FERTILIZATION

The sequence of fertilization is a complex process that involves a series of coordinated

molecular events. Typically, fertilization occurs in the ampulla of the uterine tube, although it can
occur in other parts of the tube, excluding the body of the uterus. If the oocyte (egg cell) is not
fertilized in the ampulla, it moves slowly along the tube to the body of the uterus, where it
degenerates and is resorbed.

Chemical signals, known as attractants, are secreted by the oocyte and surrounding follicular
cells. These signals guide the capacitated sperms (sperms that have undergone necessary
changes for fertilization) toward the oocyte, a process known as sperm chemotaxis.

The fertilization process itself is initiated by the contact between a sperm and an oocyte. This
interaction marks the beginning of a series of molecular events that culminate in the
intermingling of maternal and paternal chromosomes at the metaphase of the first mitotic
division of the zygote. The zygote is the unicellular embryo resulting from the fusion of the
sperm and the egg.

It's crucial for the various stages of fertilization to occur seamlessly for the zygote to develop
normally. Defects at any stage of these events may lead to the death of the zygote. The entire
fertilization process takes approximately 24 hours.

Studies involving transgenic and gene knockout animals have provided insights into the
molecular mechanisms of fertilization. Carbohydrate-binding molecules and gamete-specific
proteins on the surface of sperms play roles in sperm-egg recognition and union. Understanding
these molecular aspects is essential for comprehending the intricacies of reproductive
processes in animals.

The process of fertilization involves several distinct phases that ensure the successful fusion of
sperm and egg, leading to the formation of a genetically unique zygote. Here is a detailed
summary of these phases:

1. **Passage of a Sperm through the Corona Radiata:**

- The sperm first navigates through the corona radiata, which involves the dispersal of
follicular cells surrounding the oocyte.
- Enzymes, including hyaluronidase from the sperm acrosome, likely contribute to the
dispersal, and tubal mucosal enzymes may also play a role.
- Movements of the sperm tail are crucial for penetrating the corona radiata.

2. **Penetration of the Zona Pellucida:**

- The sperm then penetrates the zona pellucida, a critical phase in fertilization initiation.
- Enzymes such as esterase, acrosin, and neuraminidase from the sperm acrosome contribute
to the dissolution or loosening (lysis) of the zona pellucida, creating a path for the sperm.

3. **Zona Reaction:**
 - After penetrating the zona pellucida, a zona reaction occurs, changing its properties and
making it impermeable to other sperm.
- Lysosomal enzymes released by cortical granules near the oocyte's plasma membrane
contribute to the zona reaction.

4. **Fusion of Cell Membranes:**

- The plasma membranes of the oocyte and sperm fuse, creating an opening for the entry of
the sperm's head and tail into the oocyte's cytoplasm.
- The sperm's cell membrane and mitochondria, however, remain behind.

5. **Completion of the Second Meiotic Division:**

- The penetration of the oocyte by the sperm triggers the completion of the second meiotic
division.
- This results in the formation of a mature oocyte and a second polar body, with the nucleus of
the mature oocyte becoming the female pronucleus.

6. **Formation of the Male Pronucleus:**

- Within the oocyte's cytoplasm, the nucleus of the sperm enlarges to form the male
pronucleus, while the sperm's tail degenerates.
- The male and female pronuclei are morphologically indistinguishable.
- DNA replication occurs in both pronuclei, and the oocyte with two haploid pronuclei is called
an ootid.

7. **Zygote Formation:**
- The ootid becomes a zygote as the pronuclei fuse into a single diploid aggregation of
chromosomes.
- Chromosomes in the zygote align on a cleavage spindle in preparation for subsequent
cleavage.

8. **Genetic Uniqueness and Chromosomal Inheritance:**

- The zygote is genetically unique, with half of its chromosomes coming from the mother and
half from the father.
- This process forms the basis of biparental inheritance and contributes to the variation of the
human species.
- Chromosomal sex is determined at fertilization, with XX resulting in a female and XY
resulting in a male.

9. **Functional Effects of Fertilization:**

- Fertilization stimulates the penetrated oocyte to complete the second meiotic division.
- It restores the normal diploid number of chromosomes (46) in the zygote.
- Fertilization results in the variation of the human species through the mixing of maternal and
paternal chromosomes.
- The chromosomal sex of the embryo is determined by the type of sperm (X or Y) that
fertilizes the oocyte.
 - Additionally, fertilization causes metabolic activation of the ootid and initiates the cleavage of
the zygote.

The cleavage of the zygote is a crucial developmental process marked by successive mitotic
divisions, leading to a rapid increase in the number of cells known as blastomeres. Here is a
detailed summary of the cleavage process:

1. **Initiation of Cleavage:**
- Cleavage begins approximately 30 hours after fertilization, as the zygote is still within the
zona pellucida.
- The zygote undergoes repeated mitotic divisions, resulting in an increasing number of
blastomeres.

2. **Successive Cleavage Divisions:**

- Cleavage divisions continue to follow one another, leading to the formation of progressively
smaller blastomeres.
- As the cleavage progresses, the embryonic cells become smaller with each division.

3. **Change in Blastomere Shape and Compaction:**

- After the nine-cell stage, blastomeres undergo a change in shape.
- Blastomeres tightly align against each other, forming a compact ball of cells. This process is
known as compaction.
- Compaction is likely mediated by cell-surface−adhesion glycoproteins, facilitating greater
cell-to-cell interaction.

4. **Role of Compaction in Blastocyst Formation:**

- Compaction is a prerequisite for the segregation of internal cells that give rise to the
embryoblast (inner cell mass) of the blastocyst.
- The compacted ball of cells allows for increased cell-to-cell interaction and sets the stage for
the formation of distinct cell populations within the developing embryo.

5. **Hippo Signaling in Segregation:**

- Hippo signaling plays an essential role in segregating the embryoblast from the trophoblast.
This signaling pathway likely contributes to the differentiation of cell fates within the developing
embryo.

6. **Formation of the Morula:**

- When there are 12 to 32 blastomeres, the developing human conceptus is referred to as a
morula.
- The internal cells of the morula are surrounded by trophoblastic cells.
- The morula typically forms approximately 3 days after fertilization as it enters the uterus.

In summary, cleavage involves a series of mitotic divisions of the zygote, resulting in the
formation of blastomeres. Compaction, mediated by cell-surface−adhesion glycoproteins,
facilitates cell-to-cell interaction and sets the stage for the segregation of internal cells into the
embryoblast. Hippo signaling is crucial in this segregation process. The formation of the morula
marks an intermediate stage in embryonic development, with subsequent events leading to the
formation of the blastocyst.

The formation of the blastocyst is a critical stage in embryonic development, involving the
differentiation of cells and the establishment of key structures. Here is a detailed summary of the
formation of the blastocyst:

1. **Appearance of the Blastocystic Cavity:**

- Approximately 4 days after fertilization, as the morula enters the uterus, a fluid-filled space
known as the blastocystic cavity forms inside the morula.
- The fluid enters through the zona pellucida, leading to the separation of blastomeres into two
parts.

2. **Formation of Trophoblast and Embryoblast:**

- The blastocystic cavity divides the blastomeres into two distinct parts:
- A thin outer cell layer called the trophoblast, which gives rise to the embryonic part of the
placenta.
- A group of centrally located blastomeres called the embryoblast, which gives rise to the
embryo itself.

3. **Early Pregnancy Factor and Pregnancy Test:**

- Trophoblastic cells secrete early pregnancy factor, an immunosuppressant protein that
appears in maternal serum within 24 to 48 hours after fertilization.
- Early pregnancy factor forms the basis of a pregnancy test during the first 10 days of
development.

4. **Blastocyst Formation and Attachment:**

- During blastogenesis, the conceptus (embryo and its membranes) is called a blastocyst.
- The embryoblast projects into the blastocystic cavity, while the trophoblast forms the wall of
the blastocyst.
- After floating in uterine secretions for about 2 days, the zona pellucida gradually degenerates
and disappears, allowing the blastocyst to increase in size.

5. **Implantation and Trophoblast Differentiation:**

- Approximately 6 days after fertilization, the blastocyst attaches to the endometrial epithelium.
- The trophoblast proliferates rapidly and differentiates into two layers: an inner layer of
cytotrophoblast and an outer layer of syncytiotrophoblast.
- The syncytiotrophoblast consists of a multinucleated protoplasmic mass with no observable
cell boundaries.

6. **Syncytiotrophoblast Invasion and Erosion:**

 - Finger-like processes of syncytiotrophoblast extend through the endometrial epithelium and
invade the connective tissue.
- By the end of the first week, the blastocyst is superficially implanted in the endometrium,
deriving nourishment from eroded maternal tissues.
- Syncytiotrophoblast produces enzymes that erode maternal tissues, facilitating the
blastocyst's implantation and penetration into the endometrium.

7. **Formation of Hypoblast:**
- Around 7 days after fertilization, a layer of cells known as the hypoblast (primary endoderm)
appears on the surface of the embryoblast facing the blastocystic cavity.
- Comparative embryologic data suggest that the hypoblast arises through the delamination of
blastomeres from the embryoblast.

In summary, the formation of the blastocyst involves the establishment of distinct cell
populations (trophoblast and embryoblast), the appearance of the blastocystic cavity, and the
initiation of implantation with the trophoblast undergoing rapid differentiation. The
syncytiotrophoblast plays a crucial role in the invasion and erosion of maternal tissues,
facilitating the blastocyst's implantation in the endometrium. The appearance of the hypoblast
marks an important step in embryonic development.

SECOND WEEK OF DEVELOPMENT

The completion of blastocyst implantation, occurring during the second week of development, is
a highly regulated process involving trophoblast differentiation and interaction with the receptive
endometrium. Here is a detailed summary:

1. **Timing of Implantation:**
- Implantation of the blastocyst is finalized within a specific time frame, occurring 6 to 10 days
after ovulation and fertilization.

2. **Trophoblast Differentiation:**
- As the blastocyst implants, trophoblast cells contact the endometrium and differentiate into
two layers:
- Inner layer: Cytotrophoblast, which is mitotically active, forming new cells that migrate into
the syncytiotrophoblast.
- Outer layer: Syncytiotrophoblast, a rapidly expanding, multinucleated mass without
discernible cell boundaries.

3. **Syncytiotrophoblast Invasion:**
- Erosive syncytiotrophoblast invades the endometrial connective tissue.
- The blastocyst gradually embeds in the endometrium, displacing endometrial cells at the
implantation site.
- Endometrial cells undergo apoptosis (programmed cell death), facilitating invasion.
4. **Molecular Mechanisms of Implantation:**
- Synchronization between the invading blastocyst and the receptive endometrium involves
various factors:
- Microvilli of endometrial cells.
- Cell adhesion molecules (integrins).
- Cytokines, prostaglandins, hormones (human chorionic gonadotropin [hCG] and
progesterone), growth factors.
- Extracellular matrix and enzymes (matrix metalloproteinase and protein kinase A).

5. **Modulation of Syncytiotrophoblast Penetration:**

- Endometrial cells modulate the depth of syncytiotrophoblast penetration.

6. **Decidual Cell Formation and Nutrition:**

- Connective tissue cells accumulate glycogen and lipids, assuming a polyhedral appearance.
- Some cells, called decidual cells, degenerate adjacent to the invading syncytiotrophoblast.
- Syncytiotrophoblast engulfs these cells, providing a rich source of embryonic nutrition.

7. **hCG Production and Hormonal Activity:**

- Syncytiotrophoblast produces human chorionic gonadotropin (hCG).
- hCG enters maternal blood via lacunae in the syncytiotrophoblast.
- hCG maintains the hormonal activity of the corpus luteum in the ovary during early
pregnancy.
- The corpus luteum secretes estrogen and progesterone, crucial for maintaining pregnancy.

8. **Clinical Implications:**
- Highly sensitive radioimmunoassays detect hCG, forming the basis for pregnancy tests.
- Enough hCG is produced by the syncytiotrophoblast at the end of the second week, allowing
for a positive pregnancy test, even when the woman may be unaware of her pregnancy.

In summary, the completion of blastocyst implantation involves intricate molecular interactions,

trophoblast differentiation, syncytiotrophoblast invasion, and the initiation of crucial hormonal
activities necessary for maintaining early pregnancy. The process is finely regulated to ensure
the establishment of a secure connection between the developing embryo and the maternal
endometrium.

The process of blastocyst implantation leads to the formation of key structures, including the
amniotic cavity, embryonic disc, and umbilical vesicle. Here is a detailed summary:

1. **Initiation of Amniotic Cavity and Amnioblast Formation:**

- During blastocyst implantation, a small space appears in the embryoblast, which is the
primordium of the amniotic cavity.
- Amniogenic cells, known as amnioblasts, separate from the epiblast and form the amnion,
enclosing the amniotic cavity.
2. **Formation of Embryonic Disc:**
- Concurrently, morphological changes occur in the embryoblast, resulting in the formation of a
flat, almost circular bilaminar plate of cells known as the embryonic disc.
- The embryonic disc consists of two layers:
- Epiblast: A thicker layer of high columnar cells related to the amniotic cavity.
- Hypoblast: Comprising small cuboidal cells adjacent to the exocoelomic cavity.

3. **Arrangement of Epiblast and Hypoblast:**

- The epiblast forms the floor of the amniotic cavity and is continuous peripherally with the
amnion.
- The hypoblast forms the roof of the exocoelomic cavity and is continuous with the thin
exocoelomic membrane, lining the primary umbilical vesicle.

4. **Fluid-Filled Spaces and Morphogenetic Movements:**

- The embryonic disc is positioned between the amniotic cavity and the umbilical vesicle.
- Lacunae (small spaces) appear in the syncytiotrophoblast, filling with a mixture of maternal
blood and cellular debris, forming embryotroph.
- Fluid in the lacunar spaces passes to the embryonic disc by diffusion, providing nutritive
material to the embryo.

5. **Uteroplacental Circulation and Implantation:**

- Erosion of endometrial capillaries establishes the primordial uteroplacental circulation.
- Oxygen and nutritive substances pass to the embryo from maternal blood entering lacunar
networks, while poorly oxygenated blood is removed.

6. **Decidual Reaction and Surface Regeneration:**

- Endometrial connective tissue cells undergo a transformation called the decidual reaction,
swelling due to glycogen and lipid accumulation.
- Decidual reaction provides nutrition for the early embryo and an immunologically privileged
site for the conceptus.
- Surface defects in the endometrial epithelium close through the formation of a fibrin
coagulum of blood, later covered by a regenerated uterine epithelium.

7. **Formation of Intervillous Spaces and Maternal Sinusoids:**

- Syncytiotrophoblast lacunae fuse to form lacunar networks, primordia of intervillous spaces
of the placenta.
- Maternal blood flows into lacunar networks, establishing maternal sinusoids that supply
oxygen and nutrients to the embryo.

8. **Communication and Nutrient Transfer:**

- Syncytiotrophoblast erodes sinusoids, allowing maternal blood to flow into lacunar networks.
- Trophoblast absorbs nutritive fluid from lacunar networks, transferring it to the embryo.
- The slow growth of the embryonic disc results in a protrusion into the uterine cavity.
9. **Formation of Extraembryonic Coelom and Umbilical Vesicle Changes:**
- Extraembryonic mesoderm increases, and isolated coelomic spaces appear within it.
- These spaces fuse to form the extraembryonic coelom, surrounding the amnion and
umbilical vesicle.
- The primary umbilical vesicle decreases in size, and a smaller secondary umbilical vesicle
forms.

10. **Significance of Umbilical Vesicle:**

- The umbilical vesicle, although containing no yolk in humans, has essential functions,
including the origin of primordial germ cells and potential roles in nutrient transfer.

In summary, the process involves the intricate development of structures such as the amniotic
cavity, embryonic disc, and umbilical vesicle, with communication between embryonic and
maternal tissues, establishment of circulatory connections, and the formation of spaces crucial
for nutrient transfer and embryonic development.

The development of the chorionic sac involves the formation of primary chorionic villi and the
differentiation of extraembryonic mesoderm into two layers, giving rise to the chorion. Here is a
detailed summary:

1. **Appearance of Primary Chorionic Villi:**

- At the end of the second week, primary chorionic villi emerge.
- These villi are vascular processes of the chorion, forming columns covered by syncytium.
- Cellular extensions grow into the syncytiotrophoblast, with growth likely induced by the
underlying extraembryonic somatic mesoderm.

2. **Formation of Chorion and Chorionic Villi:**

- The extraembryonic coelom divides the extraembryonic mesoderm into two layers:
- Extraembryonic somatic mesoderm, lining the trophoblast and covering the amnion.
- Extraembryonic splanchnic mesoderm, surrounding the umbilical vesicle.
- The chorion, the outermost fetal membrane, is formed by the extraembryonic somatic
mesoderm and the two layers of trophoblast.
- Primary chorionic villi represent the initial stage in the development of the chorionic villi of the
placenta, an organ facilitating metabolic interchange between the embryo and mother.

3. **Chorionic Sac and Extraembryonic Coelom:**

- The chorion forms the wall of the chorionic sac, where the embryo, amniotic sac, and
umbilical vesicle are suspended by the connecting stalk.
- The extraembryonic coelom serves as the primordium of the chorionic cavity.
- Transvaginal ultrasonography is employed to measure the chorionic sac diameter, aiding in
the evaluation of early embryonic development and pregnancy outcome.

4. **Embryonic Disc Development:**

- A 14-day embryo retains the form of a flat bilaminar embryonic disc.
 - Hypoblastic cells in a specific area become columnar and form a thickened circular region
called the prechordal plate.
- The prechordal plate indicates the site of the mouth and serves as a significant organizer of
the head region.

5. **Implantation Sites of Blastocysts:**

- Implantation of blastocysts typically occurs in the superior part of the uterine endometrium in
the body of the uterus.
- Detection of blastocyst implantation is possible through ultrasonography and highly sensitive
radioimmunoassays of human chorionic gonadotropin (hCG) by the end of the second week.

In summary, the second week of development involves the establishment of primary chorionic
villi, the differentiation of extraembryonic mesoderm, the formation of the chorion and chorionic
sac, and the initial development of the embryonic disc. Additionally, the prechordal plate
becomes a key structure indicating the site of the mouth. Implantation sites are primarily in the
superior part of the uterine endometrium, and advanced diagnostic techniques can detect early
implantation events.

GASTRULATION
**Summary: Gastrulation - Formation of Germ Layers**

**Gastrulation Overview:**
- Gastrulation is a crucial process during the third week of embryonic development, transforming
the bilaminar embryonic disc into a trilaminar embryonic disc.
- This process marks the initiation of morphogenesis, establishing the three germ layers and
axial orientation in embryos.

**Cellular Changes and Signaling Molecules:**

- Gastrulation involves extensive cellular changes, including alterations in shape,
rearrangement, movement, and adhesive properties.
- Signaling molecules such as bone morphogenetic proteins (BMPs), fibroblast growth factors
(FGFs), Sonic Hedgehog (Shh), Tgifs, and Wnts play pivotal roles in orchestrating gastrulation.

**Three Germ Layers and Their Derivatives:**

1. **Embryonic Ectoderm:**
- Gives rise to the epidermis (outer skin layer).
- Forms the central and peripheral nervous systems, including the brain and spinal cord.
- Contributes to the development of eyes and internal ears.
- Produces neural crest cells, a versatile cell population involved in various tissue formations.
- Contributes to many connective tissues in the head region.

2. **Embryonic Endoderm:**
- Source of epithelial linings in the respiratory and digestive tracts.
- Includes glands opening into the gastrointestinal tract.
 - Differentiates into glandular cells of associated organs such as the liver and pancreas.
3. The embryonic mesoderm plays a pivotal role in the development of various tissues and
organs, giving rise to:

1. **Skeletal Muscles:**
- All skeletal muscles in the body originate from the embryonic mesoderm.

2. **Blood Cells and Vascular Components:**

- It is the precursor for blood cells.
- Contributes to the lining of blood vessels.
- Forms most of the cardiovascular system.

3. **Smooth Muscles:**
- Gives rise to the visceral smooth muscular coats of internal organs.

4. **Serosal Linings:**
- Contributes to the serosal linings of all body cavities.

5. **Reproductive and Excretory Systems:**

- Develops into ducts and organs of the reproductive and excretory systems.

6. **Connective Tissues:**
- In the body (trunk or torso, excluding head and limbs), it is the source of all connective
tissues.
- This includes the formation of cartilage, bones, tendons, ligaments, dermis, and stroma
(connective tissue) of internal organs.

In essence, the embryonic mesoderm is instrumental in the formation of a diverse range of

tissues, contributing significantly to the structural components of the body's trunk and internal
organs.

**Significance:**
- Gastrulation is the starting point for morphogenesis, shaping the future body form and
establishing the fundamental germ layers.
- The coordinated actions of signaling molecules guide the differentiation of cells into specific
germ layers, laying the foundation for the development of various tissues and organs in the
embryo.
**Summary: Primitive Streak and Gastrulation**

**Formation of Primitive Streak:**

- The primitive streak, a morphological sign of gastrulation, appears on the epiblast of the
bilaminar embryonic disc during the beginning of the third week.
- It is a thickened linear band in the median plane of the dorsal aspect of the embryonic disc.
- The primitive streak establishes the embryo's craniocaudal axis, dorsal and ventral surfaces,
and right and left sides.

**Development of Primitive Node and Groove:**

- The primitive streak elongates, forming the primitive node at its cranial end and a primitive
groove.
- The primitive groove is created by the invagination of epiblastic cells, leading to the formation
of a primitive pit in the primitive node.

**Mesenchyme Formation:**
- Cells leaving the deep surface of the primitive streak differentiate into mesenchyme, an
embryonic connective tissue.
- Mesenchyme contributes to supporting tissues in the embryo, including most connective
tissues and the framework of glands.
- Some mesenchyme forms mesoblast, which gives rise to intraembryonic mesoderm.

**Endoderm and Ectoderm Formation:**

- Cells from the epiblast, primitive node, and primitive streak displace the hypoblast, forming
embryonic endoderm.
- The remaining cells in the epiblast form the embryonic ectoderm.

**Signaling Molecules and Differentiation:**

- Signaling molecules such as nodal factors, Wnt3a, Wnt5a, and FGFs induce the formation of
mesoderm and specify germ cell layer fates.
- Transforming growth factor-β (nodal), T-box transcription factor (veg T), and the Wnt signaling
pathway are involved in specifying the endoderm.

**Migration and Differentiation of Mesenchymal Cells:**

- Mesenchymal cells derived from the primitive streak migrate widely and differentiate into
various cell types, including fibroblasts, chondroblasts, and osteoblasts.

**Fate of Primitive Streak:**

- The primitive streak actively forms mesoderm until the early part of the fourth week.
- Afterward, mesoderm production slows down, and the primitive streak diminishes in size.
- Normally, the primitive streak undergoes degenerative changes and disappears by the end of
the fourth week.

**Summary: Notochordal Process and Notochord**

**Formation of Notochordal Process:**

- Mesenchymal cells migrate through the primitive streak, forming a notochordal process.
- The notochordal process acquires a lumen, becoming the notochordal canal.
- It grows cranially between the ectoderm and endoderm, reaching the prechordal plate.
**Migration of Mesenchymal Cells:**
- Mesenchymal cells from the primitive streak and notochordal process migrate laterally and
cranially, reaching the margins of the embryonic disc.
- Some cells migrate around the prechordal plate, forming cardiogenic mesoderm.

**Cloacal Membrane and Fusion:**

- The cloacal membrane indicates the future site of the anus and prevents migration of
mesenchymal cells between embryonic ectoderm and endoderm.
- Intraembryonic mesoderm separates the ectoderm and endoderm, except at specific locations.

**Formation of Notochord:**
- Instructive signals induce notochordal precursor cells to form the notochord, a cellular rod-like
structure.
- Shh signaling from the floor plate of the neural tube is involved in this molecular mechanism.

**Functions of Notochord:**
- Defines the primordial longitudinal axis of the embryo, providing rigidity.
- Provides signals necessary for the development of axial musculoskeletal structures and the
central nervous system (CNS).
- Contributes to intervertebral discs between adjacent vertebrae.

**Notochordal Canal Formation and Obliteration:**

- The notochordal process elongates, forming a canal that communicates with the umbilical
vesicle.
- Openings in the floor of the notochordal process connect it to the umbilical vesicle, creating the
notochordal canal.
- The neurenteric canal forms, providing a temporary communication between the amniotic and
umbilical vesicle cavities.
- The notochordal canal disappears as the notochordal process forms the notochord.
- The notochord becomes detached from the endoderm of the umbilical vesicle, with the
neurenteric canal normally obliterated.

**Persistence and Degeneration of Notochord:**

- The notochord extends from the oropharyngeal membrane to the primitive node.
- It degenerates as the bodies of the vertebrae form, with small portions persisting as the
nucleus pulposus of intervertebral discs.

**Role as Signaling Center:**

- The notochord functions as the primary inductor in the early embryo.
- It induces the overlying embryonic ectoderm to thicken and form the neural plate, the
primordium of the CNS.

**Summary: Allantois**
**Appearance and Development:**
- The allantois emerges around day 16 as a small diverticulum from the caudal wall of the
umbilical vesicle, extending into the connecting stalk.
- Although the allantois itself remains small, the allantoic mesoderm expands beneath the
chorion, forming blood vessels crucial for placental function.
- The initial allantoic diverticulum's proximal part persists throughout development as the
urachus, forming a stalk extending from the bladder to the umbilical region.
- The urachus, in adults, is represented by the median umbilical ligament.
- Blood vessels of the allantoic stalk transform into umbilical arteries, with the intraembryonic
part of the umbilical veins having a separate origin.

**Neurulation: Formation of Neural Tube:**

- Neurulation involves the processes of forming the neural plate, neural folds, and the
subsequent closure of the folds, ultimately giving rise to the neural tube.
- Neurulation is completed by the end of the fourth week, culminating in the closure of the
caudal neuropore.

**Summary: Neural Plate and Neural Tube Formation**

**Neural Plate Development:**

- Induced by the developing notochord, the overlying embryonic ectoderm thickens and forms
the neural plate, composed of neuroectodermal cells.
- The neural plate initially corresponds in length to the notochord and extends rostrally beyond
the primitive node, dorsal to the notochord and adjacent mesoderm.
- As the notochord elongates, the neural plate broadens and extends cranially, eventually
surpassing the notochord's length.

**Neurulation Process:**
- Around the 18th day, the neural plate undergoes invagination along its central axis, forming a
neural groove with neural folds on each side.
- Neural folds become prominent, especially at the cranial end, signaling the onset of brain
development.
- By the end of the third week, neural folds begin to fuse, converting the neural plate into the
neural tube, the precursor of the brain and spinal cord.
- Neural tube separation from surface ectoderm occurs as neural folds meet, and neural crest
cells undergo an epithelial to mesenchymal transition, migrating away.
- The free edges of surface ectoderm fuse, differentiating into the epidermis.
- Completion of neurulation takes place during the fourth week.

**Neural Crest Formation:**

- As neural folds fuse, neuroectodermal cells along the inner margin lose epithelial
characteristics, forming neural crest cells.
- Neural crest cells, activated by Wnt/β-catenin signaling, create a mass, the neural crest,
between the neural tube and overlying surface ectoderm.
- Neural crest separates into right and left parts, contributing to the sensory ganglia of spinal
and cranial nerves, spinal ganglia, autonomic nervous system ganglia, and more.
- Neural crest cells play a role in the formation of pigment cells, the suprarenal medulla, and
various tissues and organs.
- Proper cell interactions within the surface epithelium and between it and underlying mesoderm
are vital for establishing neural plate boundaries and specifying sites for
epithelial−mesenchymal transformation, mediated by various signaling systems.
- Defective migration and differentiation of neural crest cells can lead to numerous human
diseases.
**Summary: Development of Somites**

**Formation of Paraxial Mesoderm:**

- Cells derived from the primitive node contribute to the formation of paraxial mesoderm.
- Paraxial mesoderm, initially appearing as a thick longitudinal column near the primitive node,
extends laterally to connect with intermediate mesoderm, which, in turn, thins into lateral
mesoderm.
- The lateral mesoderm continues as extraembryonic mesoderm covering the umbilical vesicle
and amnion.

**Differentiation into Somites:**

- Toward the end of the third week, paraxial mesoderm undergoes differentiation, condensation,
and division to form paired cuboidal structures called somites.
- Somites develop in a craniocaudal sequence on each side of the developing neural tube.
- Approximately 38 pairs of somites form during the somite period (days 20 to 30), determining
the embryo's age.
- By the end of the fifth week, the number of somite pairs increases to 42 to 44.
- Somites, prominent during the fourth and fifth weeks, serve as criteria for determining embryo
age.

**Spatial and Functional Significance of Somites:**

- Somites first appear in the occipital region of the embryo's head, extending craniocaudally.
- They give rise to the axial skeleton, associated musculature, and adjacent dermis of the skin.
- The initial pair of somites emerges near the site of the otic placode formation.
- Correct guidance of axons from the spinal cord to target muscle cells is crucial for innervation.

**Molecular Mechanisms in Somite Formation:**

- Somite formation involves the expression of NOTCH pathway genes, HOX genes, and other
signaling factors.
- Expression of forkhead transcription factors FoxC1 and FoxC2 precedes somite formation, and
the Delta-Notch signaling pathway regulates the craniocaudal segmental pattern.
- A molecular oscillator or clock mechanism is proposed to orchestrate the orderly sequencing of
somites.

**Summary: Development of Intraembryonic Coelom and Early Cardiovascular System**

**Development of Intraembryonic Coelom:**
- Intraembryonic coelom, the embryonic body cavity, originates as isolated coelomic spaces in
lateral intraembryonic mesoderm and cardiogenic mesoderm.
- These spaces coalesce to form a horseshoe-shaped cavity, dividing lateral mesoderm into
somatic (parietal) and splanchnic (visceral) layers.
- Somatic mesoderm, with overlying embryonic ectoderm, forms the embryonic body wall
(somatopleure), while splanchnic mesoderm and underlying embryonic endoderm constitute the
embryonic gut (splanchnopleure).
- During the second month, the intraembryonic coelom divides into pericardial, pleural, and
peritoneal cavities.

**Early Development of Cardiovascular System:**

- In the second week, embryonic nutrition is obtained through diffusion in the extraembryonic
coelom and umbilical vesicle.
- Blood vessel formation begins in extraembryonic mesoderm of the umbilical vesicle,
connecting stalk, and chorion at the start of the third week.
- Vasculogenesis involves the assembly of angioblasts into blood islands, forming endothelial
channels that later fuse.
- Angiogenesis is the budding and branching of new vessels from preexisting ones.
- Hemangiogenic epithelium differentiates into endothelial cells, contributing to blood vessel
formation.
- Blood cells develop from specialized endothelial cells on the umbilical vesicle and allantois at
the end of the third week.
- The heart and great vessels form from mesenchymal cells in the cardiogenic area.
- Paired endocardial heart tubes fuse to create a primordial heart tube by the end of the third
week.
- The primordial cardiovascular system, connecting the stalk, chorion, and umbilical vesicle,
becomes functional by the 21st or 22nd day.
- The embryonic heartbeat, signaling functional cardiovascular development, can be detected
using Doppler ultrasonography in the fourth week.
**Summary: Development of Chorionic Villi**

- Primary chorionic villi appear at the end of the second week and start branching shortly
afterward.
- Early in the third week, mesenchyme grows into these villi, forming secondary chorionic villi
that cover the entire surface of the chorionic sac.
- Some mesenchymal cells in the villi differentiate into capillaries and blood cells, marking the
transition to tertiary chorionic villi.
- Capillaries in the chorionic villi fuse to form arteriocapillary networks, connecting with the
embryonic heart through vessels in the chorion and connecting stalk.
- By the end of the third week, embryonic blood starts flowing through the capillaries in the
chorionic villi.
- Oxygen and nutrients from maternal blood in the intervillous space diffuse through villi walls
into the embryo's blood, while waste products diffuse in the opposite direction.
- Cytotrophoblastic cells of the chorionic villi proliferate and extend through the
syncytiotrophoblast to form an extravillous cytotrophoblastic shell, surrounding the chorionic sac
and attaching it to the endometrium.
- Stem villi (anchoring villi) attach to maternal tissues through the cytotrophoblastic shell, and
branch villi grow from the sides of stem villi.
- The main exchange of materials between maternal and embryonic blood occurs through the
walls of the branch villi, bathed in continually changing maternal blood in the intervillous space.

FOURTH TO EIGHT WEEKS OF DEVELOPMENT

Phases of Embryonic Development and Folding of Embryo**

**Phases of Embryonic Development:**

1. **Growth:**
- Involves cell division and the production of cell products.

2. **Morphogenesis:**
- Concerns the development of shape, size, and features at the organ, part, or whole body
level.
- Regulated by specific gene expression and molecular processes.
- Involves changes in cell fate, shape, and movement to facilitate the formation of tissues and
organs.

3. **Differentiation:**
- Cells organize into precise patterns of tissues and organs.
- Results in specialized functions for these tissues and organs.

**Folding of Embryo:**
- An essential event in establishing body form involves folding of the flat trilaminar embryonic
disc.
- The embryonic disc transforms into a somewhat cylindrical embryo.
- Folding occurs in the median and horizontal planes.
- Rapid growth of the embryo contributes to the folding process.
- Growth rate differences between the sides and the long axis lead to simultaneous folding at
the cranial and caudal ends and sides of the embryo.
- There is a relative constriction at the junction of the embryo and the umbilical vesicle during
folding.
**Summary: Folding of Embryo in the Median and Horizontal Planes**

**Folding in the Median Plane:**

1. **Head Fold:**
- Neural folds in the cranial region form the brain's primordium.
 - Developing forebrain overhangs the developing heart.
- Septum transversum, primordial heart, pericardial coelom, and oropharyngeal membrane
move to the ventral surface.
- Part of the endoderm of the umbilical vesicle incorporates into the embryo as the foregut.
- Septum transversum develops into the central tendon of the diaphragm.
- Affects the arrangement of the embryonic coelom.

2. **Tail Fold:**
- Caudal end folding results from the growth of the distal part of the neural tube.
- Caudal eminence (tail region) projects over the cloacal membrane (future site of the anus).
- Part of the endodermal germ layer incorporates as the hindgut (descending colon and
rectum).
- The hindgut dilates to form the cloaca, a rudiment of the urinary bladder and rectum.
- Primitive streak shifts from cranial to caudal to the cloacal membrane.

**Folding in the Horizontal Plane:**

- Lateral folding produced by the rapidly growing spinal cord and somites.
- Ventrolateral abdominal wall primordia fold towards the median plane.
- Formation of the midgut (primordium of the small intestine) as part of the endoderm is
incorporated.
- Reduction of the connection between the midgut and umbilical vesicle forms the
omphaloenteric duct.
- Ventral fusion of lateral folds reduces the communication between intraembryonic and
extraembryonic coelomic cavities.
- Amniotic cavity expansion and obliteration of most extraembryonic coelom result in the amnion
covering the umbilical cord.
**Summary: Germ Layer Derivatives**

**Ectoderm:**
1. **Central Nervous System (CNS):**
- Formation of the brain and spinal cord.

2. **Peripheral Nervous System (PNS):**

- Includes neural crest cells that contribute to various structures.
- Forms spinal cord cells, cranial nerves (V, VII, IX, X), and autonomic ganglia.

3. **Sensory Epithelia:**
- Eyes, ears, and nose.

4. **Epidermis and Appendages:**

- Hair, nails, and subcutaneous glands.

5. **Mammary Glands:**
- Development of mammary tissue.
6. **Pituitary Gland:**
- Formation of the pituitary gland.

7. **Enamel of the Teeth:**

- Contribution to tooth enamel.

**Mesoderm:**
1. **Connective Tissue:**
- Formation of various connective tissues.

2. **Cartilage and Bone:**

- Development of skeletal structures.

3. **Muscles:**
- Striated and smooth muscles.

4. **Cardiovascular System:**
- Heart, blood, and lymphatic vessels.

5. **Genitourinary System:**
- Kidneys, ovaries, testes, genital ducts.

6. **Serous Membranes:**
- Lining the body cavities (pericardial, pleural, peritoneal membranes).

7. **Spleen:**
- Formation of the spleen.

8. **Cortex of Suprarenal Glands:**

- Development of the adrenal glands.

**Endoderm:**
1. **Epithelial Lining:**
- Digestive and respiratory tracts.

2. **Tonsils:**
- Parenchyma (connective tissue framework).

3. **Endocrine Glands:**
- Thyroid and parathyroid glands.
- Thymus, liver, and pancreas.

4. **Urinary System:**
 - Epithelial lining of the urinary bladder and most of the urethra.

5. **Ear Structures:**
- Epithelial lining of the tympanic cavity, tympanic antrum, and pharyngotympanic tube.
**Summary: Control of Embryonic Development**

Embryonic development is orchestrated by genetic plans within chromosomes, and the

understanding of genes influencing human development is growing. While much knowledge is
derived from studying other organisms like Drosophila and mice, ethical concerns limit direct
human embryo studies.

Developmental processes involve the coordinated interaction of genetic and environmental

factors, employing mechanisms such as tissue interactions, regulated cell migration, controlled
proliferation, and programmed cell death (apoptosis). Growth, achieved through mitosis and
extracellular matrix production, complements the complexity attained through morphogenesis
and differentiation.

Early embryonic cells are pluripotential, capable of affecting multiple organs or tissues. As
development progresses, these cells become more restricted in their developmental potential,
guided by cues from their immediate surroundings. Choices in tissue diversification are made
based on interactions with adjacent tissues.

Inductive interactions, where tissues influence each other's development, play a crucial role.
Examples include the optic vesicle inducing lens development in the eye. Molecular defects in
mutants and studies of embryos with gene mutations reveal the molecular mechanisms of
induction. Signal transfer between tissues varies—diffusible molecules, extracellular matrices, or
physical contacts may be involved.

Developmental signaling is not always specific, and inducible tissues have a limited time frame
of receptiveness. Failed interactions, caused by delays or spatial separation, can lead to severe
consequences, such as birth defects. Understanding these control mechanisms is crucial for
comprehending normal and abnormal development.

**Summary: Fourth to Seventh Week of Embryonic Development**

**Fourth Week:**
- **Early Changes:** The embryo is straight, with 4 to 12 somites and an open neural tube.
- **Pharyngeal Arches:** First pharyngeal arch visible, contributing to the mandible and maxilla.
- **Heart Development:** Ventral heart prominence formed, and blood circulation begins.
- **Limbs and Organs:** Upper limb buds, otic pits, and lens placodes visible. Fourth pair of
pharyngeal arches and lower limb buds appear. Rudiments of organ systems, especially the
cardiovascular system, are established. Caudal neuropore usually closed by the end of the
week.
**Fifth Week:**
- **Minor Changes:** Growth of the head surpasses other regions, influenced by brain and facial
prominences.
- **Pharyngeal Arches:** Second arch grows, creating cervical sinus.
- **Mesonephric Kidneys:** Mesonephric ridges indicate the site of developing interim excretory
organs.
- **Heart-Face Contact:** Face contacts the heart prominence.

**Sixth Week:**
- **Spontaneous Movements:** Embryo shows spontaneous movements, reflex responses to
touch.
- **Upper Limbs:** Regional differentiation begins with elbow and hand plate development.
- **Lower Limbs:** Development follows upper limbs by 4 to 5 days. Auricular hillocks form
external ear, eyes become obvious.
- **Head Position:** Head is much larger, bent over the heart prominence. Intestines herniate
into the umbilical cord due to limited abdominal cavity.

**Seventh Week:**
- **Limb Changes:** Notches appear between digital rays, indicating digits.
- **Gastrointestinal Changes:** Communication between the primordial gut and umbilical vesicle
reduces.
- **Ossification:** Ossification of upper limb bones begins by the end of the week.

These weeks mark crucial stages of embryonic development with significant changes in body
form, organ development, and the initiation of limb differentiation. The intricate coordination of
these processes ensures the gradual formation of the human embryo.

**Summary: Eighth Week of Embryonic Development**

- **Early Week Changes:**

- Digits of the hand are separated but noticeably webbed.
- Notches visible between digital rays of the feet.
- Caudal eminence is stubby.
- Scalp vascular plexus forms a band around the head.

- **End of the Eighth Week:**

- Limb Development: All regions of the limbs are apparent, and digits have lengthened and are
completely separated.
- Limb Movements: Purposeful limb movements begin.
- Ossification: Primary ossification begins in the femora.
- Caudal Eminence: Disappears completely.
- Limb Proximity: Hands and feet approach each other ventrally.
- Human Characteristics: Embryo exhibits distinct human characteristics, but the head is still
disproportionately large.
 - Facial Development: Neck is established, eyelids are closing, and by the end of the week,
they begin to unite by epithelial fusion.
- Internal Organs: Intestines are still in the proximal portion of the umbilical cord.
- Sexual Differentiation: Slight sex differences in the appearance of external genitalia, but not
distinctive enough for accurate identification.

- **Estimation of Embryonic Age:**

- Methods: Estimates based on external characteristics and measurements, including
crown–rump length (CRL) for older embryos (14 to 18 weeks).
- Reliability: Size alone may be unreliable, and the rate of growth varies before death.
- Carnegie Embryonic Staging System: Internationally used for standardized comparisons,
especially in embryos recovered after spontaneous abortion.

The eighth week marks the completion of the embryonic period, with the embryo acquiring more
distinct human features, limb development, and the initiation of purposeful limb movements.
Estimation of embryonic age involves various measurements and the use of standardized
staging systems for accuracy and comparability.
**Summary: Estimation of Fetal Age**

- **Methods of Estimation:**
- **Ultrasound Measurements:** Utilizes crown–rump length (CRL), fetal head measurements,
and femur length to determine size, probable age, and expected delivery date.
- **Gestational Age Calculation:** In clinical practice, often timed from the onset of the last
normal menstrual period (LNMP).
- **Embryologic Terminology:** Important to use embryologic terminology to avoid confusion
regarding gestational age calculation.

- **Embryologic vs. Clinical Gestational Age:**

- **Embryologic Perspective:** Gestation begins at fertilization, not LNMP.
- **Clinical Perspective:** LNMP is commonly used for gestational age calculation in clinical
settings.

- **Divisions of Pregnancy:**
- **Trimesters:** Clinically divided into three trimesters, each lasting 3 months.
- **First Trimester:** By its end, major systems have developed.
- **Second Trimester:** Allows visualization of good anatomical detail.
- **Third Trimester:** Fetus may survive if born prematurely, major developmental landmark at
35 weeks.

- **Measurements and Characteristics for Estimation:**

- **CRL:** Preferred method for estimating fetal age until the end of the first trimester due to
low variability.
- **Ultrasound Measurements (Second and Third Trimesters):**
 - Biparietal diameter, head circumference, abdominal circumference, femur length, and foot
length.
- **Weight:** Useful criterion, but discrepancies may occur, especially with maternal metabolic
disturbances.
- **Fetal Dimensions:** Obtained from ultrasound closely approximate CRL measurements
from spontaneously aborted fetuses.

- **Clinical Significance:**
- **Obstetric Management:** Determination of fetal size, especially head circumference, aids
obstetricians in patient management.

Note: It is crucial to specify whether gestational age is calculated from LNMP or estimated time
of fertilization, and embryologic terminology should be used consistently to avoid confusion.
Measurements and characteristics help in accurate age estimation and provide valuable
information for obstetric management.
**Summary: Fetal Development (Twenty-Six to Thirty-Eight Weeks)**

**Twenty-Six to Twenty-Nine Weeks:**

- **Survivability:** Fetuses usually survive if born prematurely and receive intensive care.
- **Lung Development:** Lungs and pulmonary vasculature sufficiently developed for gas
exchange.
- **Central Nervous System (CNS):** Maturation allows rhythmic breathing and body
temperature control.
- **Physical Characteristics:** Open eyelids at 26 weeks, well-developed lanugo and head hair;
toenails visible; subcutaneous fat increases; fetal spleen's role in erythropoiesis ends by 28
weeks.

**Thirty to Thirty-Four Weeks:**

- **Neonatal Mortality:** Higher in low birth weight infants; eyelids open, lanugo, head hair,
toenails, subcutaneous fat well-developed; fetal spleen's role in erythropoiesis ends at 28
weeks; pupillary reflex at 30 weeks.

**Thirty-Five to Thirty-Eight Weeks:**

- **Developmental Milestones:** Grasping reflex, spontaneous orientation to light at 35 weeks;
mature nervous system for integrative functions.
- **Physical Characteristics:** Plump appearance; head and abdomen circumferences equal at
36 weeks; slowing of growth as birth approaches.
- **Full Term (38 Weeks):** CRL around 360 mm, weight approximately 3400 g; 16% of body
weight is white fat; thorax prominent, breasts protrude slightly; testes usually in scrotum in
full-term males.

**General Trends:**
- **Growth Slowing:** Growth slows as birth approaches.
- **Sex Differences:** In general, male fetuses are longer and heavier at birth than females.
- **Fat Accumulation:** Fetuses add approximately 14 g of fat per day during the last weeks of
development.