Cellular and Molecular Features of Morphogenesis

• Concept 47.2: Morphogenesis in animals

involves specific changes in cell shape,
position, and adhesion
• Morphogenesis is a major aspect of
development in both plants and animals
– But only in animals does it involve the
movement of cells

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 The Cytoskeleton, Cell Motility, and Convergent
Extension

• Changes in the shape of a cell

– Usually involve reorganization of the
cytoskeleton
– Cytoskeleton: polymeric protein complex under
the plasma membrane providing the physical
strength of cells (actin microfilament,
microtubule, intermediate filaments)

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 The formation of the neural tube
– Is affected by microtubules and microfilaments
Ectoderm

Neural
plate
1 Microtubules help elongate
the cells of the neural plate.

2 Microfilaments at the dorsal

end of the cells may then contract,
deforming the cells into wedge shapes.

3 Cell wedging in the opposite

direction causes the ectoderm to
form a “hinge.”

4 Pinching off of the neural plate

forms the neural tube.

Figure 47.19

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 • The cytoskeleton also drives cell migration, or
cell crawling
– The active movement of cells from one place
to another

• In gastrulation, tissue invagination

– Is caused by changes in both cell shape and
cell migration

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 • Cell crawling is also involved in convergent
extension
– A type of morphogenetic movement in which
the cells of a tissue become narrower and
longer – (gasturula cell migration of frog
embryo)

n ce
e
erg Exten
si o
o nv n
C
Figure 47.20

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 Roles of the Extracellular Matrix and Cell
Adhesion Molecules

• Fibers of the extracellular matrix

– May function as tracks, directing migrating
cells along particular routes
– Ex. wound healing, neuronal path finding

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 Cell-to-Subtrate (bottom plate) interaction
• Several kinds of glycoproteins, including
fibronectin
– Promote cell migration by providing specific
molecular anchorage for moving cells
EXPERIMENT Researchers placed a strip of fibronectin on an artificial underlayer. After positioning
migratory neural crest cells at one end of the strip, the researchers observed the movement of the cells
in a light microscope.

RESULTS In this micrograph, the dashed lines indicate the edges of the fibronectin layer. Note
that cells are migrating along the strip, not off of it.

Direction of migration
50 µm

CONCLUSION Fibronectin helps promote cell migration, possibly by providing anchorage for the
Figure 47.21 migrating cells.

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 Cell-to-Cell Adhesion
• Cell adhesion molecules
– Also contribute to cell migration and stable
tissue structure

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 • One important class of cell-to-cell adhesion
molecule is the cadherins
– Which are important in the formation of the
frog blastula
EXPERIMENT Researchers injected frog eggs with nucleic acid complementary to the mRNA encoding
a cadherin known as EP cadherin. This “antisense” nucleic acid leads to destruction of the mRNA for
normal EP cadherin, so no EP cadherin protein is produced. Frog sperm were then added to control
(noninjected) eggs and to experimental (injected) eggs. The control and experimental embryos that
developed were observed in a light microscope.

RESULTS As shown in these micrographs, fertilized control eggs developed into normal blastulas,
but fertilized experimental eggs did not. In the absence of EP cadherin, the blastocoel did not form properly,
and the cells were arranged in a disorganized fashion.

Control embryo

Experimental embryo
Figure 47.22 CONCLUSION Proper blastula formation in the frog requires EP cadherin.

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Induction – Cell-To-Cell Interaction & Communication
• Concept 47.3: The developmental fate of cells
depends on their history and on inductive
signals
• Coupled with morphogenetic changes
– Development also requires the timely
differentiation of many kinds of cells at specific
locations

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Two general principles
underlie differentiation during embryonic development

• First, during early cleavage divisions

– Embryonic cells must somehow become
different from one another (asymmetric division)

• Second, once initial cell asymmetries are set up

– Subsequent interactions among the embryonic
cells influence their fate, usually by causing
changes in gene expression (induction)

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 Fate Mapping
• Classic studies using frogs
– Gave indications that the lineage of cells
making up the three germ layers created by
gastrulation is traceable to cells in the blastula
Epidermis
Central
Epidermis nervous
system

Notochord

Mesoderm

Endoderm
Neural tube stage
Blastula (transverse section)
(a) Fate map of a frog embryo. The fates of groups of cells in a frog blastula (left) were
determined in part by marking different regions of the blastula surface with nontoxic dyes
of various colors. The embryos were sectioned at later stages of development, such as
Figure 47.23a the neural tube stage shown on the right, and the locations of the dyed cells determined.

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 • Later studies developed techniques
– That marked an individual blastomere during
cleavage and then followed it through
development

(b) Cell lineage analysis in a tunicate. In lineage analysis, an individual cell is injected with a
dye during cleavage, as indicated in the drawings of 64-cell embryos of a tunicate, an
invertebrate chordate. The dark regions in the light micrographs of larvae correspond to
Figure 47.23b the cells that developed from the two different blastomeres indicated in the drawings.

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 Two views of cell fate determination
• Inputs from founder (earlier generated) cells
instruct the fate of the younger cells
(Environmental Restriction)
• Restriction of developmental potential as
development proceeds (Intrinsic Restriction)

• But, all these are available only when cells

divide asymmetrically

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 Establishing Cellular Asymmetries
•To understand at the molecular level how embryonic
cells acquire their fates
– It is helpful to think first about how the basic
axes of the embryo are established

Asymmetric cell division and axis formation in the embryo.url

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 The Axes of the Basic Body Plan
• In nonamniotic vertebrates (ex. Frog)
– Basic instructions for establishing the body
axes are set down early, during oogenesis or
fertilization
– Grey crescent

• In amniotes, local environmental differences

– Play the major role in establishing initial
differences between cells and, later, the body
axes
– Gravity and pH in chick
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 Restriction of Cellular Potency
• In many species that have cytoplasmic
determinants
– Only the zygote is totipotent, capable of
developing into all the cell types found in the
adult

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 • Unevenly distributed cytoplasmic determinants
in the egg cell
– Are important in establishing the body axes

– Set up differences in blastomeres resulting

from cleavage

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 EXPERIMENT
1
Left (control): Right (experimental):
Fertilized Fertilized eggs were
salamander eggs constricted by a
were allowed to thread so that the
divide normally, first cleavage plane
resulting in the restricted the gray
gray crescent being crescent to one
evenly divided blastomere. Gray
between the two crescent
Gray blastomeres.
crescent
2 The two blastomeres were
then separated and
allowed to develop.

Belly
Normal piece Normal
RESULTS Blastomeres that receive half or all of the gray crescent develop into normal embryos, but a blastomere
that receives none of the gray crescent gives rise to an abnormal embryo without dorsal structures. Spemann called it a
“belly piece.”
CONCLUSION The totipotency of the two blastomeres normally formed during the first cleavage division depends on
cytoplasmic determinants localized in the gray crescent.
Figure 47.24
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 • As embryonic development proceeds
– The potency of cells becomes progressively
more limited in all species
– Totipotent until 16-cell stage in mammals

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 Cell Fate Determination and Pattern Formation by
Inductive Signals

• Once embryonic cell division creates cells that

differ from each other
– The cells begin to influence each other’s fates
by induction

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 The “Organizer” of Spemann and Mangold
• Based on the results of their most famous
experiment
– Spemann and Mangold concluded that the
dorsal lip of the blastopore functions as an
organizer of the embryo

Hans Spemann (1869-1941) Hilde Mangold

(The Nobel Prize in Physiology or Medicine 1935) (1898-1924)
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 • The organizer initiates a chain of inductions
– That results in the formation of the notochord,
the neural tube, and other organs
EXPERIMENT Spemann and Mangold transplanted a piece of the dorsal lip of a pigmented newt gastrula to the
ventral side of the early gastrula of a nonpigmented newt.
Pigmented gastrula
(donor embryo)
Dorsal lip of
blastopore

Nonpigmented gastrula
(recipient embryo)
RESULTS During subsequent development, the recipient embryo formed a second notochord and neural tube in
the region of the transplant, and eventually most of a second embryo. Examination of the interior of the double embryo
revealed that the secondary structures were formed in part from host tissue.
Primary embryo

Primary
structures: Secondary (induced) embryo
Secondary
Neural tube
structures:
Notochord
Notochord (pigmented cells)
Neural tube (mostly nonpigmented cells)

CONCLUSION The transplanted dorsal lip was able to induce cells in a different region of the recipient to form
Figure 47.25 structures different from their normal fate. In effect, the dorsal lip “organized” the later development of an entire embryo.

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 Formation of the Vertebrate Limb
• Inductive signals play a major role in pattern
formation
– The development of an animal’s spatial
organization
– Depending on where I am in the body and
who’re my neighbors

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 • The molecular cues that control pattern
formation, called positional information
– Tell a cell where it is with respect to the
animal’s body axes
– Determine how the cell and its descendents
respond to future molecular signals

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 • The wings and legs of chicks, like all vertebrate
limbs
– Begin as bumps of tissue called limb buds

Anterior
(a) Organizer regions. Vertebrate limbs develop from
protrusions called limb buds, each consisting of
mesoderm cells covered by a layer of ectoderm.
Two regions, termed the apical ectodermal ridge AER
(AER, shown in this SEM) and the zone of polarizing
activity (ZPA), play key organizer roles in limb Limb bud ZPA
pattern formation. Posterior

Apical
ectodermal
ridge

Figure 47.26a
50 µm

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 • The embryonic cells within a limb bud
– Respond to positional information indicating
location along three axes

(b) Wing of chick embryo. As the bud develops into a

limb, a specific pattern of tissues emerges. In the
Digits
chick wing, for example, the three digits are always
present in the arrangement shown here. Pattern
formation requires each embryonic cell to receive
some kind of positional information indicating
location along the three axes of the limb. The AER Anterior
and ZPA secrete molecules that help provide this Ventral
information. Proximal Distal
Dorsal
Figure 47.26b Posterior

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 • One limb-bud organizer region is the apical
ectodermal ridge (AER)
– A thickened area of ectoderm at the tip of the
bud
– Generation of limb-bud

• The second major limb-bud organizer region is

the zone of polarizing activity (ZPA)
– A block of mesodermal tissue located
underneath the ectoderm where the posterior
side of the bud is attached to the body
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 • Tissue transplantation experiments
– Support the hypothesis that the ZPA produces
some sort of inductive signal that conveys
positional information indicating “posterior”
EXPERIMENT ZPA tissue from a donor chick embryo was transplanted under the ectoderm in the
anterior margin of a recipient chick limb bud.
Anterior New ZPA

Donor Host
limb limb
bud bud
ZPA
Posterior
RESULTS In the grafted host limb bud, extra digits developed from host tissue in a mirror-image
arrangement to the normal digits, which also formed (see Figure 47.26b for a diagram of a normal
chick wing).

CONCLUSION The mirror-image duplication observed in this experiment suggests that ZPA cells secrete
a signal that diffuses from its source and conveys positional information indicating “posterior.” As the
Figure 47.27 distance from the ZPA increases, the signal concentration decreases and hence more anterior digits develop.

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 • Signal molecules produced by inducing cells
– Influence gene expression in the cells that
receive them
– Lead to differentiation and the development of
particular structures

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 Polydactyly (extratoe) by excessive sonic hedgehog

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