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Learning Objectives
Overall goal: understand the fundamental processes
by which the adult form is produced and the clinical
consequences that arise from abnormal development.
At the end of this lecture, you should be able to:
1. Describe fertilization and identity where it place.
2. Understand the phases of fertilization.
3. List the results of fertilization.
4. Describe the formation of blastocyst.
5. Describe the mechanism and of implantation.
6. Identify Implantation site and list site of ectopic Prenancy.
7. Describe the formation of primary chorionic villi. 3
 Fertilization
Fertilization in human
is a complex process
during which a
spermatozoon makes
contact with a mature
ovum resuting in the
successful union of the
pronuclei of the sperm
and ovum.
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 Fertilization
Creates the Zygote that
contain genetic materials
from the male and female
parents.

The zygote is a highly

specialized totipotent cell
 Fertilization
Life begins at fertilization—
the choice to honor and
protect or ignore and
terminate it is huge debate
in most parts of the world

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Site of Fertilization
•Usually in the ampulla of the
uterine tube.
•Ampulla is the longest and widest
part.
•Fertilization may occur in other
parts of tubes.
•Does not occur in the uterine cavity.
•Chemical signals from oocyte
attract the sperms.
 Phases of Fertilization
•1- Passage of sperm through corona
radiata, under the effect of :
hyaluronidase enzyme from sperms,
tubal E. and movement of tail of
sperm.
•2- Penetration of the zona pellucida
by head of sperms through acrosine
E. from acrosome of one sperm.
•3- Fusion of the plasma membrane
of the oocyte and that of the sperm.
so sperm’s plasma membrane
remains behind.
 Phases of Fertilization

•4- Completion of the second meiotic

division & formation of the female
pronucleus.
•5- Formation of the male pronucleus :
It is a swollen nucleus of the sperm.
Its tail is detached and degenerated.
Zona reaction : the change in zona
pellucida chemical properties that
makes it impermeable to other zygote

sperms.
 Phases of Fertilization

• The cortical reaction requires a high

concentration of Ca2 ions in the egg
• The reaction is triggered by a change in
Ca2 concentration
• Ca2 spread across the egg correlates
with the appearance of the fertilization
envelope

zygote
 Chromosomes in zygote
•Zygote is genetically unique.
•Half of its chromosomes come
from the father and the
other half comes from the mother.
•zygote contains 46 chromosomes
(diploid).
•New combination is formed which
is different from either of the
parents.
•This mechanism forms biparental
inheritance and leads to variation
of the human species.
 The Father Always Determine
Sex of the Embryo the Sex of the Baby!!!

•Embryo's chromosomal sex is

determined at the time of
fertilization.
•Sex is determined by the type
of sperm (X or Y)
that fertilizes the oocyte.
•So, it is the father’ gamete
that decides the sex.
 Results of Fertilization

•Stimulates the penetrated oocyte to

complete its 2nd meiotic division.
•Restores the normal diploid number
of chromosomes in the zygote (46).
•Determines the chromosomal sex of
the embryo.
•Initiates cleavage of the zygote.
 Fertilization
Following fertilization,
Cell division,
cell migration,
programmed cell death,
differentiation,
growth, and
cell
rearrangement
transform zygote into a
multicellular human being over
a period of about 9 months. 14
 Week 1: days 1-6
• Fertilization, day 1
• Cleavage, day 2-3
• Compaction, day 3
• Formation of blastocyst, day 4
• Ends with implantation, day 6

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Fertilized egg (zygote)

Fertilized egg
2 polar bodies
2 pronuclei

Day 1
0.1 mm

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 Cleavage = cell division
Cleavage
Goals: grow unicellular
zygote to multicellular embryo.

Divisions are slow: 12 - 24h each

No growth of the embryo-

stays at ~100 um in diameter

Divisions are not synchronous

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 Cleavage
Fertilization is followed by
cleavage, a period of rapid cell
division without growth
Cleavage partitions the
cytoplasm of one large cell into
many smaller cells called
blastomeres
The blastula is a ball of cells with
a fluid-filled cavity called a
blastocoel
 Cleavage
It begins about 24-30 hours
after fertilization.
Zygote divides into 2, then 4,
then 8, then 16 cells.
Zygote lies within the thick zona
pellucida during cleavage.
Zygote migrates in the uterine
tube from its lateral end to its
medial end.
Zona pellucida is translucent
under light microscope.
 2 Cell Stage

Individual cells = blastomeres

Mitotic divisions maintain

2N (diploid) complement

Cells become smaller

Blastomeres are equivalent (aka

totipotent). 20
4 cell; second cleavage

4 equivalent blastomeres

Still in zona pellucida

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8 Cell; third
cleavage

Blastomeres still
equivalent

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 Morula
•When there are 12-32 blastomeres the
developing human is called MORULA.
•The Morula reaches the uterine cavity at
this stage.
•Spherical Morula is formed about 3 days
after fertilization.
• Formation of blastocyst :
• The Morula reaches the uterine cavity by the 4th day after
fertilization, & remains free for one or two days.
Fluid passes from uterine cavity to the Morula.
• Now the Morula is called Blastocyst, its cavity is called
blastocystic cavity, its cells divided into Embryoblast &
Trophoblast.
 Embryo undergoes compaction after 8-cell stage:
first differentiation of embryonic lineages

Compaction is caused by increased cell-cell adhesion

Cells that are forced to the outside of the morula are destined to
become trophoblast--cells that will form placenta
The inner cells will form the embryo proper and are called the
inner cell mass (ICM). 25
 Formation of the blastocyst

Sodium channels appear on the surface of the outer trophoblast cells; sodium and water are
pumped into the forming blastocoele. Note that the embryo is still contained in the zona
pellucida.

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 “Hatching” of the blastocyst:
preparation for implantation

Hatching of the embryo from the zona pellucida occurs just

prior to implantation. Occasionally, the inability to hatch
results in infertility, and premature hatching can result in abnormal implantation in the uterine tube.
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 Early Pregnancy Factor

• Is an immunosuppressant protein.

• Secreted by trophoblastic cells.

• Appears in maternal serum within 24--48 hrs

• It is the basis for EPT (Early pregnancy test) in the first 10 days of
development.
 IMPLANTATION
• the process by which the Blastocyst
penetrates the superficial (compact) layer of
the endometrium of the uterus.
• Site:
• The normal site of implantation is the
posterior wall of the uterus near the
fundus.
• Time:
• It begins about the 6th day after fertilization.
• It is completed by the 11th or 12th day.
 Normal Implantation Sites
The implantation site determines
the site of formation of the
placenta
Normally it occurs in the upper
part of the body of uterus, more
often on the posterior wall
 Ectopic Pregnancy
mplantation outside the
uterus.
95 to 97% of ectopic
pregnancies occurs in the
uterine tube.
Most are in the ampulla &
sthmus.
Placenta previa :
mplantation occurs in the
ower uterine segment.
Tubal pregnancy

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by day 5, Zona pellucida degenerates & disappears to allows the
blastocyst to increase in size and penetrates the endometrium.
The embryoblast projects into the blastocystic cavity, while the
trophoblast forms the wall of the blastocyst.
By 6th day the blastocyst adheres to the endometrial epithelium.

Penetration results from proteolytic enzymes ([Link]-2)

produced by the trophoblast.

6th day
7th day
• By 7th day, Trophoblast differentiated into 2 layers:
Cytotrophblast, inner layer, mononucleated mitotically active cells.
Syncytiotrophoblast (outer multinucleated mass, with indistinct cell
boundary.
• By 8th day the blastocyst is superficially embedded in the compact
layer of the endometrium (by the end of 1st week,the blastocyst is
superficially implanted in endometrium).
• Blood-filled Lacunae appear in the
Syncytiotrophoblast
which communicate forming a network by
the 10th day or 11th day.
• Syncytiotrophoblast
erodes the endothelial lining of the
maternal capillaries which known as
sinusoids.
Now blood of maternal capillaries reaches
the lacunae so
Uteroplacental circulation
is established by 11th or 12th day.
8th day
Endometrial cells undergo apoptosis
(programmed cell death) to facilitates
invasion of endometrium by the
Syncytiotrophoblast.

Syncytiotrophoblast engulf these

degenerated cells for nutrition of the
embryo.

Implantation
can be detected by:
1- Ultrasonography.
2- hCG (human chorionic gonadotrophin
which is secreted by the
Syncytiotrophoblast) about the end of 2nd
week
 Formation of The Primary Chorionic villi

• By the day 13
Proliferation of
Cytotrophblast cells
produce extension
inside the
Syncytiotrophoblast to
form primary the
chorionic villi.
 Formation of the chorionic villi
Trophoblast proliferates rapidly and
differentiates into two layers:
inner cellular cytotrophoblast or
Langhan’s layer
outer mass of syncytiotrophoblast
(multinucleated protoplasm with
no cell boundaries)
Finger like processes of
syncytiotrophoblast extend through
the endometrium and invade the
endometrial connective tissue
 Implantation cont’d
• Small cavities, the lacunae
appear in syncytiotrophoblast,
and get filled with maternal
blood, establishing primitive
uteroplacental circulation
 Gastrulation
At gastrulation the two layered epiblast is converted into the
three primary embryonic germ layers:

• Ectoderm: outside, surrounds other layers later in

development, generates skin and nervous tissue
• Mesoderm: middle layer, generates most of the muscle,
blood and connective tissues of the body and placenta
• Endoderm: eventually most interior of embryo, generates
the epithelial lining and associated glands of the gut, lung,
and urogenital tracts
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The human embryo at gastrulation

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At gastrulation, primitive endoderm is
replaced by definitive or embryonic
endoderm then mesoderm is formed

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Cell movements during gastrulation

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Mesoderm is patterned in a cranial to caudal gradient

Axial mesoderm: passes through the

node and migrates along the midline –
forms the notochord

Paraxial mesoderm: passes just caudal

to the node and migrates slightly laterally
–forms cartilage, skeletal muscle, and
dermis

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Mesoderm is patterned in a cranial to caudal gradient

Lateral plate mesoderm: passes more

caudal and migrates more laterally –
forms circulatory system and body cavity
linings.

Extraembryonic mesoderm: passes most

caudal and migrates most laterally –
forms extraembryonic membranes and
associated connective tissue & blood
vessels. 45
 Fate of the “axial” mesoderm
The notochord and pre-chordal plate develops from mesoderm arising from cells that passed
directly through the node and migrated cranially along the midline
The notochord and pre-chordal plate are important signaling centers that pattern the
overlying ectoderm and underlying endoderm.

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Left-Right asymmetry is established at
gastrulation
Leftward beating of cilia at node moves
secreted molecules sonic hedgehog (Shh)
& FGF-8 to the left side of embryo.

Causes left side genes Nodal and PITX2 to

be expressed which then pattern
developing organs.

If cilia are defective, Shh and FGF8 can

randomly end up on right side, resulting in
reversal of symmetry, aka situs inversus
(liver on the left, spleen on the right, etc.)

Situs can be complete (everything

reversed) or partial (only some organs
reversed).
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Situs Inversus

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 What happens if there is “not enough” gastrulation?
Caudal agenesis (sirenomelia)
Premature regression of the primitive streak leads to widespread
loss of trunk and lower limb mesoderm.

VATeR association:
Vertebral defects
Anal atresia
Tracheo-esophageal fistula
Renal defects

VACTeRL association:
those above plus…
Cardiovascular defects
Limb (upper) defects

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 What happens if there is “too much” gastrulation?
Sacrococcygeal teratoma
If the primitive streak fails to
regress, multipotent primitive
streak cells can develop into
multi-lineage tumors
(containing ecto-, meso-, and
endodermal tissues).

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Multiple Pregnancy
Multiple Pregnancy/ Multifetalpregnancy
• The presence of more than one fetus in the gravid uterus is called
multiple pregnancy
• Two fetuses (twins)
• Three fetuses (triplets)
• Four fetuses (quadruplets)
• Five fetuses (quintuplets)
• Six fetuses (sextuplets)
INCIDENCE
Hellin’s Law:
Twins: 1:89
Triplets: 1:892
Quadruplets: 1:893
Quintuplets: 1:894
Conjoined twins: 1 : 60,000
Worldwide incidence of monozygotic - 1 in 250
Incidence of dizygotic varies & increasing
Demography
• Race: most common in people of African origin
• Age: Increased maternal age
• Parity: more common in multipara
• Heredity - family history of multifetal gestation
• Nutritional status – well nourished women
• ART - ovulation induction with clomiphene citrate,
gonadotrophins and IVF
• Conception after stopping OCP
Twins
Varieties:
• 1. Dizygotic twins: commonest (Two-third)
• 2. Monozygotic twins (one-third)

Genesis of Twins:
• Dizygotic twins (syn: Fraternal, binovular) -
- fertilization of two ova by two sperms.
Degree of Separation of Monozygotic Twins

Monozygotic twins (syn: Identical, uniovular):

• Up to 3 days - —diamniotic-dichorionic
• Between 4th & 7th day —diamniotic monochorionic - most
common type
• Between 8th & 12th day- monoamniotic-monochorionic
• After 13th day - —conjoined / Siamese twins.
Degree of
Separation
of
Monozygotic
Twins
Conjoined twins
Ventral:
1) Omphalopagus
2) Thoracopagus
3) Cephalopagus
4) Caudal/ ischiopagus
Lateral:
1) Parapagus
Dorsal:
1)Craniopagus,
2)Pyopagus
Superfecundation
Fertilization of two different ova released in the same cycle by
an act two sexual intercourse with the same male. This is
currently thought impossible to prove ordinarily

Heteropaternal Superfecundation
Fertilization of two different ova released in the same cycle by
an act two sexual intercourse with two different males

Superfetation
Fertilization of two ova released in different cycles
Semi-Identical (Sesquizygotic)Twins

Fetomaternal Chimerism

Tetragamatic Chimerism