Jimma University

College of Agriculture and Veterinary

Medicine

Veterinary Embryology

Instructor-Nuraddis Ibrahim/Dr
 Course description
• The course of Veterinary Embryology deals with
the study of developmental structure and
mechanisms of the body of animals. Where it
deals embryonic orgin of each organs and it is a
basis for understanding histology, gross
anatomy, physiology etc.
 Course Outline
1. Introduction
1.1. Definition and theories
1.2. Reproductive organs
2. Gametogenesis
2.1. Spermatogenesis
2.2. Oogenesis and ovulation
2.2.1. Oogenesis
2.2.2. Ovulation
2.2.3. Types of eggs
2.2.4. Accessory coverings of eggs
 Course Outline
3. Fertilization
3.1. Union of gametes
3.2. Types of zygotes
3.3. Twin formation
4. Fundamental processes and concepts in
development
4.1. Intracellular synthesis and cell division
4.2. Cell surface and cell adhesion molecules
4.3. Gene activation
4.4. Restriction and determination
 Course Outline
4.5. Differentiation
4.6. Induction
4.7. Cell movement and intercellular
communication
4.8. Cell death
5. Basic embryologic phenomena in
mammals
5.1. Cleavage and segmentation
 Course Outline
5.2. Gastrulation
5.3. Fate of the germ layers and tubulation
6. Definitive Morphogenesis
(Organogenesis)
7. Placentation in mammals
8. Embryology of birds week
8.1. Primordial Morphogenesis
8.2. Secondary Morphogenesis
8.3. Definitive Morphogenesis
 Written assignment
1. Reproductive organ in different species
2. Development of heart, reproductive organ
and urinary system
3. Compare embryonic circulatory, respiratory
and excretory system between birds and
mammals embryo
 Introduction
Embryology is the study of the
development of the fertilized egg (zygote)
through cleavage (mitotic cell divisions),
and differentiation.
Embryology is the study of growth and
differentiation undergone by an organism in
the course of its development from a single
fertilized egg into a highly complex and
independent living like the parents.
 Introduction
Embryology is about developmental biology
examines how all forms of life begin, and how
they develop into fully formed and functioning
organisms.
The higher animals start life as a single cell the
fertilized ovum called zygote.
It is formed by the fusion of a germ cell from the
male parent called spermatozoon and one from
the female called ovum.
The union of these two germ cells to form the
zygote constitutes the process of fertilization and
initiates the life of a new individual.
 Reproductive organs
• Male Reproductive Organs
• The male reproductive system has several
interconnected working parts that must
function together for successful mating to
occur.
• In the reproductive system of a male
mammal, the major organs are the pair of
testicles, Epididymis, scrotum, vas
deferens, urethra, accessory gland and
penis.
 Testicles
The testicles are located outside of the
body cavity in the scrotum (sack which
carries testicles and thermal regulatory
organ) except in birds.
Testes are components of both the
reproductive system (being gonads) and
the endocrine system (being endocrine
glands).
 Testicles
• The testicles play a major role in animal
reproduction by producing sperm, called
spermatozoa.
• They also produce a hormone,
testosterone, which causes the
appearance and behavior of the animal to
have masculine traits.
• Sperm cells enter the Epididymis attached
to each testicle. They are stored there
while they mature and until ejaculation.
 Testicles
Both functions of the testicle, sperm-
forming and endocrine, are under control
of gonadotropic hormones produced by
the anterior pituitary:Luteinizing hormone
(LH) and Follicle stimulating hormone
(FSH).
 Scrotum
• Scrotum is a two-lobed sac protects the
two testicles.
• Regulates the temperature of the
testicles, this is because of its effect on
the production and vitality of sperm.
• Temperature regulation is done by means
of a temperature sensitive layer of muscle
(cremaster muscle) located in the walls of
the scrotum, which relaxes when hot and
contracts when cold.
 Scrotum
• Relaxation increases the relative length of
the scrotum, thus moving the testicles
away from body heat.
• In cold weather just the reverse happens -
the scrotum shortens and the testicles are
held close to the warm body.
 Temperature regulation
• The problem of keeping sperm at a low
enough temperature is even greater in
birds that have a higher body temperature
than mammals.
• For this reason bird’s sperm are usually
produced at night when the body
temperature is lower and the sperm
themselves are more resistant to heat.
 Penis
The penis is the organ of insemination. The
penis deposits the semen within the female
reproductive system.
In all domestic animals it consists of two
cylindrical bodies called the corpora cavernosa
penis.
The spaces of the corpora cavernosa become
filled with blood during sexual excitement,
resulting in erection of the organ.
 Penis
The end of the penis is the glans penis.
The glans penis is richly supplied with nerves
and is the source of the sensations associated
with copulation.
 Penis
• In ram, the penis is characterized by a
filiform appendage containing the urethra.
• The penis of the stallion is vascular and
has urethra that protrudes several
centimeters from the surface of the glans
penis.
• The penis of bull, ram, and boar contains
a sigmoid flexure that is straightened
during erection and extension of the penis.
 Penis
• The sigmoid flexure is an anatomical
structure that provides the means by
which the penis is held inside the body
and sheath except during time of service.
• Strong retractor muscles serve to hold the
penis in the "S" shaped configuration.
• Retracts penis (pulls the penis back within
the sheath)
 Penis
• In dogs and most other carnivores, the
penis contains a bone called the os penis
(it has an actual bone).
• The penis of cat is distinguished by the
presence of spines.
 Vas deferens
• Serves as a transportation tube that
carries the sperm-containing fluid from
each Epididymis to the urethra.
Urethra
• Is the large, muscular canal extending
from the urinary bladder to the end of the
penis. The urethra serves as the common
passage way for the excretory products of
the two male tracts--semen of the
reproductive tract and urine of the urinary
tract.
 Accessory sex glands
• Accessory sex glands (add volume and
nutrients to the sperm-rich fluid coming
from the epididymis).
These glands produce the secretions that
make up most of the liquid portion of the
semen. In addition, the secretions activate
the sperm to become motile.
 Accessory gland
The seminal vesicles are paired glands
that produce about 60% of the semen and
transport and provide nutrients for the
sperm.
Their secretions contain fructose sugar,
ascorbic acid and prostaglandins.
 Seminal vesicles
The fructose is supplied as an energy
source for the sperm, and the
prostaglandins serve to stimulate smooth
muscle contractions in the vagina and
cervix. This is thought to facilitate the
uptake of sperm into the uterus.
Seminal vesicles are important in rats,
bulls, boars and stallions but are absent in
cats and dogs.The largest of all accessory
glands and the one producing the largest
fraction of the seminal fluid, is the seminal
 Prostate gland
Another accessory gland is the prostate
gland, which is located at the neck of the
urinary bladder where it empties into the
urethra.
The prostate is poorly developed in the
bull and does not produce a very large
volume of secretion.
The prostate gland is important in dogs
and humans. It produces an alkaline
secretion that neutralizes the acidity of the
male urethra and female vagina.
 Cowper’s glands
The third accessory gland, the Cowper's glands
(bulbourethral glands), are small, firm glands
located on either side of the urethra.
Cowper’s glands have various functions in
different species. The secretions may lubricate,
flush out urine or form a gelatinous plug that
traps the semen in the female reproductive
system after copulation and prevents other
males of the same species fertilizing an already
mated female.
 Cowper’s glands are absent in bears, dogs,
and aquatic mammals.
 The Female Reproductive Organs
• Some major organs that make up the female
reproductive tract are: pairs of ovaries,
oviducts, Uterus, Vulva, Clitoris and Vagina.
 The Female Reproductive Organs
Ovary
• The ovary is the primary reproductive
organ of the female.
• A female mammal typically has two
ovaries.
• Within each ovary are hundreds of tiny
follicles, or cavities, where the ova are
produced.
• The ovary produces the egg by a process
called oogenesis.
 Ovary
• In contrast to spermatogenesis in the bull
which is continuous, oogenesis is cyclic.
• The ova, or eggs, are the female sex cells.
• Each ovum is the largest single cell in the
body.
• The ovaries also produce the female sex
hormones, estrogen and progesterone.
 Ovary
• The secondary sex organs are, in effect, a
series of tubes which receive the semen
of the male, transport the sperm to the
egg so it can be fertilized, nourish the
fertilized egg (embryo), and expel the
offspring.
• These organs include the vagina, cervix,
uterus, uterine horns, and oviducts (also
called Fallopian tubes) which have a
funnel shaped opening called the
 Ovary
• The oviducts or fallopian tubes carry the
ova from the ovaries to the uterus.
• The funnel shaped end of each oviduct
nearest an ovary is called the
infundibulum.
• At ovulation a follicle ruptures, releasing
an ovum that is caught by the
infundibulum.
 Ovary
• After copulation, sperm moves through the
uterus to the oviduct. Fertilization of the
ovum occurs in the upper end of the
oviduct.
• The fertilized egg, known as a zygote,
moves to the uterus about three days after
fertilization.
 Uterus
• The uterus is the site where the fetus
grows until parturition, or birth.
• Uterus is a Y-shaped structure consisting
of the body, two uterine horns, and the
cervix.
• The size and shape of the uterus vary with
the species.
• The upper part of the uterus consists of
two uterine horns that progress to the
oviducts, or fallopian tubes.
 Uterus
• In most species, except the horse,
pregnancy occurs in the uterine horns.
• In the horse, however, pregnancy occurs
in the body of the uterus.
• The uterus of a mammal that normally
produces a large number of offspring at
each breeding cycle has relatively large
horns and a small body.
 Uterus
• In contrast, the uterus of a mammal that
normally produces a single offspring or
twins has smaller horns and a larger body.
• In mares and sows semen is deposited in
the uterus.
 Cervix- Mouth of the uterus
• Cervix is the lower outlet of the uterus and
is composed primarily of connective tissue
and constitutes the gateway between the
uterus and the vagina.
• It serves as a passageway for sperm
deposited in the vagina and for the fetus
at the time of birth.
• Cervix makes significant changes as the
animal goes from one estrous cycle to
another and during pregnancy or
gestation.
 Cervix- Mouth of the womb
• During pregnancy cervix becomes blocked
with a mucous plug to prevent infection
entering from the vagina.
• It has thick walls and a small opening that
is difficult to penetrate in the cow because
of overlapping or interlocking folds.
• At birth cervix stretches to allow fetus to
pass.
 Vulva
• Vulva is the external opening of the
reproductive and urinary systems.
• The exterior, or the visible parts of the
vulva, consists of two folds called the labia
majora.
• Inside the labia majora are two folds
called the labia minora.
• Vulva provides visual signs of heat.
 Clitoris
• Clitoris is the sensory and erectile organ
of the female.
• The clitoris develops from the same
embryonic tissue as the penis in the male
and produces sexual stimulation during
copulation.
 Vagina
• Vagina lies between vulva and cervix. It is
the passage between the cervix and the
vulva.
• The vagina serves as the receptacle for
the male's penis during service.
• Serves as the female organ of copulation
at mating and as the birth canal at
parturition.
 Vagina
• The lining is moist during estrus and dry
when the animal is not experiencing
estrus.
• In cows and ewes semen is deposited in
vagina.
 Spermatogenesis
• The process by which spermatozoa are
produced in the seminiferous tubules of
the testis is known as spermatogenesis.
• This process takes place by a series of
cell divisions which are followed by an
alteration in the cell structure and a
physiological maturation process.
 Spermatogenesis
• Spermatogenesis is considered to consist
of two stages: spermatocytogenesis,
which is the proliferation of cells by mitotic
and meiotic division to form spermatid and
spermiogenesis, which is the
transformation of the immature spermatid
into spermatozoon.
 Spermatogenesis
• In the adult animal spermatozoa are
formed from spermatogonia or stem cells
which arise from multiplication of the
primordial germ cells or genocytes during
puberty and lie in the basement
membrane of the seminiferous tubules.
• Spermatogenesis is a complex process
and involves varies cell divisions during
which the number of chromosomes is
halved and both nuclear and cytoplasmic
components of the cell extensively reorganized.
 Spermatogenesis
• The developing sperm cells progressively
migrate from the basement membrane to
the lumen of the seminiferous tubules.
• During this time however they remain in
contact with the sertoli cell cytoplasm
which probably nourishes them.
• The spermatogonia which rest on the
basement membrane of the seminiferous
tubules are of two types.
• Type A cells and Type B cells.
 Spermatogenesis
• Type A cells are in close contact with
basement membrane and undergo mitosis
to form additional type A cells
spermatogonia or to form type B
spermatogonia.
• Type B spermatogonia differentiate into
spermatozoa where as type A cells
spermatogonia continue to produce either
one or the other type of daughter
spermatogonia.
 Spermatogenesis
• Type B cells may be in contact with the
basement membrane but generally form a
stratum on the luminal surface of the
basilar layer.
• Type B spermatogonia also divide by
mitotic division before differentiating in to
spermatozoa.
 Spermatogenesis
• The cells formed by the mitotic division of
type B spermatogonia are referred to as
primary spermatocytes these cells are
usually larger than other cells of the
seminiferous tubules and are not in
contact with the basement membrane of
the tubules.
• In turn primary spermatocytes undergo
meiosis which involves reduction of their
chromosomes number by one-half.
 Spermatogenesis
• The daughter cells of this division are
called secondary spermatocytes.
• Secondary spermatocytes quickly divide
to form spermatid marking the end of the
process of spermatocytogenesis.
 Spermatogenesis
• The continuing differentiation of these
cells to form mature spermatozoa is called
spermiogenesis.
• The newly formed spermatids are rounded cells
containing a centrally placed nucleus.
• These cells contain a well defined golgi
apparatus surrounding a central granule known
as the acrosomic granule.
• Spermatid lines the luminal surface of the
seminiferous tubular epithelium.
 Spermatogenesis
• Spermiogenesis begins within the seminiferous
tubules but is not complete until the
spermatozoa reach the efferent duct system of
the testis.
• The transformation of spermatids in to
spermatozoa involves a series of
configurational alteration in the cell, including
the formation of a nuclear cap from the golgi
apparatus of the cell, condensation of the
nucleus, the formation of a motile flagellum and
an extensive lose of cytoplasm.
 Spermatogenesis
• During this transformation nearly all the
ribonucleic acid and considerable water and
glycogen are lost from the cell and the density
of the cell increases.
• Thus the mature spermatozoa contain high
contents of solid matter in proportion to water.
• Most somatic cells contain approximately 70%
of water but spermatozoa contain only 50
percent.
• Spermatogenesis duration varies: 34 days in
mouse; 36 days in stallion; 74 days in human.
 Oogenesis
• The process of germ cell production differ
in male and female in that the supply of
germ cell is replenished (maintained) in
the male where as in the female, germ cell
continue to decrease through the
reproductive life and germ cell number are
greatly increased during Spermatogenesis
by mitosis in the male where as mitosis
ceases at birth in the female and
Oogenesis involve the development of a
limited number of performed germ cells.
 Oogenesis
• Gamete production proceeds in the
embryonic ovary through mitotic division
of the primordial germ cells.
• Mitosis ceases at birth, with the maximal
number of oocytes that a female will ever
have being present at this time.
• If the destruction of the gametes occurs
the animal will become infertile.
 Oogenesis
• Meiosis is soon initiated by factors from
the rete ovari but is arrested at resting
stage (diplotene stage) with resumption of
meiosis not occurring until the onset of
puberty.
• In primates, the interval from the initiation
of meiosis to its resumption in the last
oocytes to be developed can be as long
as 50 to 55 years.
 Oogenesis
• The ovum during its period of growth and
the accumulation of deutoplasm is a
primary oocytes, developmentally
homologues to primary spermatocytes of
the male.
• The cells of the granular zone of the
ovarian follicle are oogonia which have
foregone their chances of becoming
mature ova.
 Oogenesis
• These cells form a protective covering
about the oogonium chosen for growth
into primary oocytes.
• When the full allotment of deutoplasm has
accumulated the investing tissue of theca
is ruptured and the oocytes are liberated
from the ovary.
 Oogenesis
• Almost coincidentally with ovulation, the
first maturation division occurs.
• In this the nuclear material of the primary
oocytes is halved between the two
resulting cells but one of them gets
practically all of the cytoplasm and it
contained all the food material.
 Oogenesis
• Both of these unequal cells are secondary
oocytes. But only the one which received
the entire deutoplasm has any chance to
becoming functional.
• The small oocytes containing practically
no cytoplasm is commonly called polar
body.
• This polar body may undergo an abortive
second maturation division. But it is
doomed to degenerate.
 Oogenesis
• Again in the second maturation division all
the stored food material goes to one cell.
• The cell which receives no deutoplasm is
called the second polar body and like the
first, destined to degenerate.
• The cell which has all the deutoplasm that
might have been divided among four
sisters ootid is the mature ready for
fertilization.
Oogenesis
 Ovulation
• Ovulation is the process of release of ova
from the ovary.
• Before rupture the egg is embedded in
solid mass of follicular cells, the cumulus
oophorus which protrudes in to the fluid
filled cavity.
• Ovulation results in the expulsion of the
ovum including the cumulus oophorus and
follicular fluid with the retention of the
remaining portion of the follicle.
 Ovulation
• In most species of animals it is a
spontaneous process and is triggered by a
balance between the two gonadotropic
hormones FSH and LH in the circulating
blood.
• In other animals such as the rabbit, cat,
mink, and ferret ovulation is induced by
stimulation of cervix by copulation or by
other means.
 Ovulation
• The rupture occurs at the apex of the
follicle and the outer most layer of the
follicle is the first to part, the minor layer
protrudes through the gap to form a
papillae or stigma.
• The stigma thins out and bulges on the
surface of the ovary and becomes
completely avascular.
 Ovulation
• The bulging stigma soon ruptures
releasing some of the thin follicular fluid.
• After a short interval the egg mass moves
towards the opening becoming elongated
as it progresses.
 Ovulation
• More fluid moves through the opening
carrying the egg whose connections with
cumulus oophorus were dissociated
during the later stage of follicular
development into peritoneal cavity.
• It is then picked up by the fimbriated end
of the oviduct.
 Ovulation
• Ovulation results in hemorrhage into the
empty cavity of the follicle and the
surrounding tissues.
• The resulting haemorrhage filled follicle is
called the corpus haemorrhagicum and
can easily be recognized on the surface of
the ovary by its red colour.
 Ovulation
• The granulose cells and the surrounding
cells of the theca interna rapidly proliferate
to form luteal cells.
• These cells develop rapidly within the
corpus haemorrhagicum to form the
corpus luteum.
 Ovulation
• As the corpus luteum develops, its
vascularity also develops from the
surrounding interstitial tissue so that it
acquires a rich blood supply.
• The cells of the corpus luteum produce a
second hormone known as progesterone.
 Ovulation
• If fertilization of ovum occurs and
pregnancy takes place, the corpus luteum
is maintained through out the pregnancy
and serves as an endocrine organ present
during pregnancy.
• If the ovum is not fertilized, the corpus
luteum will degenerate after a period by a
luteolytic factor, a prostaglandin F2α
produced in the uterus and brought to the
ovary by blood circulation.
 Ovulation
• The cells of the corpus luteum after
degeneration are replaced by connective
tissue elements resulting in a scar
formation.
• This scar formation results in the
development of corpus albicans, which
gradually becomes less evident within the
ovary with time.
Follicles
Follicle - tiny structure
that produces the ova
Image
 The types of ova
According to the amount and
distribution of the yolk within the egg
cell (ova)
The types of ova can be:
1.Isolecithal: (equal distribution) and
oligolethial (little yolk) as in mammals.
2.Telolecithal: (yolk at end) and
mediolecithal (medium yolk) as in
Amphibians.
 The types of ova
3.Telolecithal: (yolk at end) and
megalecithal (much yolk) as in birds.
4.Centrolecithal: (yolk at the center) and
megalecithal and is surrounded by clear
cytoplasma in arthropods.
 Formation of the accessory
coverings of bird eggs
• At ovulation, the ovum is surrounded by vitelline
membrane, which contains an interwoven
meshwork of relatively coarse, noncollagenous
protein fibrils.
• VITELLINE MEMBRANE -The clear casing that
encloses the yolk.
• The remainder of the accessory coverings, as
the other components of the egg are called, are
secreted about the ovum during its subsequent
passage toward the cloaca.
 Formation of the accessory
coverings of bird eggs
• First an outer vitelline membrane composed of
finer protein fibrils than those of the original
inner vitelline membrane is laid down around
the ovum when it is in the part of the adjacent to
the ovary.
• Fibrils of this layer project along the oviduct
from opposite ends of the ovum midway
between the animal and vegetal poles and
become enwrapped in albumin that is secreted
in the upper oviduct.
 Formation of the accessory
coverings of bird eggs
• Because of the spirally arranged folds in the
walls of the oviduct, the egg is rotated as it
moves toward the cloaca. This rotation twists
the adherent albumen into the form of spiral
strands projecting at either end of the yolk;
these are known as the chalazae.
• Chalazae - Opaque ropes of egg white, the
chalazae hold the yolk in the center of the egg.
Like little anchors, they attach the yolk’s casing
to the membrane lining the eggshell. The more
prominent they are, the fresher the egg.
 Formation of the accessory
coverings of bird eggs
• Additional albumen, which has been
secreted abundantly in advance of the
ovum by the glandular lining of the
oviduct, is caught in the chalazae and
during the further descent of the ovum is
wrapped around it in concentric layers.
 Formation of the accessory
coverings of bird eggs
• The albumen secreting region of the
oviduct constitutes about half its entire
length.
• One of the major albuminous proteins of
egg white is ovalbumin.
• The synthesis of ovalbumin and other
secretions by the oviduct is a striking
example of a specific response to the to
the action of hormones.
 Formation of the accessory
coverings of bird eggs
• Normally the oviduct of a chicken does not
become capable of secreting the components
of egg white until the bird becomes sexually
mature.
• The shell membranes, which consist of two
sheets of matted organic fibers, are added
farther along in the oviduct.
 Formation of the accessory
coverings of bird eggs
• The shell is secreted as the egg is passing
through the shell-gland portion of the oviduct.
• The entire passage of the ovum from the time
of its discharge from the ovary to the time when
it is ready for laying has been estimated to
occupy about 25 to 26 hours.
• If the completely formed egg reaches the
cloacal end of the oviduct during the middle of
the day, it is usually laid at once; otherwise, it is
likely to be retained until the following day.
 Formation of the accessory
coverings of bird eggs
• This over night retention of the egg is one
of the factors accounting for the variability
in the stage of development reached at
the time of laying.
 YOLK
• Two kind of Yolk color can be
differentiated, white and yellow yolk.
• The yolk is also a source of lecithin, an
effective emulsifier.
 Yellow yolk
Yellow yolk – a major source of vitamins,
minerals, almost half of the protein, and all of the
fat and cholesterol.
The Yellow yolk contains less water and more
protein than the white, some fat, and most of
the vitamins and minerals of the egg. These
include iron, vitamin A, vitamin D, phosphorus,
calcium, thiamine, and riboflavin.
 White yolk
White yolk – Also known as, the latebra is an
area of white yolk located in the center of the
yolk.
It is lower in fat and therefore stands out as
a bright white area in many Magnetic Resonance
Images.
The specific function of the latebra is
uncertain but it may act as a central structure
around which the additional layers of the yolk are
formed.
 ALBUMEN
• The egg white is known as the albumen,
which comes from albus, the Latin word
for “white.”
• Four alternating layers of thick and thin
albumen contain approximately 40
different proteins, the main components of
the egg white in addition to water.
 INNER AND OUTER MEMBRANES
• Lying between the eggshell and egg
white, these two transparent protein
membranes provide efficient defense
against bacterial invasion.
• If you give these layers a tug, you’ll find
they’re surprisingly strong. They’re made
partly of keratin, a protein that’s also in
human hair.
 SHELL
• Bumpy and grainy in texture, an eggshell is
covered with as many as 17,000 tiny pores.
• Eggshell is made almost entirely of calcium
carbonate (CaCO3) crystals.
• It is a semipermeable membrane, which means
that air and moisture can pass through its
pores.
• The shell also has a thin outermost coating
called the bloom or cuticle that helps keep out
bacteria and dust.
 AIR CELL
• An air space forms when the contents of
the egg cool and contract after the egg is
laid.
• The air cell usually rests between the
outer and inner membranes at the egg’s
larger end, and it accounts for the crater
you often see at the end of a hard-cooked
egg.
• The air cell grows larger as an egg ages.
 Germinal disk (blastoderm)
Germinal disk (blastoderm) – a small, circular,
white spot (2-3 mm across) on the surface of the
yolk; it is where the sperm enters the egg.
The nucleus of the egg is in the blastodisc.
The embryo develops from this disk, and
gradually sends blood vessels into the yolk to use
it for nutrition as the embryo develops.
 The coverings of mammalian eggs
• The zona pellucida is seen as a thick clear
girdle surrounded by the cells of the
corona radiata.
• The zona pellucida (plural zonae
pellucidae, also egg coat) is a
glycoprotein membrane surrounding the
plasma membrane of an oocyte.
 The coverings of mammalian eggs
• It is a vital constitutive part of the oocyte,
external but of essential importance to it.
• The zona pellucida first appears in
unilaminar primary oocytes. It is secreted
by both the oocyte and the follicular cells.
• In non-mammalian animals, the zona
pellucida (called vitelline layer) plays an
important role in preventing cross-
breeding of different species, especially in
species that fertilize outside of the body
(e.g. fish).
 The coverings of mammalian eggs
• The zona pellucida is commonly used to
control wildlife population problems by
immunocontraception.
• When the zona pellucida of one animal
species is injected into the bloodstream of
another, it results in sterility of the second
species due to immune response.
• This effect can be temporary or
permanent, depending on the method
used.
The coverings of mammalian eggs
• Porcine zona pellucida is used to keep
deer populations low, and this process is
commonly referred to as "spay-vac".
• There are four major zona pellucida
glycoproteins, termed ZP1-4. ZP1, ZP3
and ZP4 bind to capacitated spermatozoa
and induce the acrosome reaction.
 Zona pellucida glycoproteins
• Successful fertilization depends on the
ability of sperm to penetrate extracellular
matrix surrounding eggs.
• In the mouse:
• ZP3 allows species-specific sperm binding
• ZP2 mediates subsequent sperm binding
• ZP1 cross-links ZP2 and ZP3.
 The Oestrous Cycle
Heat is the time when a female is receptive to
the male and will allow breeding to take place
or time of day when a female will accept a male
for breeding
The oestrous cycle is the sequence of
hormonal changes that occurs through the
ovarian cycle. These changes influence the
behaviour and body changes of the female.
The first hormone involved in the oestrous cycle
is follicle stimulating hormone (F.S.H.), secreted
by the anterior pituitary gland. It stimulates the
follicle to develop.
 The Oestrous Cycle
 As the follicle matures the outer cells begin to secrete the
hormone oestrogen and this stimulates the mammary glands to
develop. It also prepares the lining of the uterus to receive a
fertilised egg.
 Ovulation is initiated by a surge of another hormone from the
anterior pituitary, luteinising hormone (LH).
 This hormone also influences the development of the corpus
luteum, which produces progesterone, a hormone that prepares
the lining of the uterus for the fertilised ovum and readies the
mammary glands for milk production.
 If no pregnancy takes place the corpus luteum shrinks and the
production of progesterone decreases. This causes FSH to be
produced again and a new oestrous cycle begins.
 For fertilisation of the ovum by the sperm to occur, the female
must be receptive to the male at around the time of ovulation.
This is when the hormones turn on the signs of “heat”, and she
is “in season” or “in oestrous”. These signs are turned off
again at the end of the oestrous cycle.
 Oestrous
 During the oestrous cycle the lining of the uterus
(endometrium) thickens ready for the fertilised ovum to be
implanted. If no pregnancy occurs this thickened tissue is
absorbed and the next cycle starts.
 In humans and other higher primates, however, the
endometrium is shed as a flow of blood and instead of an
oestrous cycle there is a menstrual cycle.
 Some species have only one heat period each year and are
called monoestrous.
 The cow is in a group that exhibits heat more than one time per
year and is called polyestrous.
 There is considerable variation in the latter group, however,
from those having estrus continuously throughout the year to
those that have only a few cycles during a restricted season
(seasonal breeders).
 The nonbreeding period is called anestrus.
 Species that are considered to be continuous breeders (such
as the cow) are not without periods of anestrus during which
the estrous cycles stop.
Reproductive cycle data on domestic species
Heat duration
Heat duration Cycle length
(hours) (days)

cattle 12 21
sheep 30 17
horses 6 21
dog 9 -
cat 5 10
swine 44 21
 Signs Of Oestrous Or Heat
 When on heat a bitch has a blood stained discharge from the
vulva that changes a little later to a straw coloured one that
attracts all the dogs in the neighbourhood.
 Female cats “call” at night, roll and tread the carpet and are
generally restless but will “stand” firm when pressure is placed
on the pelvic region (this is the lordosis response).
 A female rat shows the lordosis response when on heat. It will
“mount” other females and be more active than normal.
 A cow mounts other cows (bulling), bellows, is restless and has
a discharge from the vulva.
 Heat (Estrous Cycle) is actually divided into 4 phases of the
cycle.
 Proestrus: ovary is about to release an egg
 Estrus: female receptivity
 Metestrus: uterus prepares for pregnancy, fertilized egg
attaches to uterus
 Diestrus: longest period of cycle, inactive
 Estrous Cycles stop after conception, and begin soon after
Parturition (birth)
 Estrogen has varied effects
1) The development and functioning of the
secondary sex organs,
2) The onset of heat, or estrus, the period of
sexual receptivity,
3) it affects rate and type of growth, especially the
deposition of fat, and
4) it primes or prepares the prepuberal heifer and
post-partum cow for onset of sexual activity.
 Progesterone
Progesterone, the hormone of pregnancy,
suppresses the further development of follicles
and secretion of estrogen.
The female does not come into heat while
progesterone is being produced.
It is also necessary for preparing the uterus to
receive the fertilized egg, and maintains the
proper uterine environment for the continuation
of pregnancy.
Estrogen and progesterone are not completely
separate in their effects since both are
necessary for complete development of some
important organs.
 Progesterone
 The development of the uterus is initiated by estrogen and
completed by progesterone.
 The fertilized egg will not implant and survive in the uterus
unless that tissue has been properly prepared by the action of
estrogen and then by that of progesterone.
 Estrogen causes rhythmic contractions of the uterus.
 Progesterone, on the other hand, has a quieting effect on the
uterus so there are no contractions which might disturb
pregnancy.
 Complete development of the mammary gland also depends
upon both hormones.
 Estrogen promotes the growth of the duct system and
progesterone is necessary for the development of the clusters
of milk-secreting alveoli on the ducts.
 Thus, it can be seen in general that estrogen makes things
happen and progesterone calms them down.
 The production of the ovarian hormones is under the direct
influence of the gonadotrophic hormones produced by the
anterior pituitary gland which is located at the base of the brain.
 Fertilizable life
• Fertilizable life of the egg is the maximum
period during which it remains capable of
fertilization and normal development.
• In most species the egg is capable of
being fertilized for some 12 to 24 hours.
• It rapidly loses its fertilizablity upon
reaching the isthmus and is completely
nonfertilizable after reaching the uterine
horn.
 Fertilizable life
• If the egg is not fertilized it will fragment
into many (2 to 20) cytoplasmic segments
of unequal size and in same cases it may
even resemble a fertilized egg.
• All unfertilized eggs eventually disappear
through complete disintegration of
phagocytosis in the uterus.
 Fertilization
Fertilization is the process of fusion of two
cells, the male and female gametes to form
one single cell, the zygote.
It generally occurs in the oviduct of the farm
animals.
The ovum, after ovulation from the ovary
reaches the oviduct. Where it remain for
about 4 days in great majority of the
species.
 Fertilization
• The size of ovum in rodent is 120-180µm
and in large domestic animals it is 75 to
100 µm in most species; fertilization
begins after the first polar body has been
extruded, so that the sperm penetrate the
ovum while the 2nd reduction division is in
progress.
 Fertilization
• In the horse and dog, however sperm may
enter the ova before the 2nd reduction
division has begun.
• The site of fertilization in all farm and most
other mammals is the lower portion of the
ampulla of the oviduct.
• The ovum, in its mucoprotein coat the
zona pellucida, when enters the ampulla is
still surrounded by a cluster of granulose
cells, which are known as cumulus cells.
 Fertilization
• In most species ova that are not fertilized
degenerate within a few days but in the
horse they remain in the fallopian tube for
several months.
• The main feature of the fertilization lies in
the mingling of paternal and maternal
chromosomes, whereby genetic
information from two different sources
comes to constitute the new individual.
 Fertilization
• The process of fertilization involves, the
penetration of the spermatozoa into the
egg, the activation of egg the formation
and development of male and female
pronuclei and replacement of pronuclei by
chromosomes groups which come
together in the prophase of first cleavage
of the zygote.
 Fertilization
• In addition there are three other
consequences of sperm entry and the
depleted chromosomes number is
restored, sex is determined and through
the medium of the sperm tail paternal
cytoplasmic elements are contributed to
the embryo.
• Spermatozoa normally arrive at the site of
fertilization before the ova; they do not
take part in fertilization for some time.
 Fertilization
• Spermatozoa are incapable of fertilization
until changes in the morphological and
biochemical properties of the acrosome
have occurred.
• This phenomenon is known as
capacitation.
• This takes about 6 hours or more in rabbit
about 2 hours in rat and 1.5 hours in ewe.
• Capacitation make the sperm fit for the
task of penetration in to the cumulous
oophorus and zona pellucida of the egg.
Fertilization
Sperm Structure

nucleus actin
mitochondria acrosome
tail
 Fertilization
• Capacitation normally takes place in the
female reproductive tract.
• Transport of ovum in the oviduct is
facilitated by the ciliary action, contraction
of the oviductal musculature and the
secretion of the oviduct.
• Ampullary isthmus and uterotubal junction
determine the period of stay of the ovum
in the oviduct.
 Fertilization
• The transport of spermatozoa in the
female tract differs to a considerable
extent in different domestic species.
• In the horse and pig with intra uterine
deposition of the ejaculate, the ejaculate
itself has been found to reach the
uterotubal junction, but in most species
with vaginal deposition of the ejaculate the
spermatozoa leave the fluid fraction of the
ejaculate lower in the female reproductive
tract some time in the uterine cervix.
 Fertilization
• The sperm transport in the female
reproductive tract is a passive process, as
a result of contraction of the female
reproductive tract and the action of the
cilia.
• The contractile activity of the vagina and
myometerium play a major role in the
transport of spermatozoa in to and
through the uterus.
 Fertilization
• The pattern and rate of sperm transport
through the oviduct are controlled by
peristalsis and antiperistalsis of oviductal
musculature, complex contraction of
oviductal mucosal folds and of the
mesosalpinx, fluid movements created by
ciliary action and possibly the opening and
closing of the uterotubal junction.
 Fertilization
• In ruminants some spermatozoa reach the
site of fertilization in less than 15 minutes.
• Dead sperm and inert particles are also
quickly transported up the reproductive
tract which demonstrates the small
contribution made by sperm motility.
• The spermatozoa do not remain viable in
the female reproductive tract of most
domestic animals for more than about 24
hours.
 Fertilization
• However, they remain alive for about 5
days in the mare reproductive tract.
• In some species of bats sperm are
actually stored over winter in the uterus
and fertilize eggs which will be ovulated in
the following spring.
• The mating of sperm and eggs appears to
depend upon chance which is of about the
same order in all species, although more
spermatozoa reach the site of fertilization
in sheep and rabbit than in rats and mice.
 Fertilization
• The eggs arrive in the fallopian tube
surrounded by cumulous oophorus which
consist of large number of follicle cells
embedded in jelly like matrix composed of
hyaluronic acid protein complex.
• The spermatozoon carries probably in its
acrosome, an enzyme that is capable of
depolymerizing the hyaluronic acid protein
matrix of the cumulous known as
hyaluronidase.
 Fertilization
• It is believed that the spermatozoa with
the aid of this enzyme, diffusing from
reacted acrosomes, individually digest
path for them selves through the
cumulous and between the coronal cells.
• The cumulous is broken down later during
the course of fertilization.
• Partly through an autolytic process and
partly through the action of bicarbonate
ions in the secretion of fallopian tube.
 Fertilization
• A separate enzyme is thought to be
needed to penetrate the coronal cells and
penetration of zona pellucida is achieved
by means of an enzyme provisionally
called zonalysin.
• During this fertilization activity, the motility
of sperm is helpful in getting the cells
through the cumulous and zona pellucida.
• Only one spermatozoon after a successful
entry in to the egg is required for
fertilization.
 Fertilization
• Participation of more than one
spermatozoon in fertilization is
pathological and is called polyspermy. It
leads to early death of embryo.
• Eggs themselves have imperfect
protection against the penetration of more
than one spermatozoon.
• In most species polyspermy is blocked by
two processes.
 Fertilization
• The first is called the zona reaction, by
which the zona pellucida resist penetration
by further spermatozoa, the 2nd affects
the cell membrane and is known as block
to polyspermy.
• Both reactions are worked by attachment
of sperm head to the vitelline surface.
• The zona reaction is thought to be
induced by a substance released from the
surface of the vitellus and diffusing across
the perivitelline space.
 Fertilization
• Extra sperm which succeed in passing
through the zona pellucida in to the
perivitelline space are called
supplementary sperm.
• In sheep, dog and hamster the zona
reaction is quick and effective and
supplementary sperm are found rarely if at
all.
• In other species like rat and mouse, they
are more common.
 Fertilization
• In the pig extra sperm enter the pellucida
but do not succeed in passing right
through it.
• The rabbit show no zona reaction and up
to 200 supplementary sperm have been
observed in the perivitelline space of the
fertilized ovum.
 Fertilization - Egg structure
Jelly coat

Cortical granules

Plasma Egg cytoplasm

membrane

Vitelline layer

Sperm binding receptor

Events in Fertilization
4 5
3
2
6
1

1. Head of the sperm contacts the jelly coat of the egg

- triggers the release of hydrolytic enzymes which
dissolve the jelly coat and starts the acrosomal
reaction
Events in Fertilization
4 5
3
2
6
1

2. The Acrosomal Reaction

-hydrolytic enzymes dissolve a hole in the jelly coat
-actin fibres begin elongate and form the acrosomal
process
Events in Fertilization
4 5
3
2
6
1

3. The Acrosomal Reaction

- the acrosomal process elongates further and
binds with the sperm binding receptors
-this binding causes the vitelline membrane to
begin to break down
 Events in Fertilization
4 5
3
2
6
1

4. Membrane fusion
- membranes of the sperm and egg fuse
- fast block to
- causes depolarization of the membrane
polyspermy
which turns off the sperm binding
receptors
Events in Fertilization
4 5
3
2
6
1

5. Sperm nucleus and beginning of cortical reaction

- fusion of membranes causes an increase in Ca++ release
- slow block to - Ca++ release causes cortical granules to fuse with egg’s
polyspermy plasma membrane and empty contents into perivitelline
space
- sperm nucleus enters egg cytoplasm
Events in Fertilization
4 5
3
2
6
1

6. Continuation of Cortical Reaction and Activation of Egg

- conversion of vitelline layer into fertilization membrane

-increase in cellular respiration and protein synthesis in egg

(egg activation)
 Twin formation
• Sometimes animals give birth to more than one
offspring at a time. This is normal in litter
bearing animals such as swine, cats, dogs, etc.
• In other animals, multiple offspring are called:
2=twins 3=triplets 4= quadruplets
• There are two types of twins: identical twins
(Monozygotic-fertilized egg splits in two) and
fraternal twins (Dizygotic) -2 separate eggs are
ovulated and fertilized.
• Normally, the length of gestation is shorter for
twin than for single pregnancy.
 Twins

Dizygotic Monozygotic
Fraternal Identical

Many
Variations
 Freemartins
This condition occurs commonly in all species
of cattle and affects most females born as a
twin to a male. It is rare or unknown in other
mammals, including humans.
In cattle with multiple conceptions, the chorionic
placental blood vessels form a common
circulation between the fetuses prior to sexual
differentiation, allowing antimullerian duct
hormone and testosterone secreted by the male
to inhibit development of the female tract, and
masculinize her.
 Her sexual organs do not develop fully, and
her ovaries may even contain testicular tissue.
 Freemartins
When adult, such a freemartin is very like a normal
female in external appearance, but she is infertile, and
behaves more like a castrated male (a steer).
 In ~92% of cases of mixed-sex twins, the females are
sterile.
The male twin is not significantly affected, although (if
he remains entire) his testes may be slightly reduced
in size.
The degree of masculinization of the freemartin
depends on the stage of pregnancy at which the
placental fusion occurs – in about ten percent of such
births no fusion occurs and both calves develop
normally as in other mammals.
 Freemartins
The freemartin cattle cervix is absent.
The ovaries usually fail to develop and remain
small.
Normal and freemartin cattle can be
differentiated on the basis of length of the
vagina and on presence or absence of a cervix.
 In calves (1-4 wk old), the normal vaginal
length is 13-15 cm, while in a freemartin vaginal
length is 5-6 cm.
 Cell-division cycle
• The cell cycle, or cell-division cycle, is
the series of events that take place in a
cell leading to its division and duplication
(replication).
• In cells without a nucleus (prokaryotic),
the cell cycle occurs via a process termed
binary fission.
 Cell-division cycle
• In cells with a nucleus (eukaryotes), the
cell cycle can be divided in two periods:
interphase - during which the cell grows,
accumulating nutrients needed for mitosis
and duplicating its DNA and the mitosis
(M) phase, during which the cell splits
itself into two distinct cells, often called
"daughter cells" and the final phase,
cytokinesis, where the new cell is
completely divided.
 Cell-division cycle
• The cell-division cycle is a vital process by
which a single-celled fertilized egg develops
into a mature organism, as well as the process
by which hair, skin, blood cells, and some
internal organs are renewed.
 Cell cycle phases
• The cell cycle consists of four distinct
phases: G1 phase, S phase (synthesis),
G2 phase (collectively known as
interphase) and M phase (mitosis).
• M phase is itself composed of two tightly
coupled processes: mitosis, in which the
cell's chromosomes are divided between
the two sister cells, and cytokinesis, in
which the cell's cytoplasm divides in half
forming distinct cells.
 Cell cycle phases
• Activation of each phase is dependent on the
proper progression and completion of the
previous one.
• Cells that have temporarily or reversibly
stopped dividing are said to have entered a
state of quiescence called G0 phase.
 interphase
• After cell division, each of the daughter cells
begin the interphase of a new cycle.
• Although the various stages of interphase are
not usually morphologically distinguishable,
each phase of the cell cycle has a distinct set of
specialized biochemical processes that prepare
the cell for initiation of cell division.
 interphase
• Before a cell can enter cell division, it needs to
take in nutrients. All of the preparations are done
during the interphase. Interphase proceeds in
three stages, G1, S, and G2. Cell division
operates in a cycle. Therefore, interphase is
preceded by the previous cycle of mitosis and
cytokinesis. Interphase is also known as
preparatory phase, in this stage nucleus and
cytosol division does not occur. The cell
prepares for division.
 G1 phase

• The first phase within interphase, from the end of the

previous M phase until the beginning of DNA
synthesis is called G1 (G indicating gap). It is also
called the growth phase. During this phase the
biosynthetic activities of the cell, which had been
considerably slowed down during M phase, resume at
a high rate. This phase is marked by the use of 20
amino acids to form millions of proteins and later on
enzymes that are required in S phase, mainly those
needed for DNA replication. Duration of G1 is highly
variable, even among different cells of the same
species. It is under the control of the p53 gene.
 S phase
• The ensuing S phase starts when DNA
replication commences; when it is complete, all
of the chromosomes have been replicated, i.e.,
each chromosome has two (sister) chromatids.
Thus, during this phase, the amount of DNA in
the cell has effectively doubled, though the
ploidy of the cell remains the same. During this
phase, synthesis is completed as quickly as
possible due to the exposed base pairs being
sensitive to external factors such as any drugs
taken or any mutagens (such as nicotine).
 G2 phase

• The cell then enters the G2 phase, which

lasts until the cell enters mitosis. Again,
significant biosynthesis occurs during this
phase, mainly involving the production of
microtubules, which are required during
the process of mitosis. Inhibition of protein
synthesis during G2 phase prevents the
cell from undergoing mitosis.
 Mitosis (M Phase/Mitotic phase)

• The relatively brief M phase consists of nuclear

division (karyokinesis). The M phase has been broken
down into several distinct phases, sequentially known
as:
• prophase,
• metaphase,
• anaphase,
• telophase
• cytokinesis (strictly speaking, cytokinesis is not part of
mitosis but is an event that directly follows mitosis in
which cytoplasm is divided into two daughter cells)
 Mitosis
• Mitosis is the process by which a eukaryotic
cell separates the chromosomes in its cell
nucleus into two identical sets in two nuclei.[3] It
is generally followed immediately by
cytokinesis, which divides the nuclei,
cytoplasm, organelles and cell membrane into
two cells containing roughly equal shares of
these cellular components. Mitosis and
cytokinesis together define the mitotic (M)
phaseof the cell cycle - the division of the
mother cell into two daughter cells,
genetically identical to each other and to
their parent cell. This accounts for
approximately 10% of the cell cycle.
 Mitosis
• Mitosis occurs exclusively in eukaryotic cells, but
occurs in different ways in different species. For
example, animals undergo an "open" mitosis,
where the nuclear envelope breaks down before
the chromosomes separate, while fungi such as
Aspergillus nidulans and Saccharomyces
cerevisiae (yeast) undergo a "closed" mitosis,
where chromosomes divide within an intact cell
nucleus.[4] Prokaryotic cells, which lack a
nucleus, divide by a process called binary
fission.
 Mitosis
• The process of mitosis is complex and highly
regulated. The sequence of events is divided
into phases, corresponding to the completion of
one set of activities and the start of the next.
These stages are prophase, prometaphase,
metaphase, anaphase and telophase. During
the process of mitosis the pairs of
chromosomes condense and attach to fibers
that pull the sister chromatids to opposite sides
of the cell. The cell then divides in cytokinesis,
to produce two identical daughter cells.[5]
 Mitosis
• Because cytokinesis usually occurs in conjunction with
mitosis, "mitosis" is often used interchangeably with
"M phase". However, there are many cells where
mitosis and cytokinesis occur separately, forming
single cells with multiple nuclei in a process called
endoreplication. This occurs most notably among the
fungi and slime moulds, but is found in various groups.
Even in animals, cytokinesis and mitosis may occur
independently, for instance during certain stages of
fruit fly embryonic development.[6] Errors in mitosis
can either kill a cell through apoptosis or cause
mutations that may lead to cancer.
 Regulation of eukaryotic cell
cycle
• Regulation of the cell cycle involves
processes crucial to the survival of a cell,
including the detection and repair of
genetic damage as well as the prevention
of uncontrolled cell division. The molecular
events that control the cell cycle are
ordered and directional; that is, each
process occurs in a sequential fashion and
it is impossible to "reverse" the cycle.
 G0 phase
• The term "post-mitotic" is sometimes used to refer to
both quiescent and senescent cells. Nonproliferative
cells in multicellular eukaryotes generally enter the
quiescent G0 state from G1 and may remain
quiescent for long periods of time, possibly indefinitely
(as is often the case for neurons). This is very
common for cells that are fully differentiated. Cellular
senescence occurs in response to DNA damage or
degradation that would make a cell's progeny
nonviable; it is often a biochemical reaction; division of
such a cell could, for example, become cancerous.
Some cells enter the G0 phase semi-permanentally
e.g., some liver and kidney cells.