently accumulate small lipid droplets and attained its full size, 125 to 150 pm. Although
their cytoplasm takes on the ultrastructural the ovum grows no more thereafter, the
characteristics of steroid-secreting endocrine follicle as a whole continues to enlarge until
cells (Fig. 32-15). The cells of the theca it reaches a diameter of 10 mm or more.
858 • FEMALE REPRODUCTIVE SYSTEM

Figure 32-10. Photomicrograph of a follicle comparable with that in Figure 32-8, showing the stratified folliculi
epithelium, its prominent basal lamina, and the thick zona pellucida around the oocyte. The spherical clear areas among
the granulosa cells are Call-Exner bodies. An area similar to that in the rectangle is shown in an electron micrograph in
Figure 32-11. (Photomicrograph courtesy of E. Anderson.)

The typical small antrum follicle is lined Whether they are intra- or extracellular was
with a stratified epithelium of granulosa cells, formerly a subject of dispute, but electron
which displays a local thickening on one side micrographs clearly show them to be extra¬
called the cumulus oophorus. This thicker re¬ cellular (Fig. 32-12). They stain positively
gion protruding into the fluid-hlled cavity with the PAS reaction. Their origin and sig¬
has the oocyte in its center (Figs. 32-13, nificance are unknown.
32-14). The oocyte is surrounded by a single Mature Follicule (Graafian Follicle). In
layer of cuboidal follicular cells whose apical the human, follicles require 10 to 14 days
processes are firmly anchored in the zona from the beginning of the cycle to reach
pellucida. This cellular investment is referred maturity. As they approach their maximal
to as the corona radiata. size, they are large vesicles that occupy the
Although the lining of the follicle is de¬ full thickness of the ovarian cortex and bulge
scribed as a stratified epithelium, its granu¬ from the free surface of the organ. The
losa cells are less compact in their organiza¬ follicles appear tense, as though the liquid in
tion than the cells of most epithelia. They the follicular cavity were under considerable
may be columnar immediately surrounding pressure, but actual measurements have not
the zona pellucida, but elsewhere liquor fol¬ borne out this impression.
liculi accumulates between them, they be¬ In late stages of follicular growth, mitotic
come angular or stellate in form, and they figures gradually decrease in number among
are connected with one another by short the granulosa cells. Intercellular spaces
processes. In growing follicles, small accu¬ among the cells of the inner layers of the
mulations of densely staining material may epithelium become more prominent. The
appear among the granulosa cells. These are connection of the ovum and the associated
the Call-Exner bodies (Figs. 32—9, 32—10). granulosa cells of the cumulus oophorus with
 FEMALE REPRODUCTIVE SYSTEM • 859

Figure 32-11. Electron micrograph of an area similar to that enclosed in the rectangle in Figure 32-10, but in a younger
follicle. There are many microvilli on the oocyte and slender irregular apical processes on the follicular cells, which
extend into the substance of the zona pellucida. (Micrograph courtesy of E. Anderson.)

Figure 32-12. Electron micrograph of Call-Exner body. It

appears to consist of a cavity lined with a distinct basal
lamina and with a filigree of excess basal lamina in the
interior. There is also a sparse flocculent precipitate of
proteinaceous material. (Micrograph courtesy of E. An¬
derson.)
Figure 32-13. Photomicrograph of a secondary follicle from monkey ovary, showing a small antrum and cumulus
oophorus. A well-developed theca is seen around the periphery of the follicle.

Follicular
roi lo

pellucida

Figure 32-14. Photomicrograph of a human oocyte in an antral follicle. Where it projects into the antrum, the investmen
courtesy ofaLCZambornid)UCed *° 3 S'n9 ° ^ Wh°Se processes are anchored in the zona pellucida. (Photomicrograpt

860
 FEMALE REPRODUCTIVE SYSTEM • 861

Follicular epithelial
cell in mitosis
Cavity of follicle

Follicular
epithelium
Basal
amina

Cell of theca
interna

Blood vessel

Cell of theca
externa

Figure 32-15. Section of part of wall of large follicle, illustrating the organization of the theca folliculi. (After A. A.
Maximow.)

the rest of the epithelium is gradually loos¬ Histophysiology of the Ovarian Follicles.
ened by the development of new, liquid-filled In addition to their role in sustenance of the
intercellular spaces. In the loosening up of oocytes, the ovarian follicles have important
the cumulus the ovum, together with its zona endocrine functions. Of the several follicles
pellucida and corona radiata, are freed of that are activated and undergo varying de¬
their attachment in preparation for ovula¬ grees of development in each cycle, usually
tion. only one progresses to ovulation. Nothing is
The theca folliculi reaches its greatest de¬ known about what determines the number
velopment in the mature follicle. The theca of primordial follicles that will begin to grow
interna is composed of large, spindle-shaped or how one among them is selected to com¬
or polyhedral cells with oval or elliptical nu¬ plete its development. It is possible that those
clei and fine lipid droplets in their cytoplasm which become atretic serve some transient
(Figs. 32-15, 32-16). They are enmeshed in endocrine function unrelated to ovulation.
a network of reticular fibers that are contin¬ Certainly the maturing follicle is active as an
uous with those of the theca externa and the endocrine gland.
rest of the ovarian stroma. Although they are The granulosa cells of developing follicles
modified stromal cells and superhcally resem¬ have receptors for follicle stimulating hor¬
ble fibroblasts, the cells of the theca interna mone (FSH), luteinizing hormone (LH), es¬
in electron micrographs have cytological trogens, and testosterone. The relative num¬
characteristics similar to those of cells in other ber of each of these receptors is subject to
steroid-secreting endocrine glands. They are modulation by the varying levels of the other
principally responsible for elaboration of the hormones at successive stages of follicle de¬
steroid precursor of the female sex hor¬ velopment in the menstrual cycle.: The pri¬
mones, estrogens. Consistent with the endo¬ mary stimulus for follicular growth is pro¬
crine function of the theca interna is its rich vided by FSH from the adenohypophysis.
capillary plexus. The theca externa consists LH induces differentiation of the cells of the
of concentrically arranged fibers and fusi¬ theca interna and stimulates their secretion
form cells that do not appear to have any of testosterone, which diffuses into the folli¬
secretory function. cle where aromatizing enzymes in the gran-
862 • FEMALE REPRODUCTIVE SYSTEM

receptors in the membrane of the granulosa

cells. Responding to LH, the granulosa cells
then begin to secrete progesterone. At the
same time, the midcycle increase in plasma
LH down-regulates the number of LSH re¬
ceptors on the granulosa cells, reducing their
estrogen production as they shift to secretion
of progesterone.
There is increasing experimental evidence
that the granulosa cells also secrete a nonste¬
roidal hormone that acts back upon the hy¬
pothalamohypophyseal axis to suppress LSH
release. This substance, detectable in follicu¬
lar fluid, has yet to be isolated and fully
characterized but seems to be a relatively
small peptide. It has been called folliculostatin,
but since most preparations inhibit secretion
of both LSH and LH, some prefer the less
specific term gonadostatin. Its precise role in
the complex regulation of reproduction re¬
mains to be elucidated.

Ovulation
The process by which the follicle ruptures
and sets free the ovum is called ovulation. A
follicle matures at intervals of about 28 days
in the human female, although variations of
a week are not uncommon. Normally ovula¬
tion occurs on or about the 14th day of an
ideal 28-day cycle. Usually only one ovum is
released, but occasionally two, and rarely
more, may be discharged. Cycles of typical
duration may occur without associated ovu¬
lation. These are called anovulatory cycles. The
stages preparatory to rupture of the follicle
Figure 32-16. Photomicrograph of a portion of the wall have been extensively studied in histological
of an antral follicle, illustrating the loose organization of sections of ovaries, and the actual process of
the follicular epithelium (above) and the fusiform thecal ovulation has been directly observed and
cells (below), with multiple lipid droplets in their cytoplasm,
appearing here as clear vacuoles.
photographed in anesthetized living animals
and in humans.
The ovum and the granulosa cells imme¬
diately surrounding it are loosened from the
ulosa catalyze its conversion to the estrogen, cumulus oophorus in the last stages of follic¬
17(3 estradiol. FSH acting upon the granulosa ular maturation and float free in the liquor
cells induces this conversion of thecal andro¬ folliculi. During this period, the follicular
gen to estrogen, which in turn stimulates fluid seems to accumulate faster than the
granulosa cell proliferation and growth of follicle grows, and the part of the follicular
the follicle. Estrogen also diffuses into the wall that bulges on the surface of the ovary
capillaries of the theca interna, raising the becomes progressively thinner. The follicular
level of the hormone in the general circula¬ fluid that forms just before ovulation contains
tion. The increasing level of estrogen in the more water than that formed earlier and
blood late in the follicular phase of the cycle appears to be secreted at a rapid rate. The
acts back upon the hypothalamohypophyseal first indication of impending ovulation is the
system to induce the preovulatory surge of appearance on the outer surface of the folli¬
LH. Concurrently, the increasing concentra¬ cle of a small oval area, the macula pellucida
tion of estrogen within the follicle shortly or stigma. In this area the flow of blood slows
before ovulation induces formation of LH and then ceases, resulting in a local change
 FEMALE REPRODUCTIVE SYSTEM • 863

in color and translucency of the follicular the theca externa would be expected to com¬
wall. The germinal epithelium overlying this press the follicular contents and raise intra-
area becomes discontinuous and the inter¬ follicular pressure. Sensitive pressure record¬
vening stroma greatly thinned out. The ings have failed to demonstrate any
stigma then bulges outward as a clear vesicle significant rise in intrafollicular pressure im¬
or cone. In the rat, in which these events mediately preceding ovulation. Therefore,
have been observed in greatest detail, the the smooth muscle—like cells in the theca
formation of the stigma takes place in five probably do not play a role in this process.
minutes or less. The cone then ruptures, and They may, however, be involved in the post¬
in a minute or two the ovum and its adherent ovulatory collapse of the follicle that precedes
mass of cumulus cells pass through the ori¬ formation of the corpus luteum.
fice, followed by a gush of follicular fluid At the risk of some redundancy, it may be
(Fig. 32-17). The fluid immediately associ¬ useful to review here the endocrine control
ated with the ovum appears viscous, while of the menstrual cycle and ovulation.
that which follows is quite thin.
The turgid fronds or fimbriae of the ovi¬
Endocrine Control of Ovulation
duct, or fallopian tube, are closely applied to
the surface of the ovary at the time of ovu¬ Ovulation depends on a complex sequence
lation. Their active movements, and the cur¬ of endocrine events involving the hypothal¬
rents created in the surface film by the cilia amus, the hypophysis, and the ovary. The
on their epithelial cells, are responsible for central control of the menstrual cycle resides
drawing the ovum into the open ostium of in the arcuate region of the medial basal
the oviduct. hypothalamus. Neurons in this region have
The mechanisms involved in rupture of an inherent rhythm of activity with a peri¬
the follicle and extrusion of the ovum are odicity of 90 minutes in the human. Their
still poorly understood. It has been suggested activity results in a pulsatile release of go¬
that collagenase and other proteases may nadotropin releasing hormone (GnRH),
depolymerize collagen fibrils of the extracel¬ which is carried in the hypophyseoportal ves¬
lular matrix, contributing to the weakening sels to the anterior lobe of the hypophysis.
of the follicular wall at the stigma, but there There, the hormone stimulates synthesis and
is no compelling evidence that enzymes are release of follicle stimulating hormone (FSH)
involved. Electron microscopic studies have and luteinizing hormone (LH) by the gona-
shown that some of the fusiform cells of the dotropes. FSH carried to the ovary in the
theca externa have the ultrastructural fea¬ general circulation stimulates growth and
tures of smooth muscle cells. However, their maturation of ovarian follicles. The growing
role in ovulation is questionable. Contraction follicles, in turn, secrete progressively in¬
of circumferentially oriented muscle cells in creasing amounts of the steroid hormone
estradiol during the follicular phase of the
cycle. The rising level of estradiol is believed
to act back on the gonadotropes, inducing
them to accumulate LH and to a lesser extent
FSH. At midcycle, when the dominant follicle
attains maturity, the circulating estradiol
reaches a critical threshold level that exerts a
positive feedback on the hypophysis, altering
gonadotrope function from a phase of hor¬
mone accumulation to one of abrupt release.
The resulting surge of circulating LH trig¬
gers rupture of the follicle and discharge of
its ovum.
The ruptured follicle is rapidly trans¬
formed into a corpus luteum secreting in¬
creasing amounts of progesterone, which in¬
hibits the positive feedback of the ovary on
the hypophysis. If fertilization does not oc¬
Figure 32-17. Photomicrograph of an ovulating ovarian cur, the corpus luteum has a life span of
follicle from the rat. The cumulus with the enclosed egg
(at arrow) can be seen passing through the stigma.
about 14 days. With its involution, progester¬
(Courtesy of R. J. Blandau.) one and estrogen are no longer produced in
864 • FEMALE REPRODUCTIVE SYSTEM

appreciable amounts, and menstruation fol¬ throughout the reproductive life of the
lows. Continuing activity of the hypothala¬ woman one oocyte, as a rule, undergoes mat¬
mus then initiates a new cycle. uration and completes meiosis shortly before
It was formerly thought that the LH surge ovulation. This long interruption in conti¬
inducing ovulation depended on an in¬ nuity of the meiotic process, lasting from 12
creased release of luteinizing hormone—re¬ to 40 years, is one of the most remarkable
leasing hormone (LHRH) acting directly phenomena in reproductive biology and one
upon the hypophysis and that the target of of the least understood. Various substances
the estradiol feedback was the hypothalamus. have been postulated to account for the
Although an effect of ovarian steroids on switch of primordial germ cells from mitosis
hypothalamic function cannot be ruled out, to meiosis; the acceleration of the process to
it is now generally accepted that the feedback diplotene; and the arrest at the diplotene
of ovarian steroids acts mainly at the level of stage of prophase, but the experimental evi¬
the hypophysis rather than on the hypothal¬ dence for the existence of these substances is
amus. The inhibition of positive feedback by scant. There is some indication that cells in
progesterone is the basis for the widespread the rete ovarii derived from the mesonephros
contraceptive use of progesterone and its are responsible for induction of meiosis by
analogues to prevent ovulation. secreting a meiosis-inducing substance. The
The essential central component of the mechanism for arrest of meiosis is not under¬
system controlling the cycle in the female is stood, but the follicular fluid in small and
the hypothalamic pulse generator. The re¬ large follicles and the supernatant of granu¬
lease of pulses of LHRH at regular intervals losa cells in culture have been shown to
of 90 minutes in the human (60 minutes in inhibit resumption of meiosis and maturation
the macaque) is all that is required to initiate of large oocytes.
and maintain the menstrual cycle. In prepub¬ There has been some progress in our un¬
ertal primates, pituitary function is normal derstanding of the factors involved in re¬
in all respects except that little or no gonado¬ sumption of meiosis in the preovulatory fol¬
tropin is secreted. Pulsatile administration of licle. The trigger for completion of meiosis
LHRH to prepubertal monkeys or humans appears to be a surge in the release of LH
initiates puberty and the onset of normal occurring near the middle of the cycle. If
ovulatory cycles. In women who are arnenor- large follicles are removed from rat ovaries
rheic owing to a pathologic condition in the and placed in organ culture before the LH
hypothalamus, normal menstrual cycles can surge has occurred, the oocytes remain in
be induced by pulsatile administration of the diplotene stage. However, addition of LH
LHRH with a small computerized pump. to the culture medium, or microinjection of
dibutyryl cyclic AMP into the antrum, will
Maturation of the Oocyte induce resumption of meiosis and completion
of oocyte maturation in vitro. Thus, the LH
Before ovulation and fertilization, the surge seems to activate the prepared oocyte,
oocyte must complete meiosis, reducing the and the action of the hormone is mediated
diploid complement of chromosomes to the by the adenyl cyclase/cyclic AMP system.
haploid number. In the female, meiosis be¬ In the resulting division of the primary
gins early in fetal life. The primordial germ oocyte, the chromosomes are divided equally
cells in very early embryos are located in the between the daughter cells, but cytoplasmic
endoderm of the yolk sac. They subsequently division is exceptional, in that nearly all the
migrate along the root of the mesentery and cytoplasm remains with one daughter cell,
laterally into the germinal ridges that give the secondary oocyte. In this division the nu¬
rise to the ovaries. During their migration cleus of a primary oocyte moves to a position
they proliferate by mitosis, greatly increasing immediately beneath the oolemma. The nu¬
their number. Mitosis continues for some clear envelope is broken down and the con¬
time after the primordial germ cells take up densed chromosomes assemble on the meta¬
residence in the developing ovaries. The re¬ phase plate of a spindle that is oriented
sulting primary oocytes then enter prophase of paratangential to the oolemma (Fig. 32—18A).
the first meiotic division and then proceed I he spindle then rotates to a position per¬
to the diplotene stage. Meiosis is then ar¬ pendicular to the oolemma. A small, rounded
rested, and the oocytes remain in this state protrusion of cytoplasm appears on the sur¬
throughout the remainder of fetal life, child¬ face of the oocyte at the pole of the spindle.
hood, and puberty. Thereafter, each month At anaphase, half of the chromosomes are
 FEMALE REPRODUCTIVE SYSTEM • 865

drawn into this minute bleb of cytoplasm, to the surface of the ovary. Their active
which is pinched off to form the first polar sweeping movements over its surface, and
body, lying free in the perivitelline space be¬ the Currents ..created in the overlying him of
tween the secondary oocyte and the zona fluid by the cilia on its epithelial cells, are
pellucida (Fig. 32-185). The spindle of the responsible for drawing the ovum into the
second meiotic division forms almost imme¬ ostium of the oviduct.
diately with the chromosomes that remain in The newly ovulated tubal ovum in most
the oocyte arranged on the metaphase plate mammalian species is surrounded by a cor¬
(Fig. 32-19). From the resumption of meiosis ona radiata of adhering granulosa cells. With
until the attainment of this state occupies a the invasion of this mass of cells by sperma¬
period of about ten hours immediately pre¬ tozoa, their attachments to one another are
ceding ovulation, and the ovum remains ar¬ loosened. The gradual dispersion of the cells
rested in this condition until its fertilization of the corona radiata is attributed to a de¬
in the ampulla of the oviduct. polymerization of the intercellular substance
by enzymes released from the acrosomes of
the spermatozoa. Increased surface activity
Fertilization of the cells themselves may also be involved.
Upon reaching the zona pellucida, the
At the time of ovulation the turgid fronds
sperm head gradually penetrates this layer.
or fimbriae of the oviduct are closely applied
The details of this process are still not com¬
pletely understood. The spermatozoon re¬
mains actively motile during its penetration.
This movement may contribute to the proc¬
ess, but local lysis of the zona by enzymes of
the sperm acrosome is also believed to play
an important role in sperm penetration.
Once within the vitelline space, the move¬
ments of the sperm cease. The membrane
overlying the postacrosomal region of the
sperm head fuses with the oolemma, and the
sperm sinks into the ooplasm.
Fertilization proper consists of the entry of
the spermatozoon into the ovum. This event
somehow stimulates the ovum to complete
the second meiotic division and cast off the
second polar body. This is followed by fusion
of the egg and sperm nuclei to restore the
diploid chromosome number, and cleavage
of the zygote ensues.
If the ovum is not fertilized, it gradually
fragments and is absorbed or phagocytized.
The length of the period during which the
human ovum remains fertilizable is not pre¬
cisely known, but it is probably less than 24
hours.

Formation of the Corpus Luteum

Following ovulation and discharge of the
liquor folliculi, the wall of the follicle col¬
lapses, and its granulosa cell lining is thrown
into folds (Fig. 32-20). The basal lamina that
Figure 32-18. Photomicrographs of two stages in the
maturation of the rat ovum. A, Section of a rat egg shortly formerly separated the granulosa and the
before ovulation, showing the first polar spindle with theca interna is depolymerized. T here may
diploid chromosomes on the metaphase plate (arrow). be some associated extravasation of blood
(Courtesy of R. J. Blandau.) B, Ovulated egg recovered from the capillaries of the theca interna,
from the ampulla of the oviduct but before sperm penetra¬
tion. The first polar body lies in the perivitelline space
resulting in the formation of a central clot.
(arrow). The second maturation spindle can be seen just The cells of the plicated granulosa layer and
above it. (Courtesy of R. J. Blandau.) those of the theca interna then undergo strik-
866 • FEMALE REPRODUCTIVE SYSTEM

Figure 32-19. Photomicrograph of human follicular oocyte at completion of the first meiotic division. A first polar body
has been extruded, and the chromosomes and spindle of the second meiotic metaphase are visible. (Photomicrograph
courtesy of L. Zamboni.)

ing cytological alterations. They enlarge, ac¬ plasmic reticulum characteristic of steroid-
cumulate lipid, and are transformed into secreting cells (Figs. 32—23, 32—24). The cor¬
plump, pale-staining polygonal cells—the lu¬ pus luteum secretes the steroid hormone pro¬
tein cells. After these postovulatory changes gesterone.
have taken place, the follicle is called the While the cells of the collapsed follicular
corpus luteum. wall are undergoing luteinization, the capil¬
Early in the development of the corpus laries of the theca interna sprout and invade
luteum, two kinds of lutein cells are distin¬ the lutein tissue. Connective tissue elements
guishable. Those at the periphery that are also penetrate the developing corpus luteum
smaller and more deeply stained have been from its periphery, forming a delicate retic¬
called theca lutein cells in the belief that they ulum around the lutein cells and gradually
originate from cells of the theca interna. converting the resolving blood clot in the
Those in the interior that make up the bulk central cavity into a fibrous core.
of the lutein tissue have been called granulosa If the ovum is not fertilized, the ruptured
lutein cells (Figs. 32-21, 32-22). It is not clear follicle gives rise to a corpus luteum of menstrua¬
whether these two types are ontogenetically tion, which lasts for only about 14 days. Its
distinct or are merely transitional stages in rate of secretion of progesterone then drops
the development of a single type of lutein as it undergoes histological involution. The
cell. lutein cells become loaded with lipid and
The lipid in the lutein cells is dissolved in ultimately degenerate. In the succeeding
routine histological preparations, leaving nu¬ months the connective tissue cells become
merous vacuoles. Their vacuolated cytoplasm pyknotic, hyaline intercellular material accu¬
gives them an appearance reminiscent of the mulates, and the former corpus luteum is
cells of the adrenal cortex. In electron micro¬ reduced to a white scar, the corpus albicans
graphs they have mitochondria with tubular (Fig. 32—25). This slowly sinks deeper into
cristae and the abundant smooth endo¬ the interior of the ovary and gradually dis-
 FEMALE REPRODUCTIVE SYSTEM • 867

Theca lutein cells

Figure 32-20. Photomicrograph of section of corpus luteum from human ovary, x 11.

appears over a period of many months or to darker theca lutein cells, which are only
years. about 15 pm in diameter. In the luteinization
If ovulation is followed by fertilization, the of granulosa cells there is (1) a transforma¬
corpus luteum enlarges further and becomes tion of a dense heterochromatic pattern in
a corpus luteum of pregnancy, which persists for the nucleus to a more homogeneous nucleo¬
about six months and then gradually declines plasm with a single prominent nucleolus; (2)
up to full term. After delivery its involution a change in form of the mitochondria from
is accelerated and it undergoes changes lead¬ elongate organelles with lamelliform cristae
ing to the formation of a scar similar to that to highly pleomorphic mitochondria with a
left behind by the corpus luteum of men¬ dense matrix, with tubular cristae, and often
struation. with osmiophilic inclusions; (3) an increase in
The development of the corpora lutea has lipid droplets and in the smooth membranes
now been studied by electron microscopy in of the agranular reticulum, which take the
the human and in the rhesus monkey. In form of networks of anastomosing tubules or
addition to vascular and connective tissue concentric systems of cisternae; (4) evolution
invasion of the ruptured follicular epithe¬ of a single Golgi complex of the granulosa
lium, there is hyperplasia and hypertrophy cells to multiple small stacks of cisternae
of the cells. The two populations of lutein widely dispersed in the cytoplasm of the
cells identified with the-light microscope are lutein cells; (5) an increase in lysosomes and
also distinguishable in electron micrographs lipofuscin pigment deposits; and (6) devel¬
(Figs. 32-23, 32-24). The lighter-appearing opment of many microvilli on the surface of
granulosa lutein cells are very large, meas¬ granulosa lutein cells, that project into inter¬
uring up to 30 pm in diameter, in contrast cellular clefts and invaginations of the cell
868 • FEMALE REPRODUCTIVE SYSTEM

Granulosa iutein cells

Interstitial connective tissue

Blood
vessel

Granulosa lutein

Capsu le cel Is

of corpus
luteum

Blood vessel

Theca lutein cells

Figure 32-21. Drawing of a peripheral area of human corpus luteum of pregnancy, stained for reticular fibers by the
Bielschowsky method. The smaller darker cells at lower left are theca lutein cells, the large paler cells above are
granulosa lutein celis. (After A. A. Maximow.)

surface that are reminiscent of the intracel¬ metrium in pregnancy. It also promotes dil¬
lular canaliculi of gastric parietal cells. This atation of the cervix, and in some species
surface amplification is less apparent on the loosens the symphysis pubis, thus facilitating
theca lutein cells, which remain smaller and parturition.
generally have a somewhat less extensive
smooth endoplasmic reticulum, and more
numerous lipid droplets.
Atresia of Follicles
The cytological changes described for lu- In the human female during the early part
teinization in vivo have also been observed in of each cycle, a group of follicles starts to
tissue cultures of granulosa cells from pre¬ grow. Usually only one of these goes on to
ovulatory follicles. The acquisition of the ul- develop into a mature follicle, and all the
trastructural characteristics of lutein cells is others undergo a degenerative process called
correlated with the synthesis of increased follicular atresia. Some 99 per cent of the
amounts of progesterone. oocytes present in the ovary at birth are
Late in pregnancy, a new population of destined to degenerate. This depletion of the
small dense granules appears in the cyto¬ stock of oocytes begins in intrauterine life,
plasm of the lutein cells. These do not contain becomes prominent at birth and before pu¬
lysosomal enzymes. They are believed to rep¬ berty, and continues on a smaller scale
resent sites of storage of the polypeptide throughout reproductive life. Every normal
hormone relaxin, which has been localized to ovary, therefore, contains degenerating fol¬
the corpus luteum by fluorescent antibody licles. Why only a few follicles reach maturity
methods. The exact physiological role of this and rupture, while the great majority degen¬
hormone is still poorly understood, but it is erate at various stages of development, is not
known to inhibit contractions of the myo¬ known. The mechanism by which a single
 FEMALE REPRODUCTIVE SYSTEM • 869

Figure 32-22. A, Photomicrograph of human corpus luteum at nine weeks’ gestation. Larger cells above are granulosa
lutein cells; smaller ones below are theca lutein cells derived from the theca interna. B, Corpus luteum of the human
menstrual cycle about one day after ovulation. Cells of the theca interna have luteinized and are migrating into the
developing corpus luteum. (Photomicrographs from Crisp, T. M., et al. Am. J. Anat. 127:37, 1970.)

follicle is selected to complete its develop¬ the earliest indications of this atretic process
ment is also unknown. Nor do we have any is the invasion of the granulosa layer and
clear idea of the biological significance of this cumulus oophorus by strands of vascularized
wastage of oocytes. connective tissue. This is followed by a loos¬
Atresia may begin at any stage of devel¬ ening and shedding of the granulosa cells
opment of the follicle, even in ones that are into the follicular cavity and a hypertrophy
apparently mature. In a primary follicle of the theca interna. In follicles exhibiting
doomed to destruction, the ovum shrinks and these changes, the oocyte may still appear
degenerates, a process followed by the dis¬ normal in routine histological preparations.
solution of the granulosa cells. The resulting As the degeneration of granulosa cells ad¬
small cavity in the stroma is closed rapidly vances, the follicle collapses, its outlines be¬
without leaving a trace. In small secondary come wavy, and the cavity is filled by a large
follicles, the earliest sign of abnormality is number of fibroblasts. The remnants of the
often the eccentric location of the egg nu¬ degenerated follicular epithelium are rapidly
cleus, which goes on to develop a coarse resorbed. The folded and collapsed zona
granularity and finally becomes pyknotic. In pellucida may remain alone amid the con¬
follicles of larger size, the histological changes nective tissue elements.
in atresia become somewhat more complex The theca interna also undergoes impor¬
and variable. In the cyclic atresia of follicles tant changes. The basal lamina that separates
in the adult human ovary, the process ap¬ it from the epithelium often increases in
pears to be initiated in the follicle wall, with thickness and is transformed into a thick
secondary effects upon the oocyte. One of hyaline layer, the “glassy membrane," which
870 • FEMALE REPRODUCTIVE SYSTEM

Figure 32-23. Electron micrograph of a portion of a theca lutein cell of a human corpus luteum at nine weeks’ gestation.
The granular and agranular endoplasmic reticulum are well developed and lipid droplets are abundant. The mitochondria
are highly variable in size and have tubular cristae. (From Crisp, T. M., et al. Am. J. Anat. 727:37, 1970.)

is characteristic of follicles in advanced atre¬ Strands of fibrous connective tissue and

sia. The large cells of the theca interna in¬ blood vessels ultimately penetrate the glassy
crease further in size and are usually ar¬ membrane, and the remains of the degener¬
ranged in radial groups or strands, separated ated elements in the interior are broken
from one another by partitions of collagen¬ down. The resulting scar with its hyaline
ous fibers and smaller fusiform cells. The streaks sometimes resembles a corpus albi¬
cells acquire a typical epithelioid character cans derived from old corpora lutea, but is
and are filled with lipid droplets. They are usually much smaller, and sooner or later it
very similar in appearance to the theca lutein disappears in the stroma of the ovary. The
cells but reach a higher degree of develop¬ layer of hypertrophic theca interna cells sur¬
ment in the atretic follicle. The cavity of the rounding the atretic follicle is broken up by
atretic follicle, containing the collapsed zona the invading strands of hbrou^ tissue into
pellucida and connective tissue, is now sur¬ separate cell islands of various shapes and
rounded by a broad, festooned layer of epi¬ sizes. These islands are irregularly scattered
thelioid, lipid-containing theca interna cells, in the stroma and may persist for a time.
arranged in radial cords and provided with They contribute to the so-called “interstitial
a rich capillary network. The microscopic gland” of the ovary.
appearance of such an atretic follicle is rather
similar to that of an old corpus luteum. Such
The Interstitial Tissue of the Ovary
structures have therefore been given the mis¬
leading name corpora lutea atretica. The main The stroma of the human ovarian cortex
differences are, of course, the presence of consists of spindle-shaped cells and networks
the glassy membrane, degenerated granulosa of reticular fibers. Elastic fibers occur only in
cells, and sometimes a zona pellucida in the the walls of blood vessels. The cells bear a
center. superficial resemblance to smooth muscle but
 FEMALE REPRODUCTIVE SYSTEM • 871

Figure 32-24. Electron micrograph of a portion of human granulosa lutein cell at nine weeks’ gestation. As in other
steroid-producing endocrine glands, there is an extensive development of tubular smooth reticulum, and lesser amounts
of the granular form. (From Crisp, T. M., et al. Am. J. Anat. 127:37, 1970.)

do not have myofilaments in their cytoplasm. extensive. Cell clumps originating from the
The endocrine function and develop¬ breaking up of the hypertrophied theca in¬
mental potentialities of the layer of special¬ terna of atretic follicles persist, enlarge, and
ized interstitial tissue composing the theca fuse. Through the continuous addition of
folliculi have already been described. The new cells, a large part of the organ is ulti¬
interstitial tissue of the ovarian cortex consists mately transformed into a diffuse mass of
of reticular fibers and spindle-shaped cells large, closely packed, lipid-containing inter¬
that have potentialities distinct from those of stitial cells that are almost identical in ap¬
ordinary fibroblasts. The medulla, on the pearance to lutein cells. The follicles and the
other hand, is made up of more typical loose corpora lutea are embedded in this cell mass,
connective tissue with fibroblasts, many elas¬ and only a thin tunica albuginea separates it
tic fibers, and strands of smooth muscle cells from the germinal epithelium on the surface.
accompanying the blood vessels. The interstitial gland is less well developed
In many mammals the ovarian stroma also in the human ovary. Interstitial cells are
contains conspicuous clusters and cords of found in the greatest numbers during the
large epithelioid interstitial cells. They are first year of life, when atretic follicles are
rich in lipid and strikingly resemble lutein most numerous, and they are believed to
cells. In some species they have been shown arise from the hypertrophied theca interna
to secrete estrogen. Because of their epithe¬ of regressing follicles. The interstitial gland
lioid appearance and presumed secretory involutes at puberty with the onset of men¬
functions, these cells, dispersed in the stroma, struation and the cyclic development of cor¬
are referred to collectively as the interstitial pora lutea. In the adult human ovary, cells
gland. In animal species that have large litters, of this kind are present only in small numbers
particularly among the rodents, the devel¬ widely scattered in the stroma.
opment of the interstitial gland may be very In the hilus of the ovary and in the adjacent
872 • FEMALE REPRODUCTIVE SYSTEM

Figure 32-25. Corpus albicans of human ovary. Fixation by perfusion—hence the empty vessel (V). Dense hyaline
material separates the residual cells of the corpus luteum. The whole structure is surrounded by the stroma of the
ovary, x 135.

mesovarium, groups of another kind of large ward the oviduct and fusing with a longitu¬
epithelioid cell may be found closely associ¬ dinal canal parallel to the oviduct. All of
ated with vascular spaces and unmyelinated these tubules end blindly. They are lined
nerve fibers. These cell clusters, now simply with low cuboidal or columnar epithelium,
called hilus cells, were originally named the which is sometimes ciliated, and are sur¬
sympathicotropic hilus gland and were consid¬ rounded by a condensed connective tissue
ered to be chromaffin cells. This view is now layer containing smooth muscle. The upper
less widely accepted, since they do not always end of the longitudinal duct sometimes ends
stain with chromates. Moreover, convincing in a cystlike enlargement, the hydatid of Mor¬
evidence has been presented that they are gagni, while its other end may extend far
similar to the Leydig cells of the testis. They toward the uterus as the so-called duct of
are rich in lipid, contain cholesterol esters, Gartner. The transverse tubules and the lon¬
and lipochrome pigment, and may even have gitudinal duct of Gartner together comprise
cytoplasmic crystals apparently identical to the epoophoron. Between the epoophoron and
the crystals of Reinke (see Chapter 31). They the uterus in the tissue of the broad ligament
have the histochemical and cytological char¬ is another group of irregular fragments of
acteristics of actively secreting endocrine epithelial tubules, the paroophoron. The epoo¬
cells. They are prominent during pregnancy phoron is a rudiment of the embryonic me¬
and at the menopause. Tumor or hyperplasia sonephros and is the homologue of the duc-
of the ovarian hilus cells is accompanied by tuli efferentes and epididymis of the male.
masculinization. This clinical observation and The paroophoron is the remnant of the cau¬
their cytological and cytochemical resem¬ dal part of the mesonephros and corresponds
blances to Leydig cells suggest that they se¬ to the vestigial paradidymis of the male.
crete androgens.
In the broad ligament and in the mesovar¬
Vessels and Nerves
ium, the occurrence of small accumulations
of “interrenal tissue” corresponding to ad¬ The principal arterial supply to the ovary
renocortical tissue has also been described. is from the ovarian artery, which arises from
the aorta below the level of the renal vessels
Vestigial Organs Associated with the and reaches the ovary through the infundib-
Ovary ulopelvic ligament. Along the mesovarial bor¬
der of the ovary this vessel anastomoses with
Certain vestigial organs are found in con¬ the uterine artery, which courses upward
nection with the ovary. The most obvious of along the lateral aspect of the uterus from
these is the epoophoron. It consists of several the region of the cervix. Relatively large
parallel or divergent tubules, running in the vessels from the region of anastomosis of the
mesovarium from the hilus of the ovary to¬ uterine and ovarian arteries enter the hilus
 FEMALE REPRODUCTIVE SYSTEM • 873

of the ovary and branch profusely as they fibers penetrate into the cortex and form
course through the medulla. Because of their plexuses around the follicles and under the
tortuous course, they are called arteriae heli- germinal epithelium. It seems doubtful that
cinae, or helicine arteries. These vessels, like they penetrate through the basal lamina into
those in the corpora cavernosa penis, may the epithelium of the follicles. Sensory fibers
show longitudinal ridges on their intima. In ending in corpuscles of Pacini have been
the periphery of the medulla they form a described in the ovarian stroma.
plexus, from which smaller twigs penetrate
radially, passing between the follicles to enter
the cortex, where they break up into loose
networks of capillaries. These are continuous
THE OVIDUCT OR FALLOPIAN
with dense networks in the theca of the larger TUBE
follicles. The veins accompany the arteries.
In the medulla they are large and tortuous The oviduct or fallopian tube is the part of
and form a plexus in the hilus. the female reproductive tract that receives
Networks of lymph capillaries arise in the the ovum, provides the appropriate environ¬
cortex, especially in the theca externa of the ment for its fertilization, and transports it to
large follicles. Lymph vessels with valves are the uterus. It is a muscular tube about 12 cm
found only outside the hilus. long situated in the edge of the mesosalpinx,
The nerves of the ovary are derived from which is the upper free margin of the broad
the ovarian plexus and from the uterine ligament of the uterus. Its lumen communi¬
nerves. They enter the organ through the cates with the uterine cavity at one end and
hilus, together with the blood vessels. They is open to the peritoneal cavity at the other.
consist, for the most part, of nonmyelinated Several segments along its length are identi¬
fibers, but thin myelinated fibers are also fied by different descriptive terms (Fig.
present. The reported presence of sympa¬ 32-26). The part of the tube traversing the
thetic nerve cells in the ovary has not been wall of the uterus is called the pars interstitialis.
confirmed. The majority of the nerves supply The narrow medial third near the uterine
the muscular coat of blood vessels. Many wall is the isthmus. The expanded intermedi-

Isthmus Interstitial
segment

Infundibulum

Figure 32-26. Drawing illustrating the mucosal pattern of the various segments of the human oviduct and the
topographical relationship of the oviduct to the ovary. (From Eastman, M. J., and L. M. Heilman, eds.: Williams
Obstetrics. 13th ed. New York, Appleton-Century-Crofts, 1961. Labeling added.)
874 • FEMALE REPRODUCTIVE SYSTEM

ate segment is the ampulla> and the funnel- the ampulla, is provided with cilia that beat
shaped abdominal opening is the infundibu¬ toward the uterus (Fig. 32—30). The other
lum. The margins of the latter are drawn out cell type is devoid of cilia and is commonly
into numerous tapering, fringelike processes, considered to be secretory. The secretion
the fimbriae. may provide the ovum with nutritive mate¬
rial, and in some species, notably the rabbit,
it adds to the ovum an outer albuminous
Histological Organization
envelope. In the monotremes and some mar¬
The wall of the oviduct consists of a mu¬ supials, a shell, as well as an albuminous coat,
cosa, a muscular layer, and an external serous is formed around the ova. The two types of
coat. The mucosa in the ampulla is thick and epithelial cells are probably different func¬
forms numerous elaborately branched folds. tional states of a single cell type. In women
The lumen in cross section, therefore, is a the epithelium of the oviduct undergoes
labyrinthine system of narrow spaces between cyclic changes along with those of the uterine
profusely branching folia covered by epithe¬ mucosa. True glands are absent in the ovi¬
lium (Figs. 32-26, 32-27C). In the isthmus duct.
the longitudinal folds are much shorter and The relative proportions of ciliated and
less highly branched (Fig. 32—27B), and in nonciliated cells is under endocrine control.
the interstitial part they are reduced to low Ciliated cells are said to increase in height in
ridges (Fig. 32-27A). the human ovidtict during the follicular
The epithelium is of the simple columnar phase of the cycle and to decrease during the
variety (Figs. 32-28, 32-29A) but may some¬ luteal phase, but they do not seem to lose
times appear pseudostratihed when cut their cilia completely. The cyclic changes
obliquely. It is highest in the ampulla and have been most thoroughly studied in the
diminishes in height toward the uterus. It rhesus monkey. The changes are most
consists of two kinds of cells. One of these, marked in the Umbria and upper ampulla,
especially numerous on the fimbriae and in and diminish toward the isthmus. On the
A

Figure 32 27. Photomicrographs of the fallopian tube of a 23-year-old woman, x 30. A, The pars interstitialis; B, the
isthmus, C, the outer portion of the ampulla. Area enclosed in rectangle is shown at higher magnification in Figure
32—28.
 FEMALE REPRODUCTIVE SYSTEM • 875

other parts of the body are completely indif¬

ferent to circulating estrogens.
Steroid hormones also appear to affect the
rate of ciliary beat. A significant increase has
been reported 48 hours after copulation in
the rabbit and after progesterone treatment
of the estrogen-primed monkey oviduct.
Thus, estrogen seems to prepare the ciliated
surface destined to transport the ovum, and
progesterone accelerates the ciliary beat at
the time an ovum is available to be trans-

Figure 32-28. Photomicrograph of the branching folds

of the mucous membrane of the human fallopian tube,
x 280. For orientation, see rectangle in Figure 32-27.

fimbria, the epithelium becomes devoid of

cilia and nonsecretory in the late luteal phase
of the cycle. In the early follicular phase, the
cells hypertrophy and begin active ciliogene-
sis. There is also cytological evidence of se¬
cretory activity. These changes reach their
peak at midcycle. Dedifferentiation is so com¬
plete on the fimbriae in the late luteal phase
that one cannot distinguish deciliated cells
from atrophic secretory cells. The degree of
dedifferentiation is less marked toward the
isthmus, suggesting that this segment of the
oviduct is less estrogen dependent.
After ovariectomy the oviductal epithelium
rapidly atrophies and dedifferentiates (Fig.
32—29B). Within a few weeks the fimbriae
are almost completely deciliated and all the
cells are structurally indistinguishable (Fig. 32-
29B). In the ampulla some cells retain their Figure 32-29. A, Photomicrograph of oviductal epithelium
cilia, and in the isthmus regression is still less from a normal macaque in the follicular phase of the
marked. Similar dedifferentiation after ovar¬ cycle. Ciliated and nonciliated cells are distinguishable.
iectomy is observed in the rabbit and in other B, oviductal epithelium of a macaque six weeks after
ovariectomy. The cells are shorter and nonciliated, and
species (Fig. 32-31). Within a week after
two types of cells are no longer distinguishable. (From
administration of estradiol, there is a remark¬ Brenner, R. In Hafez, E. S., and R. J. Blandau, eds.: The
able hypertrophy, with restoration of cilia Mammalian Oviduct. Chicago, University of Chicago
and secretory activity. Ciliated epithelia in Press, 1969.)
876 • FEMALE REPRODUCTIVE SYSTEM

Figure 32-30. Scanning electron micrograph of the surface of the epithelium on the fimbria of rabbit oviduct in thf
postovulatory period, showing the dense ciliation and the convex apices of the secretory cells covered with microvilli
(From Rumery, R. E., and E. M. Eddy. Anat. Rec. 178:83, 1974.)

ported. Later in the cycle, progesterone fa¬ At the time of ovulation the oviduct ex¬
vors the loss of cilia. hibits active movements. The abdominal
The lamina propria of the mucosa in the opening of the oviduct contains large blood
oviduct consists of a network of reticular vessels in its mucosa, especially veins, and
fibers and of numerous fusiform cells. Lym¬ these extend into the fimbriae. Smooth mus¬
phocytes, monocytes, and mast cells also oc¬ cle bundles form a network between the
cur in limited numbers. The fixed cells here blood vessels. This results, in effect, in a sort
seem to have the same developmental poten¬ of erectile tissue. At the time of ovulation the
tialities as those in the stroma of the uterus. vessels are engorged with blood, and the
In cases of tubal pregnancy, some of them resulting enlargement and turgescence of the
are transformed into typical decidual cells. fimbriae, together with the contraction of
No true muscularis mucosae can be distin¬ their intrinsic muscle, brings the opening of
guished in the oviduct. The mucosa is sur¬ the tubal infundibulum into contact with the
rounded directly by the muscularis, which surface of the ovary.
consists of two layers of smooth muscle bun¬ The rhythmic contractions of the oviduct
dles. The inner layer is circular or spiral; the are probably of primary importance in the
outer is principally longitudinal, but there is transport of the ovum. Contraction waves
no distinct boundary between the two. To¬ pass from the infundibulum to the uterus,
ward the periphery, longitudinal bundles and the beat of the cilia on the mucosa is in
gradually appear in increasing numbers the same direction.
among the circular bundles. The smooth
muscle bundles are embedded in an abun¬
dant, loose connective tissue, and they extend Blood Vessels, Lymphatics, and
into the broad ligament. Toward the uterus
Nerves
the muscularis increases in thickness. The The mucosa and its folds, as well as the
peritoneal coat of the fallopian tube has the serous coat, contain abundant blood and
usual serosal structure. lymph vessels. The lymph channels within
 FEMALE REPRODUCTIVE SYSTEM • 877

Figure 32-31. Scanning micrographs illustrating the dependence of cytological differentiation of the oviductal epithelium
upon hormones. A, From the fimbria of a rabbit oviduct 16 months after ovariectomy. The epithelium is flat and smooth,
with only occasional ciliated cells. B, The other oviduct of the same rabbit after receiving estrogen replacement for ten
days. (From Rumery, R. E., and E. M. Eddy. Anat. Rec. 178:83, 1974.)

the folds of the mucosa are extensive and sary for sustenance of the embryo through¬
appear in sections as long clefts that are often out its development. In the human it is a
mistaken for artifactitious splits in the tissue, single pear-shaped organ with a thick mus¬
but careful inspection reveals their smooth cular wall (Fig. 32-32). It is slightly flattened
endothelial lining. In periods of vascular en¬ dorsoventrally and contains a corresponding
gorgement, when these lymphatics are also flattened uterine cavity. In the nonpregnant
distended with lymph, they no doubt contrib¬ condition, the uterus is about 6.5 cm long,
ute to the increased turgor of the tissue and 3.5 cm wide, and 2.5 cm thick. Several re¬
stiffen the mucosal folds. gions are distinguished. The expanded up¬
Larger nerve bundles are found accompa¬ per portion, constituting the bulk of the or¬
nying the vessels in the serous layer and in gan, is called the body or corpus uteri. The
the peripheral parts of the longitudinal mus¬ rounded upper end of the body, where the
cle. The circular muscle layer contains a oviducts join the uterus, is often referred to
dense plexus of thin nerve bundles supplying as the fundus. The slightly constricted portion
the muscle fibers and penetrating into the below the corpus is the isthmus, and the cylin¬
mucosa. drical lower part is the cervix. The portion of
the cervix that protrudes into the vagina is
the portio vaginalis. The slender cervical canal
that passes from the uterine cavity down
UTERUS through the cervix opens into the vagina at
the external os.
The uterus is the portion of the reproduc¬ A serous membrane, the peritoneum, cov¬
tive tract that receives the fertilized ovum ers the fundus and much of the posterior
from the oviduct, provides its attachment, aspect of the uterus. The peritoneum is re¬
and establishes the vascular relations neces¬ flected onto the bladder anteriorly and onto
878 • FEMALE REPRODUCTIVE SYSTEM

Myometrium Broad
ligament

Uterine
Endometrium lumen

Figure 32-32. Photomicrograph of the uterus of a macaque in transverse section, illustrating the relative thickness of
the myometrium and the late proliferative endometrium. x9. (Courtesy of H. Mizoguchi.)

the rectum posteriorly, so that this layer is muscle bundles are predominant. In the suc¬
found only on part of the surface of the ceeding stratum supravasculare, the fibers are
uterus. The greater part of the thickness of mainly circular and longitudinal. Finally, the
the uterine wall is smooth muscle, the myo¬ outermost stratum subserosum is a thin longi¬
metrium. The uterus is lined by a glandular tudinal muscle layer. The two most superfi¬
mucosa called the endometrium. cial layers send muscular bundles out into
the wall of the oviduct and into the broad
ligament and the round ligament.
Myometrium The smooth muscle cells of the myome¬
trium ordinarily have a length of about 50
The smooth muscle fibers of the muscular pm. In pregnancy, when the mass of the
layer are arranged in cylindrical or flat bun¬ uterus increases about 24 times, they hyper¬
dles separated by thin septa of connective trophy to a length of more than 500 pm.
tissue. Several layers can be distinguished in Although smooth muscle hypertrophy ac¬
the myometrium according to the direction counts for much of the enlargement of the
and disposition of the bundles. The layers gravid uterus, there also appears to be an
are not sharply demarcated, however, be¬ increase in the number of muscle fibers
cause fiber bundles cross over from one layer through division and possibly through trans¬
into another. formation of persisting embryonic connective
Immediately beneath the mucosa is a thin tissue cells into new muscular elements, es¬
layer of smooth muscle called the stratum pecially in the innermost layers of the myo¬
submucosum. Its fibers are predominantly lon¬ metrium. There is also a marked increase in
gitudinal, but some oblique and circular bun¬ the amount of connective tissue as indicated
dles may be found. This layer forms distinct by a fivefold increase in the amount of col¬
muscular rings around the intramural por¬ lagen. There is evidence that smooth muscle
tions of the oviducts and may have a sphinc¬ cells can synthesize collagen in response to
ter-like action. The next layer is the thickest estrogen stimulation. During return of the
and is called the stratum vasculare, because it uterus to normal size after delivery, the mus¬
contains many large blood vessels that give it cle cells rapidly diminish in size. It is possible
a spongy appearance. Circular and oblique that some of them degenerate.
 FEMALE REPRODUCTIVE SYSTEM • 879

The connective tissue between the muscu¬ responsiveness of the uterine muscle to oxy¬
lar bundles consists of collagenous fibers, tocin increases during the last few months of
bbroblasts, undifferentiated connective tissue pregnancy, and the rate of secretion of oxy¬
cells, macrophages, and mast cells. A typical tocin by the neurohypophysis increases at the
argyrophilic reticulum surrounds the smooth onset of labor. Thus, there is reason to be¬
muscle cells and is continuous with the col¬ lieve that this hormone plays an important
lagenous tissue septa between muscle bun¬ role in preparation of the uterus for partu¬
dles. Elastic networks are especially promi¬ rition. Oxytocin is sometimes used by the
nent in the peripheral layers of the uterine obstetrician to initiate labor or to increase the
wall. From there they extend inward between effectiveness of the uterine contractions.
the muscle bundles. The innermost layers of Another class of compounds that affect the
the myometrium contain no elastic fibers, uterine musculature is the prostaglandins. The
except those in the walls of the blood vessels. fetal membranes release prostaglandins in
The cervix is composed mainly of dense high concentration during labor and this may
collagenous and elastic fibers, among which increase the strength of the uterine contrac¬
are distributed bbroblasts and smooth muscle tions. Although the mechanism of action of
cells. The dense bbrous nature of the cervix prostaglandins on smooth muscle is not well
accounts for the very brm consistency of this understood, they are now widely used to
segment of the uterus. induce abortion.
Histophysiology of the Myometrium. The Contractility of the uterus is also increased
maintenance of normal size and cytological by mechanical factors. Stretching of the wall
differentiation of uterine smooth muscle cells of hollow viscera in general increases smooth
depends on estrogen produced by the ovary muscle contractility. The greater stretching
and carried to the uterus in the blood. In the of the uterine wall is believed to account for
absence of estrogen, uterine smooth muscle the fact that twin pregnancies terminate, on
atrophies. the average, at least two weeks earlier than
Contractions of the myometrium are very single pregnancies. Stretching or irritation of
largely responsible for expulsion of the fetus the cervix also appears to induce uterine
at parturition. In the normal nonpregnant contractions. Obstetricians take advantage of
uterus, the musculature is continually this when they rupture the fetal membrane
undergoing shallow, intermittent myogenic to induce labor. In the absence of amniotic
contractions that are not attended by any fluid, the fetal head irritates and stretches
subjective sensation. These may become ex¬ the endocervix, initiating reflexes that result
aggerated during sexual stimulation or dur¬ in contraction of the body of the uterus.
ing menstruation, resulting in cramplike
pain. The factors regulating this activity are Endometrium
poorly understood.
When pregnancy ensues, the activity of the The primary functions of the endome¬
myometrium is greatly reduced. Experiments trium are preparation for implantation of a
on animals have led to a widely held belief fertilized ovum, participation in implanta¬
that the level of progesterone produced by tion, and formation of the maternal portion
the corpus luteum of pregnancy inhibits my- of the placenta. The structural and functional
ometrial contractility, and that withdrawal of changes of the endometrium are dependent
this inhibition at the end of gestation initiates on the hormones secreted by the ovaries.
labor. This theory is now seriously chal¬ Upon removal of the ovaries, the endome¬
lenged. An unequivocal demonstration of an trium atrophies. Upon administration of es¬
effect of progesterone on the human uterine trogen, there is a rapid increase in the blood
muscle is lacking, and in several other animal flow to the uterus, the endometrium becomes
species, progesterone has been shown to have edematous, its cells begin to proliferate and
no effect. Cross circulation experiments be¬ hypertrophy, and there is a marked increase
tween pregnant and nonpregnant animals in its metabolic activity.
clearly show that some other blood-borne Beginning with puberty at 11 to 15 years
substance plays a more important role than of age and continuing until menopause at age
progesterone. There is some reason to be¬ 45 to 50, the uterine mucosa undergoes
lieve that this effect may be mediated.by the monthly cyclic changes in its structure in
hormone relaxin. response to rhythmic variations in the secre¬
Uterine contractility is increased by oxyto¬ tion of ovarian hormones. At the end of each
cin, a hormone of the neurohypophysis. The cycle there is a partial degeneration and
880 • FEMALE REPRODUCTIVE SYSTEM

sloughing of the endometrium, accompanied vessels from the other side. Branches from
by a more or less abundant extravasation of the arcuate arteries penetrate the deeper
blood. The products of these destructive layers of the myometrium to reach the en¬
changes appear as a bloody vaginal discharge, dometrium. Where they cross the myome-
the menstrual flow, which normally continues trial-endometrial junction, they give off small
for three to five days. basal arteries supplying the deepest portion of
The endometrium is approximately 5 mm the endometrium, the basalis or stratum basale
in thickness at the height of its development (the portion that is not sloughed off during
in a normal menstrual cycle. It consists of a menstruation). Continuing into the thicker
surface epithelium invaginated to form nu¬ layer commonly called the functionalis, the
merous tubular uterine glands that extend arteries are unbranched but highly con¬
down into a very thick lamina propria, usu¬ torted. These “coiled or spiral arteries” ram¬
ally referred to as the endometrial stroma. ify into arterioles that supply a rich capillary
The surface epithelium is simple columnar bed in the superficial portion of the endo¬
and is composed of a mixture of ciliated and metrium. The thin-walled veins form an ir¬
secretory cells. The epithelium of the uterine regular anastomosing network with sinus¬
glands is similar, but the ciliated cells are oidal enlargements at all levels of the
fewer. The direction of ciliary heat is said to endometrium. During most of the cycle, the
be upward in the glands and toward the coiled arteries constrict and dilate rhythmi¬
vagina on the endometrial surface. The cally, so that the surface is alternately
glands are, for the most part, simple tubules, blanched or suffused with blood. These ves¬
but they may show some bifurcation in the sels play an important role in initiating men¬
zone adjacent to the myometrium. They oc¬ struation.
casionally penetrate a short distance among
the muscle bundles. Under pathological con¬ Cyclic Changes in the Endometrium
ditions the myometrium may be extensively
invaded in this manner, a change described In the course of a normal menstrual cycle,
by the term adenomyosis. In old age, the en¬ the endometrium passes through a continu¬
dometrium atrophies and becomes thin. The ous sequence of morphological and func¬
openings of the glands may become partly tional changes, but for convenience of de¬
obliterated and they then become distended scription the cycle is divided into three
to form small cysts. recognizable stages that are correlated with
The endometrial stroma strongly resem¬ the functional activities of the ovary. These
bles mesenchyme. Its irregularly stellate cells are the proliferative, the secretory, and the men¬
have large, ovoid nuclei. The cell processes strual phases. The proliferative phase coin¬
appear to be in contact throughout the tissue cides with the period of growth of the ovarian
and adhere to a delicate framework of retic¬ follicles and their secretion of estrogenic hor¬
ular fibers. Elastic fibers are absent except in mone. The secretory phase is the period
the walls of the arterioles. There is an extra¬ when the corpus luteum is functionally active
cellular matrix, which at some phases of the and secreting progesterone, and the men¬
cycle is rich in metachromatic glycoprotein. strual phase ensues when the hormonal stim¬
In the interstices of the reticulum and stellate ulation of the endometrium by the ovaries
cells are lymphoid cells and granular leuko¬ rapidly declines (Fig. 32—33).
cytes. Macrophages are not uncommon but, The Proliferative or Follicular Phase.
for some unknown reason, they are not mo¬ During this phase, which begins at the end
bilized to phagocytize the blood extravasated of the menstrual flow, there is a two- to
in menstruation. threefold increase in thickness of the endo¬
A knowledge of the blood supply of the metrium. Mitoses are numerous in the epi¬
endometrium is of special importance for an thelium and in the stroma. The straight tu¬
understanding of the mechanisms of men¬ bular glands increase in number and in
struation and of placentation. From the uter¬ length (Fig. 32-34). Their epithelium is co¬
ine arteries that course in the broad ligaments lumnar, and the lumen narrow. The extra¬
along the sides of the uterus, branches pen¬ cellular matrix of the stroma is abundant and
etrate to the stratum vasculare of the myo¬ metachromatic. The coiled arteries are elon¬
metrium. In this layer, circumferentially ori¬ gating but are only moderately convoluted
ented arcuate arteries run toward the midline, and do not yet extend into the superficial
where they anastomose with corresponding third of the endometrium. Toward the end
 FEMALE REPRODUCTIVE SYSTEM 881

PITUITARY GLAND

PITUITARY GONADOTROPIC HORMONES <- ANT LOBE

DAYS
A A
O 3-5

.VIOUS CORPUS LUTEUM

AND
ESTROGENS

DEVELOPING FOLLICLE'

ESTROGENS PROGESTERONE ano

PR O L R ATy O,N
y3A PROGESTERONE
-RE LAX-IN

R T E FCHM E

Figure 32-33. Drawing depicting the morphological changes in the ovary and the endometrium in the course of the
menstrual cycle, and the hormones controlling these changes. (From Eastman, N. J., ed.: Williams Obstetrics. 11th ed.
Englewood Cliffs, NJ, Appleton-Century-Crofts, 1956.)

Figure 32-34. Photomicrographs of human endometrium in different days of the cycle. A, Proliferative endometrium of
the ninth day. B, Early secretory endometrium, 15th day. C, Secretory endometrium, 19th day. D, Gestational hyperplasia,
12th day of pregnancy. (Courtesy of A. T. Hertig.)
882 • FEMALE REPRODUCTIVE SYSTEM

of this phase, the glands become somewhat endometrium by ovarian hormones declines
sinuous and their cells begin to accumulate and marked vascular changes take place. The
glycogen (Fig. 32—346). coiled arteries constrict, so that the superficial
The proliferative growth of the endome¬ zone of the endometrium is blanched for
trium may continue for a day after ovulation, hours at a time. The glands cease secreting;
on about day 14 of an ideal 28-day cycle. the height of the endometrium shrinks some¬
There may be some diapedesis of erythro¬ what, owing to loss of interstitial fluid; and
cytes into the stroma beneath the surface the stroma appears more cellular and stains
epithelium, and rarely a little blood may more deeply. Many leukocytes are found in
enter the uterine lurnen and reach the va¬ the stroma at this time. After about two days
gina. Such intermenstrual bleeding is rare in the of intermittent ischemia, the coiled arteries
human, but is common in the dog. close down, making the superficial zone is¬
When the endometrium has been prepared chemic, while blood continues to circulate in
by the normal sequential action of estrogen the basal zone. After a variable number of
and progesterone, it is capable of undergoing hours, the constricted arteries open up for a
decidualization—a change in which its stromal short time; the walls of the damaged vessels
cells transform into large, pale decidual cells near the surface burst, and blood pours into
rich in glycogen. The normal stimulus for the stroma and soon breaks out into the
this transformation is an implanting blasto¬ uterine lumen. Normally such blood does not
cyst, but electrical stimulation, intraluminal clot. Subsequently, patches of blood-soaked
injection of oil, or simple mechanical trau¬ tissue separate off, leaving the torn ends of
matization of the endometrium will induce glands, arteries, and veins open to the sur¬
the same changes and result in formation of face. Blood may ooze from such veins, re¬
a mass of decidual cells, called a deciduoma. fluxing from the intact basal circulation. The
The exact function of the decidual tissue is menstrual discharge thus contains (1) altered
still a subject of debate, but there is agree¬ arterial and venous blood, with hemolyzed,
ment that it provides a favorable milieu for and sometimes agglutinated erythrocytes; (2)
nourishment of the conceptus and creates a partially intact, autolyzed epithelial and
specialized layer facilitating dehiscence of the stroma cells; and (3) the secretions of the
placenta at the termination of pregnancy. uterine and cervical glands. Sometimes there
The Secretory or Luteal Phase. In this are tissue fragments in the menstrual dis¬
phase of the cycle, some further thickening charge, but blood clots are considered abnor¬
of the endometrium occurs, but this is largely mal. The average loss of blood is 35 ml. By
attributable to edema of the stroma and to the third or fourth day of the flow, the entire
the accumulation of secretion in the uterine lining of the uterus presents a raw-appearing
glands. The glandular epithelium early in surface.
the secretory phase of the endometrium The endometrium deep to the zone of
shows a characteristic displacement of the extravasation remains intact during men¬
nuclei toward the free surface, owing to the struation, although it does shrink down. The
accumulation of a large amount of glycogen deep ends of glands typical of the secretory
in the basal cytoplasm. This appearance is phase are recognizable as such until the end
transient and is no longer seen after active of menstruation. Before the vaginal dis¬
secretion is established. The glands continue charge has ceased, epithelial cells glide out
to grow, becoming more tortuous and ulti¬ from the torn ends of the glands, and the
mately developing a marked sacculation, re¬ surface epithelium is quickly restored. The
sulting in a relatively wide lumen of irregular superficial circulation is resumed; the stroma
outline, containing a carbohydrate-rich secre¬ again becomes rich in ground substance; and
tion (Fig. 32-34C). the proliferative activity of the follicular
The elongation and convolution of the phase of the new cycle begins.
coiled arteries continues in this phase. They The typical thick secretory endometrium
extend into the superficial portion of the (Fig. 32-33) is not always attained. In some
endometrium and become more prominent cycles, the ovary may not produce a mature
in sections because of the hypertrophy of the follicle. In such anovulatory cycles the endo¬
periarterial stromal cells. metrial changes are minimal. The prolifera¬
The Menstrual Phase. About two weeks tive endometrium develops as usual, but since
after ovulation, in a cycle in which fertiliza¬ there is no ovulation and no corpus luteum
tion fails to occur, the stimulation of the is formed, the endometrium does not pro-
 FEMALE REPRODUCTIVE SYSTEM • 883

gress to the secretory phase but continues to the base of the cells, and the greater part of
be of the proliferative type until menstrua¬ the cytoplasm is filled with mucus. The mu¬
tion begins. cosa contains numerous large glands, which
The various morphological changes in the differ from those of the corpus and isthmus
normal menstrual cycle are so characteristic in that they are extensively branched and are
and reproducible that an experienced pa¬ lined with tall, mucus-secreting columnar
thologist can establish the day of the cycle cells similar to those of the surface epithe¬
with surprising accuracy from examination lium. Occasional cells are ciliated. The cer¬
of endometrial curettings or biopsies (Fig. vical canal is usually filled with mucus. In the
32-35). It is also possible from examination human, the ducts of some of the glands not
of biopsies in the second half of the cycle to infrequently become occluded, and accumu¬
determine whether a woman is having an lation of secretion then transforms these
ovulatory or an anovulatory cycle. Such ex¬ glands into cysts that may reach 5 to 6 mm
aminations are essential in clinical investiga¬ in diameter. These are called nabothian cysts.
tion of the causes of infertility, or detection The outer surface of the portio vaginalis
of dysfunction of the ovaries, or disorders of of the cervix is smooth and covered with a
menstruation. It is therefore of practical stratified squamous epithelium similar to that
value to be able to recognize the principal of the vagina (Fig. 32—49). The cells of this
phases of the endometrial cycle. The criteria, epithelium are rich in glycogen. The transi¬
in brief, are as follows. (1) Proliferative or tion between the columnar mucus-secreting
follicular phase: endometrium 1 to 5 mm thick; epithelium of the cervical canal and the strat¬
straight, narrow glands becoming wavy; the ified squamous epithelium of the portio va¬
epithelium tall, becoming vacuolated; many ginalis is abrupt. As a rule, the borderline is
mitoses in all tissues; and no coiled arteries just inside the external os of the cervix. In
in the superficial third. (2) Secretory or luteal some individuals, however, particularly after
phase: endometrium 3 to 6 mm thick; glands childbearing, patches of the columnar epithe¬
sinuous and sacculated, with wide lumina; lium of the endocervix may extend for vary¬
epithelial cells tall, with surface blebs; stroma ing distances out onto the portio vaginalis.
edematous superficially; mitoses confined to These are inappropriately called cervical
coiled arteries, which are present near the “erosions.” They are especially susceptible to
surface. (3) Premenstrual phase: endometrium inflammatory reactions and are a common
3 to 4 mm thick, greatly contorted glands cause of increased vaginal discharge, leukor-
and arteries; dense stroma with leukocytic rhea. Virtually all multiparous women have
infiltration. (4) Menstrual phase: endometrium some degree of cervical inflammation. If un¬
0.5 to 3 mm thick; superficially extravasated treated, cervical erosions and their attendant
blood; the glands and arteries appear col¬ chronic inflammation may predispose to can¬
lapsed and shortened; the stroma is dense; cer of the cervix, which accounts for 10 per
and the surface is denuded of epithelium. cent of all cancer deaths in women.
The superficial cells of the cervical epithe¬
lium are constantly being exfoliated into the
Isthmus and Cervix
vaginal fluid. These can be examined in
The mucosa of the corpus uteri passes over stained “vaginal smears,” and the discovery
abruptly into that of the isthmus, which re¬ of abnormal cells may provide a very early
mains thin and shows little evidence of cyclic diagnosis of cancer.
morphological changes. It lacks coiled arter¬ Histophysiology of the Cervix. The mu¬
ies and usually does not bleed during men¬ cosa of the cervix does not take part in the
struation. menstrual changes that are characteristic of
The cervix forms the wall of the cervical the endometrium. There are, however, cyclic
canal, which is about 3 cm in length. Its changes in the secretory activity of its mucous
mucosal lining has a thickness of 2 to 3 mm glands. The gland cells are affected by the
and a structure quite different from that of circulating levels of ovarian hormones so that
the corpus uteri. Its irregular surface consists the amount and properties of the mucus
of branching folds called the plicae palmatae. secreted vary at different times in the men¬
Its stroma is dense, and the glands, which strual cycle. There are normally about 100
are relatively sparse, are oriented obliquely crypts or aggregations of glands in the cervix,
to the axis of the cervical canal. and these secrete up to 60 mg of mucus a
The canal is lined by a tall columnar epi¬ day throughout much of the cycle. At mid¬
thelium in which the nuclei are located near cycle, there is a tenfold increase in secretion
 884 • FEMALE REPRODUCTIVE SYSTEM

EARLY MID LATE

PROLIF¬ PROLIF¬ PROLIF¬ SECRETION |
| MENSES
ERATION ERATION ERATION
) 1 13 4 3 $ T 0 v 10 II lc 13 I*
IA IT IA Ift to 21 ft 23 t4 23 26 27 23
5•
F 1 9 t0 n 12 13 14 IS
—----1-r-- r ' 1 1

GLAND MITOSES
Gland mitoses Indicate proliferation They
occur during menstruation because repair
and breakdown are progressing simultane¬
ously at that time

PSEUDOSTRATIFICATION OF NUCLEI
This Is characteristic of the prolif era five
phose but persists until active secretion
begins It is not resumed until the
glands have Involuted during menstruation

BASAL VACUOLATION
This is the earliest morphological evidence
of ovulation found in the endometrium
It begins approximately 36 to 48 hours
following ovulation

SECRETION
This curve represents visible secretion In
the gland lumen; active secretion falls off
more abruptly In the later stages the
secretion becomes inspissated

STROMAL EDEMA
This factor varies with the individual,
particularly the rise during proliferation
which may be almost absent The edema
which accompanies secretion is more con¬
stant.

PSEUDODECIDUAL REACTION
This is evident first around the arterioles
and progresses until just before mens¬
truation a superficial compact layer is
formed

STROMAL MITOSES
These ore most abundant during the pro¬
liferative phase, absent during active
secretion but reappear during the stage
of predecidual formation

LEUCOCYTIC INFILTRATION
Throughout the cycle there are always
o few lymphocytes Polymorphonuclear
infiltration begins about two days before
the onset of flow

23 24 2s zs 27 ee

EARLY MIO LATE CHAftT 0*AVM *Y

MENSES PROLIF- PROLIF- PROLIF- SECRETION O H VIilfwmi*/

34/ H0rttr4
E RAT ION E RATION E RAT ION C0m*r!4f0 34, *0H

Figure 32-35. Schematic representation of the cyclic changes in the several morphological factors that are useful in
dating the endometrium. (From Noyes, R. W., A. T. Hertig, and J. Rock. Fertil. Steril. 7:3, 1950.)
 FEMALE REPRODUCTIVE SYSTEM • 885

rate, probably as a result of increasing estro¬ metrium is prepared for reception of a fer¬
genic stimulation. There is also a change in tilized egg.
the consistency of the mucus, from the highly At the end of the follicular phase in most
viscous state that prevails during most of the mammals, morphological and neural changes
cycle, to a less viscous, more highly hydrated occur that make the female receptive to the
condition at midcycle. These changes have male at or near the time of ovulation. This is
significance for fertility in that the viscous the period of heat or estrus. Although the
cervical mucus appears to be a hostile envi¬ menstrual cycle in the human female is basi¬
ronment and a serious impediment to prog¬ cally similar to the estrus cycle of other spe¬
ress of spermatozoa throughout much of the cies, receptivity to the male is not limited to
cycle. The changes occurring at midcycle, the end of the follicular phase and there is
however, favor sperm migration. no behavioral indication that ovulation has
In pregnancy, the cervical glands enlarge, occurred or is imminent.
proliferate, and accumulate large quantities During the ensuing luteal phase of the
of mucus, and the connective tissue between cycle, the preparation of the endometrium
them is reduced to thin partitions. for reception of a fertilized ovum is more
The thick wall of the cervix is composed extensive in the primates than in many other
of very dense connective tissue, with smooth animals. If no egg reaches the uterus, the
muscle constituting only about 15 per cent of bulk of the endometrium breaks down after
its mass. In the portio vaginalis, smooth mus¬ about two weeks. Its discharge is attended by
cle cells are absent. In the very late stage of uterine bleeding—menstruation.
pregnancy, changes take place in the fibrous In a cycle in which the ovum is fertilized,
and amorphous components of the extracel¬ the secretion of gonadotropins by the troph-
lular matrix that result in softening of the oblast of the implanting ovum helps to main¬
cervix, facilitating its dilatation by the ad¬ tain the corpus luteum beyond its usual life
vancing fetal head during labor. span, and it becomes the corpus luteum of
pregnancy. The continuing function of this
corpus luteum prevents the regressive and
ENDOCRINE REGULATION OF THE ischemic changes that lead to menstruation
FEMALE REPRODUCTIVE SYSTEM in an infertile cycle. Instead of regressing,
the secretory endometrium persists and
The histology of the female reproductive undergoes further hyperplasia (Fig. 32-
system cannot be fully understood without 34D), and menstruation is suppressed for the
some overview of the interactions of the duration of pregnancy.
brain, the hypophysis, the ovaries, and the The temporal correlation of the events in
uterus in regulation of the cyclic changes the endometrium with those of the ovary is
involved in reproduction. Although some of mediated by ovarian hormones. The devel¬
these relations have already been presented, oping follicle secretes the steroid hormones
a brief recapitulation may promote under¬ estradiol and estrone, collectively described as
standing of the cyclic changes described in estrogens. These stimulate growth of the uter¬
this chapter. ine endometrium. In species exhibiting es¬
The cyclic activities of the ovary are under trus, the estrogens also act upon the central
the control of the anterior lobe of the hypo¬ nervous system to bring about sexual recep¬
physis. The secretion of follicle stimulating tivity and its associated behavioral manifes¬
hormone (FSH) is responsible for the growth tations. After ovulation, the collapsed follicle
of the follicle up to the point of ovulation. is reorganized and transformed into a corpus
Luteinizing hormone (LH), together with luteum that secretes progesterone. This hor¬
FSH, is required for ovulation and for the mone is responsible for the secretory changes
early development of the corpus luteum. The in the endometrium that are characteristic of
endometrium of the uterus exhibits two the luteal phase of the cycle.
phases of functional activity that are corre¬ The secretion of gonadotropins by the hy¬
lated with the events in the ovary—a follicular pophysis is influenced by various factors. The
(or proliferative) phase of endometrial growth, rhythm of hypothalamic stimuli carried by
coinciding with maturation of the follicles the neurohumoral pathway to the adenohy¬
and their ovulation, and a luteal (or secretory) pophysis appears to be determined by some
phase, correlated with the development of a internal clock, but it can also be influenced
corpus luteum and during which the endo¬ by psychic factors and various external stim-
886 • FEMALE REPRODUCTIVE SYSTEM

uli. Production of excess ovarian hormones

also acts back upon the hypothalamus to
diminish gonadotropin secretion. This feed¬
back mechanism is not only operative in the
regulation of the normal cycle but is also the
basis for the successes in conception control,
wherein orally administered analogues of
ovarian steroids act upon the hypothalamus
and hypophysis to suppress the surge of LH
that is essential for ovulation.

IMPLANTATION

After fertilization takes place in the upper

part of the oviduct, segmentation of the
ovum proceeds as it passes down the oviduct
(Fig. 32—36A, B). When it reaches the uterus
on about the fourth day, it consists of many
cells arranged in a hollow sphere called the
blastocyst (Fig. 32—36C). The blastocyst re¬
mains free in the lumen of the uterus for a
day or so and then attaches to the surface of
the secretory endometrium. The blastocyst
by this time has differentiated into (1) an
assemblage of cells at one pole called the
inner cell mass, which is destined to form the
embryo proper; and (2) a layer of primitive
trophoblast cells making up the rest of the wall
of the blastocyst. The trophoblast cells are
concerned with the attachment and implan¬
tation of the ovum and with the subsequent
establishment of the placenta, the organ in
which the physiological exchange of nutrients
and waste products takes place between the
embryonic and the maternal circulations.
When the trophoblast makes contact with the
surface of the endometrium, its cells prolif¬
erate rapidly, forming, at the interface be¬
tween the ovum and the maternal tissue, a
multinucleate mass of protoplasm in which
no cell boundaries are discernible. This is
called the syncytial trophoblast. This actively
erosive syncytium destroys the surface epi¬
thelium and permits the blastocyst to invade
the underlying stroma (Fig. 32-37A). By the
11th day the blastocyst is entirely within the
endometrium; the trophoblast has formed a
broad layer completely surrounding the in¬ C
ner cell mass; and the uterine epithelium has Figure 32-36. Photomicrographs of early human ova. A,
repaired the breach made in it by the im¬ Segmenting human ovum. Two-cell stage recovered from
planting blastocyst (Figs. 32-38, 32-39). This the fallopian tube. Ovulation age 1 Vz to 214 days. Notice
form of implantation, in which the embryo polar body between the two blastomeres. x500. B, Free
human blastocyst. Section of a 58-cell intrauterine blas¬
and its associated membranes become
tocyst. Segmentation cavity is just beginning to form.
embedded in and completely encapsulated Zona pellucida is disappearing. Ovulation age 4 days.
by the endometrium, is called interstitial im¬ x 600. C, Free human blastocyst. Section of a 107-cell
plantation and is characteristic of the human. blastocyst recovered from the uterine cavity. The inner
From the ninth to the 11th day, the cell mass is at the right. Ovulation age 4Vz days. x60Q.
(All three micrographs from Hertig, A., J. Rock, and E.
expanding trophoblastic shell becomes Adams. Am. J. Anat. 98:435, 1956.)
 FEMALE REPRODUCTIVE SYSTEM • 887

B
Figure 32-37. Photomicrographs of early human implantation sites. A, Human seven-day implantation. The embryo is
a simple bilaminar disc. Development of an amniotic cavity is beginning. There is a solid plaque of syncytio- and
cytotrophoblast. x300. (After Hertig and Rock, 1941. Courtesy of the Carnegie Institution of Washington.) B, Human
nine-day implantation. The embryo is a bilaminar disc. The syncytiotrophoblast now shows prominent lacunae. Notice
at arrow a maternal blood space communicating with lacuna. x25. (After Hertig and Rock, 1941. Courtesy of the
Carnegie Institution of Washington.)

permeated by a labyrinthine system of inter¬ totically active and contributes to the increas¬
communicating lacunae containing blood lib¬ ing mass of the syncytiotrophoblast by form¬
erated by erosion of maternal blood vessels ing new cells that fuse with and become part
(Fig. 32—39). This extravasated blood evi¬ of the syncytium.
dently serves as a source of nourishment for At the 11-day stage, the embryo proper
the embryo and represents the first step to¬ consists of a bilaminar disc of epithelial
ward establishment of the uteroplacental cir¬ cells—a thick plate of ectoderm and a thinner
culation on which the growth of the embryo layer of primitive endoderm. The ectodermal
will later depend. plate is continuous at its margins with a thin
Two forms of trophoblast are recognizable, layer of squamous cells that enclose a small
an inner layer of cytotrophoblast, composed of amniotic cavity. The endoderm is similarly
individual cells, and a thicker outer layer of continuous with a thin sheet of cells forming
syncytiotrophoblast. The cytotrophoblast is mi- the yolk sac. Surrounding these structures are
888 • FEMALE REPRODUCTIVE SYSTEM

solid cords of trophoblast grow outward from

the surface of the chorion to form the primary
chorionic villi. These are soon invaded at their
base by chorionic mesenchyme, which ad¬
vances toward their growing tips, converting
the primary villi into secondary villi (Figs.
32—40, 32—41). The secondary villi then con¬
sist of an outer layer of syncytial trophoblast,
an inner layer of cytotrophoblast, and a mes¬
enchymal core. They are bathed in maternal
blood that flows sluggishly through a labyrin¬
thine system of intercommunicating channels
collectively making up the intervillous space.
From the ends of the secondary chorionic
villi, solid cords of trophoblast, the cytotropho-
hlastic cell columns, extend across the intervil¬
lous space and, upon reaching the opposite
wall, spread along it, coalescing with similar
outgrowths from -neighboring villi to form a
more or less continuous trophoblastic shell, in¬
terrupted only at sites of communication of
maternal vessels with the intervillous space.
The trophoblastic shell consists mainly of
cytotrophoblast, but some areas of syncytio-
trophoblast can be found. Through its inter¬
stitial growth, the trophoblastic shell provides
a mechanism for rapid circumferential ex¬
pansion of the entire implantation site and
Figure 32-38. Human gestational endometrium with an for enlargement of the intervillous space.
11-day implantation site (arrow). The entire thickness of From the time of its formation throughout
the endometrium is shown. The glands are secretory, the the remainder of pregnancy, the intervillous
stroma is edematous, and the superficial veins are dilated, space is lined by trophoblast and traversed
x 18. (After Hertig and Rock, 1941. Courtesy of Carnegie
Institution of Washington.) by villi that are attached to the maternal tissue
via the trophoblastic shell. The villi absorb
nutriments from the maternal blood in the
large extracellular spaces traversed by ten¬ intervillous space and excrete wastes into it.
uous strands of extraembryonic mesenchyme The efficiency of this process is greatly en¬
(mesoblast). These spaces constitute the exo¬ hanced after the development of a function¬
coelom. The surrounding broad zone of ing vascular system in the embryo.
trophoblast is called the chorion. Fetal blood vessels differentiate in the mes¬
enchymal cores of the secondary villi as dis¬
continuous endothelial-lined spaces, which
later coalesce to form continuous vascular
PLACENTA channels. These become connected with the
embryonic heart via vessels that differentiate
Formation and Structure in the mesenchyme of the inner surface of
the chorion and in the body stalk or umbilical
From the 11th to the 16th day of preg¬ cord. By the 21st to the 23rd day, fetal blood
nancy the trophoblast continues to prolifer¬ begins to circulate through the capillaries of
ate rapidly, and the implantation cavity is the villi. After their vascularization, the sec¬
progressively enlarged at the expense of the ondary villi are called tertiary or definitive
surrounding maternal tissue. Invasion of the placental villi (Fig. 32—47). These radiate from
maternal blood vascular system by syncytio- the entire periphery of the chorion (Fig.
trophoblast becomes extensive. The large la¬ 32—41). In the subsequent growth of the
cunae in the syncytial labyrinth communicate placenta, the villi, which extend across the
at many places with venous sinuses in the intervillous space to the trophoblastic shell,
endometrium. From the 15th day onward, develop numerous lateral branches whose
 FEMALE REPRODUCTIVE SYSTEM • 889

Lacuna with Coiled

Endometrial gland maternal blood artery

mesoblast disk epithelium trophoblast trophoblast

Figure 32-39. Photomicrograph from the same section as Figure 32-37, magnified 160 diameters. The bulk of the
ovum consists of masses of trophoblast (syncytium) invading the endometrium. Within the syncytial trophoblast is the
cellular trophoblast with obvious cell boundaries. The cells are arranged as a simple epithelium except for the clump at
C. The cellular trophoblast immediately surrounds the primitive chorionic mesoblast in which the embryo is suspended.
(After Hertig and Rock, 1941. Courtesy of the Carnegie Institution of Washington.)

unattached tips float free in the blood of the blast, bringing the total surface area to about
intervillous space. 90 sq m—another remarkable example of the
The villi that arise from the chorionic plate strategem of increasing efficiency by ampli¬
are usually called stem villi; those branches fication of the area of physiologically impor¬
that attach to the basal plate are anchoring tant membranes.
villi (Figs. 32-44, 32-45); and those that end The villi of the early placenta have capil¬
free in the intervillous space are the terminal laries with continuous unfenestrated endo¬
villi (Fig. 32—46). The stem villi undergo thelium in a stroma of loose mesenchyme.
extensive branching—as many as 15 genera¬ Scattered in the interstices of this stellate
tions of branches have been described. The reticulum are large, globular Hofbauer cells
terminal villi are very slender and no doubt filled with clear vacuoles. Their function re¬
easily moved by movement of blood in the mains obscure but they are believed to be
intervillous space. They present a very large macrophages whose vacuolated appearance
surface area for diffusion of nutrients and early in pregnancy is due to uptake of extra¬
metabolites between the maternal and fetal cellular fluid by pinocytosis. As pregnancy
circulations. The villous surface of the ma¬ progresses, they lose much of their vacuola-
ture human placenta is estimated to be about tion and take on an appearance more typical
10 sq m. This is further amplified by the of tissue macrophages.
numerous microvilli on the syncytiotropho- The core of the villus is enclosed by two
890 • FEMALE REPRODUCTIVE SYSTEM

Amniotic
cavity
Figure 32-40. Human embryo of ovulation age 18 to 19
days. The curved embryonic disc lies between a large
yolk sac and a smaller crescentic amniotic cavity. The
body stalk bends back and blends with the chorionic
mesoblast. Many secondary villi project into an extensive
intervillous space, x 15. (Courtesy of A. T. Hertig.)

the ertire^surface'oHh^cho^ion.^McKa^D.^GTc1 CmRoby(AaTeH^rt!g°amfN(^VSR<?chn^d^^aCeA*a' ^[iPr°jec*!<n9 ^rom

69:735. 1955. Courtesy of the Carnegie institution oTwashington.) 9 R'^ardson. Am. J. Obstet. Gynecol.
 FEMALE REPRODUCTIVE SYSTEM • 891

layers of trophoblast—the inner cytotrophoblast

and the outer syncytial trophoblast, directly ex¬
Chorion
posed to the maternal blood. The cytotro-
Decidua Villi phoblastic cells have a euchromatic nucleus
capsularis Decidua
and a relatively pale-staining cytoplasm poor
Decidua basalis
vera
in cell organelles. They function mainly as
stem cells for the overlying syncytium. Early
in pregnancy they divide frequently, trans¬
form, and fuse with the expanding syncy¬
tium. Their number therefore decreases, and
from the fifth month onward they are rela¬
tively few and no longer form a continuous
inner layer. The thickness of the syncytium
gradually comes to vary, thick areas contain¬
ing clusters of nuclei alternating with thin
areas devoid of nuclei. In these thin areas,
dilated capillaries of the villus core are closely
applied to the inner aspect of the attenuated
syncytium so that the diffusion barrier be¬
tween the bloodstreams is little more than 2
pm—a relationship reminiscent of the thin
blood-air diffusion barrier in the lung.
In contrast to the relatively undifferen¬
tiated cytotrophoblast, the syncytial tropho¬
blast is highly specialized for its multiple
functions associated with the uptake of nu¬
trients and synthesis, and secretion of both
steroid and protein placental hormones. The
free surface is irregular in contour and pro¬
vided with abundant microvilli. Between the
bases of the microvilli are numerous coated
pits and coated vesicles involved in receptor-
mediated endocytosis of macromolecules.
The cytoplasm is strongly basophilic, and in
electron micrographs has an extensive devel¬
opment of rough endoplasmic reticulum and
abundant free polyribosomes. Multiple Golgi
complexes are distributed at intervals in the
syncytium. The numerous long mitochondria
have foliate and tubular cristae. The ultra-
structural appearance of the syncytium is
predominantly that of cells actively synthesiz¬
ing protein for export. The smooth endo¬
plasmic reticulum that is the most conspicu¬
ous organelle in other steroid-secreting
endocrine glands is not evident in the syn¬
cytium, and one must assume that enzymes
of this biosynthetic pathway are incorporated
in the membrane of the rough endoplasmic
reticulum. The presence of numerous cho¬
lesterol-rich lipid inclusions is consistent with
steroid synthesis, as are mitochondria with
tubular cristae.
Figure 32-42. Drawings of successive stages of human The products of conception occupy only a
pregnancy, showing the gradual obliteration of the uterine portion of the entire endometrium of preg¬
lumen, the disappearance of the decidua capsularis, and
the establishment of the definitive discoid placenta. (Draw¬
nancy (decidua). Different regions are identi¬
ings by Brodel, M., from Williams, J. Am. J. Obstet. fied by separate terms descriptive of their
Gynecol. 73:1, 1927.) topographical relation to the implantation
892 • FEMALE REPRODUCTIVE SYSTEM

Figure 32-43. Drawing of the disposition of the fetal

membranes in the later months of pregnancy. The amnion
and chorion have come into contact and have become
adherent to each other and to the decidua vera. (Drawings
by Brodel, M., from Williams, J. Am. J. Obstet. Gynecoi.
73:1, 1927.)

Chorionic plate Stem villi

Figure 32 44. Scanning micrograph of a portion of the placental disc of a macaque at 22 days’ gestation. Side branches
from the stem villi are just beginning to appear (small arrows). (Micrograph from King, B. Anat. Embryo!. 765:361, 1982.)
 FEMALE REPRODUCTIVE SYSTEM • 893

Figure 32-45. Scanning micrograph of a portion of the placental disc of a macaque at 31 days’ gestation. Free villous
side branches have continued to develop near the chorionic plate. Longitudinally branched central villi and cell columns
appear to reunite before merging with the basal plate (at arrowhead). (Micrograph from King, B. Anat. Embryol. 765:361,
1982.)

Figure 32-46. Scanning micrograph of a segment of the placental disc of a macaque at 60 days’ gestation. Large-stem
villi can be seen at upper left emerging from the chorionic plate. The greater part of the placenta is now made up of
long, slender, branching terminal villi. (Micrograph from King, B. Anat. Embryol. 765:361, 1982.)
894 • FEMALE REPRODUCTIVE SYSTEM

chorion frondosum and a concomitant ex¬

pansion of the intervillous space. During the
fourth and fifth months the placental disc is
partitioned into 15 to 20 cotyledons by the
formation of incomplete septa that project
from the decidual plate into the intervillous
space. There are also changes during this
period in the histological organization of the
villi.

Placental Circulation
Blood poor in oxygen is carried from the
fetus to the placenta in the umbilical arteries
of the umbilical cord. At the junction of the
cord with the placenta, the umbilical arteries
divide into a number of radially disposed
Figure 32-47. Section through placental villus from a 2-
placental arteries that branch freely in the
cm human embryo. The brush border (b) on the syncytial chorionic plate. Numerous branches from
trophoblast is barely visible. Beneath it is the continuous these pass downward into the stem villi and
layer of cellular trophoblast. The vessels in the mesen¬ ramify in the arborescent pattern of subsidi¬
chyme are filled with primitive erythrocytes. One mesen¬
ary villi down to the capillary networks of the
chymal cell is in mitosis. x450.
terminal villi. The oxygen-rich venous blood
is collected into thin-walled veins, which re¬
turn the blood through vessels of increasing
site. The portion that underlies the implan¬ caliber that follow the course of the arteries
tation site and forms the maternal compo¬ to the chorionic plate. There they join veins
nent of the placenta is the decidua basalis. The that converge upon the single umbilical vein,
thin superficial portion between the implan¬ which carries the blood through the umbilical
tation site and the lumen is the decidua cap- cord to the ductus venosus, whence it enters
sularis, and that lining the remainder of the the inferior vena cava near its point of con¬
uterus down to the internal os is the decidua fluence with the right atrium.
vera (Figs. 32-42, 32-43). On the maternal side, blood from the ar¬
Up to about the eighth week, the villi are cuate branches of the uterine arteries is car¬
equally numerous around the entire surface ried by the coiled arteries through openings
of the chorion (Fig. 32-41), but as pregnancy in the basal plate of the placenta into the
advances the villi adjacent to the decidua intervillous space. The flow from the mater¬
basalis enlarge and rapidly increase in num¬ nal arterioles is pulsatile and is delivered at
ber, while those facing the decidua capsularis a pressure considerably higher than that pre¬
degenerate, leaving this surface of the cho¬ vailing in the intervillous space. It therefore
rion smooth and relatively avascular after the spurts from the basal plate deep into the
third month. This region is thenceforth intervillous space in jets (Fig. 32-48). As its
called the chorion laeve, while the villous por¬ pressure is dissipated, it flows back around
tion toward the base is called the chorion and over the surface of the placental villi,
frondosum. This latter becomes confined to a permitting exchange of metabolites with the
circular area that goes on to form the defin¬ fetal blood. Since the human has a hemochorial
itive discoid placenta. placenta, the trophoblast of the villi is exposed
As the volume of the conceptus increases directly to maternal blood, and the diffusion
and it bulges further into the lumen, the barrier in the mature placenta consists only
decidua capsularis becomes greatly atten¬ of the thin layer of syncytiotrophoblast, its
uated. Its vascularity is jeopardized, and it basal lamina, and the wall of the subjacent
degenerates. By four and a half months, the fetal capillaries.
decidua capsularis has disappeared and the The pressure of the incoming blood and
chorion laeve has fused with the decidua vera its fountain-like distribution tend to force the
of the opposite wall, largely obliterating the blood back toward the basal plate, where it is
uterine lumen (Fig. 32-43). The later devel¬ drained away through numerous communi¬
opment of the placenta involves a steady cations between the intervillous space and
growth in size and length of the villi of the dilated veins in the decidua basalis.
Figure 32-48. Placenta, showing structure and circulation. The head of maternal blood pressure drives entering blood
FEMALE REPRODUCTIVE SYSTEM

toward the chorionic plate in fountain-like spurts. As the head of pressure is dissipated, lateral dispersion of blood
occurs. Inflowing arterial blood pushes venous blood out into the endometrial veins. (After Ramsey, E. M., and J. W.
Harris. Contributions to Embryology, No. 261, Vol. 38, 1966. Courtesy of the Carnegie Institution of Washington. Drawing
895

by Ranice Davis Crosby.)

896 • FEMALE REPRODUCTIVE SYSTEM

Hisiophysiology of the Placenta carbohydrate and fat metabolism of the

mother, which makes more glucose available
The placenta is a transient organ consisting for nutrition of the fetus. The bulk of avail¬
of both maternal and fetal components so able evidence indicates that all the placental
structured as to bring fetal and maternal hormones are products of the syncytial
blood into close proximity to permit ex¬ trophoblast. The messenger RNA for soma¬
change of gaseous metabolites, nutrients, and tomammotropin has recently been localized
waste products. In the early months of preg¬ in the syncytium by in situ hybridization.
nancy it also has an important storage func¬ The syncytiotrophoblast is unique among
tion, accumulating carbohydrate in the form absorptive epithelia in that substances cannot
of glycogen that can later be released as traverse it by a paracellular route. Owing to
glucose. Similarly, proteins, calcium, and iron its syncytial nature all substances, from ions
are stored for use later in pregnancy. Thus, to macromolecules, entering or leaving the
for the fetus the placenta has some of the fetal blood must pass through it. Oxygen,
same functions as the lung, intestine, liver, carbon dioxide, fatty acids, steroids, lipid-
and kidney in postnatal life. soluble vitamins, and electrolytes can traverse
In addition, the placenta is a major endo¬ it by passive diffusion. Sugars cross the syncy¬
crine gland delivering directly into the ma¬ tium more rapidly than expected on the basis
ternal blood hormones that are essential for of passive diffusion, and it is likely that a
the continuance of pregnancy. The hormone carrier molecule is involved in their facilitated
chorionic gonadotropin is first secreted by the diffusion. The fetus synthesizes its own pro¬
trophoblast when the blastocyst is implanting teins from amino acids that cross the placenta
six to eight days after fertilization, and in¬
by active transport. Polypeptides and larger
creases rapidly thereafter, declining to low protein molecules are transported across the
levels by the fifth month. This glycoprotein
syncytium by coated vesicles formed by recep¬
hormone is very similar to luteinizing hor¬
tor-mediated endocytosis. The syncytium has re¬
mone in chemical structure and function. It
ceptors for insulin, transferrin, the Fc por¬
prevents the involution of the corpus luteum
tion of immunoglobulin, and probably for
of ovulation that would otherwise occur, and
certain other macromolecules. The vesicular
stimulates the corpus luteum of pregnancy
transport of IgG is, in part, the basis for the
to grow and secrete more progesterone.
passive immunization of the baby against
The placenta also secretes the steroid hor¬
certain pathogens.
mones estrogens and progesterone in the latter
half of pregnancy at a rate hundreds of times
that of a normal cycle. The estrogen pro¬
duced is predominantly estriol, which has rel¬ VAGINA
atively low estrogenic potency. Nevertheless,
the total estrogenic activity of that produced The vagina is a distensible muscular tube
is some 30 times normal. These high levels extending from the vestibule of the female
of estrogen contribute to growth of the external genitalia to the cervix of the uterus.
uterus and to gestational development of the The lower end of the vagina in the virgin is
mammary glands. marked by a transverse semicircular fold or
The tenfold increase in the rate of proges¬ fenestrated membrane, the hymen. The wall
terone secretion in the course of pregnancy of the vagina consists of three layers: the
stimulates differentiation and proliferation mucosa, the muscular coat, and the adventi¬
of decidual cells in the endometrium of the tial connective tissue.
gravid uterus, and contributes to develop¬ The adventitial coat is a thin layer of dense
ment of the mammary glands. connective tissue, which merges into the loose
Another placental hormone was initially connective tissue joining the vagina to the
called placental lactogen because in some spe¬ surrounding structures. In this connective
cies it had an effect on the mammary glands tissue can be found an extensive venous
similar to that of prolactin. An effect on plexus, nerve bundles, and small groups of
lactation has not been demonstrated in the nerve cells.
human and it is now commonly called soma¬ The interlacing smooth muscle bundles of
tomammotropin. Its chemical structure resem¬ the muscular layer are arranged circularly
bles that of the growth hormone somatotro¬ and longitudinally. The longitudinal bundles
pin, and it has some growth-promoting are far more numerous, especially in the
action. More significant is its action upon the outer half of the layer. Striated fibers of the
 FEMALE REPRODUCTIVE SYSTEM • 897

Mucosa of cervix Muscle coat

of cervi:

W&WSmv*’ Muscle coat

ot vagina

Mucosa of vagina

Cylindrical
epithelium

Externa
orifice
of uterus

Cervical glands
Beginning of stratified
squamous epithelium
with papillae
Figure 32-49. Sagittal section through posterior half of the portio vaginalis uteri and the fornix vaginae of a young
woman. xIO. (After von Ebner.)

bulbocavernous muscles form a kind of infiltrate the intercellular spaces from the
sphincter around the ostium of the vagina. base of the epithelium but do not penetrate
The mucosa consists of a surface epithe¬ the intercellular clefts of the superficial lay¬
lium and an underlying lamina propria. The ers.
epithelium is a typical stratified squamous The vagina is devoid of glands, and much
epithelium 150 to 200 (im in thickness, con¬ of the lubricating fluid is contributed by se¬
sisting of about 45 layers of cells during the cretion from glands of the cervix. It is gen¬
follicular phase of the cycle and about 30 in erally agreed, however, that there is a true
the luteal phase. The superficial cells may vaginal fluid that increases in abundance dur¬
contain keratohyaline granules, but they re¬ ing sexual stimulation. This is believed to
tain stainable nuclei and undergo little kera- arise as a transudate from capillaries of the
tinization. Their cytoplasm is filled with gly¬ lamina propria and to move through inter¬
cogen, especially at midcycle. In electron cellular channels of the epithelium to the
micrographs the cells are joined by numerous lumen. The discrepancy between this inter¬
desmosomes and occasional gap junctions. pretation and the demonstration of a perme¬
The latter are most numerous in the deeper ability barrier to electron-opaque tracers has
layers, and diminish toward the surface. yet to be resolved.
Tight junctions of limited extent and lamel¬ The intercellular spaces of the epithelium
lar intercellular deposits of a lipid material are accessible to mononuclear leukocytes.
have also been described near the free sur¬ Lymphocytes normally breach the basal lam¬
face. These apparently constitute a per¬ ina, open the desmosomes, and invade the
meability barrier for large water-soluble enlarged intercellular channels so formed. It
molecules. Electron-opaque tracers readily has recently been shown that, like the epi-
898 • FEMALE REPRODUCTIVE SYSTEM

dermis, the vaginal epithelium includes a pH of the vagina rises, favoring growth of
population of Langerhans cells, which appear the protozoan parasite Trichomonas vaginalis.
as scattered clear cells occupying expanded Thus, estrogen can be administered as an
intercellular channels in its basal and inter¬ adjunct to antimicrobial therapy for this and
mediate layers. In the epidermis the Langer¬ other vaginal infections.
hans cells are known to participate in antigen The lamina propria is a moderately dense
presentation and cooperate with T lympho¬ connective tissue. Toward the muscular layer
cytes in immunological surveillance. They no it becomes looser, and this layer may be
doubt play a similar role in the vagina. considered a submucosa. In the anterior wall
Superficial cells are continually shed from of the vagina, papillae associated with the
the surface of the vaginal epithelium deep surface of the epithelium are few and
throughout the cycle, but this desquamation small, but in the posterior wall the lamina
is greater late in the luteal phase and during propria sends numerous papillae far into the
menstruation. Glycogen from the exfoliated covering epithelium. Immediately under the
cells is a rich substrate for certain members epithelium there is a dense network of fine
of the bacterial flora, which break it down to elastic fibers. From there, fine fibers run
lactic acid, lowering the pH of the vagina. downward to the muscular layer. Accumula¬
Since the amount of glycogen in the epithe¬ tions of lymphocytes are numerous, and
lium is controlled by estrogen, the pH of the sometimes lymph nodules are present. Lym¬
vaginal fluid is lowest at midcycle. With less phocytes are always found migrating into the
estrogen being secreted in the luteal phase epithelium. The deeper layers of the lamina
of the cycle, less glycogen is formed and the propria contain a dense plexus of small veins.

r mucous cells Stratified columnar

lium

Blood vessel

Interstitial
connective tissue

Large mucous cells

Figure 32 50. Section of gland of Bartholin. A large duct with patches of stratified columnar epithelium gives off smaller
branches lined with columnar mucous cells and continuing into tubuloalveolar terminal portions, which are lined with
large mucous cells, x 185. (After A. A. Maximow.)
 FEMALE REPRODUCTIVE SYSTEM • 899

EXTERNAL GENITALIA Young, W. C. ed.: Sex and Internal Secretions. Vol.

2. 3rd ed. Baltimore, Williams & Wilkins Co., 1955.
The external genital organs of the female Blandau, R. J., and K. Moghissi: The Biology of the
Cervix. Chicago, University of Chicago Press, 1973.
comprise the clitoris, the labia majora and
Finn, C. A., and D. G. Porter: The Uterus. Handbooks
minora, and certain glands that open into the of Reproductive Biology. London, Paul Elek Ltd.,
vestibule, the space flanked by the labia mi¬ 1974.
nora. Hafez, E. S. E., and R. J. Blandau: The Mammalian
The clitoris corresponds embryologically to Oviduct. Chicago, University of Chicago Press, 1969.
Reid, L. N., ed.: Neuroendocrine Aspects of Reproduc¬
the dorsal part of the penis. It consists of two tion. ORPRC Symposia on Primate Reproductive
small, erectile, corpora cavernosa ending in Biology. New York, Academic Press, 1983.
a rudimentary glans clitoridis. The vagina and Yen, S. S. C., and R. B. Jaffe: Reproductive Endocrinology:
the urethra open into the vestibule, which is Physiology, Pathophysiology and Clinical Manage¬
ment. 2nd ed. Philadelphia, W. B. Saunders Co., 1986.
lined with stratified squamous epithelium.
Around the opening of the urethra and on OVARY
the clitoris are several small vestibular glands Anderson, E., and H. W. Beams: Cytological observa¬
(glandulae vestibulares minores). They resemble tions on the fine structure of the guinea pig ovary
with special reference to the oogonium, primary
the glands of Littre in the male urethra and
oocyte and associated follicle cells. J. Ultrastruct.
contain mucous cells. Res. 5:432, 1960.
Two larger glands, the glands of Bartholin Baker, T. G., and L. L. Franchi: The fine structure of
(glandulae vestibular es major es), each about 1 oogonia and oocytes in human ovary. }. Cell Sci.
cm in diameter, are located in the lateral 2:213, 1967.
Corner, G. W., Jr.: The histological dating of the human
walls of the vestibule and open on the inner
corpus luteum of menstruation. Am. J. Anat.
surface of the labia minora. They are of the 98:337, 1956.
tubuloalveolar type, closely corresponding Crisp, T. M., and C. Channing: Fine structural events
structurally to the bulbourethral glands of correlated with progestin secretion during luteiniza-
tion of rhesus monkey granulosa cells in culture.
the male and secreting a similar lubricating
Biol. Reprod. 7:55, 1972.
mucus (Fig. 32—50). Crisp, T. M., D. A. Dessouky, and F. R. Denys: The fine
The labia minora are covered with stratified structure of the human corpus luteum of early
squamous epithelium and have a core of pregnancy and during the progestational phase of
spongy connective tissue permeated by fine the menstrual cycle. Am. J. Anat. 127:37, 1970.
Gillim, S. W., A. K. Christensen, and C. E. McLennan:
elastic networks. Blood vessels are very nu¬ Fine structure of human granulosa and theca lutein
merous. The epithelium contains pigment in cells at the stage of maximum progesterone secre¬
its deeper layer and has a thin keratinized tion during the menstrual cycle. Anat. Rec. 163:189,
layer on the surface. Numerous large seba¬ 1969.
Creep, R. O.: Histology, histochemistry and ultrastruc¬
ceous glands are found on both surfaces.
ture of adult ovary. In Smith, D. E., ed.: The Ovary.
There are no associated hairs. Baltimore, Williams & Wilkins Co., 1962.
The labia majora are folds of skin contain¬ Hertig, A. T.: The primary human oocyte: some obser¬
ing a large amount of subcutaneous adipose vations on the fine structure of Balbiani’s vitelline
tissue and a thin layer of smooth muscle, body and the origin of the annulate lamellae. Am.
J. Anat. 122:107, 1968.
corresponding to the tunica dartos of the Hertig, A. T., and E. C. Adams: Studies on the human
scrotum. The outer surface is covered with oocyte and its follicle. 1. Ultrastructural and histo-
hair; the inner is smooth and hairless. Seba¬ chemical observations on the primordial follicle
ceous and sweat glands are numerous on stage. J, Cell Biol. 54:647, 1957.
Long, J. A.: Corpus luteum of pregnancy in the
both surfaces.
rat—ultrastructural and cytochemical observations.
The outer genital organs are richly sup¬ Biol. Reprod. 8:87, 1973.
plied with sensory nerve endings. Meissner Mossman, M. H., M. J. Koering, and D. Ferry, Jr.: Cyclic
corpuscles are scattered in the papillae of the changes of interstitial gland tissue of the human
epithelium, and genital corpuscles are pres¬ ovary. Am. J. Anat. 7 75:235, 1964.
Ryan, K. }., and R. V. Short: Formation of estradiol by
ent in the subpapillary layer. Pacinian cor¬ granulosa and theca cells of the equine ovarian
puscles have been found in the deeper parts follicle. Endocrinology 76:108, 1965.
of the connective tissue of the labia majora Strassman, E. O.: The theca cone and its tropism toward
and in the cavernous bodies of the clitoris. the ovarian surface, a typical feature of growing
human mammalian follicles. Am. J. Obstet. Gynecol.
47:363, 1941.

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Austin, C. R.: The Mammalian Egg. Oxford, Blackwell Decker, A.: Culdoscopic observations on the tubo-ovar-
Scientific Publications, 1961. ian mechanism of ovum reception. Fertil. Steril.
Blandau, R. J.: Biology of Eggs and Implantation. In 2:253, 1951.
900 • FEMALE REPRODUCTIVE SYSTEM

Doyle, J. B.: Exploratory culdotomy for observation of eleven and twelve days respectively. Carnegie Con¬
tubo-ovarian physiology at ovulation time. Fertil. trib. Embryol. 29:127, 1941.
Steril. 2:474, 1951. Hertig, A. T., and J. Rock: Two human ova of the
previllous stage, having a development age of about
OVIDUCT seven and nine days respectively. Carnegie Contrib.
Brenner, R. M.: Electron microscopy of estrogen effects Embryol. 57:67, 1954.
on ciliogenesis and secretory cell growth in rhesus Hertig, A. T., J. Rock, E. C. Adams, and W. J. Mulligan:
monkey oviduct. Anat. Rec. 757:218, 1967. On the preimplantation stages of the human ovum;
Brenner, R. M.: The biology of oviductal cilia. In Hafez, a description of four normal and four abnormal
E. S. E., and R. J. Blandau, eds.: The Mammalian specimens ranging from the second to the fifth day
Oviduct. Chicago, University of Chicago Press, 1969. of development. Carnegie Contrib. Embryol.
55:199, 1954.
UTERUS Johnson, L. W., and C. H. Smith: Monosaccharide trans¬
port across microvillous membrane of human pla¬
Bartelmez, G. W.: Histological studies on the menstruat¬
centa. Am. J. Physiol. 238 (Cell Physiol. 7):C160, 1980.
ing mucous membrane of the human uterus. Car¬
Kaufman, P., D. K. Sen, and G. Schweikhart: Classifi¬
negie Inst. Contrib. Embryol. 24:141, 1933.
cation of human placental villi. I. Histology. Cell
Markee, J. E.: Menstruation in intraocular endometrial
Tissue Res. 200:409, 1979.
transplants in the Rhesus monkey. Carnegie Con¬
King, B. F.: Localization of transferrin on the surface
trib. Embryol. 28:219, 1940.
of the human placenta by electron microscopic im-
Markee, J. E.: The morphological and endocrine basis
munocytochemistry. Anat. Rec. 786:151, 1976.
for menstrual bleeding. Progr. Gynecol. 2:63, 1950.
Metcalfe, J., H. Barterls, and W. Moll: Gas exchange in
Schmidt-Matthiesen, H.: The Normal Human Endo¬
metrium. New York, McGraw-Hill Book Co., 1963. the pregnant uterus. Physiol. Rev. 47:782, 1967.
Nelson, M. D., A. C. Enders, and B. F. King: Cytological
CERVIX events involved in glycoprotein synthesis in cellular
and syncytial trophoblast of human placenta. An
Blandau, R. J., and K. Moghissi: The Biology of the
electron microscope autoradiographic study of
Cervix. Chicago, University of Chicago Press, 1973.
(3H)galactose incorporation. J. Cell Biol. 76:418,
Danforth, D. N.: The fibrous nature of the human
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cervix, and its relations to the isthmic segment in
Nelson, D. M., A. C. Enders, and B. F. King: Cytological
gravid and non-gravid uteri. Am. J. Obstet. Gynecol.
events involved in protein synthesis in cellular and
55:541, 1947.
syncytial trophoblast of human placenta. An elec¬
Flukman, C. F.: The glandular structures of the cervix
tron microscope autoradiographic study of
uteri. Surg. Gynecol. Obstet. 106:515, 1958.
(3H)leucine incorporation. J. Cell Biol. 76:400, 1978.
Moghissi, K. S.: The function of the cervix in fertility.
Okleford, C. D., and J. M. Clint: The uptake of IgG by
Fertil. Steril. 25:295, 1972.
human placental chorionic villi: a correlated auto¬
Vickery, B. H., and J. P. Bennett: The cervix and its
radiographic and wide aperture counting. Placenta
secretions in mammals. Physiol. Rev. 483135, 1968.
7:91, 1980.
IMPLANTATION AND PLACENTA Schneider, H., K. El. Mohlen, and J. Dancis: Transfer
of amino acids across in vitro perfused human
Amoroso, E. C.: Placentation. In Parker, A. 3., ed.: placenta. Pediatr. Res. 75:236, 1979.
Marshall’s Physiology of Reproduction. Vol. 2. 3rd Wislocki, G. B., and H. S. Bennett: The histology and
ed. London, Longmans, Green & Co. Ltd., 1952. cytology of the human and monkey placenta, with
Amoroso, E. C.: Histology of the placenta. Br. Med special reference to the trophoblast. Am. J. Anat.
Bull. 77:81, 1961. 75:335, 1943.
Austin, C. R.: The Mammalian Egg. Oxford, Blackwell
Scientific Publications, 1961. VAGINA
Baker, T. G.: A quantitative and cytological study of Averette, H. E., G. D. Weinstein, and P. Frost: Autora¬
oogenesis in the rhesus monkey. T. Anat. 100-161 diographic analysis of cell proliferation kinetics in
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human genital tissues I. Normal cervix and vagina.
Dancis, J., W. L. Money, S. Springer, and M. Levitz: Am. J. Obstet. Gynecol. 108:8, 1970.
Transport of amino acids by placenta. Am. 1. Obstet. Burgos, M. H., and C. E. Roig de Vargas-Linares: Cell
Gynecol. 101:820, 1968. junctions in the human vaginal epithelium. Am. J.
Enders, A. C.: Formation of syncytium from cytotro- Obstet. Gynecol. 798:565, 1970.
phoblast in the human placenta. Obstet. Gynecol Burgos, M. H., and C. E. Roig de Varga-Linares: Ultra¬
25:378, 1965.
structure of the vaginal mucosa. In Hafez, E. S. E.,
Enders, A. C.: Fine structure of anchoring villi of the and T. N. Evans, eds.: Human Vagina. Amsterdam,
human placenta. Am. J. Anat. 722:419, 1968. Elsevier/North Holland Biomedical Press, 1978.
Enders, A. C., and A. G. Hendrickx: Morphological King, B. F.: Ultrastructure of the non-human primate
basis of implantation in the rhesus monkey. In vaginal mucosa: epithelial changes during the men¬
Hubinont, P. O., C. A. Finn, A. Psychoyos, and F. strual cycle and pregnancy. J. Ultrastruct. Res. 82:1,
Leroy, eds.: Blastocyst Endometrium Relationship. 1983.
Basel, S. Larger, 1980.
King, B. F.: The permeability of non-human primate
Enders, A. C., and B. F. King: The cytology of Hofbauer
vaginal epithelium: a freeze-fracture and tracer per¬
cells. Anat. Rec. 767:231, 1970.
fusion study J. Ultrastruct. Res. 85:99, 1983.
Hamilton, W. J., and J. D. Boyd: Development of the
Roig de Vargas-Linares, C. E., and M. H. Burgos:
human placenta in the first three months of gesta¬
Migration ol lymphocytes in the normal human
tion. J. Anat. 94:297, 1960.
vagina. Am. J. Obstet. Gynecol.792:1094, 1968.
Hertig, A. T.: Gestational hyperplasia of the endome¬
Roig de Vargas-Linares, C. E., and M. H. Burgos:
trium. Lab. Invest. 75:1153, 1964.
Langerhans cells in the intercellular channels of
Hertig, A. T., and j. Rock: Two human ova of the
normal human vagina. Microsc. Electron. Biol. Cell.
previllous stage, having an ovulation age of about 7:93, 1983.
 ....—— 33
MAMMARY GLAND

The mammary glands are specialized ac¬ creases during pregnancy. An elaborate pat¬
cessory glands of the skin that have evolved tern of bundles of smooth muscle disposed
in mammals to provide for the nourishment longitudinally along the lactiferous ducts and
of their offspring, which are born in a rela¬ circumferentially both within the nipple and
tively immature and dependent state. They around its base (Fig. 33-1) are responsible
are paired glands that are laid down in the for the erection of the nipple in response to
embryo along two lines called the mammary certain stimuli. At other times it is flat. In the
lines, extending from the axilla to the groin areola are the accessory areolar glands of Mont¬
on either side of the midline on the ventral gomery, which are intermediate in their struc¬
aspect of the thorax and abdomen. Mammary ture between sweat glands and true mam¬
glands may arise anywhere along these lines. mary glands. Along the margin of the areola
The number formed and their location vary are large sweat glands and sebaceous glands,
with the species. In humans, only two nor¬ which usually lack associated hairs.
mally develop, but additional accessory nip¬ The skin at the tip of the nipple is richly
ples or glandular masses are not uncommon. innervated with free nerve endings and Meis¬
In their structure and mode of develop¬ sner’s corpuscles in the dermal papillae.
ment, mammary glands somewhat resemble There are also superficial nerves and nerve
sweat glands. Their differentiation during end organs on the sides of the nipple and on
embryonic life is similar in the two sexes. In the areola. The skin peripheral to the areola
the male, however, little additional develop¬ has neural plexuses around hair follicles, as
ment occurs in postnatal life, whereas in the well as nerve endings resembling Merkel’s
female the glands undergo extensive struc¬ discs and Krause’s end bulbs. Pacinian cor¬
tural changes correlated with age and with puscles may also be found deep in the dermis
the functional condition of the reproductive and in the glandular tissue. The sensory
system. The greatest development of the fe¬ innervation of the nipple and areola are of
male breast is reached in about the twentieth great functional importance because their
year, with atrophic changes setting in by the stimulation by the suckling infant initiates
age of 40 and becoming marked after the the train of neural and neurohumoral events
menopause. In addition to these gradual that result in ejection of milk and mainte¬
changes, there are variations in the size of nance of secretion of prolactin from the pi¬
the breasts correlated with the menstrual tuitary, which is essential for continued lac¬
cycle, and striking changes in the amount tation.
and functional activity of the glandular tissue
during pregnancy and lactation. Resting Mammary Gland
The mammary gland is a compound tubu-
Nipple and Areola
loalveolar gland consisting of 15 to 25 irreg¬
The nipple is surrounded by a circular ular lobes radiating from the mammary papilla,
pigmented area of skin called the areola. The or nipple. The lobes are separated by layers
base of the epidermis of the nipple and areola of dense connective tissue and surrounded
is invaded by unusually long dermal papillae, by abundant adipose tissue. Each lobe is pro¬
whose capillaries bring blood close to the vided with a lactiferous duct 2 to 4.5 mm in
surface, imparting a pinkish color to this diameter and lined by stratified squamous
region in immature and blonde individuals. epithelium. Each duct opens on the nipple
The epidermis becomes pigmented at pu¬ and has an irregular angular outline in cross
berty, and the degree of pigmentation in¬ section. Deep to the areola, each of the ducts
901
902 • MAMMARY GLAND

Sebaceous glands
Apex of nipple Epidermis without hairs

C onnective tissue stroma

Figure 33-1. Nipple of female breast in perpendicular section, x 6. (After Schaffer.)

converging upon the nipple has a local dila¬ ponent consists only of ducts and their
tation, the sinus lactiferus. Distal to this the branches. Other authors, however, insist that
duct narrows again and emerges at the end a few small alveoli are present at the ends of
of the nipple as a separate opening 0.4 to 0.7 the duct system and form miniature lobules.
mm in diameter. The question remains unresolved, owing in
Each lobe is subdivided into lobules of part to the rarity of opportunities to obtain
various orders. The smallest consist of elon¬ normal human breast tissue at known phases
gated tubules, the alveolar ducts, covered by of the menstrual cycle. Certainly the total
small saccular evaginations, the alveoli. The mass of the epithelial elements is greatly
interlobular connective tissue is more dense reduced early in the cycle. A lumen is not
than the intralobular connective tissue. The evident and the cells seem to form more or
latter is more cellular and contains fewer less solid cords. In this condition, it is not
collagenous fibers and almost no fat. This easy to distinguish alveoli from primary
loose intralobular stroma surrounding the ducts. Late in the menstrual cycle, the epi¬
system of ducts is believed to permit greater thelial cells become cuboidal or low colum¬
distensibility when the epithelial portions of nar, a lumen is apparent, and the surround¬
the organ hypertrophy during pregnancy ing connective tissue is highly vascular. Since
and lactation. The secretory portions of the both alveolar ducts and alveoli are potentially
gland, the alveolar ducts and alveoli, consist secretory, it is not important to make this
of cuboidal or low columnar secretory cells distinction in the nonlactating breast. It is
resting on a basal lamina and underlying sufficient to note that there are microscopi-
processes of myoepithelial cells. The highly ally detectable cyclic changes in the epithelial
branched myoepithelial cells enclose the glan¬ portions of the mammary gland, but these
dular alveoli in an open-meshed, basket-like are relatively slight. The more obvious
network. They usually lie between the secre¬ changes in breast size, and the sense of en¬
tory cells and the basal lamina. The presence gorgement experienced by some women in
of myoepithelial cells is further evidence that certain phases of the cycle, are attributable
mammary glands are morphogenetically re¬ to increased blood flow and a slight associated
lated to sweat glands. edema of the connective tissues of the breast.
There are differences of opinion as to Unlike many other glands, the mammary
whether alveoli are present in the nonlactat- gland does not have a single excurrent duct.
ing human breast. According to most descrip¬ Each lobe is an independent compound al¬
tions of the inactive gland, its epithelial com¬ veolar gland whose primary ducts join and
 MAMMARY GLAND • 903

rejoin to form larger ducts that converge During the first half of gestation, there is a
upon a single lactiferous duct, and lactiferous rapid growth and branching from the ter¬
ducts of the several lobes open separately at minal portion of the duct system of the gland
the tip of the nipple. Thus, the mammary and a proliferation of alveoli. The growth of
gland is a conglomerate made up of a variable epithelial components of the gland takes
number of independent units. place, at least in part, at the expense of the
The myoepithelial cells are stellate in form, interstitial adipose tissue of the breast, which
with long branching processes that occupy regresses concurrently with the growth of the
recesses in the bases of the glandular cells. glandular tissue. In this period of growth,
The processes of neighboring myoepithelial there is also an increasing infiltration of the
cells join to form a cellular network that interstitial tissue with lymphocytes, plasma
completely envelops the alveolus. These cells cells, and eosinophils. In the later months of
lie between the epithelial cells and the basal pregnancy, the hyperplasia of the glandular
lamina. Their form and topographical rela¬ tissue slows down, and the subsequent en¬
tionships are seen to advantage in scanning largement of the breasts is mostly a conse¬
electron micrographs of glands exposed dur¬ quence of enlargement of the parenchymal
ing specimen preparation to collagenase to cells and distention of the alveoli with a
remove the basal lamina and associated retic¬ secretion rich in lactoproteins but relatively
ular fibers (Figs. 33-2, 33-3). poor in lipid. This constitutes the colostrum,
the first milk that comes from the breasts
The Active Mammary Glands after birth. It has special laxative properties
and contains antibodies that provide the new¬
Pregnancy brings about changes in the born with some measure of passive immunity.
levels of circulating hormones that result in During the first few days after delivery the
profound changes in the mammary glands. degree of infiltration of the stroma of the

»s

Figure 33-2. Scanning micrograph of acini of rodent mammary gland. The tissue was treated with enzymes during
specimen preparation to digest away collagen fibrils, extracellular matrix, and basal lamina, thus exposing the bases of
the epithelial and myoepithelial cells. (Micrograph from Nagato, T. Cell Tissue Res. 209:1, 1980.)
904 • MAMMARY GLAND

In the cytoplasm are short, rod-shaped

mitochondria, few in number in the flattened
cells but more plentiful in the taller ones.
The cells are generally acidophilic, but some
basophilic substance may be found at the
base of the cells. Droplets of lipid, often of
large size, accumulate near the free surface
and often project into the lumen (Fig. 33—6).
After the extraction of fat in preparing his¬
tological sections, large clear vacuoles remain
in place of the lipid droplets. In addition to
the accumulations of lipid, small proteina¬
ceous secretory granules can also be seen in
the apical region of the cell. Cyclic changes
in the Golgi apparatus during the different
phases of secretion have been described. The
lumen of the alveoli is filled with fine gran¬
ules and lipid droplets similar to those that
protrude from the cells (Figs. 33-6, 33-7).
The mammary gland was formerly be¬
lieved to have a mode of release of its product
that was intermediate between merocrine se¬
cretion, in which the secretory materials pass
out through the cell apex without loss of
cytoplasm, and holocrine secretion, in which
Figure 33-3. Scanning micrograph of an acinus of the
the entire cell is given up in contributing its
mammary gland, clearly showing the branching myoepi¬
thelial cells occupying grooves between the bases of the contents to the secretion. The cells of the
secretory cells. Specimen prepared as described for Fig¬ mammary gland were believed to undergo a
ure 33-2. (Micrograph from Nagato, T. Cell Tissue Res. partial disintegration in which the lipid-hlled
209:1,1980.)
apical portion of the cell, projecting into the
lumen, was described as constricting from
the base of the cell, which remained in place.
It was believed that the remainder of the cell
gland by lymphoid elements becomes less did not die but rapidly replaced the lost
intense, and the colostrum gives way to a protoplasm and reaccumulated secretion.
copious secretion of milk rich in lipid. This mode of release was called apocrine se¬
The histological appearance of different cretion. Studies with the electron microscope
parts of the active mammary gland varies have now radically changed our views as to
considerably (Figs. 33-4, 33-5). Apparently, the mechanisms of release of cell products,
different areas are not all in the same func¬ and the traditional concept of apocrine secre¬
tional state at the same time. In some places, tion is no longer applicable to the mammary
the secretory portions are filled with milk; gland.
their lumen is wide and the walls are dilated In electron micrographs the main cytolog-
and thin. In other areas, the lumen is narrow ical features of the glandular cells are in
and the epithelium relatively thick. accord with desriptions resulting from use of
The shape of the epithelial cells varies from the light microscope. The chromophilic areas
flat to low columnar. The boundary between of the cytoplasm contain numerous cisternal
them is usually indistinct in histological sec¬ profiles of the granular endoplasmic reticu¬
tions. If the cells are tall, their distal ends are lum. There are a moderate number of mi¬
often separated and project into the lumen tochondria, a large supranuclear Golgi com¬
of the alveoli as rounded or dome-shaped plex, and a few lysosomes. It is evident that
protrusions. The nucleus may be round or the cell has two distinct secretory products,
oval and is at about the middle of the cell. If formed and released by different mecha¬
the cells are short, their free surface is usually nisms. The protein constituents of the milk,
more or less smooth. like other protein secretions, are elaborated
 MAMMARY GLAND • 905

Figure 33-4. Photomicrograph of human mammary gland at eight months of pregnancy.

on the ribosome-studded membranes of the of cell membrane and, in some instances, of

endoplasmic reticulum. They first become a thin layer of cytoplasm, but the amount lost
visible as multiple dense spherical granules is certainly far less than envisioned by the
about 400 nm in diameter in vesicles asso¬ classical cytologists who introduced the con¬
ciated with the Golgi complex. They are cept.
transported to the cell surface in these mem¬ Lymphocytes are sometimes encountered
branous vesicles, which fuse with the plas- among the alveolar epithelial cells. Between
malemma and discharge their contents into the epithelial cells and the basal lamina are
the lumen of the acinus. The mode of for¬ occasional cells with pale cytoplasm rich in
mation and release of this particulate com¬ lipid droplets and vacuoles with highly het¬
ponent of the milk is identical to that of other erogeneous granular and membranous con¬
protein-secreting glands that are generally tents. These have been interpreted by some
classified as ynerocrine. as degenerating epithelial cells, but it seems
The fatty components of the milk do not more likely that they are macrophages.
develop in association with the Golgi appa¬ The myoepithelial cells lie on the epithelial
ratus but arise as small lipid droplets free in side of the basal lamina. Their processes are
the cytoplasmic matrix. These increase in size filled with parallel arrays of myofilaments 6
by fusion and move into the apical region, nm in diameter. There are spindle-shaped
where they come to project into the lumen densities among the myofilaments like those
covered by the plasmalemma. These droplets found in smooth muscle cells. The cell organ¬
are ultimately cast off, enveloped by a de¬ elles are concentrated in the perinuclear re¬
tached portion of the cell membrane (Fig. gion of the cell body, but occasional mito¬
33-8). This mode of release could be consid¬ chondria and profiles of the endoplasmic
ered apocrine in the sense that it involyes loss reticulum extend into the cell processes
906 MAMMARY GLAND

Figure 33-5. Photomicrograph of lactating human mammary gland. Notice the local variations within the gland. Alveoli
in some areas have a large lumen and abundant secretion, whereas in other areas the lumen is very small.
 MAMMARY GLAND • 907

Figure 33-7. Photomicrograph of lactating mammary

gland from a mouse, fixed in osmium tetroxide. The large
Figure 33-6. Drawing of rat mammary gland acinus,
droplets of lipid are preserved both in the apex of the
showing the relationships of the myoepithelial cells, the
cells and in the lumen of the acini. The smaller protein
structure of the secretory cells, and the presence of small
granules are not visible, x 560. (Preparation by N. Feder.)
granules of casein and membrane-bounded lipid droplets
in the lumen. (Drawing from Krstic, R. Die Gewebe des
Menschen und der Saugetiere. Berlin, Springer-Verlag,
1978.)
in proportion to the growth of the body as a
whole until puberty. Then, in the female, a
more rapid extension of the duct system
Histophysiology of the begins. This accelerated growth depends pri¬
Mammary Gland marily on estrogen and progesterone from
The functioning of the mammary glands the ovaries, but also requires prolactin and
is dependent on multiple and complex neural somatotrophin from the pituitary.
and endocrine factors. Some are involved in In pregnancy the large quantities of estro¬
the development of the glands to a functional gen secreted by the placenta stimulate fur¬
state {mammo gene sis), others in the establish¬ ther growth of the duct system. Increased
ment of milk secretion (lacto gene sis), and still levels of circulating progesterone from the
others are necessary for maintenance of lac¬ corpus luteum of pregnancy acting together
tation. Given a normal coordinated interplay with hypophyseal and adrenal hormones in¬
of all the factors concerned, the average milk duce development of alveoli and differentia¬
production of a mother breast-feeding a sin¬ tion of their cells in preparation for lactogen-
gle infant is in excess of 1100 ml/24 hr for esis. Secretion of prolactin by the pituitary
the first six months of lactation, and mothers steadily increases from the fifth week of preg¬
breast-feeding twins may produce over 2100 nancy until full term, when it may reach
ml/24 hr. levels ten times those of the nonpregnant
The duct system of the gland that develops state. Hypophyseal prolactin is the principal
in fetal life continues to grow after birth only hormone stimulating milk production. Ad-
908 • MAMMARY GLAND

Figure 33-9. Electron micrograph of a mammary epithe¬

Myoepithelial cell lial cell, showing several vacuoles containing granules of
milk protein. One (at arrow) is in process of exocytosis.
process
(Micrograph courtesy of A. Ichikawa.)
Figure 33-8. Diagrammatic representation of a cell from
a lactating mammary gland, showing large lipid droplets
being cast off enclosed in a layer of cytoplasm, and small
granules of protein secretion being concentrated in the
Golgi apparatus and released by coalescence of their mary glands, the high level of prolactin can
small vesicles with the plasma membrane. A myoepithelial
cell process is depicted in cross section between the exert its powerful lactogenic effect, and
epithelial cell and the basal lamina. (Drawing based on within a few days full lactation is established.
observations of Bargmann, W., and A. Knoop. Zeitschr. f. Prolactin secretion by the pituitary falls to
Zellforsch. 49:344, 1959.) basal levels post partum, but each time the
baby is breast-fed the stimulation of the nip¬
ples by suckling sends nervous impulses to
renal glucocorticoids and insulin also play a the hypothalamus that are relayed to the
significant role in lactation. In the absence of hypophysis, resulting in a brief tenfold in¬
insulin, the niRXA for synthesis of the milk crease in release of prolactin. These fre¬
protein casein fails to accumulate to normal quently repeated surges of the hormone stim¬
levels in the mammary epithelium. ulating the cells of the mammary gland serve
Although the mammary gland is fully de¬ to maintain milk production. Frequent breast
veloped late in pregnancy, very little milk is feeding is important to maintain lactogenesis.
formed owing to inhibition of secretion by The tendency of mothers in developed coun¬
high levels of estrogen and progesterone. tries to prolong the interval between feedings
The small amount of fluid produced, called and to abandon nighttime feeding as soon as
colostrum, is poor in lipid but rich in protein, possible contributes to their poor lactational
and contains IgA. With the elimination of performance. In contrast, women of hunter-
the placenta at birth, estrogen and progester¬ gatherer societies whose babies are always
one levels in the mother fall dramatically. with them and feed on demand throughout
Without their inhibitory action on the mam- the day and night are able to continue to
 MAMMARY GLAND • 909

where it accumulates in the intervals between

feedings. Flow of milk from the nipples re¬
quires participation of the baby. The stimulus
of suckling generates afferent nerve impulses
that ascend in the spinal cord to the hypo¬
thalamus. In addition to activating release of
prolactin from the anterior pituitary, efferent
impulses from the hypothalamus to the pos¬
terior lobe of the pituitary result in its secre¬
tion of the hormone oxytocin. Carried in the
blood to the mammary gland, oxytocin stim¬
ulates contraction of the myoepithelial cells,
resulting in ejection of milk from the alveoli
into the ducts and the initiation of flow from
the nipple.
The secretory cells of the mammary epi¬
thelium are unusual among glandular cells
in that they produce multiple products using
different biosynthetic pathways. The milk
proteins are synthesized on ribosomes; seg¬
regated in the endoplasmic reticulum; and
glycosylated, concentrated, and packaged in
vesicles in the Golgi complex. Among the
proteins produced is the enzyme lactose syn¬
thetase, which catalyzes the synthesis of the
milk sugar lactose from glucose and UDP-
galactose. The membrane of the Golgi vesi¬
cles is permeable to these precursor mole¬
Figure 33-10. Micrograph of a mammary epithelial cell cules but not to the product lactose, which
with a lipid droplet protruding into the lumen covered by accumulates and creates an osmotic gradient
a portion of the cell membrane. Secreted lipid droplets
and granules of milk protein are visible in the lumen. that draws water and monovalent ions into
(Micrograph courtesy of A. Ichikawa.) the vesicles. In other protein-secreting cells,
large dense granules are formed and en¬
closed individually in a close-fitting mem¬
brane. In the mammary gland, multiple small
breast-feed their babies for up to three and granules of milk protein are sequestered
a half years. in relatively large vesicles also containing
In women breast-feeding their babies, lactose, water, and ions, which are extracted
there is usually a concurrent suppression of in specimen preparation. In micrographs,
the menstrual cycle and ovulation—lactational therefore, the secretion vesicles appear
amenorrhea. Its mechanism is not fully under¬ empty save for a few small protein granules
stood, but it is believed that neural inputs to (Fig. 33-9). Their contents are discharged
the hypothalamus generated by suckling into the lumen by exocytosis.
stimulate release of beta endorphin, which The lipid of the milk is not synthesized in
suppresses secretion of gonadotropin releas¬ a membrane-bounded organelle but appears
ing hormone and results in diminished secre¬ as small droplets in the cytoplasmic matrix.
tion of luteinizing hormone and failure of These enlarge and coalesce into very large
ovulation. Prolonged lactation is the only droplets that bulge into the lumen, covered
method of contraception practiced by the only by the apical plasmalemma (Fig. 33-10).
great majority of women in the developing By a process of exocytosis that is unique and
countries. poorly understood, the cell membrane fuses
The production of milk is a continuous behind the projecting droplets, which are
process, but its delivery is episodic. The cells thus cast off into the lumen invested in a
of the lactating mammary gland synthesize detached portion of the plasmalemma.
and release their product continuously into The immunoglobulin of the milk follows a
the lumen of the alveoli and alveolar ducts, third transcellular path. It is synthesized by
910 • MAMMARY GLAND

Lactose Regression of the Mammary Gland

Ca + +, PO4 Lipid
If regular suckling is permitted, lactation
can be maintained for many months or even
for several years. However, if milk is not
removed, the glands become greatly dis¬
tended and milk production quickly ceases.
This is in part due to interruption of the
neurohormonal reflex mechanism for main¬
tenance of prolactin secretion, but the en¬
gorgement of the breasts may also compress
the blood vessels, resulting in diminished
access of oxytocin to the myoepithelial cells.
After a few days the secretion remaining in
the alveolar spaces and ducts is absorbed,
and the glandular elements gradually return
to the resting state. The gland, however, does
not return completely to its original state,
because many of the alveoli that had formed
during the period of pregnancy do not dis¬
appear entirely, and the remains of the se¬
cretion may sometimes be retained in the
mammary ducts for a considerable time. The
gland remains in such a resting condition
until the following pregnancy, when the same
cycle of changes is repeated.
Figure 33-11. Schematic representation of the transcel- The process of mammary gland regression
lular pathways involved in milk secretion. Casein, lactate,
has been studied mainly in laboratory ani¬
calcium, and citrate are packaged in vacuoles arising in
the Golgi complex and released by exocytosis. Water and mals, but the changes are undoubtedly very
ions diffuse freely through the cell membrane. Lipid drop¬ similar in the human. A few days after wean¬
lets enclosed in detached portions of plasmalemma are ing, the alveoli are greatly distended with
released in a unique form of apocrine secretion, immu¬ secretory products, and the epithelium is
noglobulin taken up at the basal and lateral surfaces by
receptor-mediated endocytosis is transported in small correspondingly flattened. Later there is a
vesicles across the cell and released into the lumen. gradual collapse of the alveoli and an associ¬
(Redrawn and modified after Neville, M.C., et al. In ated increase in perialveolar connective tissue
Lactation: Physiology, Metabolism and Breast-feeding. and adipose tissue. There is an increase in
New York, Plenum Press, 1983.)
macrophages in the interstitial tissue but no
true inflammatory reaction. By ten days after
weaning, the glandular tissue is largely re¬
plasma cells in the stroma of the mammary placed by connective and adipose tissue, and
gland that were exposed to intestinal patho¬ the remaining alveoli appear as scattered
gens in the lamina propria of the mother’s solid cords of epithelial cells.
intestine and carried to the gland in the Examined in electron micrographs, the al¬
blood. There, IgA is taken up by receptor- veolar cells show an early accumulation of
mediated endocytosis at the basal and lateral intracellular secretory protein in large vacu¬
surfaces of the glandular cells, transported oles. There is also a marked progressive in¬
in vesicles to the cell apex, and discharged crease in the number of intraepithelial mac¬
into the lumen (Fig. 33-11). In the fourth rophages. Whereas autophagic vacuoles are
and fifth months of lactation, a mother may rare in the alveolar epithelium of the lactat-
be secreting in her milk as much as 0.5 g of ing gland, they rapidly increase in number
antibody per day. The antibody in the lumen and size in the first few days after weaning.
of the baby’s gut combats enteric infections, I heir contents include mitochondria, gran¬
a common cause of infant mortality. Early ular reticulum, and secretory granules.
weaning and bottle feeding deprive the baby There is a concomitant increase in the het-
of this passive immunity.
erophagic activity of the intraepithelial mac-
 MAMMARY GLAND • 911

rophages. Since they may contain ingested the secretory portions. The veins drain into
secretory granules, it is concluded that they the axillary and anterior thoracic veins.
take up organelles and inclusions from re¬ The lymphatic vessels begin with capillary
gressing or degenerating epithelial cells. networks located in the connective tissue lay¬
Some of the latter clearly slough in later ers surrounding separate alveoli. They collect
stages of regression, and these are disposed along the course of the mammary ducts into
of by macrophages. a subpapillary lymphatic network. From here
Concomitant with the electron microscopic several large vessels drain the lymph mainly
appearance of increases in autophagic and into the lymph nodes in the axilla and sub-
heterophagic vacuoles in the first few days clavicular area, but they also have connec¬
after weaning, there is also a marked increase tions with the lymphatics that penetrate the
in activity of the lysosomal enzymes aryl sul- intercostal spaces to reach parasternal lymph
fatase, cathepsin D, and acid phosphatase nodes. Understanding the lymphatic drain¬
despite the fact that the activity of other age of the gland is of clinical importance
nonlysosomal enzymes is declining. Thus, owing to the necessity of removing the re¬
there is apparently a synthesis of new lyso¬ gional lymph nodes in radical mastectomy
somal enzymes during the early phases of for breast cancer.
regression.
In old age, the mammary gland gradually Innervation
undergoes involution. The epithelium of the
secretory portions, and partly also of the Norepinephrine-containing nerve fibers
excretory ducts, atrophies, and the gland are abundant among the smooth muscle cells
tends, in a general way, to return to the of the nipple and at the interface between
prepubertal condition, in which there are media and adventitia of the arteries of the
only a few scattered ducts. Equally striking breast. This is in accord with physiological
changes occur in the interstitial connective observations indicating that the efferent
tissue. This becomes decidedly less cellular; nerves of these structures are sympathetic
the number of collagenous fibrils decreases, adrenergic. There .seems to be no evidence
and the whole mass becomes more homoge¬ that cholinergic fibers supply any part of the
neous and stains much less intensely with gland.
eosin. Most mammary nerves follow the arteries
The mammary epithelium not infrequently and arterioles and supply these structures. A
is the site of pathological changes. The dis¬ few fibers leave the perivascular networks
order known as chronic cystic disease is very and lie near the walls of the ducts. They may
common in women between 30 and 50 years correspond to sensory fibers for sensing milk
of age. The terminal ducts and acini may pressure, which have been postulated on the
lose their continuity with the remainder of basis of behavioral and electrophysiological
the duct system and may form fluid-filled studies. There is no morphological evidence
cysts of varying size. The breast is the most of a nerve supply to the secretory cells or
common site of cancer among women. About myoepithelial cells.
one in every 17 newborn girls (6 per cent)
may be expected to develop breast cancer at
some time during her life—an incidence REFERENCES
three times that of cancer of the colon, the
second most common type. Bauman, D. E., and C. L. Davis: Biosynthesis of milk
fat. In Larson, B. L., and V. R. Smith, eds.: Lacta¬
tion: A Comprehensive Treatise. Vol. II. New York,
Blood and Lymphatic Vessels Academic Press, 1974, pp. 31—75.
Cowie, A. T., and S. J. Lolley: The mammary gland and
The arteries of the mammary gland arise lactation. In Young, W. C., ed.: Sex and Internal
from the anterior thoracic (internal mam¬ Secretions, 3rd ed. Baltimore, Williams & Wilkins
mary) artery, the thoracic branches of the Co., 1961, p. 590.
Dempsey, E. W., H. Bunting, and G. B. Wislocki: Ob¬
axillary artery, and the intercostal arteries. servations on the chemical cytology of the mammary
They pass mainly along the larger ducts and gland. Am. J. Anat. 67:309, 1947.
break up into dense capillary networks on Loote, L. W., and L. W. Stewart: Comparative studies of
the external surface of the basal lamina of cancerous versus non-cancerous breasts. I. Basic
912 • MAMMARY GLAND

morphological characteristics. Ann. Surg. 121:6, Mills, E. S., and Y. J. Topper: Some ultrastructural
1945. effects of insulin, hydrocortisone and prolactin on
Gardner, W. U., and G. van Wagenen: Experimental mammary gland explants. J. Cell Biol. 44:310, 1970.
development of the mammary gland in the monkey. Montagna, W.: Histology and cytochemistry of human
Endocrinology 22:164, 1938. skin. XXXV. The nipple and areola. Br. J. Derma¬
Hebb, C., and J. L. Linzell: Innervation of the mammary tol. <*?5(Suppl.):2, 1970.
gland. A histochemical study in the rabbit. Histo- Mostov, K. E., J. P. Krakenbuhl, and G. Blobel: Recep¬
chem. J. 2:491, 1970. tor-mediated transcellular transport of immuno¬
Helminen, H. J., and J. L. E. Ericsson: Studies on globulin: synthesis of secretory component as mul¬
mammary gland involution. I. On the ultrastructure tiple and larger transcellular forms. Proc. Natl.
of the lactating mammary gland. J. Ultrastruct. Res. Acad. Sci. U.S.A. 77:7257, 1980.
25:193, 1968. Neville, M. C., J. C. Allen, and C. Watters: The mecha¬
Helminen, H. J., and J. L. E. Ericsson: Studies on nism of milk secretion. In Neville, M. C., and M. R.
mammary gland involution. II. Ultrastructural evi¬ Neifert, eds.: Lactation: Physiology, Nutrition, and
dence for auto- and heterophagocytosis. J. Ultras¬ Breast Feeding. New York, Plenum Press, 1983, pp.
truct. Res. 25:214, 1968. 49-103.
Helminen, H. J., J. L. E. Ericsson, and S. Orrenius: Richardson, K. C.: Contractile tissues in the mammary
Studies on mammary gland involution. IV. Histo¬ gland, with special reference to myoepithelium in
chemical and biochemical observations on alteratons the goat. Proc. R. Soc. B 756:30, 1949.
in lysosomes and lysosomal enzymes. J. Ultrastruct. Short, R. V.: Breast feeding. Sci. Am. 250:23, 1984.
Res. 25:240, 1968. Tindal, J. S.: Hypothalamic control of secretion and
Hollmann, K. H.: Cytology and fine structure of the release of prolactin. J. Reprod. Fertil. 59:437,
mammary gland. In Larson, B. L., and V. R. Smith, 1974.
eds.: Lactation: A Comprehensive Treatise. Vol. I. Vorherr, H., ed.: Human Lactation. New York, Grune
New York, Academic Press, 1974, pp. 1-91. & Stratton, 1979.
Kon, S. K., and A. T. Cowie: Milk; The Mammary Gland Wellings, S. R., K. B. DeOme, and D. R. Pitelka: Electron
and Its Secretion. New York, Academic Press, 1961. microscopy of milk secretion in the mammary gland
Kurosumi, K., Y. Kobayashi, and N. Baba: The fine of the C3H/Crgl mouse. I. Cytomorphology of the
structure of mammary glands of lactating rats with prelactating and the lactating gland. J. Natl. Cancer
special reference to the apocrine secretion. Exp. Inst. 25:393, 1960.
Cell Res. 50:177, 1968. Wellings. S. R., B. W. Grunbaum, and K. B. DeOme:
Linzell, J. L., and M. Peaker: Mechanism of milk secre¬ Electron microscopy of milk secretion in the mam¬
tion. Physiol. Rev. 57:564, 1971. mary gland of the C3H/Crgl mouse. II. Identifica¬
Mayer, G., and M. Klein: Histology and cytology of the tion of fat and protein particles in milk and in tissue.
mammary gland. In Kon, S. K., and A. T. Cowie, J. Natl. Cancer Inst. 25:423, 1960.
eds.: Milk: The Mammary Gland and its Secretion. Wellings, S. R., and J. R. Phelp: The function of the
Vol. 1. New York, Academic Press, 1961. Golgi apparatus in lactating cells of the BALB/cCrgl
Miller, M. R., and M. Kasahara: Cutaneous innervation mouse: an electron microscopic and autoradi¬
of the human female breast. Anat. Rec. 755:153, ographic study. Zeitschr. f. Zellforsch. 67:871,
1959. 1964.
 34

The ability to react to light is a widespread light to enter. The rest of the wall of the eye
property of living matter. Plants use solar is opaque and possesses a darkly pigmented
energy for photosynthesis and exhibit pho¬ inner surface, which absorbs light rays. The
totropic responses. Primitive invertebrates posterior segment of the eye is to a great
have scattered photoreceptor cells that detect extent lined with photosensitive nervous tis¬
varying intensities of light and enable them sue, the retina, which develops as an out¬
to position themselves favorably with respect growth from the brain. The cavity of the
to light or darkness. Vertebrates have evolved eyeball is filled with transparent media ar¬
eyes—more efficient organs with a lens to ranged in separate bodies, which, together
concentrate light and focus an image on with the cornea, act as a system of convex
closely packed photoreceptors in a retina, lenses. These produce on the photosensitive
which detects light intensity, color, form, and layer of the retina an inverted and reduced
motion and encodes the various parameters image of the objects in the environment.
of the image for transmission to the brain. The wall of the eyeball is composed of
The eyes of most vertebrates are placed on three layers: the tough, fibrous, corneoscleral
the sides of the head where they provide a coat; the middle, vascular coat, or uvea; and
nearly complete panoramic view of the envi¬ the innermost layer, the photosensitive retina.
ronment. Panoramic vision, together with the The thick fibrous layer protects the delicate
evolution of muscles and reflexes for rotation inner structures of the eye and, together with
of the eyes, provided early warning against the intraocular fluid pressure, serves to main¬
predators. On the other hand, in many pred¬ tain the shape and turgor of the eyeball. It is
atory mammals and birds the orbits gradually divided into a large opaque posterior seg¬
moved forward so that the uniocular fields ment, the sclera, and a smaller transparent
of vision came to overlap in varying degree anterior segment, the cornea. The uvea is
and the sector of the environment in front concerned with the nutrition of the ocular
of the animal was seen by both eyes. Complex tissues and also provides mechanisms for
neural mechanisms developed to coordinate visual accommodation and control of the
and fuse the slightly different images, thus amount of light entering the eye. Its three
achieving binocular vision and, in some in¬ regional differentiations are the choroid, the
stances, stereoscopic vision with perception ciliary body, and the iris. The choroid is the
of depth in a three-dimensional view of the highly vascular portion of the uvea that un¬
environment essential to successful pursuit derlies the photosensitive retina. Extending
of prey. Acquisition of stereoscopic vision, forward from the scalloped anterior margin
evolution of a large brain to process the of the retina, called the ora serrata, to the
information, and freeing of the hands from corneoscleral junction is the ciliary body. It
a locomotor function enabled our hominoid forms a belt 5 to 6 mm wide around the
ancestors to develop the manipulative skills interior of the eyeball and contains the
that contributed to the ascendancy of hu¬ smooth muscle that makes this structure the
mans. instrument of accommodation, acting upon
the lens to bring light rays from different
distances to focus upon the retina. The iris
STRUCTURE OF THE EYE is a thin continuation of the ciliary body
IN GENERAL projecting over the anterior surface of the
lens, with its free edge outlining the pupil.
The anterior segment of the eye, the cor¬ The diameter of the iris is approximately 12
nea, is transparent, permitting the rays of mm. Its opening, the pupil, can be reduced

913
914 • THE EYE

Figure 34-1. Diagram of horizontal meridional section through the right eye of man.

or expanded through the contraction or re¬ the surrounding air (1.0), the cornea is the
laxation of the constrictor and dilator muscles chief refractive element of the eye. Of the
of the pupil. In this way, the iris functions as enclosed transparent media, the most ante¬
an adjustable optic diaphragm regulating the rior is the aqueous humor. It is contained in
amount of light entering the eye (Figs. 34-1, the anterior chamber, a small cavity bounded
34-2). in front by the cornea and behind by the iris
The innermost layer, the retina, contains and the central portion of the anterior sur¬
in its sensory part the receptors for light and face of the lens. The posterior chamber, also
complex neural networks that encode the filled with aqueous humor, is a narrow, an¬
visual information and send impulses nular space enclosed anteriorly by the lens,
through the optic nerve to the brain. The spot the iris, and the ciliary body and posteriorly
where the nerve enters the eyeball, the papilla by the vitreous body (Figs. 34-1, 34-2).
of the optic nerve, is a pink disc approxi¬ The next of the transparent media is the
mately 1.4 mm in diameter located about 3 crystalline lens. This is an elastic biconvex body
mrn medial to the posterior pole of the eye. suspended from the inner surface of the
The portion of the retina anterior to the ora ciliary body by a circular ligament, the ciliary
serrata and lining the inner surface of the zonule. It is placed directly behind the pupil,
ciliary muscle (ciliary portion of the retina) and between the aqueous humor of the anterior
that lining the posterior surface of the iris chamber and the vitreous body posteriorly.
(iridial portion of the retina) are not photosen¬ The lens is second in importance to the
sitive. These will be discussed with the uvea cornea as a refractive element of the eye, and
and iris. is the dioptric organ of accommodation.
I he transparent dioptric media include the The greater portion of the cavity of the
cornea and the contents of the cavity enclosed eye, situated between the posterior surface
by the tunics of the eye. Because of the of the lens and ciliary body anteriorly and
considerable difference between the index of the retina posteriorly, is the vitreal cavity,
refraction of the cornea (1.376) and that of filled with a viscous transparent substance,
 THE EYE • 915

Cornea Anterior
Chamber

Posterior
Ciliary Chamber
Body

Optic
Retina Nerve

Choroid

Sclera

Figure 34-2. Photomicrograph of a meridional section of the eye of a rhesus monkey, x iO. (Courtesy of H. Mizoguchi.)
916 • THE EYE

the vitreous humor or vitreous body. It permits made up of loose connective and fatty tissue,
light to pass freely from the lens to the muscles, fasciae, blood and lymphatic vessels,
photoreceptors. nerves, and a gland. This soft tissue cushion
The retina is transparent in the living state. permits the eye to move freely around its
Only its outermost layer, the pigment epithe¬ center of rotation. The eye is connected to the
lium, is opaque and forms the first barrier to general integument by the conjunctiva, which
the rays of light. lines the lids and continues over the eyeball
to the margins of the cornea. The lids are a
mechanical protection against external nox¬
ious agents.
DIMENSIONS, AXES,
PLANES OF REFERENCE

The adult human eyeball is a roughly FIBROUS TUNIC

spherical body about 24 mm in diameter and
weighing 6 to 8 g. The center of the cornea
Sclera
is the anterior pole; the posterior pole is located
between the optic papilla and the fovea, a The sclera is 1 mm thick at the posterior
thin depression in the retina providing the pole, 0.4 to 0.3 mm at the equator, and 0.6
most distinct vision. The line connecting the mm toward the edge of the cornea. It consists
two poles is the anatomical axis. The visual axis of flat collagenous bundles that run in various
is the line drawn from the center of the fovea directions parallel to the surface. Between
to the apparent center of the pupil (Fig. these bundles are networks of elastic fibers.
34-1). The equatorial plane is vertical and The cells of the sclera are flat, elongated
perpendicular to the visual axis, passing fibroblasts. Melanocytes also can be found in
through the greatest width of the eyeball, the the deeper layers, especially in the vicinity of
equator. Other planes passing through the the entrance of the optic nerve.
axis determine the meridians of the eye. The The tendons of the eye muscles are at¬
two most important are the vertical and the tached to the outer surface of the sclera,
horizontal meridians. The vertical passes which in turn is connected with a dense layer
through the fovea and divides the eyeball, of connective tissue—the fascial sheath of the
including the retina, into nasal and temporal eye, or capsule of Tenon—by an exceedingly
halves. The plane of the horizontal meridian loose system of thin collagenous membranes
divides the eyeball and retina into an upper separated by clefts—the episcleral space, or
and a lower half. These two planes divide space of Tenon. The eyeball and the capsule
the eyeball and the retina into four quad¬ of Tenon rotate together in all directions on
rants, an upper nasal, an upper temporal, a a bed of orbital fat.
lower nasal, and a lower temporal. Between the sclera and the choroid is a
The anteroposterior diameter along the axis layer of loose connective tissue with elastic
of the eye is 24 mm, or a little more. The networks and numerous melanocytes and fi¬
inner axis, the distance between the inner broblasts. When these two tunics are sepa¬
surface of the cornea and the inner surface rated, part of this loose tissue adheres to the
of the retina at the posterior pole, measures choroid and part to the sclera as its supracho¬
a little less than 22 mm. The optical axis passes roid lamina.
through the optical centers of the refractive
media and is almost identical with the ana¬
Cornea
tomical axis. The visual axis, where it touches
the retina, is from 4 to 7 degrees lateral and The cornea is slightly thicker than the
3.5 degrees below the optical axis. sclera, measuring 0.8 to 0.9 mm in the center
The radius of curvature of the large poste¬ and 1.1 mm at the periphery. In the human
rior segment at the fundus measures some¬ the refractive power of the cornea, which is
what less than 13 mm, and gradually de¬ a function of the index of refraction of its
creases toward the corneoscleral junction. tissue (1.376) and of the radius of curvature
The cornea has the smallest radius of curva¬ of its surface (7.8 mm), is twice as high as
ture, approximately 7.8 mm (outer corneal that of the lens.
surface). In a cross section through the cornea, the
The eyeball is lodged in a soft tissue cush¬ following layers can be seen: (1) the corneal
ion Filing the bony orbit of the skull and epithelium, (2) the membrane of Bowman,
 THE EYE • 917

(3) the stroma, or substantia propria, (4) the the wound. A few mitoses can be found in
membrane of Descemet, and (5) the endothe¬ the basal cell layer under normal conditions.
lium (Fig. 34-3). Bowman’s Membrane. The corneal epithe¬
Epithelium. The epithelium is stratified lium rests on a faintly fibrillar lamina 6 to 9
squamous, with an average thickness of 50 (Jim thick. This structure is not actually a
fim. It consists, as a rule, of five layers of membrane but the outer layer of the substan¬
cells. The outer surface is quite smooth and tia propria of the cornea, from which it
is composed of large squamous cells. As in cannot be separated. It is nevertheless distin¬
other types of stratified squamous epithe¬ guishable with the optical microscope because
lium, the cells are connected with one an¬ its fibers are not so well ordered. With the
other by many short interdigitating processes electron microscope it is seen to consist of a
that adhere at desmosomes. The cytoplasm feltwork of randomly arranged collagen fi¬
contains numerous mitochondria and scat¬ brils, about 18 nm in diameter, which may
tered profiles of granular endoplasmic retic¬ show a periodic banding. It does not contain
ulum in a cytoplasmic matrix filled with ran¬ elastin and ends abruptly at the margin of
domly oriented fine filaments. the cornea. Bowman’s membrane is not pres¬
The epithelium of the cornea is extremely ent in all mammals; in rabbits the corneal
sensitive and contains numerous free nerve epithelium rests on a simple basal lamina.
endings. It is endowed with a remarkable Stroma or Substantia Propria. This layer
capacity for regeneration. Minor injuries heal forms about 90 per cent of the thickness of
rapidly by a gliding movement of the adja¬ the cornea (Fig. 34—3). It is a transparent,
cent epithelial cells to fill the defect. Mitoses regular connective tissue whose bundles form
in the basal epithelial cells appear later and thin lamellae arranged in many layers. In
may be found at considerable distances from each layer the direction of the bundles

Corneal epithelium

Descejnet's membrane
m

*********
Ifc
* * •
i
*

Endothelium
Figure 34-3. Photomicrograph of a section of human cornea, x 160. (After Kuwabara, T. In Greep, R. O., ed.:
Histology. 2nd ed. New York, McGraw-Hill Book Co., 1966.)
918 • THE EYE

Figure 34-4. Electron micrograph of a part of a keratocyte in the cornea and the surrounding layers of collagen fibrils
oriented at right angles to one another. (Micrograph courtesy of M. Jakus.)

changes and those in successive layers cross the surface of the trabeculae of the limbus.
at various angles (Figs. 34-3, 34-4). The In its structure it is essentially a very thick
lamellae everywhere interchange fibers and basal lamina elaborated by the corneal en¬
thus are kept tightly together. The collagen dothelium, which rests upon it (Figs. 34-3,
fibrils are somewhat thicker than those in 34—5). It appears homogeneous under the
Bowman’s membrane, measuring about 23 light microscope, but when examined with
nm on the average. Between the fibrils, the the electron microscope, Descemet’s mem¬
bundles, and the lamellae, there is a meta- brane of older individuals may show an ap¬
chromatic ground substance. The molecules parent cross striation, with bands about 107
responsible for its metachromasia are chon- nm apart, connected by filaments less than
droitin sulfate and keratan sulfate. The cells 10 nm in width and about 27 nm apart (Fig.
of the stroma are long slender fibroblasts 34-6A). Tangential sections reveal a two-
(keratocytes) lodged in narrow clefts among dimensional array of nodes, about 107 nm
the parallel bundles of collagen fibrils. In apart and connected by filaments to form
addition, the stroma always contains a num¬ hexagonal figures (Fig. 34-65). The diagram
ber of lymphocytes, which migrate from the in Figure 34—7 shows the relationship be¬
blood vessels of the corneal limbus. In inflam¬ tween the images seen in the two planes.
mation, enormous numbers of neutrophilic Histochemical data, chemical analyses, and
leukocytes and lymphocytes penetrate be¬ x-ray diffraction studies support the conclu¬
tween the lamellae. sion that the filaments forming this hexago¬
Membrane of Descemet and Corneal En¬ nal array are an atypical form of collagen. In
dothelium. This homogeneous-appearing la¬ young individuals, Descemet’s membrane is
mella, 5 to 10 pm thick, can be isolated from more homogeneous in appearance. It is sug¬
the posterior surface of the substantia pro¬ gested that the hexagonal pattern of fibers
pria. At the periphery of the cornea, Desce- forms with advancing age by aggregation of
met’s membrane continues as a thin layer on collagen that is normally dispersed in the
 THE EYE • 919

Figure 34-5. Electron micrograph of the endothelium and underlying Descemet’s membrane from a human eye.
(Courtesy of T. Kuwabara.)

Figure 34-6. Electron micrographs of Descemet’s mem¬

brane, showing the unusual configuration of collagen,
characteristic of this layer. A, Cross section with its striated
appearance. B, Tangential section, illustrating hexagonal
arrangement of nodes connected by filaments. (Courtesy
of M. Jakus.)
920 • THE EYE

neic recipients. One possible explanation for

this phenomenon is that the lack of blood
vessels protects the transplanted cornea from
the host’s immune system.

The Limbus
The limbus, or sclerocorneal junction, is an
important region of the eye because it rep¬
resents a valuable landmark for the ophthal¬
Figure 34-7. Diagram of structure of Descemet’s mem¬
brane based on electron micrographs. See text for expla¬ mologist and contains the apparatus for the
nation. (Courtesy of M. Jakus.) outflow of the aqueous humor (Figs. 34—8,
34-9). About 1.5 to 2 mm wide, its outer
amorphous ground substance as tropocolla- surface displays a shallow depression called
gen. This and other atypical forms of colla¬ the external scleral sulcus, where the gently
gen occur in the membrane at the periphery curving sclera is continuous with the more
of the cornea, where randomly oriented fi¬ convex cornea. On its inner aspect, the scler¬
brous bands with a 100-nrn periodicity are ocorneal stroma is marked by a circular
frequently encountered. These are particu¬ depression, the internal scleral sulcus, which is
larly common in Hassall-Henle bodies or warts, filled in by the trabecular meshwork and the
dome-shaped protrusions from the periph¬ canal of Schlernm, specialized tissues constitut¬
ery of Descemet’s membrane into the ante¬ ing the outflow system for the aqueous hu¬
rior chamber, which occur with increasing mor. On the posterior lip of the internal
frequency in human eyes after the age of 20. scleral sulcus, the scleral stroma projects to¬
The inner surface of the membrane of ward the interior of the eye, forming a small
Descemet is covered by a layer of large squa¬ circular ridge, the scleral spur; this affords
mous cells (Fig. 34—5); the intercellular attachment to the trabecular meshwork an¬
spaces between these endothelial cells permit teriorly and. to the ciliary muscle posteriorly.
free exchange of fluid between corneal At the limbus, there is a gradual transition
stroma and anterior chamber. of the corneal epithelium into that of the
conjunctiva of the bulb (Fig. 34-8). The
Histophysiology of the Cornea membrane of Bowman terminates and is re¬
placed by the conjunctival stroma and the
The transparency of the cornea is great, anterior margin of the capsule of Tenon. In
although less than that of the aqueous hu¬ the connective tissue underlying the epithe¬
mor. It is due, at least in part, to the uniform lium, the conjunctival vessels form arcades
diameter and regular spacing of its collagen that extend radially into the cornea for about
fibrils, so that scattered rays cancel each other 0.5 mm beyond the limbal edge. These ves¬
by destructive interference. Proteoglycans of sels nourish the periphery of the cornea and
the ground substance may be responsible for are the source of the occasional lymphocytes
the orderly arrangement of the fibrils. An found in the corneal stroma. The blood ves¬
increase in the amount of the interfibrillar sels that invade the corneal stroma in chronic
fluid, such as occurs in swelling, causes clou¬ inflammation arise from these loops. When
diness of the cornea. the limbus is examined in a living subject
The cornea is avascular, and its central with the slit-lamp microscope, aqueous veins,
region depends on diffusion from the veins containing aqueous humor instead of
aqueous humor for its nourishment. The blood, may be seen emerging from the limbal
blood vessels of the limbus supply the pe¬ stroma and contributing to the plexus of the
ripheral cornea by diffusion and account for episcleral veins. At the limbus, the collage¬
the presence, in the corneal stroma, of leu¬ nous sclera gradually continues into the cor¬
kocytes and substances that are excluded neal stroma, and its collagenous bundles pro¬
from the aqueous humor. Oxygen for the gressively acquire the uniform small diameter
corneal epithelium comes directly from the and orderly arrangement typical of the cor¬
atmosphere. The contribution of tears to nea. Deep to the stroma of the limbus, Des¬
corneal nourishment seems to be negligible. cemet’s membrane ends and gives way to the
The cornea is one of the few organs that spongy tissue of the trabecular meshwork, situ¬
can be successfully transplanted into alloge¬ ated between the anterior chamber, the root
 THE EYE • 921

Corneal epithelium

Cornea
Canal of Sch/emm

Bulbar conjunctiva
Anterior chamber

Ciliary muscle

Dilator of pupil
Sphincter of pupil
Sclera

Posterior chamber

Ciliary
Ciliary process Ora S?
serrata

Vitreous

Nuclear zone of lens

Hyaloideo-capsular ligament

Figure 34-8. Part of meridional section of human eye. x 14. (Modified from Schaffer.)

Figure 34-9. Photomicrograph of the sclerocorneal angle of a normal human eye. (Courtesy of T. Kuwabara.)
922 • THE EYE

of the iris, the limbal stroma, and the scleral thelium varies greatly in thickness with dif¬
spur (Figs. 34-9, 34-10). The trabecular ferent techniques of specimen preparation
meshwork is composed of a large number of and may display large intra- or intercellular
flattened, fenestrated connective tissue sheets vacuoles. Great importance has been attrib¬
and branching and anastomosing beams or uted to these “giant vacuoles,” for it is be¬
trabeculae. These are completely invested by lieved that they are involved in the process
an attenuated endothelium, continuous with of aqueous humor reabsorption from the
the corneal endothelium (Fig. 34—11). They anterior chamber.
bound a labyrinthine system of minute pas¬ The lumen of the canal does not commu¬
sages, the intertrabecular spaces, which com¬ nicate directly with the spaces of the trabec¬
municate with the anterior chamber and are ular meshwork but is separated from them
filled with aqueous humor. by the following layers: (1) the endothelium
Interposed between the trabecular mesh¬ that invests the internal wall of the canal; (2)
work and the limbal stroma is the canal of the connective tissue adventitia of the canal,
Schlemm, a flattened vessel that extends which here becomes especially rich in stromal
around the entire circumferene of the lim¬ cells and is usually referred to as juxtacanal-
bus. The canal of Schlemm has a varicose icular connective tissue; and (3) the endo¬
outline and in places breaks up into irregular thelial lining of the trabecular spaces.
branches that coalesce again. The wall of the From the outer wall of the canal, 25 to 35
canal consists of endothelium, a discontin¬ collector channels arise, which join the deep
uous basal lamina, and a thin layer of con¬ veins of the limbus; these in turn pass to the
nective tissue. On the outer wall of the surface of the limbal stroma and empty into
canal—that is, toward the limbal stroma—the the episcleral veins.
endothelium is extremely attenuated (Fig. The aqueous humor contained in the an¬
34—12); on the inner wall of the canal— terior chamber permeates the maze of min¬
toward the trabecular meshwork—the endo¬ ute intercommunicating passages of the tra-

Figure 34-10. Diagram of the outflow system of the aqueous humor. Star indicates scleral spur. (After Hogan, M. Y.,
J. A. Alvarado, and J. E. Weddell: Histology of the Human Eye. Philadelphia, W. B. Saunders Co., 1971.)
 THE EYE • 923

Figure 34-11. Electron micrograph of a beam of the trabecular meshwork in a monkey. A thick basal lamina separates
the connective tissue core of the beam from the investing endothelium. The intertrabecular spaces are crisscrossed by
processes of the endothelial cells, x 8600. (From Raviola, G. Invest. Ophthalmol. 13:828, 1974.)
924 • THE EYE

Figure 34-12. Canal of Schlemm in a monkey. The external wall of the canal consists of an attenuated endothelium, a
discontinuous basal lamina, and an adventitial layer of flattened fibroblasts. The collagen fibrils of the limbal stroma are
seen in cross section, x 6200. (From Raviola, G. invest. Ophthalmol 73:828, 1974.)

becular meshwork; thence, it reaches the Between the sclera and the choroid is a po¬
lumen of the canal of Schlemm and is finally tential cleft, the perichoroidal space, which is
drained by the episcleral veins. The precise traversed by thin lamellae that run obliquely
pathway followed by the aqueous humor from the choroid to the sclera and form a
from the intertrabecular spaces to the lumen loose, pigmented tissue layer—the supracho¬
of the Schlemm canal is poorly understood. roid lamina. This is composed of fine, trans¬
The Schlemm canal usually contains aqueous parent sheets, with fibroblasts on their sur¬
humor, but it may rarely fill with blood when face and with a rich network of elastic fibers.
there is stasis and back pressure in the venous Large flat melanocytes are scattered every¬
system. Obstruction to the filtration of where between and within the connective
aqueous humor through the intertrabecular tissue lamellae. In the suprachoroid, as in the
spaces or to its drainage via the canal of rest of the uvea, there are also scattered
Schlemm results in the rise in intraocular macrophages. The lamellae of the supracho¬
pressure characteristic of the serious eye dis¬ roid pass without a distinct boundary into
ease glaucoma. the substance of the choroid proper. This
tunic can be subdivided into three main lay¬
ers; from outside inward, they are (1) the
vessel layer, (2) the choriocapillary layer, and
THE VASCULAR TUNIC: (3) the glassy membrane, or Bruch’s membrane
THE UVEA (Fig. 34-13).
Vessel Layer. This layer consists of a mul¬
titude of large and medium-sized arteries and
Choroid
veins. The spaces between the vessels are
filled with loose connective tissue rich in
The choroid is a thin, soft, brown layer melanocytes. The lamellar arrangement here
adjacent to the inner surface of the sclera. is much less distinct than in the suprachoroid.
 THE EYE • 925

Figure 34-13. Photomicrograph of choroid and outermost layers of the retina. (After Kuwabara, T. In Greep, R. O., ed.:
Histology. 2nd ed. New York, McGraw-Hill Book Co., 1966.)

According to some, the vessel layer contains Ciliary Body

strands of smooth muscle that are independ¬
ent of the walls of blood vessels. If the eyeball is cut across along its equator,
Choriocapillary Layer. This is a capillary and its anterior half is inspected from within
network arranged in one plane. In places this after removal of the vitreous, a sharply out¬
layer is connected with the vessel layer. The lined, dentate border is seen running around
individual capillaries have a large and some¬ the inner surface of the wall in front of the
what irregular caliber; toward Bruch’s mem¬ equator (Fig. 34-14). This is the ora serrata
brane their endothelium is fenestrated. The or ora terminalis of the photosensitive retina.
layer is thicker and the capillary network The zone between the ora and the edge of
denser in the region underlying the fovea. the lens is the ciliary body, a thickening of the
Anteriorly it ends near the ora serrata. vascular tunic. Its surface is covered by the
Bruch’s Membrane (Glassy Membrane). darkly pigmented, nonphotosensitive ciliary
This is a refractile layer 1 to 4 pm thick portion of the retina. In a meridional section
between the choroid and the pigment epithe¬ through the eye bulb, the ciliary body ap¬
lium of the retina. The electron microscope pears as a thin triangle with its small base
has shown that this so-called membrane is facing the anterior chamber of the eye and
not a homogeneous structure but consists of attached by its anterior and outer angle to
five different components: (1) the basal lam¬ the scleral spur. The long, narrow posterior
ina of the endothelium of the capillaries of angle of its triangular section extends back¬
the choriocapillary layer; (2) a first layer of ward and merges with the choroid (Fig.
collagen fibers; (3) a network of elastic fibers; 34-8). The inner aspect of the ciliary body is
(4) a second layer of collagen fibers; and (5) divided into a narrow anterior zone, the
the basal lamina of the pigment epithelium ciliary crown, and a broader posterior zone,
of the retina. the ciliary ring. Seen in surface view, the inner
926 • THE EYE

Temporal
Nasal side
side

T'Ora serrata
Cireumlental space / retinae

Posterior surface of Ciliary ring

the lens with the
lens star
ry crown

Figure 34-14. Anterior half of right eye, seen from within, x 3. (After Salzmann.)

surface of the ring has shallow grooves, ciliary ocytes (Fig. 34—9). The latter become espe¬
striae, which run forward from the teeth of cially numerous toward the sclera.
the ora serrata. On its inner surface, the The inner, vascular layer of the ciliary body
ciliary crown has 70 radially arranged ridges, consists of connective tissue and numerous
the ciliary processes (Figs. 34-8, 34-9, 34-14). blood vessels. In the ciliary ring it is the
1 he bulk of the ciliary body, exclusive of direct continuation of the same layer of the
the ciliary processes, consists of the muscle choroid. In the region of the ciliary crown it
of accommodation, the ciliary muscle. It is covers the inner surface of the ciliary muscle
smooth muscle and is composed of three and forms the core of the ciliary processes.
portions. Closest to the sclera is the muscle of The vessels are almost exclusively capillaries
Briicke, whose bundles are deployed chiefly and veins of varying caliber. The correspond¬
in the meridional direction. This outer part ing arteries ramify in the peripheral layers
of the ciliary muscle stretches the choroid of the ciliary body. The capillary endothe¬
and is also called the tensor muscle of the choroid. lium is fenestrated and freely permeable to
In the next inward portion of the ciliary plasma proteins. The connective tissue is
muscle, the bundles of muscle cells radiate dense, especially near the root of the iris,
fanlike from the region of the scleral spur and contains abundant elastic fibers. In old
toward the cavity of the eyeball. This is the age it often shows hyaline degeneration.
radial or reticular portion of the ciliary muscle. The ciliary portion of the retina continues
The third or circular portion of the ciliary forward beyond the ora serrata as the ciliary
muscle (Mullers muscle) is usually absent in epithelium investing the inner surface of the
the newborn, appearing in the course of the ciliary body. Its function is the production of
second or third year. The contraction of this the aqueous humor. The ciliary epithelium
portion relaxes the tension on the lens and consists of two layers of cells, an inner layer
thus is important in accommodation for near of nonpigmented elements bounding the
vision. The classical subdivision of the ciliary posterior chamber, and an outer pigmented
muscle into three portions has been chal¬ layer, which rests on the stroma of the ciliary
lenged as too schematic, for, in fact, the body (Fig. 34—15). Toward the root of the
muscle fibers seem to be interwoven in a iris, the cells of the inner epithelial layer
tridimensional network but with regions gradually accumulate pigment granules. Be¬
where fibers of meridional, radial, and cir¬ cause of the embryonal origin of the ciliary
cular orientation predominate. The intersti¬ epithelium from the edge of the double-
ces between the muscular bundles are filled walled optic cup, the pole of the nonpig¬
with a small amount of connective tissue mented cells directed toward the interior of
containing abundant elastic fibers and melan¬ the eye is usually referred to as the cell base,
 THE EYE • 927

Figure 34-15. Ciliary epithelium in a monkey that was injected intravenously with horseradish peroxidase. The ciliary
epithelium consists of two cell layers, one nonpigmented (above), which bounds the posterior chamber; the other
pigmented (below), which rests on the stroma of the ciliary body. The tracer escaped through the permeable walls of
the vessels of the ciliary body and has permeated the intercellular clefts between pigmented and nonpigmented cells,
but its further progression toward the posterior chamber is blocked by an impermeable tight junction, which connects
the apices of the nonpigmented cells, x 6400. (From Raviola, G. invest. Ophthalmol. 13:828, 1974.)

whereas the base of the pigmented cells is oped Golgi apparatus, a centriole, and occa¬
the end that adjoins the stroma of the ciliary sionally a cilium, which protrudes from the
body. Thus, the apices of the pigmented and cell surface in a channel bounded by the
nonpigmented epithelial cells face each plasma membrane. The cytoplasm is per¬
other. At intervals, they are separated by meated by flat cisternae of the granular en¬
discontinuous intercellular spaces called cili¬ doplasmic reticulum, tubules of the agranu¬
ary channels. lar reticulum, and bundles of filaments
A basal lamina invests both surfaces of the radiating from the desmosomes that join ad¬
ciliary epithelium; that toward the stroma of jacent cells. The mitchondria are not espe¬
the ciliary body is continuous with the basal cially numerous, nor do they appear to be
lamina of the pigment epithelium of the arranged in an orderly manner with respect
retina; the other is continuous with the inner to the plasmalemmal invaginations, as is the
limiting membrane of the retina. case for the convoluted tubules of the kidney
The basal and lateral regions of the non¬ or the striated ducts of the salivary glands.
pigmented cells are occupied by a labyrinth Especially prominent in the pigmented epi¬
of interdigitating processes formerly de¬ thelial cells are the melanin granules, which
scribed as “membrane infoldings” (Fig. completely fill the cytoplasm, leaving but little
34-16). In this respect, the nonpigmented space for a moderate number of mitochon¬
cells resemble other epithelia actively en¬ dria and thin bundles of filaments. The nu¬
gaged in transport of ions and water. Be¬ cleus is located toward the apex of the cell,
tween the central nucleus and the cell apex being separated from it by a small Golgi
is the cell center, consisting of a well-devel¬ apparatus. The plasma membrane at the base
928 • THE EYE

Figure 34-16. Electron micrograph of the ciliary epithelium of a rabbit. The base of the nonpigmented cells is occupied
by a complex labyrinth of interdigitating processes. The basal lamina that separates the ciliary epithelium from the
content of the posterior chamber is barely visible, x 18,000. (Courtesy of G. Raviola.)

of the cell is repeatedly invaginated, but the Aqueous Humor and

basal labyrinth is not as complex as in non¬ the Blood-Aqueous Barrier
pigmented cells.
The ciliary epithelium is exceptional Proper eye functioning requires a precise
among actively transporting epithelia because spatial arrangement of the retina with respect
it consists of two layers of cells, both provided to the refractive media and a special chemical
with a basal labyrinth of interdigitating proc¬ composition of the intraocular fluids, opti¬
esses. These structural specializations suggest mally adjusted to the metabolic needs of the
that the ciliary epithelium represents a retina, lens, and cornea. The aqueous humor
unique biological device, consisting of two subserves both of these functions. An accu¬
pumps working in series. This might result rate balance between its rates of production
in a considerable amplification of the trans¬ and reabsorption is responsible for mainte¬
port efficiency, but it requires accurate syn¬ nance of the intraocular pressure, which con¬
chronization of the cells’ activity. Gap junc¬ fers mechanical stability upon the ocular
tions probably ensure such a precise structures. Its specific composition, differing
coordination of the function of the myriad from plasma and somewhat resembling cere¬
independent cell units; they connect adjacent brospinal fluid, cooperates with the blood-
pigmented cells, adjacent nonpigmented retinal barrier in generating an extracellular
cells, and the confronted apices of the pig¬ environment best suited to the functional
mented and nonpigmented cells. Further¬ requirements of the cells of the retina, lens,
more, the lateral surfaces of the nonpig¬ and cornea. Finally, it nourishes the lens,
mented cells are connected to each other by which lacks a blood supply.
an elaborate zonula occludens, a zonula ad¬ d he aqueous humor is a clear, watery fluid
herens, and a few desmosomes. of slightly alkaline reaction with an index of
 THE EYE • 929

refraction of 1.33, contained in the anterior margin of the iris connected with the ciliary
and posterior chambers of the eye. In its body is called the ciliary margin, or the root
chemical composition, the aqueous humor of the iris. The pupil is surrounded by the
differs from blood plasma in its lower content pupillary margin of the iris. The iris diminishes
of proteins; higher content of ascorbate, py¬ in thickness toward both margins. Besides its
ruvate, and lactate; and lower content of urea individually varying color, the anterior sur¬
and glucose. Also, its electrolyte content is face of the iris presents certain distinct mark¬
slightly different from that of the plasma. ings. About 1.5 mm from the pupil, a jagged
Continuously secreted by the ciliary epithe¬ line concentic with the pupillary margin sep¬
lium, probably through a process of active arates the anterior surface into a pupillary
transport, it fills the posterior chamber, nour¬ zone and a wider ciliary zone. Near the pupil¬
ishes the lens, and permeates the vitreous lary and the ciliary margins the anterior sur¬
body. From the posterior chamber, it flows face has many irregular excavations, the
into the anterior chamber through the pupil crypts, which may extend deep into the tissue.
and is finally drained through the trabecular In addition, there are oblique, irregularly
meshwork and canal of Schlemm. The flow arranged contraction furrows, which are es¬
of the aqueous humor is determined by the pecially marked when the pupil is dilated.
difference in pressure between the fluids The main mass of the iris consists of a
within the eye (about 20 mm Hg) and the loose, pigmented, highly vascular connective
pressure in the episcleral veins (about 13 mm tissue. The anterior surface of the stroma is
Hg). In turn, the intraocular pressure is gen¬ lined with a discontinuous layer of fibroblasts
erated by an accurate adjustment of the rate and melanocytes. A thin layer of stroma im¬
of aqueous humor secretion by the ciliary mediately beneath this cell investment, the
epithelium and its ra.e of reabsorption at the anterior stromal sheet or lamella, is devoid of
limbus. When this balance is disrupted, as in blood vessels. Deep to this is a layer contain¬
glaucoma, the intraocular pressure increases, ing numerous vessels; their walls consist of
with devastating effects on the function of endothelium, pericytes, and an unusually
the eye. thick connective tissue adventitia. The pos¬
Secretion of a fluid such as the aqueous terior surface of the iris is covered with a
humor, with a composition different from double layer of heavily pigmented epithe¬
that of the plasma, is possible only if free lium, the iridial portion of the retina (Figs.
diffusion of solutes between blood and the 34-9, 34-17).
chambers of the eye is prevented. This is the The anterior stromal sheet or lamella con¬
role of the so-called blood-aqueous barrier, the tains a few collagenous fibers and many fi¬
peculiar physiological mechanism that limits broblasts and melanocytes in a homogeneous
the exchange of materials between the vas¬ ground substance. The color of the iris de¬
cular compartment and the interior of the pends on the quantity and the arrangement
eye. When an ultrastructural tracer, such as of the pigment and on the thickness of the
horseradish peroxidase, is injected into the lamella. If this layer is thin and its cells
bloodstream, it rapidly diffuses across the contain little or no pigment, the black pig¬
permeable walls of the vessels of the ciliary ment epithelium on the posterior surface, as
body, permeates the stroma underlying the seen through the colorless tissue, gives the
ciliary epithelium, and is finally blocked by iris a blue color (Fig. 34—18). An increasing
the tight junctions that connect the apices of amount of pigment brings about the differ¬
the nonpigmented cells (Fig. 34—15). These ent shades of gray and greenish hues. Large
junctions, which limit free movement of mol¬ amounts of dark pigment cause the brown
ecules between ciliary body stroma and pos¬ color of the iris. In albinos, the pigment is
terior chamber, are therefore believed to absent oF scanty, and the iris is pink because
represent the major anatomical site of the of its rich vascularity.
blood-aqueous barrier. The epithelial pigment layer on the poste¬
rior surface of the iris is a direct continuation
of the ciliary portion of the retina and, like
Iris it, originally consists of two layers of epithe¬
The posterior surface of the iris near the lium. The inner, nonpigmented layer of the
pupil rests on the anterior surface of the ciliary portion of the retina becomes heavily
lens; in this way the iris separates the anterior pigmented in the iridial region with dark
chamber from the posterior chamber. The brown melanin granules that obscure the cell
930 • THE EYE

Stroma of the iris

Pigment
epithelium
' Melanocytes
Figure 34-17. Photomicrograph of a transverse section of human iris. (Preparation by T. Kuwabara.)

outlines. The posterior or inner surface is smooth muscle fibers are arranged in thin,
covered by the limiting membrane of the iris, a circumferentially* oriented bundles (Fig.
typical basal lamina. The outer or anterior 34—17). The dilator of the pupil opens the pupil
pigmented layer becomes less pigmented. and consists of radially arranged myo¬
These outer epithelial cells derived from the epithelial elements, which form a thin mem¬
outer wall of the embryonic optic cup brane between the vessel layer and the pig¬
undergo a remarkable transformation into ment epithelium (Fig. 34—19).
contractile elements—the myoepithelium of the The innervation of the two muscles is quite
dilator papillae. different. The dilator is innervated by sym¬
Being an adjustable diaphragm, the iris pathetic postganglionic neurons located in
contains two muscles that keep the mem¬ the superior cervical ganglion Their axons
brane stretched and hold it against the sur¬ pass to the trigeminal ganglion and thence
face of the lens. The contraction of the cir¬ into the ophthalmic branch of the latter, and
cular sphincter of the pupil reduces the finally reach the dilator muscle through the
diameter of the pupil. It is a thin, flat ring long ciliary nerves. The sphincter muscle is
surrounding the margin of the pupil. Its innervated by parasympathetic fibers from
breadth changes, according to the contrac¬ postganglionic neurons located in the ciliary
tion of the iris, from 0.6 to 1.2 mm. Its ganglion, and their axons reach the sphincter

Figure 34-18. Sections of human iris. A, Posterior part of a radial (meridional) section of a dark human iris, from an
enucleated eyeball. FL, Fibriiiae of the dilator muscle in longitudinal section; P, pigment epithelium of the inner (posterior)
layer of the iridial portion of the retina; SZ, pigment-containing connective tissue cells (melanocytes) of the vascular layer;
ZK, pigment-containing cell bodies of the dilator muscle (outer or anterior layer of the iridial portion of the retina). B,
Tangential section of a light human iris. FQ, Fibers of the dilator muscle in cross section; G, blood vessel in the stroma;
other symbols as in A. x 380. (After Schaffer.)
 THE EYE • 931

The blood vessels of the iris in the rhesus

monkey, and possibly in the human, are
unusual in that their walls have the same
structure irrespective of their diameter and
cannot be classified according to the tradi¬
tional morphological criteria for distinguish¬
ing arterioles, capillaries, and venules (Fig.
34-20). No smooth muscle is found in any
of these vessels. Their wall consists of a con¬
tinuous layer of endothelium on a thin basal
lamina; pericytes enclosed between two layers
of the basal lamina; and an adventitia of
fibroblasts, melanocytes, and occasional mac¬
rophages. The intercellular clefts of the en¬
dothelium are closed by tight junctions that
prevent paracellular escape of macromole¬
cules from the blood, and plasmalemmal ves¬
icles do not transport any significant amount
of blood-borne peroxidase across the endo¬
thelium. However, if this tracer is perfused
through the anterior chamber, it freely enters
the iridial stroma and reaches the lumen of
the blood vessels by transcellular vesicular
transport. The luminal and abluminal plasma
membranes of the endothelium seem to bear
a different electrical charge, for anionic mol¬
Figure 34-19. Photomicrograph of transverse section ecules are readily transported to the lumen
through the posterior surface of albino rabbit iris, showing
the pale cuboidal iris epithelium and the underlying dark-
but cationic substances are excluded. It is
staining myoepithelium of the dilator pupillae muscle, suggested that in the iridial vessels there is a
x 480. (After Richardson, K. C. Am. J. Anat. 114:173, unidirectional vesicular transport that selec¬
1964.) tively moves anionic organic substances from
the anterior chamber to the bloodstream.
The importance of this pathway in aqueous
with the short ciliary nerves. The sympathetic humor dynamics has yet to be evaluated.
and parasympathetic divisions of the auto¬
nomic nervous system thus have opposite
effects upon the pupil. On the other hand, REFRACTIVE MEDIA
the sphincter and the ciliary muscles, which
are both innervated by the short ciliary
OF THE EYE
nerves, work in concert. When the eye accom¬
modates for near vision by contraction of the The cornea and the anterior and posterior
ciliary muscle, there is always a simultaneous chambers of the eye have been described.
contraction of the pupillary sphincter. The other components of the refractive ap¬
In electron micrographs, the axons among paratus of the eye are the crystalline lens and
the contractile elements of the sphincter pu¬ vitreous body.
pillae are seen to be packed with synaptic
vesicles typical of cholinergic axons. Axons Lens
associated with the dilator muscle contain a
mixture of dense-cored and agranular vesi¬ The lens is a transparent, biconvex body
cles typical of the endings of adreneric sym¬ situated immediately behind the pupil. Its
pathetic nerve fibers. shape changes during the process of accom¬
An accurate adjustment of the size of the modation. Its outer form varies somewhat in
pupil modifies the amount of light entering different persons and also with age. Its di¬
the eye, thus permitting useful vision over a ameter ranges from 7 mm in a newborn to
wide range of light intensities. Furthermore, 10 mm in an adult. Its thickness is approxi¬
as the pupil constricts in brilliant light, the mately 3.7 to 4 mm, increasing during accom¬
depth of focus of the dioptric media is in¬ modation to 4.5 mm and more. The posterior
creased and aberrations are minimized. surface is more convex than the anterior, the
932 • THE EYE

Figure 34-20. A whole mount of an injected preparation of the iris of Macaca muiatta in moderate mydriasis. A series
of large radial vessels gives rise to a complex network of intermediate and small vessels throughout the length of the
iris. A continuous arcade of small vessels is seen at the pupillary margin. (Photomicrograph from Freddo, T., and G.
Raviola. Invest. Ophthalmol. Vis. Sci. 22:279, 1982.)

respective radii of curvature being 6.9 and prismatic fibers of the cortical zone of the
10 mm. The index of refraction is 1.36 in lens are hexagonal in cross section. The cell
the peripheral layers and 1.4 in the inner surfaces are about 15 nm apart and attached
zone. The lens weighs 0.2 g and is slightly by numerous gap junctions. The cells of the
yellow. lens epithelium and the fibers of the equa¬
The lens is covered with a homogeneous, torial region both exhibit complex interdigi-
highly refractive capsule, an 11- to 18-pm tations of their surfaces. This is particularly
coating, rich in collagen and proteoglycans, marked at the “sutures,” where cortical fibers
which invests the outer surface of the layer from opposite sectors of the lens converge
of cuboidal cells that make up the epithelium (Fig. 34—23). Since these interdigitations oc¬
of the lens (Figs. 34-21, 34-22). Toward the cur principally in the anterior curvature,
equator of the lens these cells approach a periaxial zone, and equator—those regions
columnar form and become arranged in me¬ that undergo the greatest dimensional
ridional rows. Becoming progressively elon¬ changes—it has been suggested that their
gated, the cells at the equator are trans¬ presence may be associated with changes of
formed into lens fibers that constitute the bulk fiber shape in the mechanism of intracapsular
of the substance of the lens. In this transi¬ accommodation.
tional or nuclear zone the cells have a charac¬ The lens fibers have a finely granular cy¬
teristic arrangement. The epithelial cells are toplasm, with a few small vesicles scattered
of prime importance for the normal metab¬ through the ectoplasmic region of the cell
olism of the lens. The capsule covering the and occasional mitochondria in the vicinity
posterior surface of the lens has no under¬ of the sutures, but in general the organelles
lying epithelium. and inclusions are exceedingly sparse. Their
In the human lens, each cell, commonly cytoskeleton consists of abundant intermedi¬
called a lens fiber, is a six-sided prism, 7 to ate filaments (Fig. 34-24).
10 mm long, 8 to 12 pm wide, and only 2 The lens is held in position by a system of
pm thick (Fig. 34-23). In the region of the fibers constituting the ciliary zonule. The zon¬
nucleus, the thickness may reach 5 pm. The ule fibers (Fig. 34—8) arise from the epithe-
 THE EYE • 933

Bow area Fibers of

of lens zonule

Lens
capsule

Figure 34-21 Figure 34-22

Figure 34-21. Photomicrograph of the bow area of the human lens, where the epithelial cells become greatly elongated
to form lens fibers. (Courtesy of T. Kuwabara.)

Figure 34-22. Photomicrograph at higher magnification of human lens stained with the periodic acid—Schiff reaction.
The lens capsule overlying the epithelium stains strongly. Zonule fibers merge with the capsule. (Preparation by T.
Kuwabara.)

lium of the ciliary portion of the retina. Near tical to the microfibrils embedded in the
the ciliary crown they fuse into thicker fibers elastic fibers of other organs. Where the
and finally form about 140 bundles. At the vitreous body touches the lens capsule, it
anterior margin of the ciliary processes they forms the hyaloideocapsular ligament.
leave the surface of the ciliary body and The radii of curvature of the surfaces of
radiate toward the equator of the lens. The the several dioptric media of the normal eye,
larger ones are straight and reach the capsule especially of the lens, and their indices of
in front of the equator of the lens (anterior refraction are such that light rays coming
zonular sheet). The thinner fibers assume a from a remote point form an inverted and
slightly curved course and are attached to real image of the object in the layer of the
the posterior surface of the lens (posterior photoreceptive cones and rods in the retina.
zonular sheet). All zonular fibers break up into If the object is approaching, the light rays
a multitude of finer fibers, which fuse with diverge more and more, and the image
the substance of the outermost layer of the moves backward in the retina. A change of
lens capsule (Fig. 34—22). With the electron position of an object from infinite distance to
microscope, the zonular fibers appear as bun¬ about 5 m causes the image to shift about 60
dles or sheets of exceedingly fine filaments, fxm backward in the retina. Since this image
11 to 12 nm in diameter, which have a hollow is still within the outer segments of the rods
appearance in cross section (Fig. 34—25). and cones, accommodation is not needed.
They are digested with elastase, but not by For nearer distances, accommodation is nec¬
collagenase, and have an amino acid compo¬ essary.
sition different from collagen. They are iden¬ In a camera, the focusing of objects that
934 • THE EYE

Figure 34-23. Schematic drawing of the arrangement of

lens fibers in rows and their prevailing hexagonal cross-
sectional form, except in the suture area, where there
may be considerable irregularities and interdigitation of
fibers converging from opposite sectors of the lens. (After
Wanko, T., and M. Gavin: The Structure of the Eye. New
York, Academic Press, 1961.)

mmm
3'

. !.■ > - ' i

■ .?! v S' ■
a - y- ' *

:• i , • • <- v.. .... .<>%& ;

Ssisill mmmmi
, r, ■

i:-'

fsasp
WmMmm
w* - ' - 11

Figure 34-24 Electron micrograph of cortical fibers of the human lens. A few profiles of granular endoplasmic reticulum,
occasional mitochondria, and numerous polyribosomes are distributed in an otherwise homogeneous and concentrated
cytoplasmic matrix. (Courtesy of T. Kuwabara.) a
 THE EYE • 935

Figure 34-25. Electron micrograph of ciliary zonule in monkeys. A, B, Zonular fibers, which consist of exceedingly fine
fibrils, 11 to 12 nm in diameter. When fibrils are tightly packed (A), the zonular fibers display a cross striation. A,
x 40,000; 8, x 160,000. C, In cross section, the fibrils have a hollow appearance. C, x 200,000. (From Raviola, G.
Invest. Ophthalmol. 70:851, 1971.)

move nearer to the lens is effected by moving ina, and the connection is especially firm at
the ground glass plate away from the lens. the ora serrata. Farther forward, it gradually
In the higher vertebrates and in the human, recedes from the surface of the ciliary por¬
the curvature of the lens is changed. When tion of the retina.
the eye is at rest, the lens is kept stretched The fresh vitreous body is a colorless,
by the ciliary zonule in the plane vertical to structureless, gelatinous mass with a glasslike
the optic axis. When the eye has to focus on transparency. Its index of refraction is 1.334.
a near object, the ciliary muscle contracts—its Nearly 99 per cent of the vitreous body
meridional fibers pull the choroid and the consists of water. A liquid and a solid phase
ciliary body forward, whereas its circular fi¬ can be distinguished in it; the liquid phase
bers, acting as a sphincter, move the ciliary contains hyaluronic acid in the form of long,
body toward the axis of the eye. This relieves coiled molecules enclosing large amounts of
the tension on the zonule; the lens gets water; the solid phase is collagen in the form
thicker, and its surface, especially at the an¬ of thin fibrils that lack the usual 64-nm pe¬
terior pole, becomes more convex. This in¬ riodicity and are arranged in a random net¬
creases the refractive power of the lens and work. The hyaluronate of the liquid phase is
keeps the focus within the photoreceptor joined to the collagen network by weak
layer. bonds. The peripheral region of the vitreous
body or cortex has more collagen fibrils and
hyaluronate than the central region and con¬
Vitreous Body
tains cells, called hyalocytes, which may be
The vitreous body fills the vitreal cavity concerned with the synthesis of collagen and
between the lens and the retina. It adheres hyaluronic acid. Macrophages also are occa¬
everywhere to the optical portion of the ret¬ sionally found.
936 • THE EYE

Extending through the vitreous body from elements of the retina are in the fovea, where
the papilla of the optic nerve to the posterior they are accumulated in greatest numbers.
surface of the lens is the hyaloid canal (canal In the retinal periphery, the elements are
of Cloquet). It is a residue remaining after fewer, larger, and less evenly distributed.
the resorption of the embryonic hyaloid ar¬ When detached from the pigment epithe¬
tery. It has a diameter of 1 mm and is filled lium, the fresh retina is almost perfectly
with liquid. In the living, especially in young transparent. It has a distinctly red color be¬
persons, it is visible with the help of the slit cause of the presence in its rod cells of visual
lamp microscope. purple, or rhodopsin. Light rapidly bleaches
the visual purple; in darkness the color grad¬
ually reappears. The fovea and its immediate
vicinity contain yellow pigment and are called
THE RETINA the macula lutea. Large blood vessels circle
above and below the central fovea, whereas
The retina is the innermost of the three only fine arteries, veins, and capillaries are
coats of the eyeball and is the photoreceptor present in it. In the very center of the fovea,
organ. It arises in early embryonic develop¬ in an area measuring 0.5 mm across, even
ment from a bilateral evagination of the pro¬ the capillaries are absent, greatly increasing
sencephalon, the primary optic vesicle. Later it its transparency.
is transformed by local invagination into the Only the portion of the image of an exter¬
secondary optic vesicle. Each optic cup remains nal object that falls upon the fovea is seen
connected with the brain by a stalk, the future sharply. Accordingly, the eyes are moved so
optic nerve. In the adult, the derivatives of as to bring the object of special attention into
the bilaminar secondary optic vesicle consist this central part of the visual held. Photore¬
of an outer pigmented epithelial layer, the ceptors are absent from the optic papilla.
pigment epithelium, and an inner sheet, the This is the “blind spot” of the visual held.
neural retina or retina proper. The latter con¬
tains elements similar to those of the brain,
Pigment Epithelium
and it may be considered to be a specially
differentiated part of the brain. This sheet of heavily pigmented epithelial
The optical or functioning portion of the cells is derived from the outer layer of the
retina lines the inner surface of the choroid cuplike outgrowth of the embryonic nervous
and extends from the papilla of the optic system that gives rise to the retina, and it has
nerve to the ora serrata anteriorly. At the traditionally been included as one of the
papilla, where the retina is continuous with layers of the retina (Figs. 34-27, 34-29).
the tissue of the nerve, and at the ora serrata, Bruch’s membrane, on the other hand, has
the retina is firmly connected with the cho¬ been considered part of the choroid. The
roid. In the retina, exclusive of the fovea, the demonstration that this latter structure in¬
papilla, and the ora serrata, ten parallel layers cludes the basal lamina of the pigment epi¬
can be distinguished from outside inward thelium makes it illogical to assign the pig¬
(Figs. 34—27, 34-28): (1) the pigment epithe¬ ment epithelium to the retina and its basal
lium; (2) the layer of rods and cones; (3) the lamina to the choroid. Some authors there¬
outer limiting membrane; (4) the outer nu¬ fore prefer to consider the pigment epithe¬
clear layer; (5) the outer plexiform layer; (6) lium as a component of the choroid. Al¬
the inner nuclear layer; (7) the inner plexi¬ though the cells of this layer extend processes
form layer; (8) the layer of ganglion cells; (9) that interdigitate with the retinal rods and
the layer of optic nerve fibers; and (10) the cones, there is no actual anatomical connec¬
inner limiting membrane. About 2.5 mm tion between the photosensitive and the pig¬
lateral to the border of the optic papilla, the mented layers, except at the head of the optic
inner surface of the retina shows a shallow, nerve and at the ora serrata. An artifactitious
round depression, the fovea (Figs. 34-26, separation is found between the two layers
34-45). This is surrounded by the central in most histological preparations, and in the
area, distinguished by the great number of “retinal detachment” that is a common cause
ganglion cells and by the general refinement of partial blindness, the separation occurs
and even distribution of the structural ele¬ along this plane of cleavage between the
ments, especially of the rods and cones. The photosensitive elements of the retina and the
smallest and most precisely ordered sensory pigment epithelium.
 THE EYE • 937

Inner Layer of
nuclear ganglion cells
layer
Outer Nerve
nuclear fiber
layer Vessels Lamina
layer
/cribrosa
Central fovea

Pigment layer

Choroid

Sclera

Optic nerve

Pial sheath

Dural sheath
of optic nerve

Figure 34-26. Central fovea and place of entrance of the optic nerve as seen in a horizontal meridional section of an
enucleated human eye. e, Excavation, x 18. (Redrawn and slightly modified from Schaffer.)

Pigment epithelium /

Photoreceptor layer

Outer limiting membrane m

Outer nuclear layer

Outer plexiform layer ^)

Inner nuclear layer Q

Inner plexiform layer ^

Layer of ganglion cells $

Layer of optic nerve fibers Q

Inner limiting membrane m

Figure 34—27. Layers of adult human retina. Left half of figure stained routinely, about 400 x. Right half of figure is a
schematic reconstruction from sections stained with Golgi’s method. (Slightly modified from Polyak.)
938 • THE EYE

Rod and cone

outer segments

Figure 34-28. Photomicrograph of cat retina. (Courtesy

of A. J. Ladman.)

IjflKj plexiform layer

dnglfon cell layer

 THE EYE • 939

Figure 34-29. Light micrograph of the outermost retinal layers in a monkey. Cones are easily distinguished from rods,
for their inner segment is larger and the ellipsoid intensely stained. CH, choroid; PE, pigment epithelium; OS, outer
segments; IS, inner segments; OLM, outer limiting membrane; ONL, outer nuclear layer. (Courtesy of J. Rostgaard.)

Pigment epithelial cells have a remarkably occupy the interstices between the photore¬
regular shape, appearing as hexagonal ceptors. The cell nucleus is displaced toward
prisms about 14 pim wide and 10 to 14 |xm the cell base, and numerous mitochondria
tall; toward the ora serrata, they increase in intervene between the nucleus and basal lab¬
diameter. The cell base, which rests on yrinth of interdigitating processes. The most
Bruch s membrane, displays the labyrinth of prominent feature of the apical cytoplasm is
interdigitating processes typical of actively the presence of numerous melanin granules,
transporting epithelia, whereas the lateral cell elliptical or rounded in shape. A second im¬
surface has only a slightly undulating course. portant component of the apical cytoplasm
Adjacent cells are connected to each other by consists of residual bodies filled with lamellar
a junctional complex consisting of apical gap debris, which represents the partially di¬
junctions, followed by an elaborate tight junc¬ gested residue of the phagocytized tips of the
tion and a zonula adherens. The cell apex, rod outer segments. Another prominent fea¬
which faces the rods and cones, gives rise to ture of the cytoplasm of these cells is a highly
two sorts of processes: cylindrical sheaths, developed agranular endoplasmic reticulum
which invest the tip of the photoreceptor in the form of a rich network of branching
outer segments, and slender microvilli, which and anastomosing tubules that permeate the
940 • THE EYE

interstices among the melanin granules and Photoreceptor Cells

residual bodies. Cisternae of the granular
There are two kinds of visual cells, the rod
endoplasmic reticulum and a supranuclear
cells and the cone cells. Their outer segments
Golgi apparatus complete the list of the cy¬
are the parts sensitive to light, and the light
toplasmic organelles.
rays, before reaching them, must first pene¬
The pigment epithelium has many impor¬
trate most of the retina.
tant functions. The tight junctions that seal
Rod Cells. The rod cell is a long, slender,
the intercellular spaces between adjoining
highly specialized cell with its outer portion
epithelial cells protect the retina proper from
vertical to the retinal layers (Figs. 34-29,
undesirable metabolites that may be present
34—30, 34—31). The parallel arrangement of
in the stroma of the choroid. The pigment
these elements is responsible for the regular
granules absorb light after it has traversed
striation of the layer of rods and cones. The
the photoreceptor layer, thus preventing its
scleral part of the rod cell, the rod proper, is
reflection from the external ocular tunics. In
situated between the outer limiting mem¬
retinas of lower vertebrates, it has been
brane and the pigment epithelium, its out¬
shown that the pigment granules migrate
ward third being embedded in the pigment-
along cell processes among the photorecep¬
tors upon illumination, thus effectively containing processes of the pigment epithe¬
screening scattered light, and they return to lial cells. The vitreal end of the rod proper
the cell body in the dark. The pigment epi¬ extends through.the so-called outer limiting
thelial cells participate in the turnover of the membrane into the outer nuclear layer. The
photoreceptors, continuously engulfing and rods are fairly uniform in appearance, al¬
digesting the growing tips of the rod outer though their dimensions vary somewhat from
segments. Finally regeneration of rhodopsin region to region. Their thickness in the cen¬
after exposure to light occurs only if photo¬ tral area is 1 to 1.5 pm, gradually increasing
receptors maintain an intimate relationship to 2.5 or 3 pm near the ora serrata. Their
with the pigment epithelium. Vitamin A, a length decreases from approximately 60 pm
precursor of rhodopsin and one of the prod¬ near the fovea to 40 pm in the far periphery.
ucts of rhodopsin degradation upon light Each rod proper consists of an outer and an
absorption, moves to the pigment epithelium inner segment. The outer segment is a slender
after maximal light adaptation and returns cylinder of uniform thickness, which appears
to the photoreceptors during dark adapta¬ homogeneous in the fresh condition. It pos¬
tion. Vitamin A is a fat-soluble hydrocarbon sesses a peculiarly brillant refractility and is
and may be stored in the membranes of the positively birefringent in polarized light. The
agranular endoplasmic reticulum found inner segment contains the usual col851ement
throughout the cytoplasm of the cells of the of cytoplasmic organelles.
pigment epithelium. The finer structure of the rod cells has
been greatly clarified by electron microscopy.
The outer segment in longitudinal section is
Neural Retina seen to be composed of a very large number
of parallel lamellae oriented transverse to the
The retina proper contains six types of axis of the rod (Figs. 34-32A, 34-33). Each
neurons: (1) photoreceptor cells, (2) horizontal lamella is in fact a closed, membrane-limited
cells, (3) bipolar cells, (4) amacrine cells, (5) sac flattened into a disc approximately 2 pm
interplexiform cells, and (6) ganglion cells. Pho¬ in diameter and about 14 nm thick. In sec¬
toreceptor, horizontal, and bipolar cells syn¬ tion, therefore, the profile of each lamella or
apse with each other in the outer plexiform disc appears as a pair of parallel membranes
layer, bipolar, amacrine, and ganglion cells continuous with one another at the ends and
synapse with each other in the inner plexiform enclosing an exceedingly narrow cavity about
layer. Interplexiform cells provide a centrif¬ 8 nm across. The outer segment is joined to
ugal pathway for transfer of signals from the inner segment by a slender stalk, which
inner to outer plexiform layer. The axons of contains nine longitudinally oriented doublet
the ganglion cells leave the retina to become microtubules terminating in a centriole or
fibers of the optic nerve. The retinal neurons basal body in the distal end of the inner
are supported by neuroglial elements called segment. In transverse sections, the stalk has
radial cells of Muller. the appearance of a defective cilium, having
 THE EYE • 941

Figure 34-30. Light micrograph of a cross section through the outer segments of the photoreceptor cells in the retinal
periphery of a monkey. The large diameter of cone inner segments keeps the rods at a distance from cone outer
segments. Toluidine blue, printed as a negative, x 800. (Courtesy of E. Raviola.)
942 • THE EYE

added at the vitreal end of the outer seg¬

ments, the old ones move sclerally toward
the tip of the rods till they are phagocytized
and destroyed by the cells of the pigment
epithelium (Fig. 34—35). In the rat, rod outer
segments are totally renewed in about ten
days.
The rest of the rod Gell is made up of the
outer fiber, the cell body, the inner fiber, and the
rod synaptic ending or spherule (Fig. 34-31).
The rod outer fiber is a slender protoplasmic
process, 1 [xm or less in thickness, that ex¬
tends from the base of the inner segment of
the rod proper deep into the outer nuclear
layer. Here it joins a spherical cell body
containing the rod nucleus, which is smaller
and stains more intensely than the cone nu¬
cleus. The rod nuclei represent the majority
of the nuclei of the outer nuclear layer in all
retinal regions except in the fovea, where
rods are few, and in its center, where they
are absent. The rod inner fiber, rich in mi¬
crotubules, connects the body to a pear-
Cone Rod shaped spherule, rich in synaptic vesicles,
Figure 34-31. A cone cell and rod cell from a rabbit retina located in the outer plexiform layer. In the
stained with the Golgi technique and drawn with a camera
central area, the rod inner fibers assume a
lucida. a, Outer segment; b, inner segment; c, cell body;
d, inner fiber; e, synaptic ending (pedicle in cone; spherule slanting-to-horizontal course, while in the
in rod). (Courtesy of G. Sacchi and E. Raviola.) more peripheral retinal regions they are ver¬
tical. The rod proper, along with the outer
fiber, is the homologue of a dendrite of a
the nine peripheral doublets but lacking the neuron; the inner rod fiber corresponds to
central pair of singlet microtubules. Studies the axon, and through its terminal spherule
on the development of the photoreceptors this fiber is connected to bipolar and horizon¬
have shown that the outer segments of the tal cells.
rods do in fact develop by modification of All rod cells, except those in a zone 3 to 4
cilia. The inner segment consists of two por¬ mm wide at the ora serrata, contain visual
tions, the ellipsoid toward the sclera and the purple or rhodopsin, the substance respon¬
myoid toward the vitreous. The ellipsoid con¬ sible for absorption of light. The rhodopsin
tains a great number of mitochondria. Cross- molecules are localized in the interior of the
striated fibrous rootlets may extend down¬ membrane of the outer segment discs, where
ward from the basal body of the rod outer they appear as intramembrane particles in
segment among the mitochondria. The freeze-fractured specimens (Fig. 34—34).
myoid contains the Golgi apparatus, free ri¬ As there are only a few rods in the periph¬
bosomes, and cisternae of both agranular and ery of the fovea and none in its center, this
granular endoplasmic reticulum. Microtu¬ area appears devoid of rhodopsin. When the
bules are abundant throughout the rod inner retina is exposed to light, rhodopsin breaks
segment. down, but it is constantly produced anew.
Autoradiographic studies with the light This regeneration occurs only as long as the
and electron microscopes demonstrate that close relation of the rods with the pigment
the outer segments of rod cells are being epithelium is preserved.
renewed constantly. Protein is first synthe¬ Cone Cells. These neurons (Figs. 34—29,
sized on the ribosomes of the myoid; after 34-30, 34-31) are made up of essentially the
moving through the Golgi apparatus, it mi¬ same parts as the rod cells, but they differ in
grates to the base of the outer segment, certain details. There is no visual purple in
where it is used in the assembly of the mem¬ the cones but instead there are different
branous discs. As new discs are continuously types of pigments sensitive to blue, green,
 THE EYE • 943

Ciiium
:K* v

Basal
body

Rootlet

Figure 34—32. A, Electron micrograph of a portion of the outer and inner segment of a rod, showing the connection of
the two by a modified ciiium. B, Corresponding region of a cone. Notice that some of the discs are open to the
extracellulr space (arrows). (Courtesy of T. Kuwabara.)
944 • THE EYE

possibly significant difference between rods

and cones.
The cone outer segment is also connected
to the inner segment by an eccentrically
placed modified cilium, terminating in a basal
body set in the distal end of the inner seg-
mept. The other member of the diplosome
is usually oriented at a right angle to the
basal body, and striated rootlets extend from
it downward among the longitudinally ori¬
ented mitochondria that crowd the ellipsoid.
Cone inner segments resemble in fine struc¬
ture those of rod cells (Figs. 34-36, 34-37).
The cones vary considerably in different
regions of the retina. In the central fovea,
they measure 75 pm or more in length and
from 1 to 1.5 pm in thickness. Their length
gradually decreases to 45 pm in the periph¬
ery. The relative length of the outer and the
inner segments is usually 3:4. In the fovea
the two segments are approximately the same
length. The proximal end of the inner cone
segment occupies an opening in the “outer

Figure 34-33. Electron micrograph showing profiles of

the membranous discs of the outer segment of a rod cell
from frog retina. They exhibit a compact granular fine
structure (arrow) when prepared with very-low-tempera-
ture and osmium fixation (osmium-cryofixation), low-tem¬
perature dehydration, and embedding. (Courtesy of H.
Fernandez-Moran.)

and red light. Instead of a slender cylinder,

the cone outer segment is a long conical
structure, considerably wider than a rod at
its base and tapering down to a blunt
rounded tip. As in the rod, the outer segment
is made up of a large number of discs stacked
one above the other. Each of these consists
of a pair of membranes. In most of the discs
the two membranes are continuous at their
margins and enclose a narrow space. In the
discs close to the inner segment, the two
membranes are continuous with the plasma
membrane, and the narrow cleft between Figure 34-34. Freeze-fracture appearance of a rod outer
segment in a monkey. The membranes of the discs
them is open to the extracellular space (Fig.
contain a large number of particles, which probably are
34-32B). This appears to be a consistent and rhodopsin. x 82,500. (Courtesy of E. Raviola.)
 THE EYE • 945

region do the cones have an outer fiber. But

from the body of all cones, a stout, smooth
inner fiber descends to the middle zone of the
outer plexiform layer, where it terminates
OS with a thick triangular or club-shaped syn¬
aptic ending, the cone pedicle. Up to a dozen
short, barblike processes emanate from the
base of each pedicle, except in the fovea,
cc where there are usually none. These out¬
e growths are deployed horizontally in the
m outer plexiform layer. The length and course
of the inner cone fibers may vary consider¬
n ably, depending on the region, the longest
(600 pm) and most nearly horizontally placed
being those in the central area, where the
inner rod and cone fibers form a thick fiber
layer at the boundary between outer nuclear
A B C D and outer plexiform layers, called the outer
Figure 34-35. Diagram illustrating the turnover of rod fiber layer of Henle. The inner cone fibers have
outer segments. After injection of a radioactive amino all the characteristics of an axon, while the
acid, electron microscope autoradiography shows that the cone pedicle has those of the synaptic ending
label is first concentrated in the myoid of rod cells (A), of a neuron and makes synapses with bipolar
where ribosomes and the Golgi apparatus are contained.
Later, labeled protein moves to the membranous discs at and horizontal cells.
the base of the outer segments (B) and ascends progres¬ The number of cones in the human retina
sively toward the tip of the rods (C). Finally, it disappears is estimated at 6 to 7 millions. The ratio of
from rod cells (D) and becomes localized in the residual the number of nerve fibers of the optic nerve
bodies contained in the cytoplasm of the pigment epithe¬
(438,000) to the number of cones of one eye
lium cells. OS, outer segments; CC, connecting cilium; e,
ellipsoid; m, myoid; n, nucleus. (From Young, R. W., and is 1:6 or 1:7.
D. Bok. J. Cel! Biol. 42:392, 1969.) The relative number and distribution of
the rods and cones in different vertebrates
present great variations, depending on the
limiting membrane,” and protrudes slightly mode of life. In diurnal birds the cones are
into the fourth layer. more numerous than the rods. In most diur¬
In teleostean fishes and amphibians the nal reptiles, rods are exceedingly rare. In
inner cone segment is contractile. It shortens many nocturnal vertebrates, only rods are
in bright light and stretches in dim light or present, although in others a few rudimen¬
darkness. The displacement of these cones tary cones can be found among numerous
is, accordingly, opposite in direction to that rods. On similar comparative data M.
of the rod. Schultze (1866) based his assumption that
In contrast to rods, the turnover of cone there is a difference in function of the two
outer segments involves neither continuous kinds of photoreceptors.
movement of the discs toward the pigment Outer Limiting Membrane. The dense
epithelium nor phagocytosis of the growing staining line traditionally called the outer lim¬
tips by the pigment epithelium cells. Proteins iting membrane is not a membrane at all. In¬
found in the inner segment are inserted stead, it is found in electron micrographs to
randomly into the disc membrane through¬ be a row of zonulae adherentes where the
out the outer segment. The reason for this photoreceptor cells are attached to the Muller
difference is not clear. cells, which surround and support all the
Proximal to the outer limiting membrane, neural elements (Fig. 34-38). Distal to this
the inner cone segment merges with its body, row of zonulae adherentes, tufts of microvilli
containing a nucleus, which is larger and project from the free surface of the Muller
paler-staining than the rod nucleus. The bod¬ cells into interstices between the rod and cone
ies and nuclei of the cones, in contrast to inner segments.
those of the rods, are arranged in a single
row immediately beneath the outer limiting Horizontal Cells
membrane. Exceptional in this regaid aie
the cones in the outer fovea, whose nuclei These cells are typical neurons whose bod¬
are accumulated in several rows. Only in this ies form the uppermost one or two rows of
946 • THE EYE

Figure 34-36

Figure 34-37
Figure 34-36. Vertical section of the inner segments of a cone and several neighboring rods in the human retina,
illustrating the larger size of the cone and the high concentration of longitudinally oriented mitochondria in the ellipsoid.
(Courtesy of T. Kuwabara.)

Figure 34-37. Horizontal section through the inner segments of the photoreceptor elements in rat retina. Notice the
larger size of the cone ellipsoid and its great number of mitochondria. (After Marchesi, V. T.. M. L. Sears and R. J.
Barrnett. Invest. Ophthalmol. 3:1, 1964.)
 THE EYE • 947

Figure 34-38. Electron micrograph of the region of the “outer limiting membrane’’ of the retina, showing that it is not a
membrane, as it appears to be with the light microscope (inset), but a row of zonulae adherentes between the rod and
cone cells and the surrounding Muller cells. (Courtesy of T. Kuwabara.)

the inner nuclear layer. From the scleral end takes a horizontal course in the outer plexi¬
of the body arise short dendritic twigs, which form layer, and its terminal twigs come into
produce several tufts deployed in the outer contact with rod spherules (Fig. 34-39). In
plexiform layer. Each dendritic tuft is con¬ mammals such as the cat and rabbit, horizon¬
nected to a single cone pedicle. The axon tal cells without an axon are also found.

Figure 34-39. Neuronal types in the retina of mammals as they appear with the light microscope after Golgi’s chromo-
arqentic impregnation, r, Rod cells (dog); c, cone cells (dog); b, bipolar cells (dog); h, horizontal cells (dog); a, amacrine
cells (dog and ox); g, ganglion cells (ox). In rod and cone cells, the photoreceptor proper is not represented. (After
Cajal, 1893, modified.)
948 • THE EYE

Figure 34-40. a, Bipolar cell; b, ganglion cell; c, amacrine cell impregnated by the Golgi technique. The bipolar and
ganglion cells are from the retina of Macaca mulatta; the amacrine cell is from rabbit retina. (Photomicrograph courtesy
of E. Raviola.)

Bipolar Cells zontal cells. The synaptic terminals of cone

cells or pedicles are large pyramidal endings
These neurons extend from the outer to
containing synaptic vesicles and mitochon¬
the inner plexiform layer and therefore stand
dria, whose flattened base is invaginated at
approximately upright with respect to the
many points to enclose the tips of the den¬
retinal layers (Figs. 34-39, 34-40). Their
drites of the horizontal and invaginating
body is located in the inner nuclear layer and
midget bipolar cells. The remaining free sur¬
gives rise to one or more primary dendrites,
face of the base of the pedicle makes
which ascend to the outer plexiform layer,
hundreds of superficial or basal contacts with
where they branch and connect with the
the dendrites of the flat midget and diffuse
photoreceptor cell terminals. The single, in¬
cone bipolars. With a high degree of consis¬
wardly directed axon of the bipolar cells
tency and geometrical order, each of the 12
ramifies in the inner plexiform layer, where
to 25 synaptic invaginations of a cone pedicle
it is synaptically related to ganglion and ama¬
contains the tip of two horizontal cell den¬
crine cells. Four types of bipolar cells can be
drites and one dendrite of an invaginating
distinguished in the primate retina: (1) rod
midget bipolar cell (a “triad”). The horizontal
bipolar cells, which connect to rod cells; (2)
cell dendrites may contain synaptic vesicles,
and (3) invaginating midget bipolar cells and flat
are deeply inserted, and lie on either side of
midget bipolar cells, each synapsing with a sin¬
a wedge-shaped projection of the pedicle,
gle cone pedicle; and (4) flat or diffuse cone
called the synaptic ridge. The dendrite of the
bipolar cells, connected to many cone pedicles.
invaginating midget bipolar cell lies centrally
The precise synaptic relationships of the
and more superficially, separated from the
axon terminals of the four types of bipolar
apex of the ridge by the cleft intervening
cells with the various types of amacrine and
ganglion cells are poorly understood. between adjoining horizontal cell dendrites.
The synaptic ridge is bisected by a dense
lamella or synaptic ribbon, surrounded by a
Outer Plexiform Layer halo of synaptic vesicles; the ribbon sits at a
right angle to the apex of the ridge, separated
This is the region of synaptic interplay from the pedicle membrane by a trough¬
between photoreceptor, bipolar, and hori¬ shaped body, the aeriform density (Fig.
 THE EYE • 949

34-415). The significance of the synaptic tion and electron microscopy, the neural in¬
ribbon is poorly understood, but it is proba¬ terconnections in the outer plexiform layer
bly instrumental in capturing synaptic vesi¬ of the primate retina have been worked out
cles and positioning them near the plasma in great detail. Both invaginating and flat
membrane. The superficial or basal contacts midget bipolars are “private” cone bipolars;
of cone pedicles do not display prominent that is, each of them is contacted by a single
junctional specializations; the synaptic cleft is cone pedicle. The invaginating variety, how¬
slightly enlarged and the adjoining mem¬ ever, sends its dendrites to the invaginating
branes of the pedicle and bipolar dendrites synapses, whereas the flat variety makes basal
bear a layer of fluffy cytoplasmic material. contacts with the cone pedicles. The diffuse
Rod spherules have a single synaptic in¬ cone bipolars, on the other hand, touch about
vagination and no basal contacts. In their six cone pedicles at superficial contacts. The
invaginating synapse, two deeply inserted ax¬ rod bipolar cells connect exclusively with rod
onal endings of the horizontal cells lie on cells; their dendritic terminals end as slightly
either side of a ridge containing a ribbon and inserted processes in the invaginations of
vesicles (Fig. 34—41A). The tips of one to four numerous spherules. Horizontal cells contact
dendrites belonging to the rod bipolar cells cone cells with their dendrites and rod cells
lie centrally and less deeply inserted. The with their axon; both dendritic and axonal
axonal endings of the horizontal cells often terminals of these cells end as deeply inserted
contain synaptic vesicles. processes in the invaginating synapses (Fig.
Using Golgi’s chromo-argentic impregna¬ 34-42).

Figure 34-41. A, Electron micrograph of the invaginating synapse of a rod spherule in the retina of a rabbit. Two deeply
inserted processes (H), probably arising from the axon of the horizontal cells, lie on either side of a wedge-shaped
projection of the spherule (synaptic ridge), which contains the synaptic ribbon, surrounded by a halo of synaptic vesicles,
and the arciform density. The dendrites of the rod bipolar cells cannot be identified in this micrograph, x 65,000. B,
Invaginating synapse of a cone pedicle in the retina of a monkey. A typical “triad” consists of two deeply invaginated
dendrites of the horizontal cells (H) and a slightly inserted, centrally positioned dendrite of an invaginating midget bipolar
cell (IMB). Notice the synaptic vesicles in the horizontal cell dendrites, x 65,000. (Micrographs courtesy of E. Raviola.)
950 • THE EYE

endings and amacrine dendrites in the inner

plexiform layer; their body is located in the
ganglion cell layer; their axon, which be¬
comes a fiber of the optic nerve, conducts to
the brain the results of the complex neural
activity that takes place in the retina. The
primate retina contains numerous varieties
of ganglion cells, classified according to the
shape of their dendritic tree and the mode
of distribution of their dendritic branches in
the inner plexiform layer. In the central area
the most common type of ganglion cell is
Figure 34-42. Diagram of the synaptic connections in the represented by the midget ganglion cell, char¬
outer plexiform layer of the primate retina. For explanation, acterized by a single dendritic shaft that as¬
see text, c, Cone cells; r, rod cells; rb, rod bipolar cell;
cends into the inner plexiform layer, where
imb, invaginating midget bipolar cell; fmb, flat midget
bipolar cell; db, diffuse cone bipolar cell; ha, horizontal it ends with a minute basket of short secon¬
cell axon; he, horizontal cell. (From Kolb, H. Phil. Trans. dary and tertiary dendrites. Diffuse ganglion
R. Soc. Lond. B 258:261, 1970.) cells send their dendrites throughout the
thickness of the inner plexiform layer,
whereas stratified ganglion cells have their den¬
dritic arborization confined to one or more
Amacrine Cells
levels of the inner plexiform layer. Interme¬
These neurons have numerous dendrites diate forms between diffuse and stratified
but lack an axon (Figs. 34-39, 34-40c). Their ganglion cells are also described.
body lies in the vitreal part of the inner
nuclear layer, and their dendrites spread in
Inner Plexiform Layer
the inner plexiform layer. They connect with
each other, with the axonal endings of the This is the region of synaptic interplay
bipolar cells, and with the dendrites of the between bipolar, amacrine, and ganglion
ganglion cells. The primate retina contains cells. Two kinds of synaptic contacts are
numerous varieties of amacrine cells; these found in this layer: the ribbon synapse, char¬
are classified as diffuse and stratified. Diffuse acterized by a dense lamella, surrounded by
amacrine cells send their dendritic branches a halo of vesicles in the presynaptic process;
throughout the thickness of the inner plexi¬ and a more conventional type of synaptic
form layer, whereas the ramifications of the contact, which lacks ribbon and is character¬
stratified amacrine cells are confined to dif¬ ized by clustering of vesicles against the pre¬
ferent sublaminae of the inner plexiform synaptic membrane (Fig. 34-43). At ribbon
layer. synapses, the axonal endings of the bipolar
cells are presynaptic to amacrine and gan¬
Interplexiform Cells glion ceil dendrites. Amacrine cell dendrites,
in turn, make conventional synapses with
Interplexiform cells have their perikaryon bipolar endings, ganglion cell dendrites, and
in the inner nuclear layer and send their dendrites of other amacrine cells. Reciprocal
processes to both plexiform layers. In the synapses between bipolar terminals and ama¬
inner plexiform layer, their processes are crine dendrites frequently occur in this layer,
both pre- and postsynaptic to amacrine cell with the bipolar contacting the amacrine cell
dendrites. In the outer plexiform layer, their as the presynaptic element of a ribbon syn¬
processes are exclusively presynaptic to hor¬ apse, and the amacrine cell returning a con¬
izontal and bipolar cells. Thus, the input to ventional feedback synapse onto the bipolar.
these cells is confined to the inner plexiform Thus, amacrine cell dendrites have the un¬
layer, whereas their output is in both plexi¬ usual property of containing synaptic vesicles
form layers. and behaving as presynaptic elements of den-
dr oaxonic and dendrodendritic synapses. The
Ganglion Cells existence of dendrodendritic synapses was
originally described in the olfactory bulb,
Ganglion cells represent the terminal link where the granule cells, which lack an axon,
of the neural networks of the retina. With establish reciprocal dendrodendritic synapses
their dendrites, they connect with bipolar with the dendrites of the mitral cells.
 THE EYE • 951

Figure 34-43. Electron micrograph of a portion of the inner plexiform layer in the retina of a monkey. An axonal ending
of a bipolar cell makes two ribbon synapses with four processes, labeled A, B, C, and D. Processes B, C, and D contain
synaptic vesicles and may represent dendrites of amacrine cells. Process B makes a conventional type of synaptic
contact with process A, whose identity is unknown. Arrows indicate the direction of the synaptic influences, x 38,400.
(Courtesy of E. Raviola.)

The precise pattern of neural interconnec¬ quadrant circle below it on their way to the
tions in the inner plexiform layer is poorly papilla. They follow the larger retinal vessels
understood. Sublayers can be distinguished fairly closely. A line connecting the fovea
in it, for the axonal arborizations of the with the temporal circumference of the retina
bipolar cells and the dendritic expansions of separates the optic nerve fibers of the upper
the stratified amacrine and ganglion cells are from those of the lower temporal quadrant.
distributed at different levels within the layer. This separation is preserved along the central
This and recent findings in nonprimate reti¬ visual pathway as far as the cortex.
nas suggest that the various types of bipolar, In primates, each retina is divided into two
amacrine, and ganglion cells establish specific halves along the vertical meridian passing
synaptic connections with each other. How¬ through the center of the fovea. The fibers
ever, the details of their “wiring” have not from the nasal half cross in the optic chiasma
yet been worked out for the primate retina. and pass to the optic tract of the opposite
side; those from the temporal half enter the
tract on the same side. Each optic tract is,
Optic Nerve Fibers in the Primate therefore, composed of fibers from the tem¬
Retina poral half of the retina of the same side and
Because of the presence of the central the nasal half of the retina of the opposite
fovea, the optic nerve fibers have a special eye. This arrangement remains in the visual
course. In general they converge radially radiation in the occipital lobes of the brain.
toward the optic papilla. However, those It accounts for the blindness in the opposite
originating in the upper temporal quadrant halves of the two fields of view (homonymous
of the retina circle above the central area, hemianopsia) when the optic tract or the
while those originating in the lower temporal visual radiation of one side is interrupted.
952 • THE EYE

Figure 34-44. Horizontal cells of rabbit retina. A, An axonless horizontal cell injected with horseradish peroxidase. The
cell has a small number of stout, tortuous dendrites, which branch sparsely and have minute terminal branchlets. At the
asterisk, a Muller cell has been stained by peroxidase that leaked from the electrode during penetration. B, The somatic
end of an axon-bearing horizontal cell. The cell has numerous wavy dendrites radiating in all directions. These carry
clusters of fine terminal branchlets. The site of electrode penetration is marked by an asterisk. Open arrows indicate
the direction to the optic nerve head. (Photomicrographs from Dacheux, R. F., and E. Raviola. J. Neurosci 2-1486
1982.) '

Inner Limiting Membrane outer to the inner limiting membrane. In the

two plexiform layers the radial pillars give
This traditional term is no longer appro¬ off many branches, which form a dense neu¬
priate. It is not a membrane but merely the roglial network in whose meshes are lodged
basal lamina of the Muller cells. It separates the ramifications of the neurons described
their inner conical ends from the vitreous earlier. In the nuclear layers, the Muller cells
body.
are beset with excavations that envelop the
bodies of the retinal neurons.
Supporting, or Neuroglial, Elements At the limit between the outer nuclear layer
of the Retina and the layer of the rods and cones, Mfiller
cells, as described, have prominent zonulae
The retina, being a modified part of the adherentes that produce the linear density
brain, contains supporting elements of neu¬ formerly interpreted as an outer limiting
roglial character. The most important are the membrane.
radial cells of Muller. These are present
throughout the central area, including the
fovea, as well as in the periphery. Central Area and Fovea (“Macula”)
Their oval nuclei lie in the middle zone of
Slightly lateral to the papilla is the place of
the inner nuclear layer. The cell body is a
most distinct vision. This region, the central
slender pillar that extends radially from the
aiea, is characterized by the presence of
 THE EYE • 953

Figure 34-45. Photomicrograph of the fovea of a macaque retina, showing the marked reduction in thickness in this
area of maximal visual acuity. (Courtesy of H. Mizoguchi.)

cones and other nervous elements in num¬ face of this dark chamber is lined with the
bers greater than elsewhere and by their photosensitive retina. The rays of light ema¬
structural specialization and synaptic perfec¬ nating from each point of an illuminated
tion. In the center of this area, the layers object impinging on the cornea are refracted
inward to the outer nuclear layer are dis¬ by it and converge on the lens. In the lens,
placed laterally, producing a shallow depres¬ the rays are further refracted and focused in
sion on the vitreal surface of the retina, called the photosensitive layer of the retina. In
the central fovea. This permits an almost free relation to the object, the retinal image is
passage of rays of light to the layer of pho¬ inverted (because of the crossing of the rays
toreceptors, and it is here that the visual axis in the pupil’s aperture); it is a real image,
touches the retina. and is very much reduced in size.
The fovea is a shallow bowl with its con¬ In the retina, the quanta of incident light
cavity toward the vitreous (Fig. 34—45). It is are converted or transduced by the photo¬
in the middle of the central area, 2 to 2.5 receptor cells into nerve signals; these are
mm on the temporal side of the papilla. In elaborated by the networks of retinal neurons
its center afloor or fundus can be distinguished, and finally translated into a code of nerve
together with the slopes and a margin of the impulses, which is conducted to the brain by
fovea. The width of the entire foveal depres¬ the fibers of the optic nerve.
sion measures 1.5 mm. The transduction process can be conven¬
In the fundus of the fovea, the cones are iently subdivided into primary and secondary
thinner and longer than elsewhere in the steps. The primary step is a photochemical
retina. The central rod-free area, where only reaction and consists in the absorption of a
cones are present, measures 500 to 550 pirn quantum of light by one of the visual pig¬
in diameter and contains up to 30,000 cones. ments contained in the discs of the photore¬
Capillaries are present in the foveal slopes ceptor outer segments, and a subsequent
to the very edge of the foveal floor, or 275 configurational change in the absorbing
pm from the very center. The avascular cen¬ molecule. The secondary step consists of
tral territory is almost as large as the rodless changes in the concentration of internal
area (450 to 500 pm). transmitters within the cytoplasm of the outer
segments, which influence the ionic permea¬
bility of the plasma membrane and cause the
hyperpolarization of the photoreceptor cell.
HISTOPHYSIOLOGY OF THE Rod and cone cells differ in their sensitivity
RETINA to intensity and wavelength of light. Rod cells
are active in dim illumination (scotopic vision)
The eye is essentially a camera obscura and contain a single visual pigment, rhodop-
provided with dioptric media: the cornea, sin, which absorbs light of various wave¬
the aqueous humor, the adjustable crystalline lengths, although most efficiently in the blue-
lens, and the vitreous body. The inner sur¬ green. In the human retina there are three
954 • THE EYE

types of cones, containing different visual icity horizontal cells, which may hyperpolarize
pigments that absorb maximally red, blue, or or depolarize, depending on the wavelength
green light; furthermore, cone cells are active of the stimulating light. In the retina of the
under the conditions of diurnal illumination turtle, the hyperpolarization of luminosity
(photopic vision). horizontal cells causes depolarization of cone
Visual pigments consist of a combination cells; it has therefore been suggested that
of vitamin A aldehyde, known as retinal, with cones receive from horizontal cells a feedback
a protein of the class called opsins. Opsins are synapse. In all vertebrates, there are two
hydrophobic proteins with one retinal group physiological classes of bipolar cells, one de¬
per molecule, a carbohydrate side chain, no polarizing, the other hyperpolarizing, upon
phospholipid, and a molecular weight of light stimulation. Bipolar cells are the first
about 27,000 daltons. They are buried within elements in the chain of retinal neurons that
the membrane of the discs of the photore¬ show a “center-and-surround” organization
ceptor outer segments. When retinal is com¬ of their receptive held; that is, the cell re¬
bined with opsin in the dark-adapted retina, sponse to stimulation of neighboring photo¬
it has a bent and twisted form (11-cis). When receptors is antagonized by the stimulation
the pigment moleule absorbs a quantum of of distant photoreceptors. Furthermore,
light, the retinal shape becomes straight (all- some bipolars are color-coded; that is, they
trans) and separates from opsin, which in respond to light of specific wavelength. The
turn undergoes a conformational change. neuronal circuitry that underlies the bipolar
This leads to activation of various enzymes activity is not well understood; the response
and the lowering of the concentration of of bipolars to stimulation of the photorecep¬
cGMP in the outer segment. At the same tors in the center of their receptive held may
time, calcium ions are released into the cy¬ be mediated by a photoreceptor-to-bipolar
toplasm. Either cGMP, calcium, or both may synapse. The “surround effect” is probably
represent the internal transmitters that reg¬ due to the activity of the horizontal cells,
ulate the patency of the ion channels in the which are activated by the photoreceptors
plasma membrane. In the dark, sodium ions and in turn antagonize the center response
are continuously pumped out of the cell in of the bipolar cells. The precise mechanism
the inner segment and reenter the cell of this interaction, however, is unknown;
through sodium channels located in the horizontal cells may feed back onto photo¬
plasma membrane of the outer segment. receptors or may influence directly the bi¬
Light diminishes or suppresses this “dark polar cells. Thus, the function of the synapses
current” by blocking the sodium channels of in the outer plexiform layer is to signal the
the outer segment, and thus causes the hy¬ intensity and color of the retinal image and
perpolarization of the photoreceptor cell. at the same time to accentuate its contrast
Meanwhile the cell is recombining retinal and through the activity of the horizontal cells.
opsin into the light-absorbing form of the The signals of bipolar cells are further
pigment molecule. processed in the inner plexiform layer by the
Most present knowledge on the electro¬ amacrine and ganglion cells before their
physiology of retinal neurons stems from transmittal to the brain in the form of
intracellular recordings in nonprimate reti¬ changes in frequency of the nerve impulses
nas. Stripped of many important details, the traveling along the fibers of the optic nerve.
complex story of the interneuronal relation¬ The function of the amacrine cells is poorly
ships in the retina is as follows. Most retinal understood; there are many varieties of
neurons generate slow, graded potentials; them, both morphological and physiological,
spikes first appear in amacrine cells and are and they modulate the transfer of signals
especially typical of the ganglion cell re¬ from bipolar to ganglion cells. In the monkey
sponse. T he photosensitive rod and cone cells retina two main types of ganglion cells were
are depolarized and release transmitter in identified by intracellular recordings, one re¬
the dark; the light-induced hyperpolarization sponding to the onset, the other to the ces¬
decreases the output of transmitter by their sation, of a light stimulus applied to the
synaptic endings. In cold-blooded verte¬ center of their receptive held; the response
brates, there are two physiological classes of was inverted upon stimulation of the periph¬
horizontal cells: luminosity horizontal cells, ery of their receptive held. Among the on-
which respond to illumination of the photo¬ center cells, some are not color-coded and
receptors by hyperpolarization; and chromat- phasic—that is, they discharge transiently to
 THE EYE • 955

sustained stimuli of any wavelength; others NERVES OF THE EYE

are color-coded and tonic—that is, they dis¬
charge continuously to sustained stimuli of These are the optic nerve, originating from
either green or red light. The tonic cells are the retina, and the ciliary nerves, supplying
thought to correspond to the midget gan¬ the eyeball with motor, sensory, and sympa¬
glion cells. It has been speculated that the thetic fibers.
neural interactions in the inner plexiform The optic nerve, an evagination of the
layer may be concerned with codification of prosencephalon, is not a peripheral nerve
the dynamic or temporal aspects of the visual like the other cranial nerves, but is a tract of
image, but much work remains to be done the central nervous system. It consists of
before the significance of the synaptic inter¬ about 1200 bundles of nerve fibers whose
actions among bipolar, amacrine, and gan¬ myelin sheaths are produced by oligoden-
glion cells is fully elucidated. droglial cells.
The meninges and the intermeningeal
spaces of the brain continue into the optic
nerve. The outer sheath of the nerve is
BLOOD VESSELS OF THE EYE formed by the dura, which continues toward
the eyeball and fuses with the sclera. The
These arise from the ophthalmic artery intermediate sheath is formed by the arach¬
and can be subdivided into two groups, which noid and the inner sheath by the pia mater.
are almost completely independent and an¬ The pia mater forms a connective tissue layer
astomose with each other only in the region that is closely adherent to the surface of the
of the entrance of the optic nerve. The first nerve and fuses with the sclera at the en¬
group, the retinal system, represented by the trance of the optic nerve. This pial layer
central artery and vein, supplies a part of the sends connective tissue partitions and blood
optic nerve and the inner retina. The second, vessels into the nerve. Inflammatory proc¬
the ciliary system, is destined for the uveal esses can extend from the eyeball toward the
tunic; through the uvea, it provides for the meningeal spaces of the brain through the
nourishment of the outer retina. Like the spaces between the sheaths.
brain, the retina is protected from circulating The optic nerve leaves the posterior pole
macromolecules by a blood-retina barrier, its of the eyeball in a slightly oblique direction
structural counterpart is chiefly represented and continues into the entrance canal of the
by the zonulae occludentes that seal the in¬ optic nerve. Just after leaving the eye
tercellular spaces between the cells of the through the openings in the lamina cribrosa,
pigment epithelium and those between the the fibers acquire their myelin sheaths. The
endothelial cells of the retinal blood vessels. central artery and central vein reach the
eyeball through the optic nerve; they pene¬
trate the nerve on its lower side at a distance
from the eyeball varying from 5 to 20 mm,
LYMPH SPACES OF THE EYE but usually 6 to 8 mm.

True lymph capillaries and lymph vessels

are present only in the scleral conjunctiva. In ACCESSORY ORGANS
the eyeball they are absent.
A mass injected into the space between the
OF THE EYE
choroid and sclera penetrates along the walls
of the vortex veins into the space of Tenon. In an early stage of embryonic develop¬
The latter continues as the supravaginal space ment the anterior segment of the eyeball
along the outer surface of the dural sheath projects freely on the surface. Later a circular
of the optic nerve to the optic foramen. fold of integument encircles the cornea.
Again, it is possible to inject into Tenon’s From its upper and lower parts the upper
space from the subarachnoid space of the and lower lids grow toward each other over
brain. From the anterior chamber the in¬ the surface of the cornea. In this way, the
jected liquid passes into the posterior cham¬ conjunctival sac is formed, which protects
ber and also into Schlemm’s canal. All these and moistens the free surface of the eye,
spaces cannot, however, be regarded as be¬ especially the cornea. I he part lining the
longing to the lymphatic system. inner surface of the lids is the palpebral con-
956 • THE EYE

junctiva, and that covering the eyeball is the stance. They are elongated and arranged in
bulbar conjunctiva. The reflection of the pal¬ one layer, parallel to one another and per¬
pebral onto the bulbar conjunctiva forms pendicular to the length of the tarsal plate.
deep recesses between the lids and the eye¬ Their openings form a single row immedi¬
ball, the superior and the inferior fornices. ately in front of the inner free edge of the
lid, at the line of transition from the skin
Eyelids into the conjunctiva.
The meibomian glands are sebaceous but
The outermost layer of the eyelids is the have lobated alveolar terminal portions. They
skin. It is thin and provided with a few are connected by short lateral ducts with a
papillae and many small hairs with sebaceous long central excretory duct lined with strati¬
and small sweat glands (Fig. 34—46). The fied squamous epithelium.
dermis contains a varyin g number of pigment The innermost layer of the lid is the con¬
cells with yellow or brown granules. The junctiva. At the inner edge of the margin of
loose subcutaneous layer is rich in hue elastic the lid, the epidermis continues into the inner
fibers, and in Caucasians is almost completely surface of the lid. Here the superficial cells
devoid of fat. Toward the edge of the lid the become thicker, the number of layers de¬
dermis becomes denser and has higher pa¬ creases, and the epithelium assumes a strati¬
pillae. fied columnar character, which is typical of
The eyelashes are large hairs obliquely in¬ the whole conjunctiva and varies only in the
serted in three or four rows along the edge thickness in different places. The superficial
of the lid. With their follicles they penetrate cells have a short prismatic form. Goblet cells
deeply into the tissue. The sebaceous glands are scattered between them (Fig. 34—47).
connected with the eyelashes are small, and At the upper edge of the tarsus the epithe¬
arrector muscles are missing. The eyelashes lium is sometimes reduced to two cell layers,
are replaced every 100 to 150 days. and its surface presents many irregular in¬
Between and behind the follicles of the vaginations. Some of them are lined with
eyelashes are peculiar sweat glands, the glands mucous cells and are described as glands. In
of Moll. Unlike ordinary sweat glands, the the conjunctiva of the fornix, the epithelium
terminal portion here is generally straight or is thicker.
only slightly coiled. The ducts open, as a rule, The lamina propria of the conjunctiva is
into the follicles of the eyelashes. The epithe¬ dense connective tissue. In the region of the
lium of the terminal portions consists of an fornix it is very loosely attached to the in¬
indistinct, outer myoepithelial layer and an traorbital fat tissue, permitting the free mo¬
inner layer of pyramidal, apocrine glandular tion of the eyeball in the conjunctival sac.
elements. The lumen is often considerably In the region of the corneal limbus the
dilated, and the glandular cells are flattened. epithelium of the conjunctiva assumes a strat¬
In the ducts the epithelium consists of two ified squamous character and continues as
distinct cell layers. The nature of the secre¬ such onto the surface of the cornea. It may
tion of these glands is not known. still contain a few scattered mucous cells.
The next layer of the eyelid inward consists The rudimentary third eyelid, or semilunar
of the thin, pale, striated fibers of the pal¬ fold (the homologue of the nictitating mem¬
pebral portion of the ring of facial muscle brane of the lower vertebrates), is formed by
surrounding the eye (orbicularis oculi). The the scleral conjunctiva at the inner palpebral
part behind the follicles of the eyelashes is commissure, lateral to the lacrimal caruncle.
the ciliary muscle of Riolan. It consists of connective tissue that contains
Deep to the orbicular muscle is a layer of smooth muscle fibers and is covered with
connective tissue, the palpebral fascia, a con¬ conjunctival epithelium, which, on the outer
tinuation of the tendon of the palpebral lev¬ surface, contains many mucous cells.
ator (levator palpebrae) muscle. In the upper
part of the upper lid, strands of smooth
muscle, the superior tarsal muscle of Muller, are
Lacrimal Gland
attached to the edge of the tarsus, a plate of Opening into the conjunctival space there
dense connective tissue that forms the skele¬ is a system of glands whose secretion mois¬
ton of the lid. In the upper lid its breadth is tens, lubricates, and flushes the surface of
about 10 mm, in the lower only 5 mm. The the eyeball and of the lids. Of these glands,
glands of Meibom are embedded in its sub¬ only the lacrimal gland reaches a high degree,
Figure 34-46. Camera lucida drawing of a slice of the upper eyelid of a newborn infant. Stained with hematoxylin.
THE EYE • 957
958 • THE EYE

Figure 34-47. Photomicrograph of the superficial portion of the conjunctival epithelium of Macaca mulatta, showing a
goblet cell and clusters of melanosomes in the neighboring epithelial cells. (Photomicrograph courtesy of G. Raviola.)

of development. It has the size and shape of On the inner surface of the lids, especially
an almond and is lodged beneath the con¬ the upper one, near the upper edge of the
junctiva at the lateral upper side of the eye¬ tarsus, there are a varying number of small
ball. It consists of a group of separate glan¬ accessory lacrimal glands—the tarsal lacrimal
dular lobules and has six to 12 excretory glands.
ducts, which open along the upper and lat¬ After having washed the conjunctival cav¬
eral quadrant of the superior conjunctival ity, tears reach the region of the inner pal¬
fornix. pebral commissure (internal canthus). Here
The lacrimal gland is of the tubuloalveolar the two eyelids are separated by a triangular
type. Its terminal portions are provided with space, the lacrimal lake, in which the secretion
a relatively large lumen and with irregular, accumulates temporarily. From here it passes
saccular outpocketings. The basal lamina is through two tiny orifices called lacrimal points,
lined with glandular cells resembling those one on the margin of each eyelid, into the
of the serous salivary glands. They have, lacrimal ducts. The latter converge medially
however, a narrower columnar shape and into the lacrimal sac, whence the nasolacrimal
contain, in addition to small lipid droplets, duct leads into the inferior meatus of the
large, pale secretion granules whose number nasal cavity.
changes according to the functional condi¬ The wall of the excretory lacrimal passages
tions. These cells are provided with secretory is formed by connective tissue lined with
canaliculi. Between their bases and the basal epithelium. The epithelium of the lacrimal
lamina are well-developed myoepithelial ducts is stratified squamous. The lacrimal sac
cells. The smallest intralobular ducts are and the nasolacrimal duct are lined with a
lined with a layer of low columnar or cuboidal pseudostratihed, tall columnar epithelium.
cells and have a few myoepithelial cells. The From the bottom of the lacrimal lake, be¬
larger intralobular ducts have a two-layered tween the two lacrimal ducts, there bulges a
epithelium. small, soft mass of tissue, the lacrimal caruncle.
 THE EYE • 959

The top is covered with a thick, squamous capillaries of the scleral conjunctiva end
epithelium in which only the uppermost lay¬ blindly near the corneal margin.
ers are flattened, although not cornihed. It The abundant supply of the conjunctiva
contains mucous cells, and gradually merges with blood and lymph capillaries accounts for
into the conjunctival epithelium. The lamina the rapid absorption of solutions introduced
propria contains bundles of striated muscles, into the conjunctival sac.
sweat glands, abortive lacrimal glands, and
tiny hairs with sebaceous glands. These are
the source of the whitish secretion that often
HISTOGENESIS OF THE EYE
collects in the region of the inner palpebral
commissure.
The stalk of the optic vesicle growing out
of the brain is transformed into the optic
Blood and Lymph Vessels nerve (Fig. 34-48). The double-walled vesicle
of the Eyelids gives rise to the retina. Where the optic
The arteries in each lid form two archlike vesicle touches the ectoderm, the latter forms
anastomoses, which run in front of the tarsus, an invagination with a greatly thickened bot¬
one near the free margin of the lid, the other tom, the primordium of the lens. It apparently
near the other margin of the tarsus. The develops as the result of inductive stimulation
palpebral conjunctiva is provided with dense, of the ectoderm by the optic vesicle. In am¬
subepithelial capillary networks that can be phibian larvae, after excision of the optic
easily studied in living condition with the aid vesicle, the lens is not formed. The lens
of the slit lamp microscope. Branches of the primordium comes to lie in the invagination
blood vessels in the scleral conjunctiva anas¬ of the optic vesicle. Simultaneously, mesen¬
tomose with the marginal blood vessels of the chyme and blood vessels grow into the cho¬
cornea and with the branches of the anterior roidal fissure, in the lower part of the optic
ciliary arteries. vesicle. Simultaneously, mesenchyme and
The lymphatics form a dense plexus in the blood vessels grow into the choroidal fissure,
conjunctiva behind the tarsus. In front of the in the lower part of the optic vesicle. These
latter there is another, thinner, pretarsal net. vessels give rise to the hyaloid and retinal
A third net can be distinguished in the skin vascular systems. The opposite margins of
and the subcutis. All these networks com¬ the fissure, which received the vessels, soon
municate with one another. The lymphatic grow together, and the secondary optic vesi-

Inner layer of
Outer layer of secondary optic vesicle
secondary
Stalk of optic vesicle

Side of anterior
.brain vesicle
Border of
optic cup
Ventricle of brain

Mesenchyme

Epithel ium Bottom of anterior

of cornea brain vesicle

Epithel ium
of lens

Lens fibers with

body
nuclear zone
Figure 34-48. Primordium of the eye of an 8-mm mouse embryo. The cavity of the primary optic vesicle is reduced to
a thin cleft, x 70. (After Schaffer.)
 960 • THE EYE

cle assumes the form of a double-walled cup, Richardson, K. C.: The hne structure of the albino
while the stalk is transformed into a solid rabbit iris with special reference to the identihcation
of adrenergic and cholinergic nerves and nerve
strand, the optic nerve.
endings in its intrinsic muscles. Am. J. Anat.
The lens primordium soon becomes de¬ 774:173, 1964.
tached from the ectoderm, and the space
CILIARY BODY
between the two is filled by a layer of mes¬
enchyme—the primordium of the substantia Fine, B. S.: Structure of the trabecular meshwork and
the canal of Schlemm. Trans. Am. Acad. Ophthal¬
propria of the cornea and of the connective
mol. Otolaryngol. 70:777, 1966.
tissue of the iris. The lens, surrounded by Inomata, H., A. Bill., and G. K. Smelser: Aqueous
vascular mesenchyme, acquires a solid, spher¬ humor pathways through the trabecular meshwork
ical form, while the original cavity disappears. and into Schlemm’s canal in the cynomolgus monkey
{Macaca irus). Am. J. Ophthalmol. 78:760, 1972.
The inner, thicker sheet of the double wall
Ishikawa, T.: Fine structure of the human ciliary muscle.
of the optic cup differentiates into the retina Invest. Ophthalmol. 7:587, 1962.
proper. It remains permanently in direct Raviola, G.: The hne structure of the ciliary zonule and
continuation with the optic nerve. The outer ciliary epithelium. Invest. Ophthalmol. 76:851,
sheet of the cup is transformed into the 1971.
Raviola, G.: The structural basis of the blood-ocular
pigment epithelium. The surrounding mes¬
barriers. Exp. Eye Res. (Suppl.) 25:27, 1977.
enchyme comes into close relation with the Raviola, G., and E. Raviola: Intercellular junctions in
optic cup and gives rise to the two outer the ciliary epithelium. Invest. Ophthalmol. Vis. Sci.
tunics of the eyeball, the uveal and fibrous 76:958,7 1978. »

tunics. The structural differentiation of the Raviola, G., and E. Raviola: Paracellular route of
aqueous outflow in the trabecular meshwork and
retina proceeds in a way similar to that of
Schlemm canal. Invest. Ophthalmol. 27:52, 1981.
the wall of the neural tube. The eyeball Toates, F. M.: Accommodation function of the human
attains full size toward the end of the first eye. Physiol. Rev. 52:828, 1972.
decade. The structure of the retina, including Tormey, J. McD.: Fine structure of the ciliary epithelium
of the rabbit with particular reference to “infolded
the central fovea, matures toward the end of
membranes,” “vesicles,” and the effects of Diamox.
the first year. J. Cell Biol. 77:641, 1963.

LENS AND VITREOUS BODY

REFERENCES
Farnsworth, P. N., S. C. Fu, P. A. Burke, and I. Bahia:
GENERAL Ultrastructure of rat eye lens hbers. Invest.
Ophthalmol. 77:274, 1974.
Davson, H.: The Physiology of the Eye. New York, Swann, D. A.: Chemistry and biology of the vitreous
Academic Press, 1972. body. Int. Rev. Exp. Pathol. 22:2, 1980.
Duke-Elder, S., and K. C. Wybar: In Duke-Elder, S., Wanko, T., and M. A. Gavin: Electron microscopic study
ed.: The Anatomy of the Visual System. System of of lens hbers. J. Biophys. Biochem. Cytol. 6:97,
Ophthalmology. Vol. 2. St. Louis, C. V. Mosby Co., 1959.
1961.
Fine, B. S., and M. Yanoff: Ocular Histology. New York, RETINA
Harper 8c Row, 1972. Dacheux, R. F., and E. Raviola: Horizontal cells in the
Hogan, M. J., J. A. Alvarado, and j. E. Weddell: Histol¬ retina of the rabbit. J. Neurosci. 2:1486, 1982.
ogy of the Human Eye. Philadelphia, W. B. Saun¬ Dowling, J. E.: Organization of vertebrate retinas. Invest.
ders Co., 1971. Ophthalmol. 9:665, 1970.
Walls, G. L.: The Vertebrate Eye and its Adaptive Polyak, S.: The Retina. Chicago, University of Chicago
Radiation. Bloomfield Hills, MI, Cranbrook Press, Press, 1941.
1943.
Raviola, E.: Intercellular junctions in the outer plexiform
CORNEA layer of the retina. Invest. Ophthalmol. 75:881,
1976.
Hay, E. D.: Development of the vertebrate cornea. Int.
Raviola, E., and N. B. Gilula: Intramembrane organi¬
Rev. Cytol. 67:263, 1980.
zation of specialized contacts in the outer plexiform
Jakus, M. A.: Studies on the cornea. II. The hne struc¬
layer of the retina. J. Cell Biol. 65:192, 1975.
ture of Descemet’s membrane. J. Biophys. Biochem.
Raviola, G., and E. Raviola: Light and electron micro¬
Cytol. (Suppl.) 2:243, 1956.
scopic observations on the inner plexiform layer of
IRIS the rabbit retina. Am. J. Anat. 726:403, 1967.
Rodieck, R. W.: The Vertebrate Retina. San Francisco,
Freddo, T. F., and G. Raviola: Homogeneous structure W. H. Freeman, 1973.
of blood-vessels in the vascular tree of Macaca mu-
Schwartz, E. A.: First events in vision: the generation of
latta iris. Invest. Ophthalmol. Vis. Sci. 22:279, 1982.
responses in vertebrate rods. 1. Cell Biol. 96:271
Gregersen, E.: The spongy structure of the human iris. 1982.
Acta Ophthalmol. 76:522, 1958.
\ oung, R. W.: Visual cells and the concept of renewal.
Raviola, G., and j. M. Butler: Unidirectional transport
Invest. Ophthalmol. 75:700, 1976.
mechanism of horseradish peroxidase in the vessels
Young, R. W., and D. Bok: Participation of the retinal
of the iris. Invest. Ophthalmol. Vis. Sci. 25 827
pigment epithelium in the rod outer segment re¬
1984.
newal process. J. Cell Biol. 42:392, 1969.
 35
THE EAR

The organ of hearing is divisible into three in the temporal bone (Fig. 35—1). It forms
parts, each of which differs from the others an S-shaped curve coursing for about 2.5 cm
not only in its gross anatomy but also in its medially and inferiorly, and is bounded at its
histology and in the functions that it sub¬ medial end by the eardrum or tympanic mem¬
serves in the translation of sound waves into brane. The skin lining the meatus is thin and
meaningful information that can be proc¬ is firmly attached to the underlying perichon¬
essed in the central nervous system. The first drium and periosteum. Numerous hairs in
part, the external ear, receives the sound the lining of the outer cartilaginous portion
waves. In the second part, the middle ear, the of the meatus tend to prevent entrance of
waves are transformed into the mechanical foreign bodies. In old age these hairs enlarge
vibrations of bony auditory ossicles. These, in considerably, as do those on the auricle. Se¬
turn, by impinging upon the fluid-filled baceous glands connected with the hair fol¬
spaces of the third part, the internal ear (or licles are exceptionally large. In the bony
labyrinth), generate specific nerve impulses portion of the meatus, small hairs and seba¬
that are conveyed by the acoustic nerve to ceous glands are found only along the upper
the central nervous system. In addition to wall. No eccrine sweat glands are present in
organs for analysis of sound, the internal ear the meatus.
contains vestibular organs, which are con¬ The external meatus contains cerumen, a
cerned chiefly with the function of maintain¬ brown, waxy secretion that protects the skin
ing equilibrium. from desiccation and presumably from inva¬
sion by insects. It is a mixture of the secretion
of the sebaceous glands and ceruminous glands
of the skin of the meatus. Ceruminous glands
EXTERNAL EAR are a special variety of coiled tubular apo¬
crine sweat gland. In cross section, they ap¬
Auricle pear to be aggregated into discrete lobules
invested by connective tissue. Each glandular
The auricle, or pinna, consists of a single,
tubule is surrounded by a thin network of
highly irregular plate of elastic cartilage, 0.5
myoepithelial cells. In the resting state, the
to 1 mm thick, overlain by a flexible peri¬
gland lumen is large and the epithelial cells
chondrium containing abundant elastic fi¬
lining it are cuboidal. In the active state,
bers. The covering skin has a distinct subcu¬
however, the cells are columnar and the lu¬
taneous layer only on the posterior surface men is constricted. Ducts of ceruminous
of the auricle and is provided with a few glands open either onto the free surface of
small hairs and associated sebaceous glands, the skin or, with the sebaceous glands, into
the latter sometimes being of considerable the necks of hair follicles.
size. In old age, especially in men, large stiff
hairs develop on the dorsal edge of the au¬
ricle and on the ear lobe. Sweat glands are
scarce, and when present are small. MIDDLE EAR
External Auditory Meatus The middle ear comprises the tympanic cav¬
The outer portion of the external auditory ity and its contents, the auditory ossicles; the
meatus is a medial continuation of the auric¬ eustachian tube; and the tympanic membrane,
ular cartilage, and the inner portion is a canal which closes the tympanic cavity externally.

961
962 • THE EAR

Figure 35-1. Schematic representation of the anatomical relations of the various parts of the human ear. (After Brodel,
M. In Malone, Guild, and Crowe. Three Unpublished Drawings of the Human Ear. Philadelphia, W. B. Saunders Co ’
1946.)

TYMPANIC CAVITY cilia. The presence of glands associated with

the lining of the cavity is generally denied.
The tympanic cavity is an irregular, air-
filled space in the temporal bone. Its lateral
wall is formed largely by the tympanic mem¬
Auditory Ossicles
brane and its medial wall by the lateral aspect Three small bones—the malleus, the incus,
of the bony wall of the internal ear (Fig. and the stapes—extend from the attachment
35-2). Anteriorly it continues into the audi¬ of the malleus on the tympanic membrane to
tory tube and posteriorly it is connected, the medial wall of the tympanic cavity, where
through the tympanic antrum, with air-filled the footplate of the stapes fits into the fenestra
cavities in the mastoid process of the tem¬ vestibuli, or oval window, a hiatus in the wall
poral bone. The cavity contains the auditory of the osseous labyrinth (Figs. 35—1, 35-2).
ossicles; the tendons of two small muscles (the The footplate is maintained in the fenestra
tensor tympani and the stapedius) connected by means of an annular fibrous ligament.
with the ossicles; and the chorda tympani nerve The three bones are connected to one an¬
(Fig. 35—2). other by means of typical diarthrodial joints
I he epithelium lining the tympanic cavity and are supported in the cavity by minute
is simple squamous, for the most part, but connective tissue ligaments. Small patches of
near the opening of the auditory tube and hyaline cartilage are usually found on the
near the edge of the tympanic membrane it manubrium of the malleus and on the foot¬
is cuboidal or columnar and provided with plate of the stapes. The mucosa lining the
 THE EAR • 963

Figure 35-2. Drawing of some of the anatomical features of the external, middle, and inner ear. (After Brodel, M. In
Malone, Guild, and Crowe. Three Unpublished Drawings of the Human Ear. Philadelphia. W. B. Saunders Co., 1946.)

tympanic cavity is reflected over the ossicles the membrane. Externally the membrane is
and is firmly attached to their periosteum. covered by a very thin (50 to 60 pm) layer of
skin devoid of hairs and other appendages.
Its inner surface is lined by the mucosa of
TYMPANIC MEMBRANE the tympanic cavity, here only 20 to 40 pm
thick and consisting of simple squamous ep¬
This oval, semitransparent membrane is ithelium overlying a lamina propria of sparse
shaped like a very flat cone with its apex collagenous fibers and capillaries. Over the
directed medially (Figs. 35—1 to 35—3). Its manubrium of the malleus is a layer of con¬
conical form is maintained by the insertion, nective tissue through which vessels and
onto its inner surface, of the manubrium of nerves reach the center of the tympanic
the malleus, which tends to pull the center membrane.
of the membrane medially. The tympanic
membrane is formed of two layers of collag¬
enous fibers and fibroblasts that are similar AUDITORY TUBE
to those of a flat tendon (Fig. 35-3). How¬
ever, there is a flaccid portion in its antero- From its origin in the anterior wall of the
superior quadrant, ShrapnelVs membrane, that tympanic cavity, the auditory, or eustachian,
is devoid of collagenous fibers. In the outer tube extends anteromedially and inferiorly
layer of the membrane the collagen fibers for about 4 cm to an opening on the poster¬
have a radial arrangement, whereas those in olateral wall of the nasopharynx. The rostral
the inner layer are disposed circularly. 4 here two thirds of the tube is supported medially
are also thin networks of elastic fibers, located by cartilage, and the portion near the tym¬
mainly in the central and peripheral parts of panic cavity is supported by bone.
964 . THE EAR

Ciliated columnar
epithelium Mucous
of tympanic membrane of
cavity tympanic cavity
9

ty

external meatus

Figure 35-3. Cross section of edge of tympanic membrane of a child. (Redrawn from von Ebner.)

The cartilage supporting the auditory tube There is considerable individual variation
lies mainly medial to the lumen, but a ridge in number and distribution of ciliated cells
of cartilage running longitudinally for most and goblet cells, and in the degree of devel¬
of the length of the tube curves superolater- opment of the glandular elements. Through¬
ally, so that in cross section the cartilage has out the lamina propria, in both portions of
the appearance of a shepherd’s crook (Fig. the tube, a great many lymphocytes can be
35-4). The cartilage is elastic throughout found, the number varying with age and
most of its length, but at the isthmus it loses from one individual to another. Near the
its elastic fibers and becomes hyaline. The pharyngeal opening there are often discrete
lumen of the tube, flattened in the vertical collections of lymphoid tissue forming the
plane, is largest at its pharyngeal end, de¬ tubal tonsils. Usually the auditory tube is
creases to a mere slit (the isthmus) at the closed. During the acts of swallowing and
junction of the cartilaginous and bony por¬ yawning, the lumen is opened for a short
tions, and then expands again in its course interval, allowing the pressure in the tym¬
through the temporal bone. The tube is lined panic cavity to equalize with that outside.
by a mucosa, of variable thickness, plicated
into rugae at both the pharyngeal and tym¬
panic ends. In the bony portion of the audi¬
tory tube, it is relatively thin and is composed INTERNAL EAR
of low columnar ciliated epithelium resting
on a thin lamina propria firmly bound to the The internal ear, called the labyrinth be¬
periosteum. The epithelium in the cartilagi¬ cause of its complex structure, is composed
nous portion of the tube is pseudostratified of a series of fluid-filled sacs and tubules in
and composed of tall columnar cells, many cavities of corresponding form in the petrous
of which are ciliated. The underlying lamina portion of the temporal bone (Fig. 35—5).
propria is much more complex here than in The canals and cavities in the bone consti¬
the bony portion. Toward the pharyngeal tute the osseous labyrinth. Occupying this sys¬
orifice, it contains many compound tubuloal- tem of cavities are the thin-walled, fluid-filled
veolar glands that secrete mucus via ducts tubules and saccules of the membranous laby¬
opening into the tubal lumen. In this vicinity, rinth, which constitute the endolymphatic system.
also, goblet cells are interspersed among the This is surrounded by the cells and fluid of
columnar epithelial cells. the perilymphatic system.
 THE EAR 965

Accessory
cartilage
Lateral
"cartilaginous
plate

Muscle bundles

Mixed glands

C i I iated
epithelium

Lumen of
Medial cartilaginous auditory tube
plate with patches
Mixed glands
of elastic cartilage
(darker spots) Membranous
lateral wall
with fat tissue

Accessory
cartilage

Figure 35-4. Transection of cartilaginous portion of the auditory tube near its opening into the pharynx, x 11. (Redrawn
from von Ebner.)

Ampulla membranacea superior

Upper end
branch of
vestibular nerve N. ampulla Ductus semi-
superior circularis
superior
Utriculus
Ductus
cochlearis
Ampul la
membranacea
lateralis
N. cochlearis

N. acusticus Ductus semi-

circularis
lateralis

Crus
commune
PiiiP
1 \ $
N. vestibularis
m Ductus semi
MIT'""•circularis
m posterior
Ganglion N. sacculus
vestibulare

Ductus
Sacculus endolymphaticus

N. ampullaris posterior Ampulla membranacea posterior

Figure 35-5. Right membranous labyrinth of an adult; medial and posterior aspects. About x 5. (Redrawn and modified
from Spalteholz.)
966 • THE EAR

THE BONY LABYRINTH within the modiolus along the inner wall of
the cochlear canal (Fig. 35-11). Their pe¬
There are two major cavities in the bony ripheral processes traverse the remaining dis¬
labyrinth: the vestibule, which houses the sac¬ tance to the cochlear hair cells that they
cule and utricle; and anteromedial to it, the innervate.
spirally coiled cochlea, which contains the or¬ The lumen of the canal of the osseous
gan of Corti (Figs. 35-1, 35—2). cochlea (about 3 mm in diameter) is divided
along its whole course (about 35 mm in the
The Vestibule human) into an upper and a lower section by
the spiral lamina. The lamina is divided into
The vestibule is an irregularly ovoid cavity two zones: an inner zone containing bone
located medial to the tympanic cavity. Its wall (the osseous spiral lamina) and a fibrous outer
facing the tympanic cavity is penetrated by zone (the membranous spiral lamina). The latter
the fenestra vestibuli, and certain recesses in is also called the basilar membrane (Figs. 35—11,
its wall produce characteristic bony protru¬ 35—13, 35—14). At the attachment of the
sions on the medial wall of the tympanic basilar membrane to the outer wall of the
cavity in relationship to the fenestra. For a cochlea, the periosteum is thickened and
more detailed description, the student is re¬ forms a structure that has been called the
ferred to a textbook of gross anatomy. Three spiral ligament, although histologically it does
semicircular canals arise from recesses in the not have the characteristics of a ligament.
wall of the vestibule and return to it. Accord¬ The cochlear canal is further subdivided by
ing to their position, they are named the a thin membrane, the vestibular membrane
superior, posterior, and lateral semicircular (Reissners membrane), which extends obliquely
canals. Two of the recesses located antero- from the spiral lamina to the outer wall of
superiorly accommodate the dilated ampullae the bony cochlea (Fig. 35-11). Thus, a cross
of the superior and lateral semicircular ducts section of the bony cochlea will show three
of the membranous labyrinth, and poste¬ compartments: an upper cavity, the scala ves-
riorly a third recess houses the posterior am¬ tibuli; a lower cavity, the scala tympana, and an
pulla. Given off from these recesses are the intermediate cavity, the scala media (Fig.
superior (or anterior), the lateral, and the 35-1 1). The latter is the cochlear duct, a por¬
posterior semicircular canals. The lateral canal tion of the endolymphatic system that con¬
curves laterally around the vestibule and re¬ nects with the vestibular part of the membra¬
joins it behind the posterior ampullary recess. nous labyrinth by way of the small ductus
The superior and posterior canals join each reuniens.
other superior to the vestibule in the recess The scala tympani and scala vestibuli are
for the crus commune, which opens into the perilymphatic spaces. The scala vestibuli ex¬
medial part of the vestibule. From the medial tends into and through the perilymphatic
wall of the vestibule a thin canal, the vestibular cistern of the vestibule and reaches the inner
aqueduct, extends to the posterior surface of surface of the fenestra ovalis. The scala tym¬
the petrous portion of the temporal bone. pani ends at the fenestra rotundum. At the
apex of the cochlea the two scalae commu¬
The Cochlea nicate through a small opening, the helico-
trerna.
The bony cochlea is anteromedial to the
vestibule (Figs. 35—1, 35—2, 35—5). It consists
of a complex bony canal that makes two and
three quarter spiral turns around an axis THE MEMBRANOUS (OR
formed by the conical pillar of spongy bone ENDOLYMPHATIC) LABYRINTH
called the modiolus. The base of the modiolus
forms the deep end of the internal acoustic The fluid-filled sacs of the membranous
meatus. Blood vessels and the central proc¬ labyrinth arise embryologically from a single
esses of nerve fibers belonging to the cochlear otic vesicle of ectodermal origin. Although
division of the eighth cranial nerve pass the semicircular ducts are derived from the
through numerous openings into the bony utricle, and the cochlear duct and the endo¬
substance of the modiolus. The cell bodies of lymphatic sac are derived from the saccule,
these bipolar afferent neurons are found in all these parts of the labyrinth are in com¬
the spiral ganglion, which courses spirally munication and all are filled with endolymph.
 THE EAR • 967

the concave side of the duct. Both the saccule

and utricle give off ducts medially, which join
and form the- slender endolymphatic duct (Fig.
35—5), which in turn courses under the utri¬
cle and then medially through the vestibular
aqueduct to end on the posterior surface of
the petrous portion of the temporal bone as
a small dilation, the endolymphatic sac. The sac
is located between layers of the meninges and
is richly surrounded by blood vessels and
connective tissue.
The epithelium lining the membranous
Figure 35-6. Diagram of membranous labyrinth with structures in the vestibule is of simple squa¬
neuroepithelial area in black, a, b, and c respectively mous type, similar to that found in the semi¬
designate the ampullae of the superior, lateral, and pos¬
terior semicircular canals. (Modified from von Ebner.)
circular ducts except in the immediate vicin¬
ity of sensory areas. The sensory areas and
cells just peripheral to them, however, are
Utricle, Saccule, and Ampullae specialized and in many respects highly com¬
plex.
In the vestibule, the oblong utricle lies Crista Ampullaris and Maculae. The epi¬
superior and then posterior to the roughly thelium in the floor of the three ampullae is
spherical saccule and communicates via five raised into a transverse ridge, the crista, which
orifices with the three semicircular ducts and is covered with the sensory epithelium and is
their ampullae (Fig. 35—5). The semicircular bounded at either end by cells of the planum
ducts are eccentrically pLced in the bony semilunatum (Figs. 35—8, 35—9). The latter are
canals and are lined by a simple squamous perpendicular to the long axis of the crista.
epithelium. Each ampulla has a flattened Sensory epithelium on the cristae is histo¬
floor and a hemispherical roof bulging on logically the same as that composing the mac-

Vestibular Secondary tympanic

membrane

Vestibular nerve
and ganglion

Sacculus with %

macula sacculi Posterior ampulla with

crista ampullaris
Figure 35-7. Photomicrograph of a section of a rhesus monkey inner ear, including the sacculus with the macula
sacculi (at arrow) and the posterior ampulla with the crista ampullaris (at arrow). (Courtesy of H. Mizoguchi.)
968 • THE EAR

those of type I hair cells. They can be found

at various levels in the cell, but usually form
a row at a higher level in the epithelium than
those of the type I hair cells and the sup¬
porting cells.
Seen with the electron microscope, the lu¬
minal border of both type I and type II hair
cells is characterized by the presence of a
single cilium and 50 to 110 straight hairs.
These are actually highly specialized micro¬
villi more appropriately described as stereo-
cilia. The hairs are covered by plasma mem¬
brane and are noticeably constricted at their
bases. They have an axial core, which is
composed of longitudinally arranged fine fil¬
aments. This core continues downward from
the narrow base of the hair and is embedded
in the thickened terminal web, which extends
across the cell immediately below its special¬
ized free border. The hairs are arranged
upon the cell surface in regular hexagonal
array, and in successive rows show a progres¬
sive increase of length from less than 1 pm
on one side to 100 pm on the other, the
longest hairs being located at the side of the
cell bearing the cilium (Fig. 35—20). The
Figure 35-8. Phase-contrast photomicrograph of an un¬ cilium originates from a basal body located
stained section in plastic of guinea pig crista ampullaris,
showing the hairs projecting from the hair cells of the in a terminal web-free area of the apical
neuroepithelium. (Courtesy of H. Enstrom.) cytoplasm. The cilium has the typical nine
outer doublet tubules and two central tubules
found in cilia elsewhere, but the central tu¬
ulae of the utricle and saccule, with variation bules end shortly after leaving the basal body.
apparently only in the relative number of A rootlet extends from the basal body into
different cell types. Classically, sensory epi- the cytoplasm on the side opposite the stereo¬
thelia in the vestibular portion of the internal cilia aggregate. Although the cilium is consid¬
ear are described as possessing two cell types, ered to be nonmotile, it is commonly called
hair cells and supporting cells. Recent investi¬ a kinocilium. The existence of a gradient of
gations have shown, however, that among stereocilia length, the eccentric location of
the hair cells two morphological types can be the kinocilium, and the arrangement of the
distinguished. rootlet result in a morphological polarization
Hair Cells. Hair cells of type I are flask¬ of the hair cell, which is reflected in its
shaped cells with a rounded base and con¬ physiological responses to stimuli. Both sen¬
stricted neck region (Fig. 35-9). The round sory cell types have a very dense terminal
nucleus is located basally and surrounded by web or cuticular plate just below the apical
a more or less dense population of mitochon¬ plasmalemma. This structure may extend 0.5
dria. Mitochondria also are found congre¬ pm or more into the cell cytoplasm, but is
gated at the apex of the cell immediately discontinuous in the area of the basal body
beneath the free surface, which bears speci¬ of the cilium.
alized microvilli, or hairs, of considerable In both types of hair cells there are scat¬
length, and a single cilium. From its con¬ tered profiles of granular endoplasmic retic¬
stricted neck inferiorly the hair cell is en¬ ulum, but smooth-surfaced tubules and vesi¬
closed in a chalice-like nerve terminal. cles are more abundant. Vesicles about 20
Hair cells of type II are simple columnar nm in diameter are present in greatest pro¬
cells innervated by numerous small synaptic fusion in the type II hair ceils. The supra¬
endings that are difficult to see with the light nuclear Golgi complex is also more exten¬
microscope (Fig. 35-9). Nuclei of type II hair sively developed in this type. Characteristic
cells are round and regular in outline, like of the type I cell is the occurrence of a great
 THE EAR • 969

TYPE I
HAIR CELL

Cilium Supporting cell

and its terminal web
basal body

0 •

Synaptic Afferent
nerve calyx bynaptic Efferent nerve
ribbon ribbon endings

Figure 35-9. Schematic representation of the principal ultrastructural features of the vestibular type I and type II hair
cells and their supporting cells. (Drawn by Sylvia Colard Keene.)

many microtubules, mostly concentrated in found in varying numbers in the hair cell
the apical cytoplasm just beneath the terminal cytoplasm immediately opposite the nerve
web. calyx. These specializations are characteristic
The nature of the synaptic contact between of chemical synapses. Additionally there are
the two types of hair cells and the termina¬ certain discrete areas where the pre- and
tions of the vestibular nerve fibers is quite postsynaptic membranes are only about 5 nm
different (Fig. 35—9). In the case of type I, apart. These resemble low-resistance contacts
the afferent nerve envelops the hair cell in a found at electrical synapses. At present, how¬
cuplike ending called the calyx. At the base ever, there are no physiological data relevant
of the cell, the intercellular space between to this question. Nearer the rim of the cuplike
the plasmalemma and the axolemma is about ending around the upper part of the hair
30 nm wide. Dense linear structures with an cell, there are in the axoplasm large numbers
associated halo of small vesicles (resembling of vesicles 50 to 200 nm in diameter, some
the so-called synaptic ribbons in the retina) are with dense cores.
970 • THE EAR

The type II hair cell is not enveloped by a cretory granules. Little is known about the
chalice-like ending, but a large number of function of the supporting cells. They may
separate terminal boutons impinge upon its contribute to the nutrition of the hair cells
surface. Synaptic ribbons are often found in or may be involved in some way in the me¬
the hair cell opposite sparsely granulated tabolism of the endolymph. Toward the pe¬
boutons, and the bouton membrane may be riphery of the sensory region, there is a
thickened at such synaptic sites. A second gradual transition to the cells making up the
type of bouton is distinguished by its dense planum semilunatum.
packing of small synaptic vesicles. It is be¬ The planum semilunatum is generally
lieved that these boutons are efferent in na¬ composed of columnar cells with slightly in¬
ture. In addition to contacting type II hair folded lateral membranes. Investigations us¬
cell bases directly, such boutons also contact ing 35S and colloidal iron implicate these cells
nerve chalices of type I hair cells, other in the elaboration and secretion of sulfated
afferent boutons, and passing nerve fibers. mucopolysaccharide components of the cu¬
However, the pre- and postsynaptic mem¬ pula.
brane thickenings traditionally considered in¬ On the sloping sides of the crista ampul-
dicative of synapses are only rarely noted at laris are very complex cells with highly in¬
points of apposition between these vesicu- folded basal membranes and a dense cyto¬
lated boutons and hair cells. They are more plasmic matrix with large vacuoles containing
common where the vesiculated efferents con¬ a flocculent material. These “dark cells” are
tact other neural structures. It is clear that reminiscent of other cells known to be in¬
the synaptic relationships are far more com¬ volved in ion movement, and it has been
plicated than was imagined from light micro¬ speculated that they maintain the high K+
scope studies, and their exact nature remains level in endolymph.
unsettled. Cupulae and Otoliths. Overlying the
The effective stimulus for vestibular hair hairs in maculae are a multitude of minute
cells is movement of the head in one or (3 to 5 jam) crystalline bodies, otoliths. These
another plane. This presumably sets up are a complex of calcium carbonate and a
movement of the endolymph, which acts in protein. In life, they are suspended within
some manner to trigger an impulse in the the jelly-like glycoprotein substance that sur¬
afferent vestibular nerve. The transducers of rounds the sensory areas of the maculae.
the original mechanical stimulus into electri¬ The cupulae are gelatinous bodies located
cal signals are presumably the hair-bearing above the cristae. In life, these are composed
surfaces of the hair cells. Exactly how a re¬ of glycoprotein that is evidently much more
ceptor potential is set up in the hair cell and viscous than the rest of the endolymph. This
how it, in turn, influences the afferent nerve component is often lost or deformed during
are unsolved problems. Some evidence seems specimen preparation, and in the past this
to indicate that bending of the hairs initiates led some otologists to question its reality.
the process, but so little is known of their That a cupula does exist is no longer
physiological properties that this is still spec¬ doubted, but there is very little substantial
ulative. information about its origin or functions.
Supporting Cells. The supporting cells
have their nuclei near the base of the sensory
Endolymphatic Sac
epithelium and extend to its free surface, but
their cell bodies are so contorted that the full The cell types encountered in different
extent of any given cell can never be seen in parts of the vestibular system are basically
a single section (Fig. 35-9). One usually sees similar from one region to the other. In the
a mosaic of sections of many cells cut at endolymphatic sac, however, one finds cells
different levels. With the light microscope that appear to be specialized for an absorp¬
little can be resolved in their cytoplasm, but tive function, and these are structurally dif¬
in electron micrographs they are found to ferent from other cells in the vestibular
possess a cytoskeleton of bundles of micro¬ membranous labyrinth. Unlike the other
tubules running from the basal cytoplasm to membranous sacs, the endolymphatic sac
a dense terminal web, which is far more usually contains cellular debris of one sort or
elaborately developed in these cells than in another. The electrolyte concentration in its
the hair cells. They have a prominent Golgi endolymph also differs from that elsewhere
complex and the cytoplasm is crowded with in the inner ear.
membrane-limited granules resembling se¬ Histologically, there is a transition from
 THE EAR • 971

the squamocuboidal cells of the endolym¬ lymphatic cells of the scala vestibuli, so atten¬
phatic duct to tall columnar cells in the sac. uated that they can scarcely be seen with the
The latter have been variously described as light microscope.
covering protruding papillae or occupying Stria Vascularis. The epithelial covering
crypts. Whichever is the case, there are two of the vestibular membrane becomes contin¬
distinct columnar cell types present. One is a uous, at the outer wall of the cochlea, with
dense cell with a large irregularly shaped the basal layer of cells in the specialized band
nucleus, a relatively unspecialized free sur¬ of striatihed epithelium called the stria vas¬
face, and a cell base with slightly infolded cularis (Figs. 35—10 to 35-12). With the light
membranes. The other is a less dense cell microscope it is possible to distinguish two
characterized by long microvilli on its surface cell types in this epithelium—a layer of light-
and many pinocytotic vesicles and vacuoles. staining basal cells and a darker-staining su¬
Basally the cell membrane is smooth, but perficial layer of marginal cells possessing nu¬
laterally it interdigitates extensively with merous mitochondria. In electron micro¬
other cells. There is good evidence that the graphs some workers have identified a third
endolymphatic sac acts as a site for absorption cell type, the intermediate cells. Although these
of endolymph and that free phagocytic cells, latter are intermediate between marginal and
which appear to be macrophages and neutro¬ basal cells in their location, they are difficult
phils, may cross the epithelium here to engulf to distinguish cytologically from basal cells.
and digest cellular debris and foreign mate¬ The convex free surface of the marginal cells
rial that may gain access to the endolymph. apparently varies with the species and may
be smooth or have microvilli, as it does in the
human. The basal portion of these cells is
The Cochlear Duct
partitioned by deep infoldings of the plas-
The cochlear duct is a highly specialized malemma into a labyrinthine system of nar¬
diverticulum of the saccule. It contains the row compartments occupied by numerous
organ of Corti—the effective organ of hear¬ mitochondria. The intermediate and basal
ing—and a number of other specialized areas cells have relatively few mitochondria and
subserving different functions and having numerous processes that interdigitate with
their own special histological characteristics. each other and with the marginal cells. As¬
This is a very complex area, and in order to cending processes of the basal cells form
facilitate understanding the different regions cuplike structures surrounding and partially
will be described individually in the following isolating each marginal cell from neighboring
order: vestibular meanbrane, stria vascularis, spi¬ areas of the epithelium (Fig. 35—12). Capil¬
ral prominence, organ of Corti, and tectorial laries penetrate into the stria vascularis and
membrane. It can be seen in Figure 35-11 that course longitudinally within the epithelium,
this order of description proceeds in a coun¬ surrounded by processes of the intermediate
terclockwise direction around the circumfer¬ and marginal cells. The stria vascularis is
ence of the cochlear duct. presumed to be involved in the secretion of
Vestibular Membrane. The vestibular the endolymph, and the resemblance of the
membrane is a delicate bilaminar structure elaborate basal compartmentation of the
extending across the tochlea from medial to marginal cells to similar basal specializations
lateral. Its inner surface is lined by cells that of other cells involved in ion transport has
are differentiated in a manner suggesting led to the suggestion that these cells may help
that they may be involved in water and elec¬ maintain the unusual ionic composition of
trolyte transport. The bulging perinuclear the endolymph.
region of the cells is readily apparent in the Spiral Prominence. The stria vascularis
light microscope, but peripherally the cell ends inferiorly, and its basal cell layer is
body is highly attenuated. Toward the scala continuous with the cells overlying the spiral
media the surface of these cells bears many prominence (Fig. 35—13). This prominence ex¬
short, clavate microvilli similar to those found tends the whole length of the cochlear duct
on cells in the choroid plexus. At the basal and rests on a very richly vascularized thick¬
surface, the membranes are highly infolded ening of the underlying periosteum. The
and interdigitate extensively with those of epithelium of the spiral prominence contin¬
the neighboring cells. A distinct basal lamina ues downward and is reflected from the outer
is found along this basal surface. Directly wall of the cochlea onto the basilar mem¬
apposed to these cells, with little or no inter¬ brane, forming at its line of reflection the
vening collagen, is a layer of squamous peri¬ external spiral sulcus. The cells here take on a
 Helicotrema
Organ
of Corii Scala Media

Scala
Vestibuli
Stria
Vascularis
Scala
Tympani

Cochlear
Nerve

Figure 35-10. Photomicrograph of the cochlea of a rhesus monkey. (Courtesy of H. Mizoguchi.)

Reissner's membrane

Scala vestibuli

Spiral limbus

Tectorial membrane

Spiral prominence

Spiral x
Organ of Corti ganglion

Scala tympani

Basilar
membrane

Keene )35_11' Schematic rePresentation of a section through one of the turns of the cochlea. (Drawn by Sylvia Colard

972
 THE EAR • 973

Figure 35-12. Electron micrograph of the stria vascularis of the cat inner ear, illustrating the intraepithelial capillaries,
the ascending process of the basal cells (at arrows), and the elaborately infolded bases of the marginal cells. (After
Hinojosa, R., and E. Rodriguez-Echandia. Am. J. Anat. 118:631, 1966.)

Vestibular
Spiral membrane
prominence

Outer
Outer
hair
cells
Tectorial
membrane
; Internal
spiral
tunnel
Ceils of tunnel
Hensen.

Inner
hair cell

Inner
tunnel

Basilar Pillars ■

membrane Outer
phalangeal Cochlear
cells nerve

Figure 35-13. Photomicrograph of the organ of Corti of a cat. The tectorial membrane has been lifted away from the
inner hair cells in specimen preparation. (Courtesy of H. Engstrom.)
 974 • THE EAR

cuboidal shape, and those continuing onto contain conspicuous tonohbrils. Although the
the pars pectinata of the basilar membrane cells are separated by large intercellular
are known as the cells of Claudius. In the basal spaces, their upper surfaces are in contact
coil of the cochlea, small groups of polyhed¬ with each other and with the hair cells to
ral cells (cells of Boettcher) are interposed be¬ form a continuous free surface for the organ.
tween the basilar membrane and the cells of This surface is called the reticular membrane.
Claudius (Fig. 35—14). These cells have large The supporting cells include inner and outer
spherical nuclei, and their cytoplasm is den¬ pillars, inner and outer phalangeal cells, border
ser than that of the adjoining cells of Clau¬ cells, and cells of Hensen.
dius. The plasma membrane of the lower Within the organ of Corti is the inner
half of these cells gives rise to numerous tunnel, a canal extending the length of the
microvilli, which may either interdigitate ex¬ cochlea and bounded below by extensions of
tensively with those of adjacent cells or pro¬ the pillar cells, which lie on top of the basilar
trude into the intercellular space. Such spe¬ membrane, and above by the bodies of the
cializations suggest that the Boettcher cells inner and outer pillar cells. The bodies of
have a secretory or absorptive function. the pillars are separated by clefts through
which the tunnel communicates with the
Organ of Corti other intercellular cavities in the organ of
Corti, including the outer tunnel or space of
Over the pars pectinata and pars arcuata Nuel.
the cells become columnar and bulge into the Inner Pillars. The inner pillars have a
cochlear duct, forming the epithelial ridge broad base that rests on the basilar mem¬
called the organ of Corti (Figs. 35-10, 35-11, brane and a conical cell body with its apex
35-13). This highly specialized complex of extending upward (Figs. 35-14, 35-16). The
epithelial cells extends throughout the length cytoplasm of the pillar cell contains the nu¬
of the cochlea and is composed of hair cells, cleus at the inner angle of the roughly tri¬
the receptors of stimuli produced by sound, angular tunnel. The most distinctive feature
and various supporting cells. of these cells is the darkly staining tonohbrils
Supporting Cells. The several types of that course from the cell base through the
supporting cells have certain characteristics cylindrical body of the pillar to end in the
in common. They are tall, slender cells ex¬ junctional complexes at the apex, where the
tending from the basilar membrane to the cell expands into a hat flange to contact
free surface of the organ of Corti, and they neighboring pillar cells and the inner hair

Outer tunnel
Cells of Hensen
m e m bran e
Cells of
Claudius
ensens
Cel Is of
Boettcher Internal

rre
<ers Inner hair cell ?/>e/
Border cell
■\\
|\ \Jnner phalangeal

Basilar r
membrane Vas spirale
Outer
phalangeal
cel Is

Cochl ear
nerue

modified from'Held f tranSeCt'0n °f the organ of Corti’ from the uPPer Part of the first coil of human cochlea. (Slightly
 THE EAR • 975

Figure 35-15. Scanning electron micrograph of guinea pig organ of Corti. For further orientation, see Figure 35-14.
(Courtesy of H. Engstrom.)

cells. The contact between inner and outer light microscope are found in electron micro¬
pillar cells is of particularly large area and graphs to be a highly organized array of
forms a structurally sound supporting mech¬ unusually large microtubules, with an outside
anism. What appear to be tonohbrils with the diameter of 27 nm and a wall thickness of
about 6 nm. Interspersed among these are
Head plate of Phalangeal process
6-nm microhlaments (Fig. 35—25). This array
inner pillar of outer pillar of tubules and filaments runs from the base
of the pillar cell to the apex of the cell at the
top of the organ of Corti.
Outer Pillars. The outer pillars are
longer than the inner ones (Figs. 35—14,
35—16). Their base is situated on the basilar
membrane at the junction of the pars pectin-
ata and pars arcuata, adjoining the base of
the inner pillar. The cell body is similar to
that of the inner pillar, but the free surface
of the cell has a somewhat different shape.
The head of the outer pillar abuts the head
of the inner pillar and sends out a phalangeal
process that forms a junction with the outer
hair cells. The outer pillars, in fact, form the
first row of phalanges.
The inner pillars number approximately
5600, the outer ones 3800. On an average,
three inner pillars are connected with two
outer pillars.
Inner Phalangeal Cells. These cells are

Figure 35-16. Diagram of inner and outer pillars of organ arranged in a row on the inner surface of
of Corti. (Modified from Kolmer.) the inner pillars and completely surround
976 • THE EAR

the inner hair cells. In contradistinction to The upper two thirds of the outer hair
the outer phalangeal cells there is no en¬ cells are not surrounded by other cells but
larged extracellular space between the sup¬ are exposed within a fluid-filled space (the
porting cells and the hair cells. Afferent and space of Nuel) that is in communication with
efferent nerve fibers travel through and are the inner tunnel through the clefts between
supported by the inner phalangeal cells. The the pillars. The fluid that bathes the hair cells
relationship between supporting cells and in¬ and occupies the space of Nuel and inner
ner hair cells is completely analogous to that tunnel is apparently separated from the en¬
between the supporting and hair cells of the dolymphatic or perilymphatic spaces, and
vestibular system. thus may be of a composition different from
Outer Phalangeal Cells (of Deiters). that of either perilymph or endolymph.
The outer phalangeal cells act as supporting Border. Cells. The inner phalangeal ceils
elements for the three to four rows of outer continue into a row of slender cells, termed
hair cells (Figs. 35—17 to 35—19). These pha¬ border cells, that delimit the inner boundary
langeal cells are columnar, with their bases of the organ of Corti (Fig. 35-14). There is
resting on the basilar membrane. Apically a gradual transition in height from these to
they surround the inferior third of the outer the squamous cells lining the inner spiral
hair cell and also enclose the afferent and sulcus.
efferent nerve bundles traveling to the hair Cells of Hensen. Adjacent to the last row
cell base. This portion of the cell does not of outer phalangeal cells are the tall cells of
reach the free surface of the organ of Corti, Hensen that constitute the outer border of
but on the side of the cell away from the the organ of Corti. They are arranged in
outer pillar cells, it gives off a slender finger¬ several rows decreasing rapidly in height and
like process internally reinforced by a bundle laterally abutting the cells of Claudius.
of microtubules. This phalanx expands at the Cochlear Hair Cells. In the cochlea, as in
surface of the organ of Corti to form a flat the vestibule, two types of hair cells are pres¬
apical plate joined at its edges to the hair cell ent (Figs. 35—11, 35—13, 35—19). The inner
that it is supporting and to the hair cell in hair cells are arranged in a single row along
the row next to it. The platelike expansion the whole length of the cochlea. The outer
at the surface also contains abundant sup¬ hair cells form three rows and are lodged
porting microtubules. between the outer pillars and the outer pha-

Figure 35-17. Scanning electron micrograph of outer hair cells and slender processes of supporting cells of Deiters.
For orientation, see Figure 35-18. (Courtesy of H. Engstrom.)
 THE EAR • 977

Phalangeal process of the cochlear nerve. Many of these run

beside the hair cell up to the level of its
nucleus, though not all the area of apposition
is synaptic. Pre- and postsynaptic membrane
thickenings and synaptic ribbons are consid¬
ered to demarcate sites of synaptic transmis¬
sion. In addition, small neuronal profiles con¬
taining many synaptic vesicles, some of which
are dense-cored, make contact with inner
hair cells, though this is rather rare. More
often, contacts with pre- and postsynaptic
membrane thickenings are seen between
these heavily vesiculated fibers and the affer¬
ent fibers just before the latter make contact
with the inner hair cell. Many such vesicu¬
lated fibers are found beneath the inner hair
cell, and they are considered efferent in na¬
ture.
Outer hair cells have a different structure
from that of inner hair cells and this has led
to speculation that the two cell types have
different functions. Outer hair cells, as pre-

Figure 35-18. Diagram of supporting cells of Deiters and

the associated outer hair cell. (Modified from Kolmer.)

langeal cells. In the human, a fourth and

sometimes a fifth row of outer hair cells may
appear toward the apex, though these super¬
numerary rows may not be as regular as the
first three.
Inner hair cells resemble type I cells in the
vestibular labyrinth in many respects. They
are relatively short, goblet-shaped cells with
a slightly constricted neck region. The sur¬
face of the cell bears hairs similar in structure
to those on vestibular hair cells, but in the
adult there is no associated cilium. However,
a basal body and an associated typical cen-
triole persist as the only remnants of the
ciliary apparatus. The hairs are arranged on
the cell surface in the form of a letter W or
U (Fig. 35-23). The rootlets of the hairs
extend down into, and at times through, the
dense terminal web or cuticular plate. The
cell body contains scattered ribosomes and
20-nm vesicles interspersed among larger
vesicular profiles, presumably representing
the smooth endoplasmic reticulum. Mito¬
chondria are aggregated under the terminal
web and at the cell base, but are scattered in
smaller numbers throughout the cytoplasm.
The synaptic area of the inner hair cell ex¬
tends from the base of the cell to the level of Figure 35-19. Schematic representation of the relation¬
the nucleus. The vast majority of the endings ship of the outer hair cells to the outer phalangeal cells,
contacting the inner hair cell are sparsely as revealed in electron micrographs. (Drawn by Sylvia
vesiculated and are considered to be endings Colard Keene.)
978 • THE EAR

viously stated, are supported on the apices of

outer phalangeal cells and receive innerva¬
tion from the cochlear nerve at their bases
(Figs. 35—14, 35—27). The hairs on the apex
of these cells form a distinctive W similar to
that of the inner hair cells, but here there
are more rows of hairs, and the length of the
hair varies from long at the perphery to short
centrally (Figs. 35-20, 35-24). Again, no cil-
ium is present, although a basal body can be
found at the base of the W. Immediately
deep to the terminal web are dense, lipid-like
inclusions interspersed with elongated,
highly convoluted elements of the granular
endoplasmic reticulum. Mitochondria are
generally aggregated in the basal cytoplasm
and line up along the sides of the cell in
relation to one or more rows of smooth-
surfaced vesicles that are aligned parallel to
the plasmalemma. A single row of such
smooth-surfaced vesicles is also found along
the nonsynaptic plasmalemma of the inner
hair cell. At present the function of these
vesicular aggregates is unknown.
The basal part of the outer hair cell re¬
ceives synapses from both efferent and affer¬
Figure 35-21. Transverse sections through the hairs on
ent nerve fibers. Flere, however, the afferent
a guinea pig outer hair cell, showing the large number of
fibers from the cochlear nerve frequently filaments in their interior. (Courtesy of H. Engstrbm.)

give rise to the smaller endings. Such syn¬

apses exhibit pre- and postsynaptic mem¬
brane thickenings and, in some species, syn¬
aptic ribbons, but they are not heavily

Cell of Claudius

Cell of Hensen

Outer
phalanges

Outer hair
cell [row 2)
Phalanx of
outer pillar
Outer hair
cell (row 1)

Head of inner
pillar

Inner hair cell

Border cell Inner phalanx

Figure 35-20. Electron micrograph showing the hairs on Figure 35—22. Diagram of the organ of Corti viewed from
a hair cell. Notice their narrow base and the continuation above, showing the relationship of the phalanges of the
of their fibrous core into the terminal web. (Courtesy of supporting cells to the hair cells. (Modified from Retzius
D. Hamilton.) Kolmer, Schaffer.)
 THE EAR • 979

Figure 35-23. Scanning micrograph of guinea pig organ of Corti, middle turn, seen from above. For orientation, see
Figure 35-22. (Courtesy of H. Engstrom.)

vesiculated. The efferent endings on the lamina bulges into the scala media as the
outer hair cells are larger than those of the spiral limbus (Figs. 35—11, 35—13, 35—14). Its
afferent fibers and contain many densely edge overhangs the internal spiral sulcus.
packed vesicles. In the hair cell cytoplasm, The two margins of the sulcus are the vestib¬
parallel to the plasmalemma and extending ular lip and tympanic lip. The collagenous
the entire length of the efferent synapse, is fibers of the limbus continue laterally, via the
found a single continuous flattened cisterna, tympanic lip, into the pars arcuata of the
the “subsynaptic cisterna.” basilar membrane. Within the body of the
Spiral Limbus. In the inner angle of the limbus the fibers are arranged vertically to
scala media, the periosteal connective tissue produce the distinctive auditoiy teeth (of
of the upper surface of the osseous spiral Huschke). Between these collagenous fibers
980 • THE EAR

remembered that similar, though less speci¬

alized, perilymphatic spaces surround the
structures in the vestibule.
Histologically the perilymphatic tissue is
described as a reticulum, and close exami¬
nation with the electron microscope shows
that this reticulum is composed primarily of
highly attenuated processes of many stellate
cells. Except close to the periosteum of the
bony labyrinth and to the membranous lab¬
yrinth, there are few extracellular fibers
associated with these reticular cells. Immedi¬
ately surrounding the membranous laby¬
rinth, however, extracellular fibers are
Figure 35-24, Electron micrograph of the W configuration formed and, in some species, make up a
of the hairs or stereocilia on the outer sensory cells of the relatively dense, stable sheath 1 or 2 jxm
human organ of Corti. x 10,500. (Courtesy of R. Kimura.)
thick, composed of multitudes of short fibers.
In the vicinity of the cells of the perilym¬
are stellate fibroblasts. Uniformly spaced phatic system, both in the cochlea and in the
along the upper margin of the limbus, be¬ vestibule, are fiber bundles that have a char¬
tween the auditory teeth, are the so-called acteristic form different from that of any
interdental cells, which secrete the tectorial other known extracellular fiber. The bundles
membrane. The bases of interdental cells are are composed of a variable number of dense
firmly embedded in the connective tissue of 10-nm filaments, which can be shown to be
the limbus, but their apices spread out over composed of four 5-nm subunits that appear
the upper surface of the limbus, interdigitat- to be helically wound around one another.
ing and joined by junctional complexes. The dense fibers that form the bundles are
These form a continuous sheet over the up¬ embedded in an amorphous matrix.
per surface of the limbus and complete the Numerous blood capillaries course
cellular investment of the cochlear duct. throughout the perilymphatic tissue destined
Tectorial Membrane. The tectorial mem¬ to supply the metabolic needs of the labyrin¬
brane is secreted from the luminal surfaces thine epithelium.
of the interdental cells and overlies these cells
as a cuticle. It extends laterally beyond the
Sca/a Vestibuli and Scala Tympani
vestibular lip of the limbus to overlie the
hairs on the hair cells of the organ of Corti The cells that line these two scalae usually
(Figs. 35—11, 35—13). Recent evidence indi¬ are extremely attenuated squamous cells with
cates that the tips of the hairs are embedded very little obvious cellular differentiation. At
within or are firmly bound to the membrane. times, however, especially in the vicinity of
If this is so, micrographs showing a space the basilar membrane, the cells do become
between the hairs and the tectorial mem¬ somewhat cuboidal, although they possess
brane must be artifactitious. The tectorial few structural features of note.
membrane is composed primarily of a pro¬ Basilar Membrane. The most elaborate
tein having a number of similarities to epi¬ specialization of perilymphatic tissue is the
dermal keratin. In fixed preparations nu¬ basilar membrane, which provides a support¬
merous fibrils are observed within it, forming ing base for the cells of the organ of Corti
patterns suggesting a highly ordered struc¬ and which by its movement presumably
ture. transmits vibrations to the hair cells. The
basilar membrane is a highly organized layer
of fibers. There is some indication that the
THE PERILYMPHATIC LABYRINTH perilymphatic cells may actively secrete the
basilar membrane, but this has not been
The perilymphatic system surrounds the clearly established. It is divided into two dis¬
whole of the membranous labyrinth and pro¬ tinct zones: one, running from the osseous
vides support for its epithelium lining. The spiral lamina approximately one third of the
distinct scalae vestibuli and tympani in the way to the outer cochlear wall, is termed the
cochlea have been mentioned, but it must be pars arcuata (tecta)\ the other, approximately
 THE EAR • 981

two thirds of the width of the basilar mem¬ It was recognized early in this century that
brane, is termed the pars pectinata and con¬ endolymph was a product of secretion, al¬
tains, as can be seen even at the light micro¬ though the actual site or sites of its elabora¬
scopic level, distinct parallel striations termed tion were not known. It was supposed that
the auditory strings. In fact, both portions of the stria vascularis, the spiral prominence,
the membrane are composed of transversely and the extrasensory cells around the mac¬
oriented filaments (8 to 10 nm thick) embed¬ ulae and cristae were primarily responsible.
ded in an amorphous matrix. In the pars Recent evidence would indicate that these
pectinata the filaments are aggregated into areas do indeed take part in elaboration of
bundles that run in two strata: one immedi¬ endolymph, but the electron microscope has
ately beneath the organ of Corti, composed made it clear that many of the cells lining the
of small bundles, and another situated more membranous labyrinth have cytological char¬
deeply in the lamina, composed of larger acteristics compatible with synthetic and se¬
bundles. At the outer wall of the cochlea, cretory activity and might therefore partici¬
these two layers again merge, to pass into the pate in endolymph metabolism. Specifically,
connective tissue of the spiral ligament. autoradiographic studies have implicated the
Blood vessels penetrate into the pars arcuata planum semilunatum in elaboration of sul-
but not into the pars pectinata. fated mucopolysaccharides, and measure¬
The term spiral ligament is an unfortunate ment with microelectrodes has shown that
designation for the lateral insertion of the the high DC potential of the scala media is
basilar membrane, because this component produced in the vicinity of the stria vascu¬
does not have the histological structure of laris, which would indicate that some sort of
ligaments found elsewhere in the body (Fig. ion secretion is taking place there.
35-11). It is merely a local differentiation of The site of absorption of endolymph has
periosteal connective tissue containing nu¬ been thought to be the endolymphatic sac.
merous fibroblasts and blood vessels. A better However, electron micrographs of cells of
term would be spiral crest. the membranous labyrinth show many in¬
stances of micropinocytotic activity, which
would suggest that absorption may be going
ENDOLYMPH AND PERILYMPH on in other areas of the labyrinth.
Although it is well established that endo¬
The spaces delimited by the membranous lymph is a secretion, the genesis of perilymph
labyrinth are filled with the viscous fluid is still being debated. Some feel that it is an
called endolymph, and the labyrinth is sur¬ ultrafiltrate of plasma, others that it is de¬
rounded by the perilymph, which occupies rived from cerebrospinal fluid. There is no
the perilymphatic spaces. The two fluids are doubt that the perilymphatic spaces are func¬
amazingly different in their chemical com¬ tionally connected to the subarachnoid space,
position. The most striking difference is in but the exact functional significance of this
their electrolyte composition. Whereas peri¬ relationship is not yet clear.
lymph to some degree resembles extracellu¬
lar fluid in general, endolymph has the char¬
acteristics of intracellular fluid in having high NERVES OF THE LABYRINTH
K+ and low Na+ concentrations (Table 35-1).
The eighth cranial nerve supplies the sen¬
sory areas of the labyrinth. It consists of two
parts of quite different functional nature and
Table 35-1. ELECTROLYTE COMPOSITION OF central connections—the vestibular and the
BODY FLUIDS (mEq/l)* cochlear nerves (Figs. 35—5, 35—28). Each is
composed of primary afferent fibers from
Endo¬
Perilymph lymph the sense organs and efferent feedback fibers
Plasma CSF
from the central nervous system. The cell
Protein 6000-8000 10-38 75-100 10
bodies of the afferent fibers are bipolar cells
K 20 12-17 15 140
Na 140 150 148 26 and form two peripheral ganglia, the spiral
Cl 600 750 120 140 or cochlear ganglion in the modiolus and the
Sugar 70-120 40-80 vestibular or Scarpa's ganglion in the internal
Mg 1.0-3.0 2.0 2.0 0.9 auditory meatus of the temporal bone.
Ca 7.0 3.0 3.0 3.0
The vestibular nerve divides into a supe¬
*From F. C. Ormerod. rior and an inferior branch. The superior
982 • THE EAR

Figure 35-25. Electron micrograph of an inner pillar cel! in transverse section, showing the highly ordered bundie of
thick walled microtubules and microfilaments. (Courtesy of H. Engstrdm.)

branch supplies the horizontal crista ampul- reaching the outer hair cells of the organ of
laris, the superior crista ampullaris, the mac¬ Corti, they turn sharply and follow a spiral
ula utriculi, and a small part of the macula course. These are the spiral fibers (Fig. 35—26).
sacculi. The inferior branch supplies the pos¬ The functional implications of these two
terior crista ampullaris and the major portion patterns of distributions are not clear. Al¬
of the macula sacculi, and it sends a small though the relationship between the periph¬
anastomosing branch to the cochlear nerve. eral receptors and acoustic neurons is not as
The bipolar cell bodies, both in the vestib¬ individualized as the monosynaptic relation¬
ular ganglion and in the cochlear ganglion, ship of the foveal cones, it is sufficiently
are invested by a thin layer of myelin, and restricted to permit the reception of localized
this continues onto the axons. The axons of stimuli impinging upon small segments of
the cochlear nerve lose their myelin as they the cochlea.
run through the openings of the osseous The vestibular nerve terminates centrally
spiral lamina beneath the inner hair cells. In in the reflex centers of the medulla oblongata
the vestibular nerve, myelin persists until the and cerebellum. Its cortical connections are
nerve enters the sensory area. unknown, although it mediates reflex move¬
The cochlear nerve contains two morpho¬ ments of the eyes through its thalamic con¬
logical kinds of afferent nerve fibers. The nections. The cochlear nerve synapses in the
more numerous ones radiate from the spiral cochlear nucleus, whence fibers ascend in the
ganglion in parallel bundles to the nearest lateral lemniscus to the medial geniculate
segments of the organ of Corti. Because of body of the thalamus and thence to the
their course, they are called the radial acoustic temporal lobe gyri of the cortex.
fibers. The second category of fibers, usually Both the vestibular and cochlear divisions
thicker and fewer than the first, are also of the eighth cranial nerve contain apprecia¬
arranged radially at the outset, but after ble numbers of efferent fibers that originate
 THE EAR • 983

Hair cells

Figure 35-27. Longitudinal section of the organ of Corti,

showing the efferent nerve fibers ending about bases of
the outer hair cells. (Courtesy of H. Engstrom.)

vestibulocochlear artery and the cochlear artery

proper.
Figure 35-26 Optical section of guinea pig organ of Corti
viewed from above, showing nerve fibers traversing the The vestibular artery supplies the upper
tunnel to reach the region of outer hair cells. Modified and lateral parts of the utricle and saccule
Maillet nerve stain using zinc iodide and osmium tetroxide. and parts of the superior and lateral semicir¬
(From Engstrom, H., H. W. Ades, and A. Andersson: cular ducts. It forms dense networks of cap¬
Structural Pattern of the Organ of Corti. Stockholm,
Almqvist and Wiksell, 1966.)
illaries in the region of the maculae; in the
thin perilymphatic tissue of these structures,
the capillary networks are relatively loose.
bilaterally from the vicinity of the superior The vestibulocochlear artery supplies, with
olive. Initially these fibers travel in the vestib¬ its vestibular branch, the lower and medial
ular nerve, but within the internal auditory parts of the utricle and saccule, the crus
meatus some efferent fibers reach the coch¬ commune, and the posterior semicircular
lear nerve by way of the anastomosis between duct. Its cochlear branch supplies the lowest
the vestibular and cochlear nerves. The pe¬ part of the first cochlear coil.
ripheral terminations of the efferent com¬ The cochlear artery proper penetrates the
ponent are presumably at the hair cells, but cavities of the modiolus, where its tortuous
incontrovertible evidence for the position branches run spirally to the apex. This is the
and mode of ending of these fibers is still so-called “spiral modiolar artery.” From it,
lacking. Stimulation of the efferent bundle branches go to the spiral ganglion and,
results in suppression of auditory nerve ac¬ through the periosteum of the scala vestibuli
tivity, and anatomical evidence derived from and the osseous spiral lamina, to the inner
sectioning the bundle indicates that the parts of the basilar membrane. Here the
“granulated” endings are efferent, for they capillaries are arranged in arcades in the
apparently degenerate after sectioning (Figs. tympanic covering layer under the tunnel
35-9, 35-19). and the limbus. The vascular stria and the
spiral crest receive their blood through
branches of the spiral modiolar artery, which
BLOOD VESSELS OF THE LABYRINTH run in the roof of the scala vestibuli. They
do not form connections with the vessels of
The labyrinthine artery is a branch of the the basilar membrane. The lower wall of the
inferior cerebellar artery. It enters the inter¬ scala tympani receives its own small arteries
nal auditory meatus and divides into two from the same source.
branches, the vestibular artery and the common The course of the veins of the labyrinth is
cochlear artery. I he latter divides into the quite different from that of the arteries.
984 • THE EAR

Figure 35-28. Diagram of distribution

of nerves in the membranous labyrinth
of the rabbit. (After deBuriet, from Kol-
mer.)

There are three main venous drainage chan¬ with the subarachnoid space. A certain
nels. In the cochlea, veins originate in the amount of drainage may be effected through
region of the spiral prominence and run perivascular and perineural connective tissue
downward and inward through the perios¬ sheaths.
teum of the scala tympani to the spiral vein,
which is found under the spiral ganglion.
Upper and lower spiral veins, belonging to
the corresponding coils of the cochlea, re¬
FUNCTIONAL
ceive branches from the osseous spiral lamina CONSIDERATIONS
and the spiral ganglion. Above the spiral vein
is the small vein of the spiral lamina, which Functions of the various structural com¬
receives a part of the blood from the spiral ponents of the ear have already been men¬
lamina and spiral ganglion and is connected tioned briefly in the descriptions under spe¬
by anastomoses with the spiral vein. These cific headings. It is impossible, of course, to
cochlear veins form a plexus in the modiolus, consider in detail the functioning of the ear
which empties the blood partly into the in¬ in a textbook of histology, but there are
ternal auditory vein and partly into the vein certain physiological considerations that sug¬
of the cochlear aqueduct, which drains into gest new problems and approaches to re¬
the jugular vein. The veins of the vestibule search on this organ.
empty into the veins of the vestibular and The external and middle ears lend them¬
cochlear aqueducts. selves quite well to physiological research and
This arrangement of the vessels in the have been intensively studied for some time.
internal ear seems to ensure the be^t possible The vibrations of the tympanic membrane
protection of the sound receptors from the are transmitted through the chain of auditory
arterial pulse wave. The arteries are arranged ossicles to the fenestra ovalis and thence to
for the most part in the wall of the scala the perilymph filling the scala tympani. The
vestibuli, while the wall of the scala tympani organ of Corti is the receptor for sound
contains the veins. The course of the spiral stimuli, but this function depends to a large
arteries in the modiolus probably also con¬ degree on the properties of the basilar mem¬
tributes to the damping of pulsations. In brane. This membrane may be compared to
certain mammals the coiling of these arteries an unstressed gelatinous plate with varying
is so prominent that the convoluted regions resistance to displacement related to its uni¬
suggest glomeruli. formly varying width. The deformation of
True lymphatics are absent from the laby¬ this membrane produced by movement of
rinth. Instead the fluid is drained into the the stapes resembles a traveling wave. Re¬
perilymphatic spaces, which are connected gions of observed maximal displacement
 THE EAR • 985

change with frequency but are rather broad. unexplained, however, for the hairs extend
As the stimulus frequency rises, the length into the endolymph with its highly unusual
of the basilar membrane responding becomes ionic composition. Thus, with equal K+ and
shorter, and progressively more of the distal Na+ inside and outside the cell, the Na+ =
area becomes inactive. The pitch-discriminat¬ K+ movements that are known to be involved
ing ability of the ear is only partly due to this in excitation in other excitable cells would
physical separation of the responding areas not take place. Nevertheless, it has been
along the basilar membrane. shown that a nerve cell membrane put into
Nerve impulses elicited by stimulation of the same ionic conditions as the hairs re¬
the maculae and the cristae play an important sponds to pressure changes by transient po¬
role in the regulation and coordination of tential changes across the membrane, and
the movements of equilibrium and locomo¬ although the mechanism for this is equally
tion. The stimuli to the vestibular end organs unclear, it is possible that this experimental
are angular acceleration for the semicircular system is analogous to the hairs. In the near
ducts and linear acceleration for the maculae. future it may help to explain how hair cells
These impulses exert their influences upon function.
coordinated muscular contraction, upon
muscular tonus, and upon eye movement
through the brain stem and cerebellum.
REFERENCES
The ear is essentially a biological trans¬
ducer. The transduction in the external and GENERAL
middle ears is relatively easily monitored.
Bast, T. H., and B. J. Anson: The Temporal Bone and
The extreme anatomical complexity of the the Ear. Springfield, IL, Charles C Thomas, 1949.
internal ear, however, has hindered attempts Bekesy, G. von: Experiments in Hearing. New York,
at understanding the transducer phenome¬ McGraw-Hill Book Co., 1960.
non by which mechanical energy (the stimu¬ Dallas, P.: The Auditory Periphery. New York, Aca¬
demic Press, 1973.
lus) is transferred into electrical energy (the
Davis, H.: Mechanisms of the inner ear. Ann. Otol.
nerve impulse). It is generally believed that 77:644, 1968.
the transduction process takes place in the Engstrom, H., H. Ades, and A. Anderson: Structural
apex of the hair cells. Indications are that Pattern of the Organ of Corti. Stockholm. Almqvist
the mechanism in vestibular and cochlear & Wiksell, 1966.
Iurato, S., ed.: Submicroscopic Structure of the Inner
hair cells probably does not differ signifi¬ Ear. New York, Pergamon Press, 1967.
cantly, even though there are considerable
EXTERNAL EAR
anatomical differences in the cells.
It has been possible to show very clearly in Perry, E. T.: The Human Ear Canal. Springfield, IL,
Charles C Thomas, 1957.
the ampulla of Lorenzini in fish, where the
Sophian, L. H., and B. H. Senturia: Anatomy and
sensory cells are in many respects anatomi¬ histology of the external ear in relation to the
cally similar to vestibular hair cells, that bend¬ histogenesis of external otitis. Laryngoscope 64:772,
ing of the hairs toward the kinocilium results 1954.
in depolarization of the cell. Bending in the EUSTACHIAN TUBE
opposite direction hyperpolarizes the cell, Graves, G. O., and L. F. Edwards: The eustachian tube.
while bending normal to these directions has A review of its descriptive, microscopic, topographic
no appreciable ionic effect. Since mammalian and clinical anatomy. Arch. Otolaryngol. 39:359,
vestibular hair cells are morphologically po¬ 1944.
Ladman, A. J., and A. J. Mitchell: The topographical
larized, the same functional responses to
relations and histological characteristics of the tub-
stimuli that bend the hair cells toward or ulo-acinar glands of the eustachian tube in mice.
away from the kinocilium would probably Anat. Rec. 727:167, 1955.
occur. The polarization of the hair cells of COCHLEA
the maculae varies regularly from one part
Bredberg, G.: Cellular pattern and nerve supply of the
of these organs to the other, covering within human organ of Corti. Acta Otolaryngol. (Suppl.)
each organ a full 360 degrees. Thus a move¬ 256:1, 1968.
ment of the head in any direction will be a Engstrom, H., H. Ades, and J. Hawkins: Structure and
sufficient stimulus to excite some of the hair function of the sensory hairs of the inner ear. J.
Acoust. Soc. Am. 34:1356, 1962.
cells and inhibit others. Furthermore, the
Fernandez, E.: The innervation of the cochlea (guinea
stepwise lengthening of the hairs may pro¬ pig). Laryngoscope 57:1152, 1951.
vide a built-in biological amplifier. How the Flock, A.: Transduction in hair cells. In Loewenstein,
transduction process takes place remains W. R., ed.: Handbook of Sensory Physiology. Vol.
986 • THE EAR

1. Principles of Receptor Physiology. Berlin, Sprin- Flock, A.: The ultrastructure of the macula utriculi with
ger-Verlag, OHG, 1971. special reference to directional interplay of sensory
Kimura, R. S.: Hairs of the cochlear sensory cells and response as revealed by morphological polarization.
their attachment to the tectorial membrane. Acta J. Cell Biol. 22:413, 1964.
Otolaryngol. 67:55, 1966. Guild, S. R.: Observations upon the structure and nor¬
Kimura, R. S., H. F. Schuknecht, and I. Sundo: Fine mal contents of the ductus and saccus endolympha-
morphology of the sensory cells of the organ of ticus in the guinea pig. Am. J. Anat. 59:1, 1927.
Corti in man. Acta Otolaryngol. (Stockh.) 56:390, Guild, S. R.: Circulation of the endolymph. Am. J. Anat.
1965. 59:57, 1927.
Rodriguez-Echandia, E. L., and M. H. Burgos: The fine Kimura, R. S., P. G. Lundquist, and J. Wersall: Secretory
structure of the stria vascularis of the guinea pig epithelial linings in the ampullae of the guinea pig
inner ear. Zeitschr. f. Zellforsch. 67:600, 1965. labyrinth. Acta Otolaryngol. 57:517, 1964.
Spoendlin, H.: The organization of the cochlear recep¬ Lundquist, P. G.: The endolymphatic duct and sac in
tor. In Advances in Oto-Rfiino-Laryngology. Vol. the guinea pig. Acta Otolaryngol. Suppl. 201:1,
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Ormerod, F. C.: The physiology of the endolymph. J.
VESTIBULAR ORGAN
Laryngol. Otol. 74:659, 1960.
Citron, L., D. Exley, and C. S. Hallpike: Formation, Wersall, J., and A. Flock: Physiological aspects on the
circulation, and chemical properties of the labyrin¬ structure of reticular end organs. Acta Otolaryngol.
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 A

Index

Note: Page numbers in italics refer to illustrations, and

page numbers followed by (t) refer to tables.

A bands, of skeletal muscle, 275, 283, 284, 285 Adipose tissue (Continued)
A cell, of intestinal mucosa, 651 multilocular, mitochondria of, 186
Absorptive cell(s), microvilli of, 644, 646, 646, vascular network of, 180
647 norepinephrine release and, 185
of intestinal epithelium, 644—648, 645—648 unilocular, 174—179, 175—178
N-Acetyl-beta-glucosaminidase A, defect in, distribution of, 178
19(t) filaments of, 175, 178
Acetylcholine, receptors for, 293, 293 pinocytosis of, 178
storage of, 290 white, 175
Acidophils, of pars distalis, 483 Adluminal cell, of eccrine sweat glands, 570
Acinar cell(s), 84, 84, 91 Adrenal glands, 516—534, 517-523, 525-527,
pancreatic, 716—719, 717—719 532
zymogen granules of, 717, 719, 720 adrenocortical cell of, 523
Acinus, of liver, 684 blood supply of, 527, 527-528
Acrosome, of sperm, 805, 805—806, 807 cells of, cortical, 522
ACTH. See Adrenocorticotrophic hormone. medullary, 525—526, 526
ACTH-RH. See Adrenocorticotrophic hormone re¬ chromaffin tissue of, 516
leasing hormone. cortex of, 516, 516—524, 517-523
Actin, 27-28 cytoplasm of, 102
filamentous, of skeletal muscle, 286, 287, fetal, 521
288 glucocorticoids of, 528
G, 28 histophysiology of, 528-530
of microvilli, 72, 73 hormones of, 524
of platelets, 116 mineralocorticoids of, 528
of skeletal muscle, 280, 282, 283 renewal of, 531, 532
of smooth muscle, 268 reproductive system and, 529
Actinin, of skeletal muscle, 284 zona fasciculata of, 518, 518, 521, 526
Action potential, 323 zona glomerulosa of, 516, 518, 519
at myoneural junction, 293 zona intermedia of, 518
Addison’s disease, 529 zona reticularis of, 518, 524
Adenohypophysis, 479, 479 histogenesis of, 531—533
endocrine cell of, 109, 110 interrenal tissue of, 516
Adenomyosis, 880 lymphatics of, 528
Adenyl cyclase, hormone action and, 107, 107 medulla of, 516, 524—527, 525, 526
Adhesion, platelet, 116 cells of, 525—526, 526
Adipose cell, 152-154, 153 chromaffin reaction of, 525
Adipose tissue, 174-187 chromogranin of, 526
adrenocortical hormone and, 183 histophysiology of, 530—531
autonomic nervous system and, 184-185 hormones of, 526
brown, 175 hyperfunction of, 530
formation of, primary, 181, 182 nervous control of, 531
secondary, 181 nerves of, 528
histogenesis of, 181 — 182, 182 phylogeny of, 533
histophysiology of, 182—187, 184, 185, 186 structure of, 104
hormonal influences on, 183—184, 184, 185 Adrenocorticotrophic hormone (ACTH), 106,
insulin and, 183-184, 184, 185 106, 486
multilocular, 178, 179—180, 179—181 Adrenocorticotrophic hormone releasing hor¬
distribution of, 180—181 mone (ACTH-RH), 106, 106
heat generation and, 185—186 Adrenocorticotropin, 491
987
988 • INDEX

Adrenogenital syndrome, 529 Arrector pili muscle, 559, 560, 561, 567
Adventitial cell, of seminiferous tubules, 798 Arteriole, 375, 375-377, 376, 382
Agammaglobulinemia, plasma cell absence Arteriovenous anastomosis, 377
and, 160 Artery (arteries), 367—381
Albinism, 558 aging and, 380-381
Albumin, 132-133 arteriovenous anastomoses of, 377
Aldosterone, 528—529, 779 capillary bud origin of, 400
renal sodium transport and, 787 conducting. See Artery, elastic.
Aldosterone stimulating factor, 528 coronary, 399
Alpha cell, of pars distalis, 483 cross section of, 391
Alveolar cell(s), 741 distributing. See Artery, muscular.
Type I, 741, 744, 745 elastic, 367-369, 368-370
Type II, 741, 744, 745, 745 histogenesis of, 399-400
Alveolar macrophage, 748-750, 749 muscular, 370-374, 371-374
Alveoli, 741-749, 741-750 elastica externa of, 371, 373
cells ol, 741, 744, 745 tunica adventitia of, 373, 374
septal, 745 sensory organs of, 377-378
Type I, 741, 744, 745 specialized, 374-375
Type II, 741, 744, 745, 745 splenic, 469-471, 470, 471
interstitium of, 745—746, 746 structure of, physiological implications of,
Kohn’s pores of, 746, 748 378-380, 379, 380
septa of, 743 transitional, 374-375
Amacrine cell, 947, 948, 950, 954 vasomotor activity of, 379, 379, 380
Ameloblast, 609 Arthus reaction, 407
ultrastructure of, 616 Arylsulfatase, 163
Amenorrhea, lactational, 909 Aspirin, gastric mucosa erosion and, 638, 638,
Amniotic cavity, 887 639
Ampulla of Vater, 711 Astrocyte(s), fibrous, 345, 346
Anaphase, of meiosis, 47 plasmatohbrous, 345-346
of mitosis, 44, 45, 46 protoplasmic, 345, 346
Anaphylaxis, 407 Atherosclerosis, 381
eosinophil chemotactic factor of, 163 Atrium (atria), of alveolar ducts, 740, 741
slow-reacting substance of, 163 Auditory meatus, external, 961, 962
Anastomosis, arteriovenous, 377 Auditory tube, 963-964, 965
Anatomy, microscopic, 1 Auerbach’s plexus, 672 , 673 , 674 , 675
Anemia, pernicious, 640 Auricle, 961, 962
sickle cell, 113 Autonomic nervous system, 342-345, 344
Angiotensin I, 777 ganglia of, 342
Angiotensin II, 777 nerve cells of, 344-345
Angiotensinogen, 777 parasympathetic division of, 343
Anisocytosis, 114 sympathetic division of, 343
Ankyrin, of erythrocyte membrane, 114, 115 Autophagy, 17, 18
Annulate lamellae, 20, 20 Axolemma, 330
Annuli hbrosi, 397 Axon ending, 337
Antibody, 128, 133, 407 Axon hillocks, 317, 317, 323
lymph node production of, 460-461 Axonal reaction, 364
production of, B lymphocytes and, 159-160 Axonal transport, 324-325
plasma cells and, 159-160 anterograde, 324
reaginic, 127 retrograde, 324
Antidiuretic hormone. See Vasopressin. slow component a of, 324
Antigen, 128, 406 slow component b of, 324
B lymphocyte response to, 419-421, 420 Axoneme
421 of cilia, 76, 77
lymph node response to, 460-461 Axon(s), 311, 312, 320, 323
presentation of, 422 arborization of, 323
T lymphocyte response to, 417-419 calyces of, 348
Anus, 663-664 collaterals of, 312
Aortic body, 378
development of, 356
Aponeurosis, 167 end bulbs of, 348
Appendices epiploicae, 663 initial segment of, 323
Appendix, 660, 661 myelin sheath of, 323
APUD (amine precursor uptake and decarbox¬ myelinated, 323, 329
ylation) cell, 108
regeneration of, 363-364
Aqueous humor, 928-929
retrograde chromatolysis of, 364
Arachnoid, 357, 357, 358, 358 unmyelinated, 323, 329
Arachnoid villus, 358, 361-362
Areola, 901, 902
Montgomery’s glands of, 901
Areolar tissue. See Connective tissue, loose. B lymphocyte(s). See Lymphocyte(s), B.
Argentaffin cell, 633-635, 634, 635
Bacterium (bacteria), neutrophilic phagocytosis
Arginine vasotocin, of pineal gland, 540 of, 123, 123-125, 124
 INDEX • 989

Band forms, 120, 250 Bone(s) (Continued)

Band of Biingner, 363 calcium in, historadiogranr of, 205
Baroreceptor, carotid sinus as, 378 calcium storage of, 233
Barr body, 39 canaliculi of, 200, 203, 204
Bartholin’s glands, 898, 899 cancellous, 201, 206
Basal body, of cilia, 76-77, 77, 78, 79 carbonate ion of, 208
Basal cell, tracheal, 735—736 cells of, 208-216, 210-215
Basal granular cell(s). See Enteroendocrine cement lines of, 205, 228-229
cell(s). citrate ion of, 208
Basal lamina. See Lamina, basal. collagen of, 205, 207-208
Basal metabolic rate, 185 collecting sinuses of, 241
Basement membrane. See Lamina, basal. compact, 199, 201
Basophil(s), 119, 126-128, 127 formation of, 228-230, 229
beta, 486 demineralized, 208
cutaneous hypersensitivity and, 128 endochondral ossification of, 218-221, 219,
delta, 486-487, 487 220
of pars distalis, 483, 484, 486-487, 487 estrogen effects on, 234
specific granules of, 119, 126, 127 formation of, age and, 230
vs. mast cells, 127 ectopic, 216, 231
Beta cell, of pars distalis, 483, 484, 486-487, in manatee, 226, 226, 227
48 7 gonadal hormones and, 234
Beta-lipotropic hormone, 491 ground substance of, 207
Bile, 679 growth hormone effects on, 234
concentration of, 712, 712—713, 713 haversian canals of, 203
hepatic secretion of, 708 haversian systems of, 200, 203, 204, 205, 206
Bilirubin, hepatic excretion of, 707-708 definitive, 228
Bipolar cell, of retina, 947, 948, 948, 954 primitive, 228
Bladder, connective tissue of, 789, 790 stages of, 229
epithelium of, 787, 787-788, 788 heterogeneous nucleation of, 221—222, 225
Blastocyst, of ovum, 886, 886 histogenesis of, 216—230, 216—231. See also
Blood, 111-135 Ossification.
clotting of, 117 histophysiology of, 231-235, 232
plasma of, 132-133 historadiogram of, 205
Blood pressure, maintenance of, 380 interstitial systems of, 200, 202
Blood-aqueous barrier, 928—929 intramembranous ossification of, 216—218,
Blood-brain barrier, 362, 390 217, 218
Blood-ocular barrier, 390 isotope incorporation in, 208
Blood-testis barrier, 833—834, 834 joints of, 235, 236
Blood-thymus barrier, 390, 443 fibrous layer of, 235
Body (bodies), aortic, 378 synovial layer of, 235
Call-Exner, 857, 858, 858, 859 lamellae of, 200, 202, 206
carotid, 377-378 circumferential, 203, 206
ciliary, 913, 915, 925—928, 926—928 lamellar, 217, 218
Hassall-Henle, 920 formation rate of, 229, 229
Hassalfs, 437, 438, 441, 442 long, 200, 201
Herring, 495, 495 development of, 220
interrenal, 533 latitudinal growth of, 225-227, 226, 227
malpighian, 464 longitudinal growth of, 222-225, 223-227
neuroepithelial, of lungs, 750-751 structure of, 199
polar, 864, 864-865, 865 macroscopic structure of, 199-200, 200, 201
Russell, 160, 414 matrix of, 207-208
ultimobranchial, 504 membrane, 216
vitreous, 935-936 metabolic, 233
Weibel-Palade, 368 minerals of, 208
Boettcher’s cell, 974, 974 nutrient arteries of, 241
Bone marrow, 199 nutritional effects on, 234-235
blood cell formation and, 239—264 osteoblasts of, 209, 210
cell migration pore of, 242, 242 osteoclasts of, 211, 213-216, 214, 215
extravascular compartment of, 242-243, 243 osteocytes of, 209-211, 211
locations of, 239 osteoprogenitor cells of, 209
reticular cells of, 242 parathyroid hormone effects on, 233—234
stem cells of, 243-245, 244 preosseous tissue of, 224
structural organization of, 241-243, 242, primary spongiosa of, 216—218, 217
243 reorganization of, internal, 228—230, 229
vascular sinuses of, 241 repair of, 230—231
Bone(s), 199-238 resorption cavities of, 228, 229, 230
alveolar, 609 resorption of, osteoclasts and, 213, 214, 215
calcification of, collagen fibers and, 221-222, spongy, 199, 201
225 cancellous, 199
mechanism of, 221-222, 223, 224, 225 structural, 233
990 • INDEX

Bone(s) (Continued) Capping, B lymphocyte stimulation and, 424

structure of, macroscopic, 199—200, 200, 201 Cardiac gland(s), esophageal, 619, 623
microscopic, 200, 202-207, 202-207 gastric, 627
surface remodeling of, 227, 227-228, 228 Cardiac muscle, 296-308
trabeculae of, 199 atrial, 303—304, 304
vascular architecture of, 241 specific granules of, 304, 304
Volkmann’s canals of, 203 conducting tissue of, 304—308, 306, 307
woven, 217, 218 contractile bands of, 296, 297
Bone matrix, 200, 202, 207-208, 224 cytology of, 296-298, 297
deposition of, 216-217, 217 gap junctions of, 301, 303
Border cell, 976 innervation of, 307, 308
Boutons en passant, 337, 347 intercalated discs of, 296, 296, 298, 300—303,
Boutons terminaux, 347, 349 302, 303
Bowman’s capsule, 757 mitochondria of, 299, 300, 301
Brain, ventricles of, 359 nerve endings in, 337
Bronchi, 738, 738—739 nuclei of, 296, 297
lobar, 738 sarcoplasm of, fine structure of, 298, 298-
primary, 738 299, 299
segmental, 738 sarcoplasmic reticulum of, 299—300, 300,
subsegmental, 738 302
Bronchiole(s), 739 T system of, 299—300, 300, 302
respiratory, 739—740, 740, 741 trabeculae carneae of, 397
Brunner’s glands, 642, 644, 658—659 Cardiac skeleton, 397
Brush border, epithelial, 72-74, 73 Cardiodilatin, *304
Buccal glands, 593 Cardionatrin, 304
Bulbourethral glands, 844, 844—845 Carotene, skin color and, 556
Bundle, atrioventricular, 305 Carotid body, 377-378
cells of, 109, 109
Carotid sinus, 378
Cartilage, 188-198
articular, 199, 225
C cell, 504 calcification of, in long bone, 220
Calcification, bone, mechanism of, 221—222, cells of, 189, 189—190
223, 224, 225 elastic, 188, 194, 195
cartilage, 196-197, 197 extracellular matrix of, 148
provisional, zone of, 197 hbro-, 188, 194-195, 196
Calcitonin, 508 histophysiology of, 197-198
bone resorption and, 213, 215 hormone influences on, 198
osteoclasts and, 216 hyaline, 188, 188—194, 189—194
Calcium, blood level of, 233 appositional growth of, 189
bone, 208 capsular territorial matrix of, 191
historadiogram of, 205 histogenesis of, 188-189, 190
bone storage of, 233 intercapsular matrix of, 191
excitable tissue response and, 107 interstitial growth of, 188
intracellular, calmodulin control of, 107 interterritorial matrix of, 191
parathyroid hormone and, 514 matrix formation in, 191, 193-194
Calmodulin, calcium control by, 107 matrix of, 189, 190-191
Canaliculus (canaliculi), bile, 700—703, 701 — of trachea, 737, 737
703 proteoglycans of, 191
of bone, 200, 202, 203 transitory phase of, 195-196
secretory, 93-94 hypertrophic, 197, 197
of oxyntic cells, 630, 631, 632 proteoglycans of, 148-149
Canal(s), haversian, 203 regeneration of, 196
Hering’s, 708 regressive changes in, 196-197, 197
interfacial, of epithelia, 65 Catecholamine(s), 530
Volkmann’s, 203 adrenal medulla storage of, 526
Canines, 602 adrenal release of, 526
Capillary (capillaries), 381-390, 382, 383, 385- reserpine depletion of, 526
387 Cavity, medullary, 199
continuous, 384, 387, 388 Cecum, 660—663
cross section of, 384, 385 Cell cycle, 47-49, 48
fenestrated, 384, 387, 388 Cell(s), 1-56
lymph, 400 A, of intestine, 651
lymphatic, 401-403, 402, 403 absorptive, of intestine, 644-648, 645-648,
anchoring filaments of, 401 666, 666-667, 667
terminal elements of, 401-402 acinar, 84, 84
muscle-type, 384, 387, 388 ACTH, 486
transendothelial exchange of, 384, 389, 389- activities of, 43-53
390 adipose, 152-154, 153
types of, 384, 387, 388 adluminal, of eccrine sweat glands, 570
visceral, 384, 387, 388 adventitial, of seminiferous tubules, 798
Cell(s) (Continued) Cell(s) (Continued)
alpha, of pars distalis, 483 Hensen’s, 976
alveolar, Type I, 741, 744, 745 hilus, ovarian, 872
Type II, 741, 744, 745, 745 Hofbauer, 889
amacrine, of retina, 947, 948, 950, 954 horizontal, of retina, 945, 947, 954
APUD (amine precursor uptake and decar¬ I, of intestinal mucosa, 651
boxylation), 108 interdigitating, 423
B, See Lymphocyte(s), B. of lymph nodes, 456, 460
basal, tracheal, 735-736 interplexiform, of retina, 950
beta, of pars distalis, 483, 484, 486—487, 487 interstitial, of pineal gland, 537
bipolar, of retina, 947, 948, 948, 954 juxtaglomerular, 777—778, 778
blast. See Lymphoblast(s). Kupffer, 157, 157, 687, 689, 689-690, 690
-blast terminology of, 15In L, of intestinal mucosa, 651
blood. See Erythrocyte(s). Langerhans, 423, 545, 554, 554—556, 555
Boettcher’s, 974, 974 Leydig, 826-828, 827-830
bone, 208-216, 210-215 light, 504
border, 976 light microscopy of, 2
brush, of trachea, 736—737 lutein, 866
C, 504 granulosa, 866, 867, 868, 869
castration, 492 theca, 866, 868, 869
chief, of gastric mucosa, 628, 629, 632-633 lymphoid, dendritic, 423
of paraganglia, 532, 533 mast, 160—164, 161—163
of parathyroid glands, 511, 512 in allergy, 162-163, 163
Clara, of bronchioles, 739 vs. basophils, 127
Claudius’, 974, 974 membrane of, 3-6, 4, 5, 6
clear, of eccrine sweat glands, 569, 573 electron microscopy of, 3, 4
cone, 939, 941-946, 942—945 fluid mosaic model of, 4
connective tissue, 151-156, 152, 153, 155, layers of, 3, 4
156 trilaminar appearance of, 4
cycle of, 47-49, 48 Merkel, 109, 545, 556, 556, 557
cytotrophoblastic, of placental villi, 891 mesenchymal, 150, 152, 240, 241. See also
D, of intestinal mucosa, 651 Cell(s), osteoprogenitor.
dark, of eccrine sweat glands, 569, 573 microglia, 157, 157
dendritic, 429 mitochondria-rich, 504
follicular, 423 modulation in, 209
differentiation of, 209 motility of, 49—50
division of, 43—47, 45, 46 mucous, 92, 92, 97
EC, 634 of gastric mucosa, 626—627, 627
of gastric mucosa, 634 of salivary glands, 588, 589, 589-590
of intestinal mucosa, 651 myoepithelial, 266
ECL, 634 of eccrine sweat glands, 569
electron microscopy of, 2 of mammary gland, 903, 903, 904, 905
endocrine, of gastric mucosa, 633-635, 634, of salivary glands, 593
635 myoid, of seminiferous tubules, 798
endothelial, 384, 385 myometrial, 878-879
of liver, 686, 688, 688-689 neck, of gastric mucosa, 629—630, 630
enteroendocrine, 634, 649-651 neuroendocrine, 108—110, 109
ependymal, 345 neuroepithelial, of taste buds, 586
epithelial, pigment, 939-940 neuroglial, 345—347, 345—347, 346
epithelioid, 155 of adenohypophysis, 109
fallopian, 874, 874, 876, 876 of adrenal gland, 521-523
fat, 152-154, 153 of bronchial epithelium, 109
of liver, 690, 690—691, 691 of carotid body, 109, 378
fixed, 136 of epididymis, 835—837, 836, 837
follicular, of pars distalis, 488—489 of human lens, 932, 934
FSH/LH, 486-487, 487 of inner ear, 109
ganglion, of retina, 950 of nasal epithelium, 109
gastrin, 109, 634, 634, 634, 634, 635, 635 of nervous system. See Neuron(s).
giant, foreign-body, 155 of pancreas, 109
glandular. See Glandular cell(s). osteoprogenitor, 206, 208, 209
glomus, 378 oxyntic, of gastric mucosa, 630—632, 630—
goblet, 92, 92 632
of intestinal mucosa, 652 oxyphilic, of parathyroid glands, 511-513,
of intestine, 648—649 512
of large intestine, 660, 662, 663 Paneth, of intestinal mucosa, 652, 652-654,
tracheal, 735 653
granule, 329 parafollicular, of thyroid, 504—505, 505
gustatory, 109 phalangeal, 975—976
hair, cochlear, 976—979, 977, 978, 979 photoreceptor, 940—945, 941—945
of ear, 968—970, 969 pillar, 974, 974-975, 975
992 • INDEX

Cell(s) (Continued) CFU-S. See Colony-forming units—spleen.

pit, of liver, 691 Chemotactic factor(s), 419
plasma, 119, 129, 158-160, 158-160, 410, eosinophil, 126
413, 413-414, 414 Chemotaxis, 49, 170
antibody response of, 421 Chiasmata, of meiosis, 47
B lymphocyte origin of, 159 Chief cell(s), 628, 629, 632-633
humoral immunity and, 159, 160 of gastric mucosa, 628, 629
of thymic cortex, 440 of paraganglia, 532, 533
Russell bodies of, 414 of parathyroid glands, 511, 512
polyploid, 36 Cholangiole, 703
prolactin, 483, 484, 485 Cholecystokinin, 651, 712, 721, 729
Purkinje, 305, 320, 321, 322, 328 Chondrocyte(s), 188, 189, 189-190, 190
pyroninophilic. See Lymphoblast(s). isogenous, 189, 191
reserve, 487-488 Chondroitin sulfate, in extracellular matrix,
reticular, 425, 426 148, 148(t)
of bone marrow, 242 in hyaline cartilage, 191
of lymph nodes, 460 Chondronectin, 136
of thymus, 437 Chorion, 888
rod, 939, 940, 941, 942, 942 Chorionic gonadotropin, 896
S, of intestinal mucosa, 651 Choroid, 913, 915, 924-925, 925
satellite, 274, 328 Bruch’s membrane of, 925
Schwann, 328, 329 choriocapillary layer of, 924
serous, of salivary glands, 588, 590, 590, 592 glassy membrane of, 925
Sertoli, 798-802, 802, 803, 804 vessel layer of, 924
stellate, of pars distalis, 488—489 Choroid plexus, 359-360, 360
of pars intermedia, 493 villi of, 360, 361
stem, 243-245, 244 Chromaffin reaction, 525
steroid-secreting, 101 of paraganglia, 533
supporting, of membranous labyrinth, 969, Chromaffin system, 533
970 Chromatid, 38
of organ of Corti, 974-976, 974-977 Chromatin, 31, 32, 35, 36
of taste buds, 586 helical structure of, 37
teloglial, 289, 290 in interphase nucleus, 32
thyroid, 100, 101, 109 nucleolus-associated, 41
thyroidectomy, 492 sex, 39
transitional, 260 Chromomere, of platelets, 115-116, 116
TSII, 486 Chromophobe, 487-488
ultimobranchial, 504 Chromosome(s), 35, 36-41, 38, 39, 40, 41
veiled, 423 acrocentric, 39, 39
of lymph nodes, 456, 460 anomalies of, 39
ventricular, 355-356 crossing over of, 47
wandering, 136, 241 cytogenetics of, 38, 38-39, 39
Cementicle, 608 deletion of, 39
Cementoblast, 608 diploid number of, 36
Cementum, 607 haploid number of, 36
acellular, 607 homologous pairs of, 36
hyperplasia of, 607 metacentric, 38, 39
Central nervous system, 311, 356-362, 357, metaphase, 38, 38-39
358, 360, 361 primary constriction of, 38
blood vessels of, 362 satellite of, 38
blood-brain barrier of, 362 secondary constriction of, 38
pituitary hormones in, 491 metaphase analysis of, 38, 38-39
Centriolar satellite, 26
nucleolus organizer region of, 41
Centriole(s), 2, 20-22, 21, 22
of dividing lymphoblast, 38, 40
cross section of, 21, 21 sex, 36, 814
microtubules of, 27, 27 submetacentric, 39, 39
origin of, 21-22, 22 telocentric, 39, 39
Centromere, 38
ultrastructure of, 39-41, 41
Centrosome, 20
uncoiled, 40-41, 42
centriolar satellites of, 21 X, 36
Cerebral thrombosis, 381 Y, 36
Cerebrospinal fluid, 360-361 Chyle, 671
Ceruloplasmin, 133
Chylomicron (chylomicra), 133, 647, 666, 668,
Cerumen, 571, 961 669
Cervix, 883, 885
Chymase, 163
glands of, 897, 897 Chyme, 624
histophysiology of, 883, 885
Ciliary body, 913, 915, 925-928, 926-928
CFU-E. See Colony-forming unit—erythroid. blood supply of, 926
CFU-GM. See Colony-forming unit—granulomono- epithelium of, 926-927, 927, 928
cyte. muscles of, 926
CFU-M. See Colony-forming unit-megakaryocyte. Ciliary zonule, 921, 932-933
 INDEX • 993

Ciliogenesis, 77-78, 79 Colon, 660-663, 661, 662, 663

centriole role in, 22 Colony-forming unit-erythroid (CFU-E), 244,
Cilium (cilia), 58, 74-78, 75-79 245
axonenre of, 76, 77 Colony-forming unit-granulomonocyte (CFU-
basal body of, 76-77, 77, 78, 79 GM), 244, 245
centriolar formation of, 22 Colony-forming unit-megakaryocyte (CFU-M),
fallopian, 874, 876 244, 245
isochronal rhythm of, 76 Colony-forming unit-spleen (CFU-S), 244, 245
metachronal rhythm of, 76 Colostrum, 903, 908
microtubules of, 27, 27, 77, 77 Complement, 50, 407
motion of, 77-78, 75 phagocytosis and, 156
of ear, 968, 978 Concanavalin A (Con A), 424
of larynx, 734 Cone cell(s), 939, 941-946, 942-945
olfactory, 733 components of, 942, 945
origin of, 77-78 length of, 944-955
tracheal, 735, 736, 752 synaptic connections of, 948-949, 949, 950
Cirrhosis, 704 Conjunctiva, 916
Cistern, perinuclear, 31 Connective tissue, 136-173
Cisterna (cisternae), of skeletal muscle, 279, adipose cells of, 152-154, 153
280, 281 cells of, 151-156, 152, 153, 155, 156
Cisterna cerebellomedullaris, 359 free, 156-164, 157-163
Clara cell(s), of bronchioles, 739 collagen fibers of, 137-143, 138-141, 142(t)
Clathrin, 51 dense, 136, 164, 165-167, 166-168
Claudius’ cell, 974, 974 irregular, 136, 164, 165, 166
Clear cell, of eccrine sweat glands, 569, 573 regular, 136, 166-167, 168
Clefts of Schmidt-Lantermann, 330, 330, 331, elastic, 169
335 elastic fibers of, 144-146, 145, 146
Clitoris, 899 eosinophils of, 160
Cloasma, 558 fibers of, 136
Clonal selection theory, 406 fibroblast of, 151-152, 152
Clotting, 117 hbronectin, 146—147
CNS. See Central nervous system,. functions of, 169-170
Cochlea, 962, 963, 965, 966, 971-974, 972, glycosaminoglycans of, 147, 148, 148(t)
972-974 ground substance of, 147-149, 148, 148(t)
hair cells of, 976-979, 977, 978, 979 histophysiology of, 169-171
spiral prominence of, 971, 973, 974, 974 hormonal effects on, 171
stria vascularis of, 971, 973, 974 inflammation of, 170
vestibular membrane of, 971, 972 laminin of, 147
Codon, signal, of secretory proteins, 85 loose, 136, 137-164, 766
Collagen, 136, 137-143, 138-141, 142(t), 165 lymphocytes of, 158
banding patterns of, 139, 141 macrophages of, 154-156, 155, 156
biosynthesis of, 139, 140 mesenchymal cells of, 152
calcification and, 221-222, 225 mucous, 167, 169
disorders of, 171 nerve endings in, 340
fibrils of, 136 repair of, 170
fractions of, 1 39 reticular, 169
acid-soluble, 139 reticular fibers of, 143, 143, 144
insoluble, 139 synthesis of, 149-151, 150
neutral-soluble, 139 fibroblasts in, 149, 150
interstitial, 141 variants of, 136
long-spacing, fibrous, 139, 141 Connexon, of epithelia, 69
segment, 139 Contour lines of Owen, 603
microscopic study of, 143 Contraction, of skeletal muscle, 283, 283, 288-
of bone, 205, 207-208 289
synthesis of, 149—151, 150 Cornea, 913, 914, 914, 916-920, 917
fibroblasts and, 149 Bowman’s membrane of, 917
fibroblasts in, 150 Descemet’s membrane of, 918-919, 919, 920
Type I, 139, 141, 142, 142(t) endothelium of, 918-920
Type II, 141, 142, 142(t) epithelium of, 917, 917
in hyaline cartilage, 191 histophysiology of, 920
Type III, 141, 142^ 142(t) stroma of, 917, 917-918, 918
Type IV, 141, 142, 142(t) substantia propria of, 167, 168, 917, 917—
in basal lamina, 71 918, 918
vs. reticular fibers, 143 Corpus albicans, 866, 872
Type V, 141 Corpus luteum, formation of, 865-868, 867—
types of, 139, 141-143, 142(t) 869
characteristics of, 142(t) of menstruation, 866
unit fibrils of, 137, 138 of pregnancy, 866
unnatural forms of, 139, 141 Corpuscle(s), genital, 342, 342
Collagenase, connective tissue repair and, 171 lingual, 342
Colliculus seminalis, 840, 840, 841 Meissner's, 342, 342, 343
994 • INDEX

Corpuscle(s) (Continued) Dentin (Continued)

of Golgi-Mazzoni, 342 development of, 612, 613, 614, 614
pacchionian, 362 Korff s fibers of, 612
thymic, 437 mantle, 603
Vater-Pacini’s, 340—342, 341 Neumann’s sheath of, 603, 604
Cortex, adrenal, 516, 516—524, 517—523 Tomes’ fiber of, 603, 604
cell, 28 Tomes’ granular layer of, 603, 603
cerebellar, cells of, 316 tubules of, 603
nerve bodies of, 328 Deoxycorticosterone, 524
neuron of, 313 Deoxyribonucleic acid (DNA), 35
cerebral, cells of, 316 length of, 41
functional areas of, 355 Dermatan sulfate, in extracellular matrix, 148,
gray matter of, 355 148(t)
white matter of, 355 Dermatosparaxis, 171
hippocampal Dermis, 543, 544, 545, 558-561, 559, 560, 561
pyramidal cell of, 313 glassy membrane of, 566
Corticotroph, of pars distalis, 484, 486 histogenesis of, 575-576
Corticotropin releasing factor, 529 nerves of, 574-575
Corticotropin releasing hormone, 492 papillae of, 543
Cortisol, 524, 529 papillary layer of, 558
Cough reflex, 753 reticular layer of, 558
Cowper’s glands, 844, 844-845 Dermoepidermal junction, 559
Crenation, erythrocyte, 112, 112, 113 Desmosine, 145
Cretinism, 508 Desmosome, 65, 66, 68, 68-69, 69
Crinophagy, 483 Deuterosome, 22
Crista ampullaris, 967-970, 968, 969 Diabetes, 726
Cryptorchidism, 826 hypothalamic tumor and, 497, 498
Crypts of Lieberktihn, 643, 644, 651-654, 652, Diaphysis, 199, 200
653 Diarthrosis (diarthroses), 235
of large intestine, 660, 662 Dictyosome, 9
paneth cell of, 652, 652-654, 653 Digestive system, 579
pericryptal fibroblasts of, 652 Digestive tube, 579
secretions of, 665 lamina propria of, 579
Crystalloids of Charcot-Bottcher, 802, 803 mucosa of, 579
Crystals of Bottcher, 849 muscularis externa of, 579, 580
Crystals of Reinke, 828, 830 muscularis mucosae of, 579
Cupula (cupulae), of membranous labyrinth, submucosa of, 579
' 970 Diploe, 200
Cushing’s disease, 529 Diplosome, 20, 21
Cyclic AMP, nonexcitable tissue response and, Diplotene, 47
107 Discs, intercalated, of cardiac muscle, 296, 296,
second messenger concept of, 107, 107 298, 300-303, 302, 303
Cysts, nabothian, 883 DNA. See Deoxyribonucleic acid.
Rathke’s, 493 Down’s syndrome. See Trisomy 21.
Cytochalasin B, endocrine hormone transport Drugs, liver response to, 706-707
and,103-104 tolerance to, 707
Cytogenetics, 38, 38-39, 39 Duct(s), alveolar, 740, 740, 741
Cytokinesis, 44 of mammary glands, 902
Cytoplasm, 1 bile, 681, 708 '
membrane partitioning of, 3 collecting
Cytoplasmic inclusions, 22-24, 23, 24 of kidneys, 774-775
Cytoplast, 30 ejaculatory, 840, 842
Cytoskeleton, 2, 24-31, 25-27, 27(t), 29(t), 29, endolymphatic, 965, 967
31 Gartner’s 872
photomicrograph of, 25 intercalary, 98
Cytotrophoblast, 887, 887 intercalated, pancreatic, 727
interlobular, 98, 98
pancreatic, 727-728, 728
intralobular, pancreatic, 727
D cells, of intestinal mucosa, 651 lactiferous, 901
Dark cells, of eccrine sweat glands, 569, 573 lobar, 98
Deciduoma, 882 lymphatic, 400, 404
Demilune(s), Giannuzzi’s, 591, 593 of exocrine glands, 98, 98
serous, 97, 97 of gallbladder, 711
Dendrite, 312 pancreatic, 727, 727-728, 728
neuronal, 320, 321, 322 Santorini’s, 728
Dendritic cell, 429 Stenson’s, 593
Dentin, 602-604, 603, 604 testicular, 834-842, 836-841
calcification of, 603-604, 604 thoracic, 404, 432
circumpulpar, 603 Wharton’s, 595
contour lines of Owen of, 603 Wirsung’s, 728
 INDEX • 995

Ductus deferens, 797, 838-842, 839-842 Endocrine gland(s) (Continued)

Ductus epididymidis, 835-838, 836-839. See hormone secretion from, 103-104
also Epididymis. hormone transport in, 103-104
Duodenum, 641 lymph system and, 104
Dura mater, 200, 357, 358 polypeptide-secreting, 100, 100
Dwarfism, pituitary, 490 steroid-secreting, 100-103, 101, 102
Dynein, arms, 27 Endocytosis, 50-53, 51, 52
ciliary motion and, 77 Endoderm,57
Endolymph, 966, 981
electrolyte composition of, 981 (t)
Ear, 961-985 Endolymphatic duct, 965, 967
external, 961, 962 Endolymphatic sac, 967, 970-971
function of, 984—985 Endometrium, 878, 879-880
hair cells of, 968—970, 969 anovulatory cycle and, 882
inner, cell of, 109, 110 blood supply of, 880
internal, 964, 965, 966-984. See also Laby¬ cyclic changes in, 880-883, 881
rinth. decidualization of, 882
endolymph of, 981, 981 (t) phases of, menstrual, 882-883, 884
perilymph of, 981, 981 (t) morphological criteria of, 883, 884
perilymphatic labyrinth of, 980—981 ovarian activity and, 885
middle, 961—964, 962—965 proliferative, 880, 881, 882
auditory ossicles of, 962, 962—963, 963 secretory, 881, 882
auditory tube of, 963-964, 965 Endomysium, 272, 272, 272
tympanic cavity of, 962—963, 963 Endoneurium, 335, 336
tympanic membrane of, 962, 963, 964 Endoplasmic reticulum. See Reticulum, endo¬
transducer function of, 984-985 plasmic.
E-BFU. See Erythroid burst-forming units. Endorphin, beta, 486, 494
EC cell, of intestinal mucosa, 651 Endosteum, 200, 207
ECF-A. See Eosinophil chemotactic factor of ana¬ Endothelial cell, 384, 385
phylaxis. of liver, 686, 688, 688-689
E-CFEJ. See Erythroid colony-forming units. Endothelium, 60
Echinocyte, 112, 112, 113 arterial, 368, 380
ECM. See Extracellular matrix. Enterochromaffin cell(s). See Endocrine cell(s),
Ectoderm, 57 of gastric mucosa.
Ectoplasm, 28 Enteroendocrine cell, 108, 634, 649-651, 650
Edema, 401 Enterogastrone, 659
E-face, 5, 5, 6 Enzymes, lysosomal, inborn defects of, 18,
Ehlers-Danlos syndrome, 171 19(t)
Elastase, elastin digestion and, 145 Eosinophil chemotactic factor of anaphylaxis,
Elastic cartilage, 188, 194, 195 163
Elastic fiber(s), 136, 144—146, 145, 146, 165 Eosinophil(s), 119, 125—126, 126, 260
microfibrils of, 145—146, 146 chemotactic factor response of, 126
networks of, 145, 146 specific granules of, 125, 126
Elastica externa, 144, 367 Ependyma, 345
Elastica interna, 144, 367, 368, 369, 371 Epicardium, 396, 397
Elastin, arterial, 370 Epidermal growth factor (EGF), 707
of elastic fibers, 145, 146 Epidermis, 543, 543-558, 544-557
Elliptocytosis, hereditary, 114 cells of, 552-557, 552-558
Embolism, pulmonary, 117 dermoepidermal junction of, 559
Embryo, uterine implantation of, 886-888, histogenesis of, 575-576
886-888 keratinizing system of, 543
Emperipolesis, 418 keratinocytes of, 552, 554
Emphysema, 753 pigmented, 553
Enamel, 604—607, 605, 606 Langerhans cells of, 545, 554, 554-556, 555
calcification of, 617 malpighian layer of, 548
inner cuticle of, 607 melanocytes of, 543, 552, 553, 556-558
lamellae of, 607 melanosomes of, 553, 554, 557-558
Retzius’ lines of, 603, 606 Merkel cell of, 109, 110, 545, 556, 556, 557
rods of, 604, 605, 605-606, 606 mucocutaneous junctions of, 558
Schreger’s lines of, 603, 606 nerves of, 574-575
Tomes’ process of, 617, 617 of body, 550, 551, 552, 552
tufts of, 607 of palms, 545—550, 546—551
Enamel sheath, 605, 605 of soles, 545-550, 546—551
Endocardium, 396 ridges of, 543
subendocardial layer of, 396 stratum basale of, 545
subendothelial layer of, 396 stratum corneum of, 546, 547, 550, 550, 551
Endocrine cell(s), of gastric mucosa, 633-635, stratum germinativum of, 545, 547, 548
634, 635 stratum granulosum of, 546, 547, 549
Endocrine gland(s), 57, 83, 99—110 cells of, 554
blood vessels and, 104 stratum lucidum of, 546, 550
control mechanisms of, 104—106, 105, 106 stratum spinosum of, 547
996 • INDEX

Epididymis, 796, 797, 835-838, 836-839 Epithelium (epithelia) (Continued)

blood supply of, 838, 838, 839 renewal of, 80-81
cells of, basal, 836, 837 seminiferous, 798-802, 799-803
contractile, 837, 837 cycle of, 820-824, 822-825
muscle, 837, 837 simple, 58
principal, 835—836, 836 specializations of, 64-72, 66-70, 72
columnar epithelium of, 74, 74 squamous, 58
motility of, 838 keratinized, 60
regions of, 797, 835 nonkeratinized, 60, 61
Epimysium, 272, 272 simple, 58, 59, 59-60, 60
Epinephrine, 530 stratified, 60, 61,63
action of, 107, 107 stratified, 58
Epineurium, 335, 336 terminal web of, 67
Epiphyseal plate, 199, 200, 220, 225 tonohlaments of, 68, 68
Epiphysis (epiphyses), 199, 200 tracheal, 736, 736, 737
closure of, 225 transitional, 58, 59, 62, 64, 64
Epiphysis cerebri. See Pineal gland. vaginal, 897
Epithelial cell, glycocalyx of, 6 zonula adherens of, 66, 67, 67-68
microvillous processes of, 4 zonula occludens of, 66, 66-67, 67
pigment, 939-940 Eponychium, 567, 567
Epithelioid cell, 155 Epoophoron, 872
Epithelium (epithelia), 57-81 Ergastoplasm. See Reticulum, endoplasmic.
attachments of, 65-71, 66-70 Erythroblast(s), basophilic, 246, 246, 249
basal surface of, 71—72 primitive, 240
bladder, 787, 787-788, 788 definitive, 239
blood vessels to, 79 orthochromatic, 247, 248, 249
bronchial, 739 polychromatophilic, 247, 247, 249
basal granular cells of, 109, 109 definitive, 241
bronchiolar, 739 primitive, 240
brush border of, 72-74, 73 primitive, 239, 240
cell polarity of, 65 Erythrocyte(s), 111-115, 112, 113, 114, 115,
classification of, 58—64, 59—64 249
columnar, 58 abnormal forms of, 114
brush border of, 72-74, 73 crenation of, 112, 112, 113, 113
pseudostratihed, 58, 61, 62 cytoskeleton of, 114-115, 115
simple, 58, 60, 61, 62 ghost membrane of, 113
sterocilia of, 74, 74 hemoglobin content of, 113, 114, 114
stratified, 58, 60, 67, 62, 63 hypochromic, 114
striated border of, 72-74, 73 membrane of, 114-115, 115
terminal web of, 72 normochromic, 114
communicating junctions of, 65-71, 66-70 polychromatophilic, 113-114, 248
connexons of, 69, 70 primitive, 241
cuboidal, 58 rouleau formation of, 112, 112
simple, 58, 60 shape of, 112, 112-113, 113
definition of, 57, 58 splenic removal of, 475-476
functions of, 57-58 Erythroid burst-forming units (E-BFL1), 245
germinal, 59 Erythroid colony-forming units (E-CFU),
ovarian, 852, 853 245
glycocalyx of, 72, 73 Erythropoiesis, 245-248, 246-249
hemidesmosomes of, 68, 69 Erythropoietin, 261
intercellular cement of, 59, 65 Esophagus, 619-624, 620-622
intestinal, 643-648, 645-650 epithelium of, 619, 620-622
renewal of, 654 glands of, 619, 622, 623
intraepithelial gland of, 93, 93 cardiac, 619, 623
keratinized, 58-59 histophysiology of, 623-624
laryngeal, 734, 735 muscularis mucosae of, 619
lymphocyte invasion of, 80 muscularis of, 619
macula adherens of, 68, 68-69, 69 submucosa of, 619
nasal, cells of, 109, 109 tunica adventitia of, 619
nerve endings in, 340 Estradiol, 885
nerves to, 79-80 ovulation and, 863
nexus of, 69, 70, 71 Estriol, placental, 896
nutrition to, 79 Estrogen(s), 885
of ciliary body, 926-927, 927, 928 bone effects of, 234
of Henle’s loop, 769, 771, 771 ovarian follicle development and, 862
of oviduct, 875, 876, 974-875 theca interna production of, 861
olfactory, 731-734, 732, 733 Estrone, 885
origin of, 57-58 Estrus, 885
photomicrographs of, 61 Euchromatin, 32, 35
pigment, of retina, 936, 937, 939, 939-940 Excretion, 83
pseudostratihed, 59 Exocoelom, 888
 INDEX • 997

Exocrine gland(s), 57, 83 Eye (Continued)

acinar, branched, 94, 94 visual axis of, 914
compound, 94—95, 96 vitreous body of, 935-9-36
simple, 94, 94 Eyelashes, 956, 957
classification of, 92-97, 92—97 Moll’s glands of, 956
compound, 94-97, 95—97 Eyelids, 956, 957
acinar, 94—95, 96 blood supply of, 959
tubular, 94, 95 conjunctiva of, 956, 958
duct system of, 98, 98 lymph vessels of, 959
histological organization of, 97-98, 98 third, 956
lobes of, 97
lobules of, 97, 98
mixed, 97
mucous, 96, 97 Factor, intrinsic, gastric, 632, 639
multicellular, 93-97, 93-97 Fallopian tube. See Oviduct.
parenchyma of, 97 Fascia (fasciae), 167
saccular, compound, 95 Fat. See Adipose tissue.
secretory control in, 98—99 dietary, digestion of, 666-667, 667—669
serous, 96, 97 fetal. See Adipose tissue, brown.
simple, 94, 94 Fat-storing cell, of liver, 690, 690—691, 691
coiled, 94, 94 Fauces, 597
tubular, 94, 94 Feldstruktur, of muscles, 299
stroma of, 97 Feminization, testicular, receptor abnormalities
tubular, branched, 94, 94 in, 107-108
coiled, 94, 94 Ferritin, 262
compound, 94, 95 Fertilization, 865
unicellular, 92, 92—93, 93 Fiber(s), collagen, 136, 137-143, 138-141,
Exocytosis, 18, 53, 90 142(t)
Exteroceptive system, 311 bone calcification and, 221-222, 225
Extracellular matrix, 136, 137-151 of bone, 205, 207-208
collagen fibers of, 137—143, 138-141, 142(t) synthesis of, 149-151, 150
elastic fibers of, 144—146, 145, 146 elastic, 136, 144—146, 145, 146
fibronectin of, 146-147 nerve. See Nerve fiber(s).
glycosaminoglycans of, 147, 148, 148(t) neuroglial, 345
ground substance of, 147-149, 148, 148(t) of skeletal muscle, 278, 278-279, 279
laminin of, 147 cytology of, 273, 273-276, 275-277
proteoglycans of, 147, 148—149 of smooth muscle, 265—266, 266
reticular fibers of, 143, 143, 144 association of, 266—267
Exudate, serous, 164 perforating, 202, 206, 206, 207
Eye, 913-960 Purkinje, 305, 306, 307
accessory organs of, 955—956 reticular, 136, 143, 143, 144
aqueous humor of, 928—929 Sharpey’s, 202, 206, 206, 207
axes of, 916 Fibrillenstruktur, of muscles, 299
blood supply of, 955 Fibrils, collagen, 136, 137, 138
blood-aqueous barrier of, 928—929 Fibrinogen, 134
choroid of, 924-925, 925 Fibroblast(s), 137, 151-152, 152, 165
ciliary body of, 925-928, 926—928 collagen synthesis and, 149, 150
cornea of, 914, 915, 916—920, 917 pericryptal, Fieberkiihn’s crypts of, 652
dimensions of, 916 tendon, 166—167, 167
dioptric media of, 914, 914, 915 Fibrocartilage, 188
fibrous tunic of, 916—924, 916-924, 917- Fibrocyte(s). See Fibroblast(s).
924, 917-924 Fibrogenesis, 149—151, 150
histogenesis of, 959, 959-960 Fibronectin, 136, 146-147
inner axis of, 916 of epithelia, 65
iris of, 929-931, 930-932 plasma, 146-147
lacrimal caruncle of, 958—959 Fields of Cohnheim, 275
lacrimal gland of, 956, 958—959 Filament(s), actin, of microvillus, 72, 73
lens of, 931—935, 932—935 contractile, of smooth muscle, 267—268, 270
limbus of, 920-924, 921-924 cvtoskeletal, 25
lymph spaces of, 955 desmin, 29(t), 29-30
nerves of, 955 of skeletal muscle, 284
optical axis, 914, 916 glial, 29(t), 30, 345
radius of curvature of, 916 intermediate, 25, 28-30, 29(t), 29
reference planes of, 916 keratin, 29, 29(t), 29
refractive media of, 931—936 lymphatic, 401
rotation center of, 916 myosin, 28
sclera of, 916 neuro-, 29(t), 30
sclerocorneal angle of, 920, 921 vimentin, 29(t), 30
structure of, 913—916, 914, 915 of skeletal muscle, 284
vascular tunic of, 924—931, 925—928, 930, Z, of skeletal muscle, 284
931 Flagellum (flagella), 78-79, 80
998 • INDEX

Follicle stimulating hormone, 486, 491 Gastric mucosa (Continued)

ovarian changes and, 885 glands of, cardiac, 627
ovarian follicle development and, 861 gastric, 627-635, 628-635
ovulation and, 863 lamina propria of, 636
spermatogenesis and, 833 muscularis mucosae of, 636
Follicle(s), hair, 545, 561, 561 pyloric glands of, 635, 635
papilla of, 560, 561, 561, 565 rugae of, 624
ovarian, 852, 853, 854—862, 855-863 Gastric submucosa, 636
atresia of, 868-870 Gastrin, 99, 627, 634, 651
Follicular dendritic cell, 423 secretion of, 639
Folliculostatin, 862 Gastrin cell, 109, 109, 634, 634, 635
Foramen of Magendie, 359 Gastrin glands, secretion of, 99
Foramina of Luschka, 359 Gastroenteropancreatic (GEP) endocrine cell,
Foreign-body giant cell, 155 108
Fovea, 936, 957, 953, 953 Gastrointestinal tract, blood vessels of, 668-
Freeze-fracture method, membrane analysis 669, 670
by, 5, 5 evaginations of, 579
FTS. See Thymic serum factor. lamina propria of, 580, 580
a-Fucosidase, 19(t) lymph vessels of, 669, 671
nerves of, 671-676, 672-675
walls of, 580
Gelatin, 137
Genes, 39
a-Galactosidase, defect in, 19(t) GERL. See Golgi-associated endoplasmic reticulum
(3-Galactosidase, defect in, 19(t) from which lysosomes form.
Gallbladder, 708-713, 709-713 Germinal center(s), of lymph nodes, 455, 456
bile concentration of, 712, 712-713, 713 of lymphoid tissue, 427-430, 428, 429
blood supply of, 712 of spleen, 465, 467
ductlike structure of, 711 GFA. See Glial fibrillary acidic protein.
epithelium of, 709, 710, 711 Giant cell, foreign-body, 155
histophysiology of, 712, 712-713, 713 Gigantism, pituitary, 483, 490
lymphatics of, 712 Gingiva, 608-609
mucosa of, 708, 709, 710 Gland(s). See also specific glands, e.g., Endo¬
nerves of, 712 crine gland(s); Exocrine gland(s).
Ganglion, nodose, cell of, 316 acinar, branched, 94, 94
Ganglion cell, of retina, 950 simple, 94, 94
Ganglion (ganglia), collateral, 342 adrenal, 516-534, 517-523, 525-527, 532.
prevertebral, 342 See also Adrenal glands.
terminal, 342 axillary, at premenstruum, 572, 572
vertebral, 342 Bartholin’s, 898, 899
Gap junction. See Nexus, of epithelia. Bowman’s, of nose, 734
Gap junction(s), of cardiac muscle, 301, 303 Brunner’s, 642, 644-648, 645-648, 658-659
of osteocytes, 210, 211 buccal, 593
of smooth muscle, 270 bulbourethral, 844, 844-845
P-face of, 70 cardiac, of gastric mucosa, 627
Gastric gland(s), blood supply of, 637, 637, 638 ceruminous, 571, 961
cells of, 627, 628 cervical, 897, 897
chief, 627, 628, 628, 629, 629, 632-633 endocrine. See Endocrine gland(s).
endocrine, 627, 633-635, 634, 635 exocrine. See Exocrine gland(s).
neck, 629-630, 629-630, 630 gastric, 627-635, 628-635
oxyntic, 627, 630-632, 630-632 glossopalatine, 595
parietal, 627 hilus, sympathicotropic, 872
zymogenic, 627 labial, 589, 593
of gastric mucosa, 627-635, 628-635 lacrimal, 956, 958-959
Gastric intrinsic factor, 632, 639 lingual, 595-596, 596
Gastric mucosa, 624-636, 625-635 Littre’s, 792, 792
blood supply of, 636-637, 637 meibomian, 568, 956
cells of, aspirin erosion of, 638, 638, 639 Moll’s, 572, 956
chief, 628, 629, 632-633 Montgomery’s, of areola, 901
EC, 634
of skin, 568-572, 569-570, 572-574
ECL, 634
of tongue, 595-596
endocrine, 633-635, 634, 635 of von Ebner, 585, 596
G, 634, 634, 635 palatine, 596
mucous, 626-627, 627
parathyroid, 511-515, 512-514. See also
neck, 629-630, 630
Parathyroid glands.
oxyntic, 630-632, 630-632 parotid, 593
renewal of, 637-638, 638
pineal, 535-542, 536-541. See also Pineal
repair of, 638, 638 gland.
epithelium of, 622, 626-627 preen, 569
foveolae of, 624, 625, 626
prostate, 840, 841, 842-844, 843
 INDEX • 999

Gland(s) (Continued) Golgi complex (Continued)

pyloric, 635, 635 secretory product packaging of, 9-13, 10, 12
sebaceous, 560, 561, 568-569 transport vesicles of, 11-13, 12
hair and, 567 vertical section of, 10
sublingual, 595, 596 Golgi-associated endoplasmic reticulum from
submucous, tracheal, 737 which lysosomes form (GERF), 11, 12, 13
sweat, eccrine, 569, 569-572, 570, 572, 573 Gonadostatin, 862
thyroid, 500-510, 501-507. See also Thyroid Gonadotrophs, of pars distalis, 486-487, 487
gland. Gonadotropin releasing hormone, 492
types of, 83 ovulation and, 863
uropygial, 569 Gonadotropins, 491
uterine, 880 G-phase, of cell cycle, 48, 48
von Ebner’s, 596 Granule(s), azurophil, 120-121, 122, 125
Glandular cell(s), condensing vacuoles of, 90 basophil, 119, 126, 127
Golgi complex function in, 9—11, 10 Birbeck, 554, 555
Golgi complex of, 88, 89, 89-91 eosinophil, 125, 126
secretory granules of, 90, 90 keratohyalin, of stratum granulosum, 547,
transport vesicles of, 89, 89 549
Gians penis, 846 lamellated, of stratum spinosum, 547
Glaucoma, 924 membrane-coating, of stratum spinosum, 547
Glial fibrillary acidic protein (GFA), 30, 345 of Fangerhans cells, 554, 555
Glisson’s capsule, 703 of monocytes, 132, 132
Globulin(s), beta, 133 of neurons, 319—320
cold-insoluble, 146-147 of neutrophils, 119, 120—121, 122
gamma, 133 secretory, 2
immune, 133 of glandular cells, 90, 90
Glomerulus, 758, 762, 764, 765 vermiform, 554, 555
Glomerulus (glomeruli), filtration barrier of, Granulocyte, 243
762, 765\ 783 Granulocyte-macrophage colony-forming cell(s)
renal, 757, 759, 759 (GM-CFC), 262
Glossopalatine glands, 595 Granulomere, of platelets, 115, 116, 116, 118
Glucagon, 651 Granulopoiesis, 248—254, 250-255
Glucocorticoid(s), 528 Granulopoietin, 262
control of, 529 Granulosa lutein cell, 866, 867, 868, 869
inflammatory response and, 529 Graves’ disease, 508
Glucose, renal reabsorption of, 784 receptor abnormality in, 108
a-Glucosidase, defect in, 19(t) Gray matter, 325, 326
(3-Glucosidase, defect of, 19(t) nuclei of, 325
[3-Glucuronidase hexosaminidase, 163 Ground substance, amorphous, 147
Glvcocalyx, 6 of connective tissue, 147—149, 148, 148(t)
of epithelia, 72, 73 Growth hormone, 490
Glycogen, alpha particles of, 22, 23 bone effects of, 234
beta particles of, 22, 23 Growth hormone inhibiting hormone, 492
in hepatocytes, 696, 698 Gustatory cell, 109, 109
Glycosaminoglycans, 136
of extracellular matrix, 147, 148, 148(t)
GM-CFC. See Granulocyte-macrophage colony-
forming cell(s). H bands, of skeletal muscle, 276, 283
Goblet cell(s), 92, 92 Hair(s), 561—567, 562—565
life span of, 652 club, 565
of intestine, 648-649 cuticle of, 561, 563, 564
of large intestine, 660, 662, 663 distribution of, 566, 568
tracheal, 735 follicles of, 545, 560, 561, 561, 564, 565
Goiter, 508 active, 561
colloid, 508 keratogenous zone of, 561
exophthalmic, 508 quiescent, 561
Goitrogen, 508 growth of, 561, 561, 565
Golgi complex, 2, 7, 10, 11, 12, 13 mosaic pattern of, 566
cisternae of, 9—10, 10 wave pattern of, 566
condensing vacuole of, 10, 12 Henle’s layer of, 565, 566
electron micrograph of, 10 histogenesis of, 576
forming face of, 11, 12 Huxley’s layer of, 564, 566
of glandular cells, 88, 89, 89-91 keratinization of, 566
of neurons, 317, 318 longitudinal section of, 564
of neutrophils, 121 melanin granules of, 563
of osteocytes, 210, 212 pigmentation of, 565, 566
of promyelocyte, 249 root of, 560
of skeletal muscle, 275 root sheath of, 561, 566
of steroid-secreting cells, 101 external, 566
secretory face of, 11, 11 internal, 564, 566
1000 . INDEX

Hair(s) (Continued) Hepatocyte(s) (Continued)

sebaceous glands and, 567 Golgi complex of, 696, 699
shaft of, cortex of, 561 lipid in, 696
medulla of, 561 peroxisomes of, 699, 700, 701
Hair cell(s), cochlear, 976-979, 977, 978, 979 Herring body, 495, 495
of ear, 968-970, 969 Hertwig’s epithelial root sheath, 607
stimulation of, 970 Heterochromatin, 32, 35
terminal boutons of, 970 Heterogametic sex, 36
type I, 968-969, 969 Heterophagy, 16, 7 7
type II, 968-970, 969 Hiatus hernia, 623
Hassall-Henle bodies, 920 Hibernation, 185—186, 186
Hassall’s bodies, 437, 438, 441, 442 Hilus cell, ovarian, 872
Haustrae, of large intestine, 663 Histamine, 162, 634
Haversian canal(s), 203 Histiocyte, 154
Haversian system(s), 200, 203, 204, 205, 206, Histology, 1
230 Histone H1; 35, 37
definitive, 228 Hofbauer cell, 889
formation of, rate of, 229, 229-230 Homogametic sex, 36
stages of, 229 Horizontal cell, of retina, 945, 947, 954
primitive, 228 Hormone(s), adipose tissue and, 183-184, 184,
secondary, cement lines of, 228-229 185
Heart, 396-399, 397, 398 adrenocortical, adipose tissue and, 183
annuli hbrosi of, 397 adrenocorticotropic, 486
aortic valve of, 398 adrenocorticotropin, 491
atrioventricular bundle of, 398 adrenomedullary, 530
atrioventricular node of, 398 antidiuretic. See Vasopressin.
atrioventricular valve of, 398, 398 beta-lipotropic, 491
atrium of, 396 connective tissue and, 171
blood supply to, 399 follicle stimulating, 486, 491
impulse-conducting system of, 398-399 ovarian changes and, 885
lymphatics of, 399 ovarian follicle development and, 861
mitral valve of, 398, 398 ovulation and, 863
nerves of, 399 spermatogenesis and, 833
nodulus Arantii of, 398 gonadal, bone effects of, 234
pulmonic valve of, 398 gonadotropin releasing, ovulation and, 863
septum membranaceum of, 397 growth, 490
sinoatrial node of, 398 bone effects of, 234
supporting structure of, 397 lipotrophic, 486
trigona fibrosa of, 397 luteinizing, 486, 491
valves of, 396 ovarian changes and, 885
ventricle of, 396 ovarian follicle development and, 861
Heat, generation of, adipose tissue and, 185— testosterone production and, 832
186 luteinizing hormone-releasing, ovulation
Hemal node, 462 and, 864
Hemidesmosome, 68, 69 melanocyte stimulating, 491, 493
Hemocytoblast, 241. See also Lymphoblast(s). ovarian, 885
extravascular, 240 parathyroid, 508, 513-515, 514
Hemoglobin, 113 bone formation and, 233-234
degradation of, spleen and, 476 bone resorption and, 213, 215-216
Hemoglobin A, 113 osteoclasts and, 213, 215-216
Hemoglobin F, 113 osteolysis and, 211
Hemoglobin S, 113 pituitary, cartilage growth and, 198
Hemolysis, 113 pre-proparathyroid, 514
Hemophilia, 117 proparathyroid, 514
Hemopoiesis, 239 receptors for, 106-108, 107
extramedullary, 241 secretion of, 103-104
hepatic phase of, 239 storage of, 103-104
humoral influence in, 261-263 thyroid, 503
mesoblastic phase of, 239 thyroid stimulating, 486, 491
microenvironment influence on, 260-261 Howship’s lacuna (lacunae), 211
myeloid phase of, 240
Hyaline cartilage. See Cartilage, hyaline.
prenatal, 239-241, 240, 241
Hyaline membrane disease, 753
regulation of, 260-263 Hyalocyte, 935
Hemosiderin, 24
Hyalomere, of platelets, 115, 116, 118
Hensen’s cell, 976
Hyaluronidase, bacterial, 147
Heparan sulfate, in extracellular matrix, 148
Hyaluronic acid, in extracellular matrix, 147,
148(t)
148(t), 148
Heparin, 162
Hydatid of Morgagni, 872
Hepatocyte(s), cytology of, 694-699, 694-699 Hymen, 896
gap junctions of, 702, 703
Hyperadrenocorticism, 529
glycogen in, 696, 698
Hyperbilirubinemia, 707
 INDEX • 1001

Hyperglobulinemia, plasma cell excess and, Immunity (Continued)

160 humoral, 128, 407
Hyperinsulinism, 727 plasma cells and, 159,^760
Hyperparathyroidism, 234 lymph node function in, 458—462
primary, 515 primary response of, 128
secondary, 515 secondary response of, 128
Hypersensitivity, basophil, cutaneous, 128 Immunoblast. See Lymphoblast(s).
cell-mediated, 127 Immunoglobulin, 407—408
immediate, 127 in primary immune response, 421
Hyperthyroidism, 508 Immunoglobulin A (IgA), 408
Hypoadrenocorticism, 529 biliary, 706, 707
Hypodermis, 543, 544, 545, 559, 561, 575 intestinal production of, 656
Hyponychium, 567, 568 mammary gland secretion of, 910, 910
Hypophyseoportal system, 395 secretory piece complex with, 597
Hypophysis, 479-481, 479—499, 484-490, Immunoglobulin D (IgD), 408
495-498 Immunoglobulin E (IgE), 127, 162, 163, 408
acidophils of, 483, 492 Immunoglobulin G (IgG), 407
alpha cells of, 483 Immunoglobulin M (IgM), 407-408
anatomy of, 479, 480, 481 Implantation, interstitial, 886
anterior lobe of, 481 of ovum, 886-888, 886-888
basophils of, 483, 484, 486-487, 487, 492 Incisors, 602
blood supply of, 489, 489-490 Incisures of Schmidt-Lantermann, 330, 330,
cells of, castration, 492 331, 335
stellate, 493 Inclusion(s), 1—2, 2
thyroidectomy, 492 crystalline, 24
chromophobes of, 487—488 cytoplasmic, 22-24, 23, 24
corticotrophs of, 486 of alveolar macrophages, 748, 749
gonadotrophs of, 486-487, 487 Incus, 962, 962, 963
histophysiology of, 493—494 Infarction, myocardial, 381
hypothalamus and, 481 Inflammation, glucocorticoids and, 529
in hypothalamic diabetes insipidus, 497 of connective tissue, 170
mammotrophs of, 483, 484, 485 Inhibin, 833
nerves of, 489 Insulin, 726
pars distalis of, 479, 482-492, 484-490 adipose tissue and, 183-184, 184, 185
somatotrophs of, 483, 484, 485 liver pathway of, 707
Hypothalamohypophyseal tract, 495, 496 negative feedback control of, 105, 105
Hypothalamus, magnocellular neurosecretory secretion of, 100
system of, 495 Intercalated disc(s), 296, 296, 298
parvicellular system of, 494—495 of cardiac muscle, 300-303, 302, 303
Hypothyroidism, 508 Interdigitating cell, 423
Interferon, 129
Interoceptive system, 311
Interphase, 44
1 bands, of skeletal muscle, 275, 283, 284 Interplexiform cell, of retina, 950
I cells, of intestinal mucosa, 651 Interrenal bodies, 533
IgA. See Immunoglobulin A. Interstitial cell, of pineal gland, 537
IgD. See Immunoglobulin D. Interstitial system, of bone, 200, 202
IgE. See Immunoglobulin E. Intestinal epithelium, cells of. See also Intestinal
IgG. See Immunoglobulin G. mucosa, cells of.
IgM. See Immunoglobulin M. secretory component of, 657
Ileocecal sphincter, 659 Intestinal mucosa, 641-648, 641—660, 650,
Ileum, 641 652, 655-659
Immotile cilia syndrome, 752 Brunner’s glands of, 642, 658-659
Immune response, 406 cells of, A, 651
cellular, 461-462 D, 651
lymph node antibody production in, 460— EC, 651
461 enteroendocrine cells of, 649—651, 650
macrophages in, 422-423 goblet, 648-649, 652
primary, 406 I, 651
secondary, 406 L, 651
splenic, 476—477 renewal of, 654
Immune system, 406—435 S, 651
cells of, 409-425, 410-417, 420, 421 turnover of, 654
histophysiology of, 414-425, 415—417, epithelium of, 643-648, 645—650
420, 421 absorptive cells of, 644-648, 645-648
histophysiology of, 430-432, 431 Kerckring’s valves of, 641, 641, 642
Immune-phagocytosis, 123 lamina propria of, 643, 644, 654
Immunity, 128, 407 Lieberktihn crypts of, 643, 644, 651—654,
cell-mediated, 129 652, 653
cellular, 407 lymphoid nodules of, 654—655, 655
thymus and, 445 muscularis mucosae of, 658, 659, 659
1002 • INDEX

Intestinal mucosa (Continued) Keratan sulfate, in extracellular matrix, 148

Paneth cells of, 652, 652—654, 653 in hyaline cartilage, 191
Peyer’s patches of, 655, 655 Keratin, 543
plicae circulares of, 641, 641 filaments of, 29, 29(t), 29
serosa of, 660 Keratinizing system, 543
submucosa of, 658 Keratinocyte, 552, 554
villi of, 642, 643, 643, 645 cytomorphosis of, 545
Intestine(s), 641-678 pigmented, 553
absorptive surface of, 664, 664 Kidneys, 755—787. See also Nephron(s).
cells of, absorptive, 666, 666—667, 663 aldosterone effects on, 787
cholinergic neurons of, 676 antidiuretic hormone and, 786
enzyme secretions of, 665 arteries of, 779, 780
histophysiology of, 664—668, 664—669 blood pressure regulation by, 776—779, 778,
large, 660—664, 661—663 779
anus of, 663—664 blood supply of, 779-780, 780, 781
appendices epiploicae of, 663 calyces of, 755, 756
appendix of, 660, 661 cells of, juxtaglomerular, 777-778, 778
cecum of, 660-663 collecting ducts of, 774-775
colon of, 552, 660—663, 661, 663 cortex of, 755, 757, 766
goblet cells of, 660, 662, 663 interstitial cells of, 775
haustrae of, 663 distal tubule of, 772, 774, 774
Lieberktihn’s crypts of, 660, 662 macula densa of, 778, 779
rectum of, 663 filtrate of, 782
taenia coli of, 663 functions of; 782—787, 783, 785, 786
nerve cells of, 676 glomerular filtration barrier of, 783
nerves of, 671—676, 672—675 glucose reabsorption of, 784
plexus of, muscular, 674 hemodynamics of, 780
myenteric, 672, 675, 674, 674, 675 Henle’s loop of, countercurrent multiplier
submucous, 674 mechanism of, 785, 785—786
subserous, 674 hilus of, 755, 756
secretions of, 665 histophysiology of, 782-787, 783, 785, 786
small, 641-648, 641-660, 650, 652, 655-659 interstitial cells of, 775—776, 776, 777
Brunner’s glands of, 658—659 juxtaglomerular cells of, 778, 778-779
contractions of, 659 juxtaglomerular complex of, 776-780, 778-
enteroendocrine cells of, 649-651, 650 781
goblet cells of, 648-649 lymphatics of, 780-782
layers of, 641, 641 medulla of, 755, 756, 757
secretory immune system of, 655-657, interstitial cells of, 775-776, 776, 777
656, 657, 658 zones of, 771, 772, 773
Intraepithelial glands, 93, 93 medullary rays of, 756, 756, 757
Inulin, nephron transport of, 783 nephron of, 757-774, 758-774
Iris, 913, 914, 915, 929-931, 930-932 nerves of, 782
blood supply of, 931, 932 proximal tubule of, 763, 766-769, 766-769
color of, 929, 930 pyramids of, 755
Iron, metabolism of, spleen and, 476 reabsorption activity of, 784
Iron deficiency anemia, 262 renal sinus of, 755, 756
Islet cell, 109 rete mirabile of, 780, 781
Islets of Langerhans, 721-725, 722-726 secretory activity of, 784—785
Isodesmosine, 145 ureter of, 755, 756
Isotopes, bone-seeking, 208 urine excretion from, 787—793, 787—793
Isthmus, uterine, 883 uriniferous tubules of, 756—775, 758—774
vasa recta of, 779, 781
medullary osmolality and, 786
Kinetochore. See Centromere.
Jaundice, 707 Kinocilium (kinocilia), 74
Jejunum, 641 of ear, 968, 968, 969
Joints, bone, 235, 236 Klinefelter’s syndrome, 39
Junction(s), gap. See Nexus, of epithelia. Korffs fibers, 607, 608, 612
occluding, of Sertoli cells, 833-834, 834 Krebs cycle, 14
tight. See Zonula occludens. Kupffer cell(s), 157, 157
Juxtaglomerular complex, 776-779, 778, 779 of liver, 687, 689, 689-690, 690

Kallikrein, 170 L cell, of intestinal mucosa, 651

Kartagener’s disease, 752 Labia majora, 899
Karyokinesis, 44 Labia minora, 899
Karyoplasm. See Nucleoplasm. Labial glands, 589, 593
Karyosome, 31 Labyrinth, 964-984
Karyotype, 39, 40 blood supply of, 983-984
 INDEX • 1003

Labyrinth (Continued) Leukocyte(s) (Continued)

bony, 962, 963, 966 neutrophilic, drumstick of, 120, 727
basilar membrane of, 966, 972, 973, 974 nuclear chromatin of„ 120, 727
cochlea of, 962, 963, 965, 966, 972 phagocytic action of, 121, 123, 123-125,
vestibule of, 966 124
membranous, 966-980, 967—969, 972—980 pseudopods of, 123, 123
cochlea of, 971-974, 972-974 specific granules of, 7 79, 120—121
crista ampullaris, 967-970, 968, 969 nongranular, 117, 779
cupulae of, 970 polymorphonuclear, 117. See Leukocyte(s),
endolymphatic duct of, 965, 967 neutrophilic.
endolymphatic sac of, 967, 970—971 pools of, circulating, 262
hair cells of, 968-970, 969 marginated, 262
organ of Corti of, 972, 973, 974-980 Leukopoietin, 262
otoliths of, 970 Leukorrhea, cervical, 883'
supporting cells of, 969, 970 Leydig cell(s), 826-828, 827-830
utricle of, 965, 967-970, 968, 969 Reinke’s crystals in, 828, 830
nerves of, 981—983, 983, 984 testosterone production of, 832
perilymphatic, 980-981 Ligaments, 167
basilar membrane of, 980—981 Ligands, 50, 423
scala tympani of, 980—981 Light cell, 504
scala vestibuli of, 980—981 Limbus, 920-924, 921-924
Lactase, 665 aqueous veins of, 920
Lactogen, placental. See Somatomammotropin. Schlemm’s canal of, 922, 924
Lactogenesis, 907 scleral spur of, 922, 922
Lacuna (lacunae), Howship’s, 211 spiral, 979-980
of bone, 200, 203 Lines of Retzius, 603, 606
Lamella (lamellae), annulate, 20, 20 Lines of Schreger, 603, 606
circumferential, of bone, 203, 206 Lip, 581, 581
of bone, 200, 202, 206 glands of, 593
Lamina, basal, 71-72 Lipid(s), 22—23, 24
zones of, 71 droplets of, 2
basement, of epithelia, 66 in liver cells, 696
elastic, external, 367, 369 intestinal absorption of, 666—669, 667—669
internal, 367, 368, 369, 371 liver transpor-pof, 705, 706
external. See Lamina, basal. mammary gland production of, 909, 909
fibrous, of nuclear envelope, 32, 33, 35 transport of, 185
Lamina epithelialis, 359 Lipoblast, 181
Lamina lucida, 71 Lipofuscin, 18, 23, 25
Lamina propria, 136 of neurons, 320
Lamina rara, 71 Lipoma, types of, 181
Laminin, 136, 147 Lipoprotein(s), beta, 133-134
Langerhans cell, 423, 554, 554-556, 555 low-density, 133—134
Larynx, 734—735, 735 liver pathway of, 707
Lathyrism, 146 serum, 133—134
collagen defects in, 171 very-low-density, 133, 705, 706
Lattice, microtrabecular, 25, 30—31, 31 Lipoprotein lipase, lipid transport and, 185
Lectin, 423 Lipotropic hormone, 486
Lens, 914, 931-935, 932-935 Liver, 679-708
capsule of, 932, 933 acinus of, 684
ciliary zonule of, 921, 932-933 arterioles of, 684, 684
fibers of, 932, 934 bile canaliculi of, 700-703, 701-703
hyaloideocapsular ligament of, 933 bile ducts of, 708
Lenticular nucleus, putamen of, cell of, 313 bile secretion of, 708
Leptomeninges, 357 bilirubin excretion of, 707-708
Leptotene, 47 blood clotting factors and, 706
Leucine aminopeptidase, 665 blood flow in, 682, 683
Leukocyte(s), 117-132 blood glucose level and, 705
band forms of, 120 blood supply of, 679, 684, 685
basophilic, 119, 126—128, 127 cells of, endothelial, 686, 688, 688-689
cutaneous hypersensitivity and, 128 fat-storing, 690, 690—691, 691
specific granules of, 119, 126, 127 Kupffer, 657, 689, 689-690, 690
classification of, 120 parenchymal, 694—699, 694—699
eosinophilic, 119, 125—126, 126 pit, 691
specific granules of, 125, 126 central veins of, 679, 681
granular, 117 cholangioles of, 703
marginated pool of, 133 cirrhosis of, 704
mononuclear, 117 connective tissue stroma of, 703—704
neutrophilic, 119, 120-125, 121-124 Disse s space of, 691—693, 692, 693
azurophil, 120-121, 722 drug interaction with, 706—707
cytoplasm of, 7 79, 120-121, 722 functional unit of, 682, 683, 683—684
1004 • INDEX

Liver (Continued) Lymph node(s) (Continued)

functions of, 705-708 medulla of, 449, 450, 456
Glisson’s capsule of, 703 medullary cords of, 452
hepatic artery of, 684, 685 nerves of, 457
histological organization of, 679-684, 680— postcapillary venules of, 456-457, 457
683 reticular cells of, 460
histophysiology of, 705—708 sinuses of, 451-454, 451-454
lipid transport and, 705, 706 cells of, 452-453, 454
lobules of, 679—684, 680—683 endothelial cells of, 453
classical, 683 interdigitating cells of, 460
portal, 683 macrophages of, 453
portal canal of, 680, 681 marginal, 451, 451, 452
zonation of, 699-700 medullary, 451, 452, 453, 458
lymph spaces of, 704 medullary cords of, 452
periductal plexus of, 684, 685 subcapsular, 451, 451, 452
perisinusoidal space of, 691—693, 692, 693 veiled cells of, 460
plasma protein synthesis by, 706 subcapsular sinus of, 451
portal vein of, 684 veiled cells of, 456, 460
regeneration of, 704-705 vessels of, afferent, 449, 450
sinusoids of, 679, 684, 685, 686—691, 687— efferent, 449, 450
691 Lymphatic ducts, 404
Lobes, thymic, 436, 437 Lymphatic vessel(s), 400-404, 402, 403
Loop of Henle, 758, 759 afferent, 400
countercurrent multiplier mechanism of, cardiac, 3914
785, 785-786 efferent, 400
long, 769, 770, 771 larger, 403-404
short, 769, 770, 771 valves of, 403
thin limbs of, 769, 771, 771 Lymphoblast(s), development of, 420
Lung(s), alveolar ducts of, 740, 740, 741 proliferating activity of, 421
alveolar macrophages of, 748—750, 749 splenic production of, 477
blood supply of, 751 T lymphocyte transformation to, 412
gas exchange of, 753 Lymphocyte(s), 128-131, 129, 130, 158, 165,
histophysiology of, 752—753 409-410, 410, 412
innervation of, 750-751 antibody response of, 460-461
lymphatics of, 751-752 antigen-dependent proliferation of, 409
neuroepithelial bodies of, 750-751 antigen-independent proliferation of, 409
nonrespiratory functions of, 753 B, 128, 408
respiratory bronchioles of, 739-740, 740, antibody binding of, 415, 415-416, 416
741 antibody production and, 159-160
respiratory portion of, 735, 736 antigen response of, 419-422, 420, 421
Lutein cell, 866 feedback control of, 263
Luteinizing hormone, 486, 491 generation of, 131
ovarian changes and, 885 mitogen stimulation of, 424
ovarian follicle development and, 861 origin of, 260
testosterone production and, 832 surface properties of, 414-417, 415-417
Luteinizing hormone-releasing hormone, ovu¬ circulation of, 430-432, 431
lation and,864 cytolytic, 407
testosterone production and, 832 epithelial invasion by, 80
Luteotropin, 491 functional properties of, 425
Ly antigen, 417 in vitro stimulation of, 128-129, 130
Lymph, 400 intrathymic life span of, 446
Lymph node(s), 449-663, 450-458 lymphoid organs for, 408-409
blood vessels of, 456-457, 457 macrophage immune response and, 422-423
capsule of, 449, 450, 456 marrow site of, 243
cell filtration of, 458 migration of, 131, 430-432, 431
compartmentalization of, 459 motility of, 425
cortex of, 449, 450, 454-456, 455, 456 nonspecific stimulation of, 423-424
cells of, 455-456 null, 408
germinal centers of, 455, 456 of thymic cortex, 439, 440
interdigitating cells of, 456 plaque-forming, 420, 420
primary nodules of, 455 plasma cell relationship to, 420
dendritic cells of, 460 primary lymphoid organ for, 408
germinal centers of, 451, 455, 456, 459 recirculation of, 431, 432
histological organization of, 449-457, 450- secondary lymphoid organs for, 409
457 size of, 409, 411
histophysiology of, 458-462 splenic transit of, 476
immune response of, 458-462 surface antigens of, 446
interdigitating cells of, 456, 460 T, 128, 408"
lymphocyte migration in, 457, 457 antigen recognition by, 417-419
lymphocytopoietic activity of, 459 antigen response of, 417-419
marginal sinus of, 451, 451 helper, 129, 408, 418
 INDEX • 1005

Lvmphocyte(s) (Continued) Mammary gland (Continued)

T, Ly antigens of, 417 fat production by, 905, 907, 908, 909, 909
lymphoblast transformation of, 412 histophysiology of, 907-910, 908—911
memory, 418 IgA secretion of, 910, 910
mitogen stimulation of, 424 lactiferous duct of, 901
origin of, 260 lactiferous sinus of, 902
rosette formation of, 416 lymphatic vessels of, 911
suppressor, 129, 408, 419 merocrine secretion of, 905
surface properties of, 414-417, 415-417 nerves of, 911
target cell interaction of, 419 oxytocin stimulation of, 909
Thy-1 determinant of, 416—417 protein production by, 905, 908, 909, 909
thymectomy and, 445 regression of, 910—911
thymus-derived, 446 resting, 901-903
transmural migration of, 242, 242 epithelium of, 902
Lymphoid dendritic cell, 423 secretory mechanisms of, 904—905, 908
Lymphoid nodule(s), 427—430, 428, 429 stimulation of, 907-909
germinal centers of, 427-430, 428, 429 Mammogenesis, 907
primary, 427 Mammotrophs, 484, 485
secondary, 427 of pars distalis, 483
Lymphoid tissue, 425—430, 426, 428, 429 secretory pathway of, 486
diffuse, 425—427 Manatee, bone formation in, 226, 226, 227
germinal centers of, 427-430, 428, 429 MAP. See Microtubule associated proteins.
nodular, 425, 426 Marrow, bone. See Bone marrow.
Lymphokines, 129, 407, 418 Mast cell(s), 160—164, 161—164, 165
Lymphopoiesis, 259-260 in allergy, 162—163, 163
Lysosome(s), 16-20, 17, 18, 19, 19(t) vs. basophils, 127
enzymes of, 16 Matrix, extracellular. See Extracellular matrix.
inborn defects of, 18, 19(t) Medulla, adrenal, 516, 524-527, 525, 526
micrograph of, 17 Megakaryoblast(s), 256
multivesicular bodies and, 19 Megakaryocyte(s), 256, 257
primary, 18 development of, 256
origin of, 18 marrow site of, 243
secondary, 18 platelet demarcation membranes of, 258,
Lysozyme, function of, 597 258
platelet-forming, 256
residual, 259
Meibomian glands, 568
M bands, of skeletal muscle, 276 Meiosis, 43, 47
Macrocyte, 114 in oocytes, 864—865, 865
Macrophage(s), 154-156, 155, 156, 165 in spermatocytes, 810, 812
activated, 158 Meissner’s corpuscles, 342, 342, 343
alveolar, 748—750, 749 Melanin, 23, 543, 553, 556, 558
elicited, 158 of hair, 563
fixed, 154 of neurons, 320
free, 154 Melanocyte stimulating hormone, 491, 493
immune response of, 422—423 alpha, 493
lamellipodia of, 155 beta, 494-495
marrow site of, 243 Melanocyte(s), 23, 543, 552, 552, 553
monocyte transition to, 155—156, 156 cytocrine secretion of, 557
of lymph node sinuses, 453 of epidermis, 552, 553, 556-558
of thymic cortex, 440 Melanosome, 23, 24, 24, 553, 554, 557-558
phagocytic enhancement of, 156 Melanotropin, 494
resident, 157 Melatonin, 538
Macrophage inhibitory factor (MIF), 419 Membrana perforata, 612
Macrophage-activating factors, 419 Membrane(s), basement. See Lamina, basal.
Macropinocytosis, 50 basilar, 980-981
Macula (maculae), of crista, 967—968 Bowman’s, 917
Macula densa, of kidney, 778, 779 Bruch’s, 925
Macula lutea, 936 cell, 3-6, 4, 5, 6
Major histocompatibility complex, 418 electron microscopy of, 3, 4
Malleus, 962, 962, 963 fluid mosaic model of, 4
Malpighian body (bodies), 464 freeze-fracture analysis of, 5, 5
Maltase, 665 layers of, 3, 4
Mammary gland, 901—912 trilaminar appearance of, 4
active, 903-907, 905-907 cytoplasmic function of, 3
alveolar ducts of, 902 Descemet’s, 918—919, 919, 920
apocrine secretion of, 905 fenestrated, of elastic connective tissue, 169
blood supply of, 91 1 periodontal, 608
cells of, epithelial, 904 postsynaptic, 351, 352
myoepithelial, 903, 903, 904, 905 presynaptic, 351, 352
chronic cystic disease of, 91 1 of myoneural junction, 292
1006 • INDEX

Membrane(s) (Continued) Mitosis, 43, 44, 45, 46, 47

serous, 164, 165 Mixed glands, 97, 98
synovial, 235-237, 236 serous demilunes of, 97, 97
tectorial, 980 Modulation, cell, 209
tympanic, 961, 962, 963, 964 Molars, 602
vestibular, 971, 972 Moll’s glands, 572
Meninges, 359 Monoblast, 254
Menopause, 879 Monocyte(s), 119, 131—132
Merkel cell(s), 109, 110 azurophil granules of, 132, 132
of epidermis, 556, 556, 557 circulating, 154
Meromyosin, heavy, of skeletal muscle, 286, macrophage transition of, 155-156, 156
257 mononuclear phagocyte system and, 156—
light, of skeletal muscle, 286, 287 158, 157, 158
Mesaxon, 331, 334, 334 Mononuclear phagocyte system, 156-158, 157,
Mesenchymal cell, 150, 152, 240, 241. See also 158
Osteoprogenitor cell. Monopoiesis, 254-255
Mesothelial cell, 165 Motility, cell, 49-50
Mesothelium, 60 Motor end plate, 289, 290
Metabolic rate, 185 of skeletal muscle, 338, 339
Metamyelocyte, 250, 250, 253 Motor unit, of skeletal muscle, 338
Metaphase Mucigen, 92, 589
of meiosis, 47 Mucin, 92
of mitosis, 44, 45, 46 Mucocutaneous junction, 543, 558
Metaphysis, 199 Mucosa, gastrit, 624-636, 625—635. See also
in bone formation, 224, 224 Gastric mucosa.
Metaplasia, myeloid, 468 intestinal, 641-660, 641—648, 650, 652, 655-
Microbody. See Peroxisome(s). 659. See also Intestinal mucosa.
Microfibrils, of elastic fibers, 145—146, 145, Mucous cell(s), 92, 92, 97
146 of gastric mucosa, 626—627, 627
Microhlaments, 25, 27-28 of salivary glands, 588, 589, 589-590
Microglia, 157, 157, 346, 346 Mucous glands, 96, 97
Micropinocytosis, 50, 384 Mucous neck cell(s), of gastric mucosa, 629—
Microsome fraction, 7 630, 630
Microtubule associated proteins (MAP), 26, Mucus, 92
27 (t) of goblet cell, 648-649
Microtubule(s), 25, 26, 26-27, 27, 27(t) tracheal, 737
centriolar satellites of, 26 Multivesicular bodies, 19
fusal, 38 Muscle spindle(s), 293-295, 294, 295, 339
of ciliary axonemes, 77, 77 annulospiral endings of, 295, 295
of platelets, 116, 118 components of, 339
protofilaments of, 26 extrafusal fibers of, 339
Microvillus (microvilli), actin filaments of, 72, flower-spray endings of, 295
73 intrafusal fibers of, 293, 294, 339
epithelial, 66, 73 motor terminals of, 295
of intestinal absorptive cells, 644, 646, 646, nuclear bag fibers of, 293, 294
647 periaxial space of, 295
terminal web of, 72 Muscle(s), arrector piii, 559, 560, 561, 567
MIF. See Macrophage inhibitory factor. fast, 299
Milky spots, 164 of facial expression, 559
Mineralocorticoids, 528 skeletal, 272—296. See also Skeletal muscle.
Mitochondrial matrix, 14 slow, 299
Mitochondria-rich cell(s), 504 smooth, 265-272, 266, 268-271. See also
Mitochondrion (mitochondria), 2, 13, 13—16, Smooth muscle.
15 tonic, 299
cristae of, 13, 13 twitch, 299
DNA of, 14-15, 15 Muscular tissue, 265-310
electron micrograph of, 13 Myasthenia gravis, acetylcholine receptors and,
evolution of, 14 293
inner membrane of, 13 Myelin, 323
inner membrane subunits of, 14 Myelin sheath, 323, 329, 331-335, 533, 334,
intercristal space of, 13-14 335
intracristal space of, 14 development of, 332, 334
matrix granules of, 15 intraperiod line of, 331
membrane space of, 14 major dense line of, 331
of adipose tissue, 186 Myelocyte(s), basophilic, 254
of cardiac muscle, 299, 300, 301 eosinophilic, 251, 254, 254
of neurons, 319 neutrophilic, 250, 250-251, 252, 253
of Aenopus oocytes, 15 Myeloid metaplasia, 468
outer membrane of, 13 Myelopoiesis, 259
Mitogens, 423-424 Myocardial infarction, 381
 INDEX • 1007

Myocardium, 396-397. See also Cardiac Nervous system (Continued)

muscle. autonomic. See also Autonomic nervous system.
Myoepithelial cell, 266 adipose tissue and, 184-185
of eccrine sweat glands, 569 Nervous tissue, 311-366. See also Neuron(s).
Myofibril(s), of skeletal muscle, 275-276, 276, development of, 355-357
277 Neural crest, 355
of smooth muscle, 266 Neural tube, 355
substructure of, 280-288, 282-287 Neurilemma, 330n
Myohlament(s), of skeletal muscle, 280, 282 Neurilemmal sheath. See Sheath of Schwann.
of smooth muscle, 268-269 Neurobiotaxis, 356
Myoglobin, 275 Neuroblast, 356
Myoid cell, of seminiferous tubules, 798 Neuroepiphysin, 536
Myometrium, 878 Neurofibril, 316, 317
Myoneural junction, 289-293, 290-293 Neurohlament, 29(t), 30
action potential at, 293 Neuroglia, 311, 345-347, 346
active zones of, 290, 292 ependyma of, 345
presynaptic membrane of, 293 Neuroglial cell, 345-347, 346
synaptic clefts of, primary, 289 Neurohumor, 530
secondary, 290 Neurohypophysis, 479, 479, 494-498, 495-498
synaptic troughs of, 289 histophysiology of, 496-498, 497, 498
synaptic vesicles of, 290, 292 pituicytes of, 495-496
Myosin, 27-28 Neurokeratin, 336, 337
in smooth muscle, 268 Neuromuscular spindle, of skeletal muscle, 339
of platelets, 116 Neuron(s), 312
of skeletal muscle, 280, 282 action potential of, 323
molecular structure of, 286, 287 axon hillock of, 317, 317
Myxedema, 508 bipolar, 325
centrioles of, 319
cholinergic, of intestine, 676
cytoskeleton of, 317, 324, 514, 516
Nail matrix, 568, 568 degeneration of, 353—354
Nail(s), 567, 567-568, 568 primary, 363
cross section of, 568 secondary, 363
histogenesis of, 576 wallerian, 363
Neck cell, of gastric mucosa, 629-630, 630 dendrites of, 320, 321, 322
Negative feedback, endocrine control by, 105, development of, 355—357
' 105 distribution of, 325—329, 326—328
Nephron(s), 757-774, 758-774 diversity of, 325-329, 326-328
blood supply of, 781 doctrine of, 312, 353
Bowman’s capsule of, 757, 757, 759 final common pathway of, 354
collecting ducts of, 774—775 Golgi complex of, 317, 318
cortical, 759 Golgi Type I, 325
distal tubule of, 772, 774, 774 Golgi Type II, 325, 329
filtration slits of, 760, 760, 761, 762, 765 impulse transmission to, 323
glomerular filtrate of, 758 inclusions of, 319-320
glomeruli of, 762, 764, 765 injury of, 362-364
glomerulus of, 757, 758 inter, 329
Henle’s loop of, 758, 759 interrelationships of, 354-355
inulin transport of, 783 lipofuscin of, 320
juxtamedullary, 759 melanin of, 320
macula densa of, 774 mitochondria of, 319
podocytes of, 759, 760, 760—762 motor, 317, 329
proximal tubule of, 757, 759 final common pathway of, 329
brush border of, 768, 768 Nissl bodies of, 317, 317, 319
peptide degradation in, 768 nucleus of, 312, 312, 314, 315
renal corpuscle of, 757, 759—763, 759—761 peptide synthesis of, 350
thin segment of, 769-772, 769—773 perikaryon of, 312, 314, 316—325, 316—319,
tubules of, 757—758 321, 322, 324
collecting, 758, 759 shape variation in, 325
proximal, 763, 766—769, 766-769 postganglionic, 344—345
Nerve hber(s), 329—342 processes of, 320—325, 321, 322, 324
myelin sheath of, 331—335, 333, 334, 335 pseudounipolar, 325, 328
sheath of Schwann of, 330, 330—331, 331 retinal, 940
Nerve growth factors, 357 sensory, 329
Nerve(s), cardiac, 399 development of, 356
peripheral, 335—337, 336—338 star-shaped, 328
somatic, 344 stellate, 328
splanchnic, 342, 344 structure of, 312
visceral, 342, 344 synapse of, 347—355, 348, 349, 352, 353
Nervous system. See also Central nervous system; types of, 312, 313
Peripheral nerve(s); Peripheral nervous system. unipolar, 325
1008 • INDEX

Neuropeptide, 108 Obesity, hypertrophic, 181

Neurophysins, 103 Odontoblast, 612, 613
Neuropil, 329 ultrastructure of, 615
Neurotensin, 305 Oligodendrocyte, 346, 346
Neurotransmitter, 347 Olivary nucleus, neuron of, 313
chemical, 108 Omentum, 164, 165
Neutrophil(s), 119, 120—125, 121-124 milky spots of, 164
azurophil granules of, 120-121, 122, 125 Oocyte(s), 852
chromatin of, 120, 121 corona radiata of, 858
cytoplasm of, 119, 120-121, 122 first polar body of, 864—865, 865
drumstick of, 120, 121 growth of, 854
Golgi complex of, 121 maturation of, 864—865, 865, 866
phagocytic action of, superoxide anions and, meiosis in, 864-865, 865
123, 125 secondary, 864
phagocytic activity of, 121, 123, 123-125, Opsins, 954
124 Opsonins, 123
polymorphonuclear, differentiation of, 255 Opsonization, 170, 407, 423
pseudopods of, 123, 123 Optic nerve, 914, 914, 915, 951
specific granules of, 119, 120—121, 125 Ora serrata, 913, 914, 915
Nexus, of epithelia, 69, 70, 71 Oral cavity, 579-581, 581
Nipple, 901, 902 blood vessels of, 580
Nissl body (bodies), 317, 317, 319 ducts of, 588—589
Node(s), atrioventricular, 305 branching, 588
cardiac, 398—399 intercalated, 589
hemal, 462 glands of, 587-597, 589-592, 594, 595, 596
lymph, 400, 449-663, 450-458. See also lamina propria of, 580
Lymph node(s). lip of, 581, 581
sinoatrial, 305 mucous membrane of, 580-581
Nodes of Ranvier, 330, 330, 335 soft palate of, 581
Nodose ganglion, cell of, 316 Organ of Corti, 972, 973, 974-980, 974-980
Nodules, lymphoid, 427-430, 428, 429 cells of, border, 976
Noduli Arantii, 398 hair, 976-979, 977, 978, 979
Norepinephrine, 108, 530 Hensen’s, 976
release of, adipose tissue and, 185 inner phalangeal, 975-976
Normoblast, 241, 247, 248, 249 inner pillar, 974, 974-975, 975
Nose, 731-734, 732, 733 outer phalangeal, 976, 976, 977
accessory sinuses of, 734 outer pillar, 974, 975
Bowman’s glands of, 734 supporting, 974-976, 974—977
epithelium of, 731-734, 732, 733 spiral limbus of, 979-980
basal cells of, 733, 733 tectorial membrane of, 980
olfactory cells of, 733, 733 Organ system, 1
supporting cells of, 731, 733, 733 Organelles, 1—2, 2
hlia olfactoria of, 733 Organs, 1
mucosa of, 731, 732 Ossicles, auditory, 962, 962-963, 963
nerves of, 733 Ossification, endochondral, 196-197, 197, 216,
sensory stimulation of, 734 218-221, 219, 220
Notochord, tissue of, 196 center of, 218, 219
Nuclear bag, 339 centers of, 224-225
Nuclear chain, 339 primary, 219, 225
Nuclear envelope, 2, 31-35, 32, 33, 34 secondary, 219, 225
fibrous lamina of, 32, 33, 35 collar of, 219, 220
vs. annulate lamellae, 20, 20 ectopic, 231
Nuclear pore channels, 31, 32 epiphysis closure and, 225
Nuclear pore complex, 32, 33 in long bone, 220
annulus of, 32, 33 nretaphysis of, 224, 224
plug of, 32, 34 periosteal band of, 219, 220, 221
spokes of, 32, 34 periosteal collar of, 221
Nucleation, heterogeneous, 221-222, 225 zone of hypertrophy of, 223
Nucleolonema, 41, 43 zone of maturation of, 223
Nucleolus, 2, 31, 40-43 zone of proliferation of, 223
ribosomal RNA in, 41 zone of provisional calcification of, 223
ribosome processing in, 43 zones of, 223-224, 224
Nucleoplasm, 1 intramembranous, 216-218, 217, 218
Nucleosome, 35, 36 primary spongiosa of, 216-217, 217
Nucleus, 31-43, 31-34, 36-43 Osteitis fibrosa, 234
cell, 1 Osteoblast, 206, 208, 209, 210
envelope of, 31-35, 32, 33, 34 Osteoclast(s), 208, 211, 213-216, 214, 215
interphase, chromatin, 32 bone resorption and, 213, 214, 215
nuclear pore channels of, 31, 32 calcitonin and, 213, 215, 216
 INDEX • 1009

Osteoclast(s) (Continued) Ovum (ova) (Continued)

formation of, 216 fertilization of, 865, 865
parathyroid hormone and, 213, 215-216 release of, 862-863, 863
ruffled border of, 211, 214, 215 Oxyhemoglobin, skin color and, 556
Osteocyte(s), 208, 209-211,277 Oxyntic cell(s), of gastric mucosa, 630-632,
endoplasmic reticulum of, 210, 212 630-632
formation of, 210-211 tubulovesicular system of, 631, 633
gap junction of, 210, 27 7 Oxyntic gland(s). See Gastric gland(s).
Golgi complex of, 210, 272 Oxytocin, 103, 495, 496, 498, 498, 879
Osteoid. See Bone matrix. mammary gland stimulation by, 909
Osteolysis, 210-211 release of, 105, 105
Osteomalacia, 224, 233, 235
bone minerals in, 208
Osteon, 200, 203, 204, 205 Pacchionian corpuscles, 362
Osteopetrosis, 216 Pachymeninx. See Dura mater.
Osteoprogenitor cell, 208, 209 Pachytene, 47
Otolith, 970 Palate, soft, 581
Oval window, 962, 962, 963 Palatine gland(s), 596
Ovary (ovaries), 851-873, 853 Pancreas, 716-730, 717-729
blood supply of, 872-873 arterial supply of, 728
endocrine control of, 885 cells of, acinar, 727
endometrial phases and, 885 alpha, 724, 724
follicles of, 852, 854-862, 855-863 beta, 724, 725
antral, 857-858, 860 centroacinar, 727
atresia of, 854, 868-870 delta, 725, 726
Call-Exner bodies of,*857, 858, 858, 859 ducts of, 727, 727-728, 728
cumulus oophorus of, 858, 860 endocrine, 721-725, 722-726
estrogen and, 862 alpha cells of, 722, 724, 724
follicle stimulating hormone and, 861 beta cells of, 722
graafian, 858, 861, 861 cells of, 722-725, 723-726
histophysiology of, 861—862 delta cells of, 722
luteinizing hormone and, 861 histophysiology of, 725-727, 726
macula pellucida of, 862 pancreatic polypeptide cell of, 724
mature, 858, 861, 861 exocrine, 716-721, 717-721
primary, 854, 856, 857, 857 acinar tissue of, 716-720, 718-721
primordial, 854, 855 digestive enzymes of, 720
rupture of, 862—863, 863 histophysiology of, 720-721
secondary, 857-858, 860 secretions of, 720-721
testosterone and, 861 in diabetes, 727
theca, 854, 857, 858, 861, 861 insulin secretion of, 100
unilaminar, 854, 855 islet cell of, 109, 110
germinal epithelium of, 852, 853 lymphatics of, 728
hilus cells of, 872 nerve supply of, 728-729, 729
interstitial gland of, 871 photomicrograph of, 96
interstitial tissue of, 870-872 Pancreatitis, 721
liquor folliculi of, 857 Paneth cell, Lieberktihn’s crypts of, 652, 652-
luteinizing hormone and, 885 654, 653
nerves of, 872-873 Panniculus carnosus, 559
tunica albuginea of, 852, 853 Papilla (papillae), circunrvallate, 582, 582
vestigial organs of, 872 dermal, 543
zona pellucida of, 854, 856 filiform, of tongue, 582, 582, 583, 584, 584
Oviduct, 873-877, 873-877 foliate, 582, 582, 585, 586
blood supply of, 876—877 fungiform, 582, 582, 584, 584
cells of, ciliated, 874, 876 of epithelia, 79
secretory, 874, 876 of hair follicles, 561, 561, 564
contractions of, 876 of tongue, 582-586, 583-586
epithelium of, 75, 874—875, 875, 876 Paracrine glands, 83
ovariectomy and, 875, 875 Paraganglion (paraganglia), 532, 533
histological organization of, 874-876, 874— cells of, 532, 533^
876 ' Paranasal sinuses, 734
isthmus of, 873, 874 Paraneurones, 108-110, 109. See also Paracrine
lymphatics of, 876—877 glands.
muscularis of, 876 types of, 109
nerves of, 877 Parapineal organ, 538
pars interstitialis of, 873, 874 Parathyroid glands, 511-515, 512-514
Ovulation, 862-863, 863 age and, 513
absence of, 882 capillary network of, 514
endocrine control of, 863—864 cells of, 511—513, 512
Ovum (ova). See also Ovary (ovaries), follicles of. chief, 511, 512
blastocyst of, 886, 886 oxyphilic, 511—513, 572
1010 • INDEX

Parathyroid glands (Continued) Peritoneum, 164, 579, 660

histophysiology of, 513—515, 514 Peritonitis, 660
principal cells of, 511, 512 Perivascular cell, 165
transplantation of, 514 Pernicious anemia, 640
Parathyroid hormone, 508, 513-515, 514 Peroxisome(s), 19, 19-20
bone effects of, 233-234 electron micrograph of, 19
bone resorption and, 213, 215-216 nucleoid inclusion of, 18, 18
osteoclasts and, 213, 215-216 Peyer’s patches, 80, 655, 655
osteolysis and, 211 P-face, 5, 5, 6
Paroophoron, 872 PHA. See Phytohemagglutinin.
Parotid glands, 588, 593 Phagoctye, mononuclear. See Macrophage(s).
duct of, 595 Phagocytosis, 16, 7 7
nerves of, 596-597 Phagocyte, 50, 154
Pars distalis, 479, 479, 482-492, 484—490 Phagocytin, 121
acidophils of, 483 Phagocytosis, 50
beta cells of, 483, 484, 486-487, 487 complement enhancement of, 156
blood supply of, 489, 489-490 enhancement of, 170, 423
cells of, 482-489, 484-488 immune, 123
acidophilic, 482 mechanism of, 50, 51
basophilic, 482 neutrophilic, 121, 123, 123-125, 124
chromophilic, 482 opsonization and, 170
chromophobic, 482 Phagosome, 16, 17, 50
terms for, 482-483 Phalangeal cell, inner, 975-976
chromophobes of, 487—488 outer, 976r, 976, 977
control of, 492 Pharynx, 599-600, 600
corticotrophs of, 484, 486 wall of, 600
gonadotrophs of, 486-487, 487 Phenylethanolamine-A'-methyl transferase, glu¬
histophysiology of, 490, 490—492 cocorticoid concentration and, 527
mammotrophs of, 483, 484, 485 Phosphovitellin, 234
nerves of, 489 Phytohemagglutinin (PHA), 424
somatotrophs of, 483, 484, 485 Pia arachnoid, 357, 358
thyrotrophs of, 486 Pia mater, 357, 357, 358-359
Pars infundibularis, 479, 479, 494 Pigment, 23-24, 24
Pars intermedia, 479, 479, 492-494 lipochrome, 23, 25
histophysiology of, 493-494 visual, 954
secretory products of, 493-494 wear-and-tear, 18, 23, 25
stellate cells of, 493 Pillar cell, inner, 974, 974-975, 975
Pars tuberalis, 479, 479, 494 outer, 974, 975
Patching, B lymphocyte stimulation and, 424 Pineal gland, arginine vasotocin of, 540
Penis, 845, 845-848, 847 brain sand of, 536, 537
circumflex veins of, 847, 847 corpora arenacea of, 536, 537
erectile mechanics of, 846-848, 847 histogenesis of, 537
lymphatics of, 848 histological organization of, 535-538
nerves of, 848 histophysiology of, 538, 538-541, 540-542
polsters of, 847, 847 innervation of, 537-538
tunica albuginea, 845-846 of lower vertebrates, 538
Tyson’s glands of, 846 pinealocytes of, 535-536, 539
Pepsin, 632 reproductive process and, 540, 541, 542
Pepsinogen, 632 Pinealocyte, 535, 539
Periarterial lymphoid sheaths, immune re¬ avian, 109, 110
sponse of, 476-477 juxtanuclear area of, 541
Pericardium, 164, 396 synaptic ribbon fields of, 536
Perichondrium, 188 synaptic ribbons of, 535
chondrogenic layer of, 188 Pinna, 961, 962
Pericranium, 200 Pinocytosis, 50
Pericyte, 381, 383 fluid-phase, 51, 57
Perikaryon, of neurons, 312, 314, 316-325, receptor-mediated, 51, 57, 52
316-319, 321, 322, 324 clathrin basket of, 51, 52
shape variation in, 325 Pit cell, of liver, 691
Perilymph, 981, 981 (t) Pituicyte, 495-496
Perimysium, 272 Pituitary gland. See Hypophysis.
Perineurium, 335, 336 Placenta, 886, 888-896, 889-895
Periosteum, 199, 206 cells of, cytotrophoblastic, 891
Peripheral nerve(s), 335-337, 336-338 syncytial trophoblast of, 891, 896
afferent, 337 chorion frondosum of, 894
efferent, 337 chorion laeve of, 894
endings of, 337-342, 339, 341-343 circulation of, 894, 895
spinal roots of, 336 decidua basalis of, 891, 894
Peripheral nervous system, 3 11 decidua capsularis of, 894
Peripolesis, 418
decidua vera of, 891, 892, 894
Peristalsis, 659
establishment of, 891
Placenta (Continued) Proelastin, 146
formation of, 888—896, 890—894 Proerythroblast, 246
histophysiology of, 896 Progesterone, 866, 885
hormone secretion of, 896 ovulation and, 863
structure of, 888—896, 889—894 Proinsulin, 726
villi of, anchoring, 889, 892, 893 Prolactin, 483, 490—491
chorionic, 888—889, 890 lactogenesis and, 907
primary, 888, 890 testosterone production and, 832
secondary, 888, 890 Prolactin cell, 484, 485
stem, 889 Prolactin inhibiting hormone, 492
terminal, 889, 893 Promegakaryocyte, 256
tertiary, 888, 894 Promonocyte, 254
Plaques, fibrous, 381 Promyelocyte, 246, 249, 250—252
Plasma, 132—133 Pro-opiocortin, 491, 494
fibronectin of, 146—147 Pro-opiomelanocortin, 491, 494
Plasma cell(s), 119, 129, 158—160, 158—160, Proparathyroid hormone, 514
410, 413, 413-414, 414 Prophase, of meiosis, 47
antibody response of, 421 of mitosis, 44, 45, 46
B lymphoctye origin of, 159 Proplasmacyte, 414, 421
humoral immunity and, 159, 160 Proprioceptive system, 311
lymphocyte relationship to, 420 Prostaglandins, 879
of thymic cortex, 440 Prostate gland, 840, 841, 842—844, 843
Russell bodies of, 414 concretions of, 843, 843
Plasma membrane, 2 hyperplasia of, 843
Plasmablast, 414, 421 morphology of, 844
Plasmacyte, 410, 413, 413-414, 414 tumors of, 843
antibody response of, 421 Protease, immunoglobulin A, 657
Protein(s), androgen-binding, of Sertoli cells,
of thymic cortex, 440
Plasmalemma. See Cell, membrane of. 833
Plasminogen activator, 170 carrier, 106
Platelet(s), 115—117, 116 integral, 3, 4
adhesion of, 116 mammary gland production of, 909, 909
aggregation of, 116 membrane, peripheral, 5
alpha granules of, 116 microtubule associated, 26
contractile material of, 116 plasma, liver synthesis of, 706, 707
demarcation system of, 258, 258 receptor, ribosome, 85
formation of, 258—259, 259 secretory, mRNAs for, 85
functions of, 116-117 signal codons of, 85
glycogen particles of, 118 signal hypothesis of, 85, 88
life span of, 116 transmembrane, 5
microtubules of, 116, 118 Proteoglycans, in hyaline cartilage, 191
release of, 259 of cartilage, 147, 148-149
secretory granules of, 118 of connective tissue, 136
thrombus of, 117 Prothrombin, 134
Plate(s), epiphyseal, 220, 225 Protoplasm, 1
Pleura, 164, 750 Protropin, 494
Plexus, Auerbach’s, 673, 674, 674, 675 Pseudoeosinophil, 121
intestinal, 672, 673, 674, 674, 675 Pulmonary surfactant, 741, 745, 746
Meissner’s, 674 Pulp, 607-608
myenteric, 672, 673, 674, 674, 675 Pupil, 913
Pneumonocyte. See Alveolar cell(s). dilator of, 930, 931
Podocalyxin, 782 sphincter of, 930, 930
Purkinje cell, 320,321,322, 328
Podocyte, 782
of nephron, 759, 760, 760—762 Purkinje fibers, 305, 306, 307
PWM. See Pokeweed mitogen.
Poikilocytosis, 114
Pokeweed mitogen, 424 Pyloric gland(s), 635, 635
Polyribosome(s), 7
Polysomes, 7
Pore, nuclear. See Nuclear pore complex.
Rathke’s cysts, 493
Pores of Kohn, 746, 748
Reaginic antibody, 127
Portal system, 395
Receptor(s), acetylcholine, 293, 293
Predentin, 612, 614
hormone, 106—108, 107
Preen gland, 569
on phagocytic cell, 50, 51
Pre-proparathyroid hormone, 514
sensory, 311
Prepuce, 846
Rectum, 663—664
Prismatic rod sheath, 605, 605
Reflex, cough, 753
Procentriole, 21
Procollagen, collagen synthesis and, 150 sneeze, 753
Relaxin, 868, 879
Procollagen peptidase, 171
Remodeling, bone, 227, 227-228, 228
collagen synthesis and, 150
1012 • INDEX

Reproduction, adrenal cortex function and, Rhodopsin, 936, 942, 944

529 Ribonucleic acid (RNA), messenger (mRNA),
pineal gland and, 540, 541, 542 31
Reproductive system. See also Penis; Testis. of secretory proteins, 85
female, 851-900. See also Ovary (ovaries); ribosomal (rRNA), classes of, 43
Uterus. nucleolar processing of, 41, 43
endocrine regulation of, 885-886 transfer (tRNA), 31
male, 796—850 Ribosome, 7, 9
accessory glands of, 842-845, 843 nucleolar processing of, 42, 43
Residual bodies, 18 Ribosome fraction, 7
Residual body of Regnaud, 820 Rickets, 224, 235
Respiratory distress syndrome, 753 bone minerals in, 208
Respiratory system, 731—754 cartilage changes in, 197
conducting portion of, 735-739, 736-738 Ridges
histophysiology of, 752—753 epidermal, 543
Rete mirabile mRNA. See Ribonucleic acid, messenger.
of kidney, 780, 781 rRNA. See Ribonucleic acid, ribosomal.
Reticular cell(s), 425, 426 tRNA. See Ribonucleic acid, transfer.
adventitial, of bone marrow, 242 Rod cell(s), 939, 940, 941, 942, 942
of lymph nodes, 460 components of, 942, 942
of thymus, 437 synaptic connections of, 949, 949, 950
Reticular fibers, 136, 143, 143, 144 Rokitansky-Aschoff sinuses, 710
vs. Type IV collagen, 143 Rosettes, T lymphocyte formation of, 416
Reticulocyte, 113—114, 248, 249 Rouleaux, 112, 112
Reticuloendothelial system, 157, 157 Russell bodies, 160, 414
Reticulum, endoplasmic, 6—7
cisternae of, 87
microsome fraction of, 7
mRNA on, 87 S cell, of intestinal mucosa, 651
of osteocytes, 210, 212 Sac, endolymphatic, 967, 970-971
polyribosomes of, 85, 86, 86 Saliva, 587, 588, 597
ribosome fraction of, 7 corpuscles of, 588, 598
rough, 2, 7, 8 Salivary gland(s), 97, 587-588
smooth, 7, 8 blood vessels of, 596
three-dimensional configuration of, 9 cells of, mucous, 588, 589, 589-590
tubules of, 87 myoepithelial, 591, 593
sarcoplasmic, of cardiac muscle, 299-300, seromucous, 590
300, 302 serous, 588, 590, 590, 592
of skeletal muscle, 275, 279, 280, 281, 281 demilunes of Giannuzzi of, 591, 593
Retina, 913, 914, 914, 915, 936-953, 937-939, ducts of, 593, 594, 595
941-953 histophysiology of, 597
cells of, amacrine, 947, 948, 950, 954 lymphatic vessels of, 596
bipolar, 947, 948, 948, 954 mixed, 588
cone, 939, 941-946, 942-945 cells of, 590, 591
horizontal, 945, 947, 954 mucous cells of, 588, 589-590
photoreceptor, 940-945, 941-945 nerves of, 596-597
rod, 939, 940, 941, 942, 942 secretions of, 597
central area of, 952-953 serous cells of, 588
fovea of, 936, 937, 953, 953 Santorini’s duct, 728
ganglion cells of, 950 Sarcolemma, 273, 273
histophysiology of, 953-955 Sarcomere, of skeletal muscle, 276, 285
inner limiting membrane of, 952 Sarcoplasm, 266, 267, 269
inner plexiform layer of, 951, 951-952 cardiac, fine structure of, 298, 298-299, 299
interplexiform cells of, 950 of skeletal muscle, 274-275
layers of, 936, 937, 938 ultrastructure of, 279-288, 280-287
macula lutea of, 936 Sarcotubules, of skeletal muscle, 279, 280, 281
neural, 940 Satellite cells, 328
neuroglial elements of, 952 of skeletal muscle, 274
neurons of, 947 Scala tympani, 980-981
electrophysiology of, 954 Scala vestibuli, 980-981
optic nerve of, 951 Schwann cells, 328, 329
outer limiting membrane of, 945, 947 Schweigger-Seidel sheath, 470, 471
outer plexiform layer of, 948-949, 949, 950 Sclera, 916
pigment epithelial cells of, 939-940 Scurvy, 235
pigment epithelium of, 936, 937, 939, 939- cartilage changes in, 197
940 collagen defects and, 171
supporting elements of, 952 Sebaceous glands, 560, 561, 568-569
synaptic connections in, 948-949, 949, 950 Secretin, 651, 721, 729
visual cells of, 109, 110
Secretion(s). See also Protein(s), secretory.
visual pigments of, 954 apocrine, 92
Retinal, 954 definition of, 83
Secretion(s) (Continued) Skeletal muscle (Continued)
glandular, 83-91 muscle spindles of, 293-295, 294, 295
cellular producdon of, 83-91, 84—91 myofibrils of, 273, 275-276, 276, 276
holocrine, 92 substructure of, 280-288, 282-287
merocrine, 91 myofilaments of, 280, 282
Semen, 848—849 myoglobin of, 275
Seminal vesicles, 840, 841, 842 myoneural junction of, 289-293, 290-293
Seminiferous tubule(s), 822 myosin of, 280, 282
cells of, Sertoli, 798-802, 802, 803, 804 molecular structure of, 286, 287
spermatogenic, 798 nerve endings in, 338—340, 559
supporting, 798 neuromuscular spindles of, 339
epithelium of, 798-802, 799-803 red, 276, 278, 278
cycle of, 820-824, 822-825 sarcolemma of, 273
lamina propria of, 798 sarcomere of, 285
Septum membranaceum, 397 sarcoplasm of, 273, 274-275
Serotonin, 162, 634 ultrastructure of, 279-288, 280-287
Serous cells, of salivary glands, 590, 590, 592 sarcoplasmic reticulum of, 275, 279, 280,
Serous glands, 96 281
Sertoli cell(s), 798-802, 802, 803, 804 sarcotubules of, 279, 280, 281
androgen-binding protein of, 833 satellite cells of, 274
Charcot-Bottcher’s crystalloids of, 802, 803 sensory nerve endings in, 338-340
occluding junctions of, 833-834, 834 T system of, 280
ultrastructure of, 804 T tubule of, 279, 280, 281
Sharpey’s fibers, 202, 206, 206, 207 terminal cisternae of, 279, 280, 281
cement lines of, 203 triads of, 279, 280, 281
Sheath of Hertwig, 617 use hypertrophy of, 273
Sheath of Neumann, 603, 604 vimentin filaments of, 284
Sheath of Schwann, 329, 330, 330-331, 331 white, 276, 278, 278
Sickle cell anemia, 113 Z filaments of, 284
Signal recognition protein (SRP), 88 Z line of, 276, 283
Sinus(es), carotid, 378 Skin, 543—578. See also Dermis; Epidermis.
of lymph nodes, 451-454, 451-454 blood supply of, 572, 574, 574
paranasal, 734 glands of, 568-572, 569-570, 572-574
Rokitansky-Aschoff, 710 histogenesis of, 575-576
Sinusoids, 390—391 lymphatic vessels of, 574
hepatic, 684, 685, 686-691, 687-691 nerves of, 574-575
Skeletal muscle, 272-296 sebaceous glands of, 560, 561, 568-569
A bands of, 275, 283, 284, 285 Skull, diploe of, 200
actin of, 280, 282, 283 dura mater of, 200
alpha-actinin of, 284 growth of, 228
blood supply to, 272, 273, 274 inner table of, 200
Cohnheim’s helds of, 275 outer table of, 200
contraction of, 283, 283 pericranium of, 200
nerve excitation and, 289 Slow-reacting substance of anaphylaxis (SRS-
sliding filament hypothesis of, 288-289 A), 163
cytological heterogeneity of, 276, 278, 278- Smooth muscle, 265-272, 266, 268-271
279, 279 ' actin in, 268
cytology of, 275, 273-276, 275-277 cell-to-cell relations of, 269-270
desmin filaments of, 284 contractile system of, 267—268, 270
diagram of, 282 contraction of, 270-272
disuse hypertrophy of, 273 rhythmic, 271
endomysium of, 272, 272 tonic, 271
epilemmal terminations of, 339 cytoskeleton of, 269
epimysium of, 272, 272 fibers of, 265—266, 266
fibers of association of, 266—267
cytology of, 275, 273-276, 275-277 fine structure of, 267—269, 268, 269
fast, 278 gap junctions of, 270
intermediate, 278 longitudinal section of, 266
red, 278, 275 myofibrils of, 266
slow, 278 myosin in, 268
white, 278, 278 nerve endings in, 337
Golgi complex of, 275 transverse section of, 266
H bands of, 276, 283 vascular, 271
heavy meromyosin of, 286, 287 visceral, 271
histological organization of, 272, 272-273 Sneeze reflex, 753
I bands of, 275, 283, 284 Soft palate, 581
interstitial terminations of, 339 Somatomammotropin, 896
light meromyosin of, 286, 287 Somatomedians, 490
M bands of, 276, 283 Somatostatin, 492, 651
motor end plate of, 289, 290 Somatotropin, 483
motor unit of, 338 Somatotrophs, of pars distalis, 483, 484, 485
1014 • INDEX

Somatotropin. See Growth hormone. Spleen, 464-478, 465-474

Space of Disse, 691-693, 692, 693 arteries of, 469-471, 470, 471
Spectrin, of erythrocyte membrane, 114, 115 capsule of, 464, 465, 468-469
Sperm. See Spermatozoon (spermatozoa). cell storage of, 476
Spermatic cord, 838 central artery of, 469
Spermatid(s), 798, 799, 802, 804 circulation of, closed, 473-475, 474
acrosomal granules of, 816, 817 open, 473-475, 474
spindle bridges of, 814, 814-815, 815 erythrocyte removal by, 475-476
Spermatocyte(s), 798, 804 filtering function of, 475-477
meiosis in, 812-813 germinal centers of, 465, 467
primary, 812 hemoglobin degradation and, 476
proacrosomal granules of, 817 histological organization of, 464-475, 465-
secondary, 802 474
Spermatocytogenesis, 810 histophysiology of, 475-477
Spermatogenesis, 798-802, 799-804, 810-826, immune response of, 476—477
814-816 iron metabolism and, 476
clonal nature of, 815, 815 lymphatic vessels of, 475
meiosis in, 812-813 lymphoblast production of, 477
spindle bridges in, 814, 814-815, 815 malpighian bodies of, 464
stage I of, in guinea pig, 823 marginal zone of, 468
stage VII of, in guinea pig, 823 nerves of, 475
stages of, 810 penicillar arteries of, 469-470
in guinea pig, 821, 822 periarterial lymphoid sheaths of, 464-465,
in humans, 824, 824, 825 465
synaptonemal complexes of, 812 immune response of, 476-477
temperature and, 826, 831 red pulp of, 466, 467, 467-468, 468
Spermatogonium (spermatogonia), 798 cells of, 467-468
Type A, 811 reticular fibers of, 466
Type B, 811 Schweigger-Seidel sheath of, 470, 471
Spermatozoon (spermatozoa), 80, 796, 800, sheathed capillaries of, 470
802-810, 805-811 trabeculae of, 469, 469
acrosomal cap of, 803, 805 vascular tree of. 470
acrosome reaction of, 806 veins of, 471-473
annulus of, 809, 809 venous sinuses of, 471-473, 472, 473
degeneration of, 825-826 venous-arterial union in, 473-475, 474
end piece of, 810, 813 white pulp of, 464, 465, 467
head of, 805, 805-807, 807 Splenic cords of Billroth, 464
acrosomal cap of, 805, 805-806, 807 Spot(s), milky, 164
equatorial segment of, 806, 807 SRP. See Signal recognition protein.
implantation fossa of, 807, 819 SRS-A. See Slow-reacting substance of anaphylaxis.
posterior ring of, 807, 808 Stapes, 962, 962, 963
middle piece of, 808, 808-809, 809, 813 Stem cell(s), 243-245, 244
annulus of, 809, 809 cell lines of, 244
mitochondrial sheath of, 809, 809, 813 committed, 243
neck of, 807-808, 819 development of, thymus and, 446
proximal centriole of, 808, 819 differentiation of, 408-409
nuclear vacuoles of, 807 pluripotential, 243, 244
postacrosomal dense lamina of, 807, 808 unipotential, 243
principal piece of, 809-810, 810-813 Stenson’s duct, 593
fibrous sheath of, 809-810, 810-813 Sterocilium (sterocilia), 74, 74
regeneration of, 824-825 of ear, 968
sex chromosomes of, 814 Steroidogenesis, 102
tail of, 812 Stomach, 624-635, 624-640, 637-639. See
development of, 818, 819 also Gastric mucosa.
movement of, 810 blood supply of, 636-637, 637, 638
Spermiation, 820, 821 body of, 624, 624
Spermiogenesis, 810, 816, 817-820, 818, 819, cardia of, 624, 624
820 fundus of, 624, 624
flagellum of, 818 histophysiology of, 638-640
manchette of, 818, 820 layers of, 624
stages of, 817, 818 mucosa of, 624-636, 625-635
S-phase, of cell cycle, 48, 48 muscularis externa of, 636
Spherocytosis, hereditary, 114 of ruminants, 624
Sphincter, anal, 664 pyloric sphincter of, 636
ileocecal, 659 pylorus of, 624, 624
Sphincter of Oddi, 711 regions of, 624, 624
Sphingomyelinase, defect in, 19(t) secretory activity of, 639-640
Spindle(s), neuromuscular. See Muscle spin- pH and, 640
dle(s). serosa of, 636
Spine, central canal of, 359 submucosa of, 636
Spiral limbus, 979-980 Stratum basale, 545
 INDEX -1015

Stratum corneum, 546, 547, 550, 550, 551 T lymphocyte. See Lymphocyte(s), T.
Stratum germinativum, 545, 547, 548 Taeniae coli, 663
Stratum granulosum, 547, 549 Tanning, 558
cells of, 554 Taste buds, 582, 584, 585, 586-587, 586-
keratohyalin granules of, 547, 549 588
Stratum lucidum, 546, 550 cells of, 586
Stratum spinosum, 547 Taste pores, 586, 587, 588
Stress, neuroendocrine response to, 105-106, Tectorial membrane, 980
106 Teeth. See Tooth.
Stria vascularis, 971, 973, 974 Tela choroidea, 359
Subarachnoid space, 359 Teloglial cell, 289, 290
Sublingual glands, 588 Telophase, of meiosis, 47
Submandibular gland(s), 98, 98, 588, 595 of mitosis, 44, 45, 46
cells of, 594, 595 Tendon(s), 166-167, 167
duct of, 594 fibroblasts of, 166-167, 167
mucous cells of, 591 sensory nerve endings of, 295-296
nerves of, 597 sensory nerve endings to, 340
serous cells of, 591, 592 Tendon organs, 295-296
terminal portion of, 590 Terminal bulbs of Krause, 342
Wharton’s duct of, 595 Testis (testes), 796-832
Substance-P, 305 blood permeability barrier of, 833-834, 834
Substantia compacta, 199 blood supply of, 828, 831, 831-832
Substantia propria, corneal, 167, 168 ductuli efferentes of, 797, 835
Substantia spongiosa, 199 ductus deferens of, 838-842, 839-842
Succus entericus, 665 ductus epididymidis of, 835-838, 836—839
Suckling reflex, neuroendrocrine control of, ejaculatory ducts of, 842
105, 105 endocrine function of, 832
Sucrase, 665 excretory ducts of, 834-842, 836-841
Sulcus, scleral, 920, 921 exocrine function of, 833
Sulfatidase, defect in, 19(t) histophysiology of, 832-834, 834
Supporting cells, of membranous labyrinth, interstitial tissue of, 826-828, 827-830
969, 970 Leydig cells of, 796
of organ of Corti, 974-976, 974-977 lymphatics of, 832
Surfactant, pulmonary, 741, 745, 746 rete testis of, 797, 835
Sweat gland(s), apocrine, 571-572, 572, 573 seminiferous tubules of, 796, 797, 798-802,
eccrine, 569, 569-572, 570, 572, 573 799-803
adluminal, 570 tubuli recti of, 834—835
cells of, 569-570 Testosterone, Leydig cell production of, 832
clear cells of, 569—570, 573 ovarian follicle development and, 861
coiled duct of, 573 Theca, of goblet cell, 92, 92
dark cells of, 569-570, 573 Theca externa, 857, 861
histophysiology of, 571 Theca folliculi, 854, 858, 861, 861
myoepithelial, 569—570 Theca interna, 857, 861
histogenesis of, 576 Theca lutein cell, 866, 868, 869
Sweating, control of, 571 Thermogenesis, nonshivering, 186
Synapse(s), 312 shivering, 186
active zone of, 351, 352 Thoracic duct, 404, 432
axoaxonic, 347, 348 Thrombin, 134
axodendritic, 347, 348 Thrombocyte, 115
axosomatic, 347, 348 Thrombocytopathia, 117
chemical, 347 Thrombocytopenia, 117, 259
electrical, 347 Thrombocytopenic purpura, 117
Gray Type I, 350 Thromboplastid. See Platelet(s).
Gray Type II, 350 Thromboplastin, 134
neuronal, 347-355, 348, 349, 352, 353 platelet, 117
neurotransmitter secretion by, 351 tissue, 117
postsynaptic membrane of, 351 Thrombopoiesis, 255-259, 257, 258
presynaptic membrane of, 351, 352, 352 kinetics of, 259
synaptic cleft of, 348 Thrombosis, cerebral, 381
synaptic vesicles of, 348, 349 coronary, 117
vesicle membrane recycling at, 353, 354 Thrombus, 381
vesicles of, 350 platelet, 117
agranular, 350 Thy-1, 446
granular, 350 Thy-1 determinant, 416—417
Synapsis, of meiosis, 47 Thymic humoral factor, 447
Synarthrosis, 235 Thymic serum factor (FTS), 447
Synchondrosis, 235 Thymocyte, 446
Syncytiotrophoblast, 887, 887 Thymosin, 447
Syndesmosis, 235 Thymus, 437-442, 437-447, 444
Synostosis, 235 capillary of, 444
Synovial membrane, 235—236, 236 cell-mediated immunity and, 445
1016 • INDEX

Thymus (Continued) Tooth (teeth), 602-606, 602-618, 608, 610-

cortex of, 437-440, 438, 439 617
capillary of, 444 alveolar bone support of, 609
lymphocytes of, 439-440, 440 alveoli of, 602
macrophages of, 440 apical foramen of, 602
plasma cells of, 440 cementum of, 602, 602, 607
histogenesis of, 443 crown of, 602, 604
histology of, 436-443, 437-442 deciduous, 602
histophysiology of, 445-447 dentin of, 602, 602-604, 603, 604
involution of, 443, 445 dentinoenamel junction of, 607
accidental, 445 enamel knot of, 609, 611
lobes of, 436, 437 enamel of, 602, 602, 604-607, 605, 606
medulla of, 441-442 enamel organ of, 609
nerves of, 442-443 gingiva of, 602, 602
reticular cells of, 437 Gottlieb’s epithelial attachment of, 608
stem cell development and, 446 growth stages of, 610
vessels of, 442-443, 444 Hertwig’s sheath of, 617
Thyrocalcitonin. See Calcitonin. histogenesis of, 609-617, 610-617
Thyroglobulin, 503 Korff s fibers of, 607, 608
hydrolysis of, 507 membrana perforata of, 612
iodination of, 506-507 neck of, 602
release of, 104 odontoblasts of, 615
secretion of, 100, 101, 507, 507 papilla of, primordium of, 609, 611
synthesis of, 506 periodontal membrane of, 602, 602, 608
Thyroid, 500-510, 501-507, 512 predentin of, 612, 614
cells of, 503-505, 504 pulp cavity of, 602, 603
epithelial, 503-504, 504 pulp of, 602, 602, 607-608
parafollicular, 504-505, 505, 508-509 sac of, 609
principal, 505-506 stellate reticulum of, 609, 611
control of, 508 structure of, 602, 602
follicles of, 500, 501, 502, 503 succedaneous, 602
gelatinous colloid of, 500, 501, 503 tooth germs of, 609, 610, 612
histological organization of, 500-505, 501- Weil’s zone of, 607
505 Trabeculae carneae, 397
histophysiology of, 505-509, 506, 507 Trachea, 735-738, 736, 737
hormones of, 503, 503 cells of, basal, 735-736
metabolic rate control by, 508 brush, 736-737
parafollicular cell of, 109, 110 goblet, 735
thyroglobulin secretion of, 100, 101 cilia of, 752
Thyroid stimulating hormone, 486 epithelium of, 76, 736, 736-737
Thyrotoxicosis, receptor abnormality in, 108 histophysiology of, 752
Thyrotroph, 486 hyaline cartilages of, 737, 737
Thyrotropic hormone, thyroid control by, muscle of, 737, 737
508 submucous glands of, 737
Thyrotropin, 491 Tractus solitarius, ovoid cell of, 313
Thyrotropin releasing factor, 492 Transcytosis, 389
thyroid secretion and, 508 Transferrin, 133, 262
Thyrotropin releasing hormone, 492 Translocation, chromosome, 39
Thyroxin, 503 Transneuronal degeneration, 353-354
Tight junction. See Zonula occludens. Tricarboxylic acid cycle, 14
Tissue(s). See specific tissues, e.g., Blood; Con¬ Trigeminal nucleus, spinal, gelatinosa ceil of,
nective tissue; Epithelium; Skeletal muscle; 313
Smooth muscle. Trigona fibrosa, 397
Tomes’ process, 617, 617 Triiodothyronine, 503
Tongue, 582-588, 582-589 Trisomy 21, 39
glands of, 595-596, 595-596 Trophoblast, forms of, 887, 887
anterior, 595 syncytial, 886, 887, 891, 896
mucous, 596 Tropocollagen, 139
posterior, 596 Tropoelastin, 146
nerves of, 587 Tropomyosin, 287, 288
papillae of, 582-586, 582-586 Troponin, 287, 288
taste buds of, 585, 586-587, 586-588 Tubule(s), dentinal, 603
von Ebner’s glands of, 585
seminiferous, 796, 797, 798-802, 799-803
Tonohbril, 65
T, of skeletal muscle, 279, 280, 281
Tonohlaments, of epithelia, 68, 68 uriniferous, 756-775, 758-774
Tonsil(s), 597-599, 598 Tubulin, 26, 27, 27(t)
crypts of, 598, 598 alpha, 26
lingual, 586 beta, 26
crypts of, 586
Tunic, fibrous, of eye, 916-924, 917-924
palatine, 598, 598, 599
vascular, of eye, 924-931, 925-928, 930,
pharyngeal, 598-599 931
 INDFV • 1017

Tunica adventitia, 367, 368, 369 Vein(s) (Continued)

Tunica albuginea, ovarian, 852, 853 of medium caliber, 393
penile, 845-846 of small caliber, 391—393, 391-393
Tunica intima, 367, 368 portal, 395
of elastic arteries, 368 pulmonary, 394
Tunica media, 367, 368 special types of, 394-395
Turner’s syndrome, 39 valves of, 395, 395
Tympanic cavity, 962-963, 963 Vena cava, 394
Tympanic membrane, 961, 962, 963, 964 Ventricles, of brain, 359
Ventricular cell, 355-356
Venule(s), 382, 391-393, 392, 393, 394
permeability of, 393
Ultimobranchial body, 504 postcapillary, of lymph nodes, 456-457, 457
Ultimobranchial cell, 504 wall of, 392
Urethra, 790-793, 791, 792, 793 Vesicle(s), matrix, in cartilage calcification, 197
female, 792-793, 793 Vestibular membrane, 971, 972
male, 790-792, 791, 792 Vestibule, 966
Littre’s gland of, 792, 793 Villin, 72
Uricase, 699 Villus (villi), arachnoid, 358, 361-362
Urinary bladder, 787, 787—788, 788 chorionic, 888-889, 890
connective tissue of, 789, 790 intestinal, 642, 643, 643, 645
Urinary system, 755—795 of choroid plexus, 360, 361
Urine, excretion of, 787-793, 787—793 placental, 888-889, 890, 892, 893
Urobilinogen, 707 Vision. See also Eye; Retina.
Urogastrone, 658 binocular, 913
Uropygial glands, 569 electrophysiology of, surround effect and,
Uterus, 877-888, 878, 881, 884, 886-888 954
contractility of, 879 panoramic, 913
endometrium of, 878, 879-880 Visual purple, 936
epithelium of, 80 Vitamin A, dehciency of, 235
isthmus of, 883 rhodopsin production and, 940
myometrium of, 878, 878—879 Vitamin C, dehciency of, 235
ovum implantation in, 886-888, 886-888 Vitamin D, dehciency of, 235
Utricle, 965, 967-970, 968, 969 Vitreous body, 935-936
Utriculus masculinus, 840 Vocal cords, 734-735, 735
Utriculus prostaticus, 843 Volkmann’s canals, 203
Uvea, 913, 924-931, 925-928, 930, 931
choroid of, 924—925, 925
ciliary body of, 925—928, 926—928
Wear-and-tear pigment, 18, 23, 25
Weibel-Palade bodies, 368
Wharton’s duct, 595
Vacuole(s), autophagic, 18, 19 White matter, 325, 327, 328
condensing, of glandular cells, 90 tracts of, 325
heterophagic, 18 Wirsung’s duct, 728
phagocytic, 16, 17, 50, 123, 124
Vagina, 896—899, 897
epithelium of, 897
Valve(s), aortic, 398 X chromosome, 36
atrioventricular, 398 Xenopus, oocytes of, DNA of, 15
cardiac, 396, 398, 398
lymphatic, 403
mitral, 396, 398
pulmonic, 398 Y chromosome, 36
semilunar, 396 Yolk sac, 887
tricuspid, 396
Valves of Kerckring, 641, 641, 642
Vas deferens, 797
Vasa recta, 779, 781 Z line, of skeletal muscle, 276, 283
Vasa vasorum, 369 Zona fasciculata, 516, 516, 518, 518, 520
Vasoactive intestinal polypeptide, 651 Zona glomerulosa, 516, 518, 519
Vasopressin, 103, 108, 495, 496, 497, 498 Zona intermedia, 518
body fluid osmolarity and, 497, 498 Zona pellucida, ovarian, 854, 856
renal duct permeability and, 786 Zona reticularis, 516, 518
Vasotocin, arginine, of pineal gland, 540 cells of, 524
Vein(s), 391-395, 391-396 Zone of Weil, 607
capillary bud origin of, 400 Zonula adherens, 66, 67, 67-68
cross section of, 391 Zonula occludens, 66, 66-67, 67
femoral, cross section of, 395 Zygote, 47
histogenesis of, 399—400 Zygotene, 47
of large caliber, 393-394 Zymogen, in acinar cells, 91
r