Fertilization details:

Fertilization begins with gamete fusion (zygote formation). The fusion of a spermatozoon with a
secondary oocyte takes place in the uterine tube, near the ovary:
— to begin, a spermatozoon binds to a specific glycoprotein on the zona pellucida that
surrounds the oocyte [this recognition process precludes union with foreign sperm];
— the spermatozoon releases degradative enzymes (acrosomal reaction) that allows the
sperm cell to penetrate the zona pellucida;
— spermatozoon and oocyte plasma membranes fuse (the secondary oocyte completes meiosis);
— the oocyte precludes fusion with other sperm by immediately canceling its
membrane potential (via Ca++ influx) and then by denaturing its zona pellucida
(via enzymes released by exocytosis from oocyte cytoplasmic granules);
— male & female haploid pronuclei make contact, lose their nuclear membranes, and begin
mitosis (mitosis begins 12 hours after sperm fusion; DNA synthesis takes place before mitosis)
Fertilization ends with the initiation of zygote cell division (the start of cleavage)

Cleavage:
This term refers to the series of mitotic divisions by which the large zygote is fractionated into
numerous “normal size” cells. Each daughter cell of the cleavage process is termed a blastomere.
— cleavage begins with a zygote, progresses through compaction to a morula stage and
terminates at the start of the blastocyst (blastula) stage
— the first eight blastomeres are undifferentiated and have identical potential in domestic mam-
mals; thereafter, blastomeres differentiate into inner & outer cells with different missions
Note: The first cleavage division occurs 1 to 5 days following ovulation (depending on species),
thereafter cells divide about once every 12 hours;
As many as eight generations of mitoses may occur without intervening cell growth (cytoplasmic
increase). Thus, e.g., one 150 micron diameter zygote can becomes a collection of 256
cells, each about 7 microns in diameter.

First Second
Cleavage Cleavage Blastula
Division Division Morula (Blastocyst) inner
outer cell
zona mass
blastomeres
pellucida

blastocoele

blastomeres
inner
blastomeres trophoblasts

A morula [L.= small mulberry] is a solid ball of blastomeres, within a zona pellucida. A
morula typically consists of 16 to 64 blastomeres = four to six cell divisions. Blastomeres become compacted;
cells packed on the inside differentiate from those along the surface of the morula:
— outer blastomeres become flattened and form tight junctions (resulting in reduced permeabil-
ity to fluids); they develop the capacity to secrete fluid (internally); they are destined to
become trophoblasts which form the chorion & amnion (fetal membranes);
— inner blastomeres form gap junctions to maximize intercellular communication; they are des-
tined to become inner cell mass which forms the embryo (plus two fetal membranes).

Gastrulation:
Gastrulation is the morphogenic process that gives rise to three germ layers: ectoderm, meso-
derm, and endoderm. (In a gastrula [Gr.= little stomach] one can see evidence of primitive gut formation.) Gas-
trulation includes the following sequence, beginning with a blastocyst:

— A thickened embryonic disc becomes evident at the blastocyst surface, due to cell prolifera-
tion of the inner cell mass cells. Trophoblast cells overlaying the inner cell mass degenerate in
domestic mammals (in some mammals, e.g., mouse and human, trophoblast cells overlaying the inner cell
mass separate and, instead of degenerating, become amnionic wall.)

— From the inner cell mass, cells proliferate, break loose (delaminate), and migrate to form a
new cell layer inside the trophoblast layer. The new layer of cells is called the hypoblast; it
forms a yolk sac. The remaining inner cell mass may henceforth be called epiblast.

— On the epiblast surface, a primitive streak forms as differential cell growth generates a pair of
ridges separated by a depression. [NOTE: The primitive streak defines the longitudinal axis
of the embryo and indicates the start of germ layer formation.]

— The separation of the hypoblast layer from the epiblast establishes a space (coelom/celom)
deep to the primitive streak. Subsequently, the coelom is temporarily filled by mesoderm that
undergoes cavitation to restablish the coelom that gives rise to body cavities.

— Epiblast cell proliferation along primitive streak ridges becomes the source of a cellular migra-
tion through the streak depression. The migrating cells form endoderm & mesoderm layers.

Hypoblast Formation (three stages)

embryonic disc
embryonic disc
magnified degenerating
trophoblast

blastocoele
inner
cell mass delaminating
trophoblast hypoblast cells
layer

hypoblast
layer

epiblast epiblast

coelom trophoblast
layer

yolk sac
(primitive gut)
coelom hypoblast
layer

yolk sac
(primitive gut)

 — Initial migrating cells join the Dorsal View of Embryonic Disc
hypoblast layer, forming embry-
onic endoderm. (The hypoblast notochord
constitutes yolk sac endoderm.)
— The majority of migrating cells
enter the coelom as primary primitive
node
mesenchyme and become meso-
derm. The primary mesenchyme primitive
migrates laterally and cranially streak
(but not along the midline region
primary
directly cranial to the primitive
mesenchyme
streak where notochord will
form). Note: Mesoderm divides
into: paraxial, intermediate, and NOTE: Arrows indicate the spread of primary
mesenchyme through the primitive streak
lateral mesodermal regions. and between the epiblast and hypoblast
— Cavitation re-establishes a coelom (hoseshoe-shaped) within the lateral mesoderm. The
mesoderm splits into two layers bordering the coelom—somatic mesoderm is attached to the
ectoderm and splanchnic mesoderm is joined to endoderm.

— The remaining epiblast becomes ectoderm which forms skin epidermis & nervous system.

primitive streak
epiblast (ectoderm)

hypoblast endoderm primary mesenchyme (mesoderm)

Formation of the notochord:

The notochord is a rod-shaped aggregate of cells located cranial to the primitive streak of
the embryo. It occupies the midline coelomic space between ectoderm and endoderm that was not
invaded by migrating primary mesenchyme.
The notochord is important because it induces formation of the head, nervous system devel-
opment, and somite formation. It marks the future location of the vertebral column and the base of
the cranium. Its ultimate fate is to become the nucleus pulposus of intervertebral discs.
The notochord develops from the primitive node located at the cranial end of the primitive streak. From the
node, mesoderm-forming cells proliferate and migrate forward into the future head region where they become the rod-
shaped notochord.
Longitudinal Section Through Primitive Node and Notochord

primitive streak notochord cells head process

ectoderm
primitive node

endoderm mesoderm cells


Early Formation of the Nervous System (Neurulation):
Neurulation refers to notochord-induced transformation of ectoderm into nervous tissue. The
process begins during the third week in the region of the future brain and then progresses caudally
into the region of the future spinal cord.

The following steps are involved in Neurulation

neurulation: paraxial mesoderm
— ectodermal cells overlaying neuroectoderm intermediate mesoderm
the notochord become tall
columnar (neuroectoderm); lateral mesoderm
they form a thickened area somatic
designated the neural plate. notochord splanchnic
Other ectodermal epithelium is endoderm
flattened.
neural groove coelom
ectoderm
— a neural groove is formed as
edges of the neural plate be-
come raised on each side of somite
a midline depression. (Apical
ends of individual neuroectoder-
mal cells constrict.)

— a neural tube then forms as neural tube neural crest

the neural groove undergoes
midline merger of its dorsal
edges. The tube separates
from non-neural ecto-
derm which unites dorsal
to it. (Tube formation begins
in the cranial cervical region
of the central nervous system
and progresses cranially and caudally until anterior and posterior neuropores, the last openings, finally close.)

— bilaterally, where the neural groove is joined to non-neural ectoderm, cells detach as the
neural groove closes; the cells proliferate and assume a position dorsolateral to the neural
tube—forming neural crest.

NOTE:
Neural tube becomes the central nervous system, i.e., the brain and spinal cord.

Neural crest cells are remarkable for the range of structures they form. Some cells mi-
grate dorsally and become pigment cells in skin. Other cells migrate ventrally and be-
come neurons and glial cells of the peripheral nervous system, or adrenal medulla cells.
In the head, neural crest forms mesenchyme (ectomesenchyme) which becomes menin-
ges, bone, fascia, and teeth.

Note: Each organ system has a critical period during development when it is most sensi-
tive to external agents (teratogens) that produce birth defects.

Somite formation:
Somites are blocks of mesoderm
located just lateral to the notochord. Gener- otic placode
ally, there are a pair of somites for every
vertebra and a half dozen somite pairs in the
head. The number of somites in an embryo
is indicative of age because somites develop
chronologically, in craniocaudal order. optic
placode
Note: The ventromedial portion of a somite
develops into a sclerotome (which forms
vertebrae, ribs, & basal bones of the pharyngeal
skull), the lateral portion becomes a arches
dermatome (skin dermis), and the rest
of the somite forms a myotome (skeletal heart
muscle).
umbilical
Somites develop as follows: stalk
— mesoderm accumulates on each
side of the notochord; this medially
positioned mesoderm is designated
paraxial mesoderm
— progressing from rostral to caudal
somites
over time, transverse fissures divide
the paraxial mesoderm into blocks
— each block is a somite (epithelioid cells
within a somite block re-orient 90°, from
transverse to the notochord to longitudinal)
— head (occipital) somites develop
from proliferation of local mesenchyme lateral to the cranial end of the notochord
— rostral to the notochord, mesenchyme forms less-developed somites, called somitomeres,
which migrate into pharyngeal arches and form muscles of the jaw, face, pharynx, & larynx.

NOTE:
Mesoderm can exist in two morphologic forms: mesenchyme and epithelioid:
Mesenchyme features aggregates of stellate cells within an abundant extracel-
lular matrix composed of fluid and macromolecules (polymers).
Epithelioid refers to organized cells having distinct apical and basal surfaces;
the latter commonly rests on a basal lamina produced by epithelioid secretion.
Mesoderm can transform from a mesenchyme to epithelioid and vice versa: The
mesoderm that streams through the primitive streak is primary mesenchyme.
Somatic, splanchnic, and somite mesoderm can be temporarily epithelioid.
The temporary epithelioid transforms to a secondary mesenchyme which ulti-
mately forms muscle and connective tissue (including cartilage, bone, liga-
ments, tendons, dermis, fascia, and adipose tissue).
Thus, the term “mesenchyme” refers to the morphologic appearance of embryonic tis-
sue. Although most mesenchyme is mesoderm, the other germ layers can also
form mesenchyme, e.g., ectomesenchyme from neural crest ectoderm.


Somite formation:
Somites are blocks of mesoderm
located just lateral to the notochord. Gener- otic placode
ally, there are a pair of somites for every
vertebra and a half dozen somite pairs in the
head. The number of somites in an embryo
is indicative of age because somites develop
chronologically, in craniocaudal order. optic
placode
Note: The ventromedial portion of a somite
develops into a sclerotome (which forms
vertebrae, ribs, & basal bones of the pharyngeal
skull), the lateral portion becomes a arches
dermatome (skin dermis), and the rest
of the somite forms a myotome (skeletal heart
muscle).
umbilical
Somites develop as follows: stalk
— mesoderm accumulates on each
side of the notochord; this medially
positioned mesoderm is designated
paraxial mesoderm
— progressing from rostral to caudal
somites
over time, transverse fissures divide
the paraxial mesoderm into blocks
— each block is a somite (epithelioid cells
within a somite block re-orient 90°, from
transverse to the notochord to longitudinal)
— head (occipital) somites develop
from proliferation of local mesenchyme lateral to the cranial end of the notochord
— rostral to the notochord, mesenchyme forms less-developed somites, called somitomeres,
which migrate into pharyngeal arches and form muscles of the jaw, face, pharynx, & larynx.

NOTE:
Mesoderm can exist in two morphologic forms: mesenchyme and epithelioid:
Mesenchyme features aggregates of stellate cells within an abundant extracel-
lular matrix composed of fluid and macromolecules (polymers).
Epithelioid refers to organized cells having distinct apical and basal surfaces;
the latter commonly rests on a basal lamina produced by epithelioid secretion.
Mesoderm can transform from a mesenchyme to epithelioid and vice versa: The
mesoderm that streams through the primitive streak is primary mesenchyme.
Somatic, splanchnic, and somite mesoderm can be temporarily epithelioid.
The temporary epithelioid transforms to a secondary mesenchyme which ulti-
mately forms muscle and connective tissue (including cartilage, bone, liga-
ments, tendons, dermis, fascia, and adipose tissue).
Thus, the term “mesenchyme” refers to the morphologic appearance of embryonic tis-
sue. Although most mesenchyme is mesoderm, the other germ layers can also
form mesenchyme, e.g., ectomesenchyme from neural crest ectoderm.


Development of a Cylindrical Body:
The early embryo is flat, but the vertebrate body plan features a cylindrical theme—various
cylindrical structures (derivatives of the gut, neural tube, notochord, etc.) enclosed within a cylindri-
cal body. Transition from a flat embryo to a cylindrical one involves the following developments:

Head Process Formation: Three Stages of

• The cranial end of the embryo grows dorsal- Head Process Formation
ly and forward so that it projects above the region (longitudinal views)
originally in front of the embryo.
• The cylindrical head process elongates
by growth from its base, located in front of the ectoderm
primitive node. Consequently, the most anterior part of mesoderm
the embryo is the oldest. The elongation incorporates the endoderm
most anterior half-dozen somites into the future head. yolk sac
• Within the head process, endoderm is
reflected ventrally upon itself, forming a blind-
ended foregut (future pharynx). head
process
Tail Fold Formation:
• At the caudal end of the embryo, a cylindri-
cal tail fold is formed in a manner similar to that
of the head process.
• Folded endoderm encloses a blind hindgut . oral plate

Lateral Body Folds:

• As the head process elongates upward &
forward, a subcephalic pocket (space) is formed head
ventral to the head process, between the head process
process and extra-embryonic tissue. The bi-
lateral margins of this pocket are lateral body subcephalic
pocket
folds—which constitute the continuity between
the elevated embryo and the relatively flat extra- pharynx
embryonic tissue.
Dorsal
View elevated • Similar folds
head
exist caudally in as-
process
sociation with the tail
lateral process. yolk sac
body pericardium
fold • As the embryo
grows and is elevated dorsally, lateral body folds adduct and join to-
gether ventrally, establishing a tubular embryo separated from flattened
extra-embryonic tissue.
primitive
• Progressing caudally from the head process and cranially from the
node
tail fold, ventral fusion of lateral body folds stops at the umbilicus—leav-
primitive ing a ventral opening in the body wall that allows vessels and the yolk
streak sac and allantois to enter the embryo (and communicate with the gut).

10
 • Ventral fusion of lateral body folds Mesoderm
distinguishes the embryo from extra-embry- = somite neural tube
onic tissue (fetal membranes): = intermediate notochord
= lateral foregut
Embryonic coelom (future body cavities
of the trunk) is distinguished from extra- Lateral
embryonic coelom within fetal membranes. Body
Somatopleure (somatic mesoderm + Folds
ectoderm) that forms body wall is distin- embryonic
coelom
guished from that forming fetal membranes mesentery
(chorion and amnion).
Splanchnopleure (splanchnic mesoderm
+ endoderm) merges bilaterally to form gut
somatopleure extra-embryonic
and mesentery, differentiated from extra- coelom
splanchnopleure yolk
embryonic yolk sac (and allantois). sac

Pharyngeal (Branchial) Arches:

In the head region, anterior to the embryonic coelom, mesenchyme between ectoderm and
endoderm forms a series of dorso-ventral arches demarcated by grooves (clefts). Only the first three
pharyngeal arches are externally evident in mammals. Each arch contains a vessel (aortic arch). Within each
arch, ectomesenchyme (derived from neural crest) gives rise to bone and fascia. Myotomes of somitomeres migrate to
pharyngeal arches to provide skeletal muslculature. Each arch is innervated by one cranial nerve.
The first pharyngeal arch develops into upper and lower jaws and muscles of mastication.
The second into hyoid bones and muscles of the face. The remaining pharyngeal arches form hyoid
bones, larynx and associated muscles. Each arch is innervated by a particular cranial nerve.
The pharynx (foregut) develops five bilateral diverticula that internally demarcate the pharyn-
geal arches. These pharyngeal pouches develop into auditory tube, parathyroid glands, thymus, etc.

NOTE: In fish, five or six branchial [Gr. = gill] arches are well developed. Cells degenerate where
branchial clefts and pharyngeal pouches meet so that the pharynx communicates with the
outside (this occurs only temporarily between the first two arches in mammals). The first arch
forms the jaw apparatus and the rest form gill arches separated by gill slits.

Flexures:
The tube-shaped embryo undergoes three flexures that make it C-shaped. The first occurs
in the future midbrain region, the second in the future neck region, and the third occurs in the tail
region.

Cardiovascular system:
• The cardiovascular system develops early (in the third week after the start of the nervous
system), as the embryo enlarges and diffusion alone becomes inadequate for tissue preservation.
• Angiogenesis (formation of blood vessels) begins in splanchnic mesoderm of the yolk sac,
in the form of blood islands composed of mesenchyme and hemocytoblasts. The latter forms blood
cells and the mesenchyme forms vesicles lined by endothelium. The vesicles coalesce to form vascu-
lar channels and then blood vessels (the latter are formed by budding, fusion, & enlargement).
• Vessels are formed first in extra-embryonic tissue: vitelline (yolk sac) and umbilical (allan-
toic) vessels appear first.
• Ventral to the pharynx, bilateral vessels merge to form a tubular heart; dorsal and ventral aortae are connected
by aortic arches. Also, cranial and caudal cardinal veins return embryonic blood to the heart and umbilical veins return
placental blood to the heart. None of these vessels will persist as such in the adult.
11
 • Ventral fusion of lateral body folds Mesoderm
distinguishes the embryo from extra-embry- = somite neural tube
onic tissue (fetal membranes): = intermediate notochord
= lateral foregut
Embryonic coelom (future body cavities
of the trunk) is distinguished from extra- Lateral
embryonic coelom within fetal membranes. Body
Somatopleure (somatic mesoderm + Folds
ectoderm) that forms body wall is distin- embryonic
coelom
guished from that forming fetal membranes mesentery
(chorion and amnion).
Splanchnopleure (splanchnic mesoderm
+ endoderm) merges bilaterally to form gut
somatopleure extra-embryonic
and mesentery, differentiated from extra- coelom
splanchnopleure yolk
embryonic yolk sac (and allantois). sac

Pharyngeal (Branchial) Arches:

In the head region, anterior to the embryonic coelom, mesenchyme between ectoderm and
endoderm forms a series of dorso-ventral arches demarcated by grooves (clefts). Only the first three
pharyngeal arches are externally evident in mammals. Each arch contains a vessel (aortic arch). Within each
arch, ectomesenchyme (derived from neural crest) gives rise to bone and fascia. Myotomes of somitomeres migrate to
pharyngeal arches to provide skeletal muslculature. Each arch is innervated by one cranial nerve.
The first pharyngeal arch develops into upper and lower jaws and muscles of mastication.
The second into hyoid bones and muscles of the face. The remaining pharyngeal arches form hyoid
bones, larynx and associated muscles. Each arch is innervated by a particular cranial nerve.
The pharynx (foregut) develops five bilateral diverticula that internally demarcate the pharyn-
geal arches. These pharyngeal pouches develop into auditory tube, parathyroid glands, thymus, etc.

NOTE: In fish, five or six branchial [Gr. = gill] arches are well developed. Cells degenerate where
branchial clefts and pharyngeal pouches meet so that the pharynx communicates with the
outside (this occurs only temporarily between the first two arches in mammals). The first arch
forms the jaw apparatus and the rest form gill arches separated by gill slits.

Flexures:
The tube-shaped embryo undergoes three flexures that make it C-shaped. The first occurs
in the future midbrain region, the second in the future neck region, and the third occurs in the tail
region.

Cardiovascular system:
• The cardiovascular system develops early (in the third week after the start of the nervous
system), as the embryo enlarges and diffusion alone becomes inadequate for tissue preservation.
• Angiogenesis (formation of blood vessels) begins in splanchnic mesoderm of the yolk sac,
in the form of blood islands composed of mesenchyme and hemocytoblasts. The latter forms blood
cells and the mesenchyme forms vesicles lined by endothelium. The vesicles coalesce to form vascu-
lar channels and then blood vessels (the latter are formed by budding, fusion, & enlargement).
• Vessels are formed first in extra-embryonic tissue: vitelline (yolk sac) and umbilical (allan-
toic) vessels appear first.
• Ventral to the pharynx, bilateral vessels merge to form a tubular heart; dorsal and ventral aortae are connected
by aortic arches. Also, cranial and caudal cardinal veins return embryonic blood to the heart and umbilical veins return
placental blood to the heart. None of these vessels will persist as such in the adult.
11
 amnionic cavity
chorion
(somatopleure)
hindgut

amnion extra-embryonic coelom

(somatopleure) (lined by mesoderm)
heart
allantois
embryonic (splanchnopleure)
vitelline membrane
coelom
(splanchnopleure)
yolk
sac
allantoic
anllantoic
cavity
cavity

somites

posterior
neuropore

umbilical
head stalk
process heart tail
process
branchial
arch

caudal
cranial otic placode notochord dorsal aorta cardinal vein
cardinal
vein somites

stomach

hindgut
umbilical
heart
aortic arch stalk
optic cup ventral aorta
pharyngeal umbilical allantois
pouch vein
yolk sac
13
Placentation:
The placenta is the area(s) of apposition between uterine lining and fetal membranes where
metabolites are exchanged for sustaining pregnancy.
Apposition areas (placental types) may be diffuse (pig), zonary (carnivore), discoid (primates
& rodents), or involve placentomes. A placentome is a discrete area of interdigitation between a ma-
ternal caruncle and a fetal cotyledon. Equine placentas are microcotyledonary (microplacentomes are
distributed diffusely). Ruminant placentas consist of rows of relatively large placentomes.
Placentas (placentae) may also be classified according to the tissue layers separating fetal and maternal blood.
Uterine epithelium, uterine connective tissue and uterine endothelium may be eroded, giving rise to four placental types:
epitheliochorial (swine, equine, cattle); synepitheliochorial, formerly called syndesmochorial, (sheep, goats); endothelial
chorial (carnivore); and hemochorial (primates & rodents).

chorion
allantois
embryo amnion

somatopleure coelom
gut

splanchnopleure yolk sac

Fetal membranes:
Four fetal membranes develop in a conceptus. Two arise from the trophoblast layer of the
blastocyst (and are continuous with the somatopleure of the embryo). Two arise from the inner cell
mass of the blastocyst (and are continuous with splanchnopleure of the embryo); these two splanch-
nopleure membranes are vascular. The four fetal membranes are:
1. Chorion — from trophoblast, forms the outer boundary of the entire conceptus.
2. Amnion — from trophoblast, is formed by folds of chorion in domestic animals (in humans,
amnion forms by caviation deep to a persistent trophoblast). The amnion encloses the embryo within a fluid-
filled amnionic cavity.
3. Allantois — from the inner cell mass, develops as an outgrowth of hindgut splanchno-
pleure. The allantois grows to fill the entire extra-embryonic coelom, with fluid-filled allantoic
cavity. The outer surface of allantois binds to the inner surface of chorion and the outer surface of
amnion. The allantois is highly vascular and provides the functional vessels of the placenta, via um-
bilical vessels.
4. Yolk sac — from the inner cell mass, develops early (with hypoblast formation) and is
continuous with midgut splanchnopleure. Supplied by vitelline vessels, yolk sac is most important in
egg laying vertebrates. It forms an early temporary placenta in the horse and dog.

Note: The term conceptus refers to the embryo or fetus plus its fetal membranes.

Implantation
Following zona pellucida rupture, the blastocyst is innitially free in the uterine lu-
men (nourished by uterine glands). Implantation of the mobile blastocyct is a gradual
process beginning with apposition leading to adhesion (or invasion in the case of the
human & Guinea Pig). Approximate implantation times are: one week (human); two weeks (dog,
cat, sheep), 3-5 weeks (cattle), 3-8 weeks horse; or delayed up to 4 mons (deer, bears).
12
 Fetal Components of Placentae

Porcine Chorionic Surface

(chorion without allantois)

(folds; diffuse placental contact)

necrotic tip

cervical star
(region over cervix)

Equine Chorionic Surface

(microcotyledons)
Bovine Chorionic Surface
(rows of cotyledons)

Carnivore Chorionic Surface

(zonary placental contact)

Human/Rodent Chorionic Surface

(discoid placental contact)
marginal
hematoma
marginal
hematoma

14