Ferterlization and Implantation
Ferterlization and Implantation
Ferterlization and Implantation
IMPLANTATION
REPRODUCTIVE HEALTH I
CLINICAL MEDICINE CLASS
INTRODUCTION
The Sperm:
• The spermatozoa leave the testis carrying 23 chromosomes but not
yet capable of fertilization.
• Their maturation is completed through their journey in the 6 meters
of the epididymis and when mixed with the seminal plasma from the
epididymis, seminal vesicle and prostate gland.
• After semen is ejaculated, the sperms reach the cervix by their own
motility within seconds leaving behind the seminal plasma in the
vagina.
INTRODUCTION
• At time of ovulation, the cervical mucous is in the most favourable
condition for sperm penetration and capacitation as:
• It becomes more copious, less viscous and its macromolecules arrange in
parallel chains providing channels for sperms passage.
• Its contents from glucose and chloride are increased.
• The sperms ascent through the uterine cavity and Fallopian tubes to
reach the site of fertilization in the ampulla by:
• its own motility,
• uterine and tubal peristalsis which is aggravated by the prostaglandins in
the seminal plasma.
INTRODUCTION
• The sperms reach the tube within 30-40 minutes but they are capable of
fertilization after 2-6 hours.
• This period is needed for sperm capacitation.
• Capacitation of sperms is the process after which the sperm becomes able
to penetrate the zona pellucida, that surrounds the ovum and fertilize it.
• The cervical and tubal secretions are mainly responsible for this
capacitation.
• Capacitation is believed to be due to:
a. increase in the DNA concentration in the nucleus
b. increase permeability of the coat of sperm head to allow more release of
hyaluronidase
INTRODUCTION
The ovum:
• The ovum leaves the ovary after rupture of the Graafian follicle,
carrying 23 chromosomes and surrounded by the zona pellucida and
corona radiata.
• The ovum is picked up by the fimbriated end of the Fallopian tubes
and moved towards the ampulla by the ciliary movement of the cells
and rhythmic peristalsis of the tube.
INTRODUCTION
OVULATION
• Ovulation is a process whereby a secondary oocyte is released from the ovary
following rupture of a mature Graafian follicle and becomes available for
conception.
Changes in the follicle
• There is preovulatory enlargement of the Graafian follicle due to accumulation of
follicular fluid and measures about 20 mm in diameter.
• The cumulus oophorus separates from the rest of the granulosa cells and floats
freely in the antrum.
• The inner layer of the cells surrounding the oocyte is arranged radially corona
radiata
• The follicular wall near the ovarian surface becomes thinner.
INTRODUCTION
Changes in the oocyte
• Cytoplasmic volume is increased along with changes in the number,
distribution of mitochondria and in the Golgi apparatus.
• Completion of the arrested first meiotic division occurs with
extrusion of first polar body, each containing haploid number of
chromosomes (23, X).
INTRODUCTION
CAUSES
1. Endocrinal the combined LH/FSH midcycle surge is responsible for the
final stage of maturation, rupture of the follicle and expulsion of the oocyte.
• LH surge: Sustained peak level of estrogen for 24–36 hours in the late
follicular phase → LH surge occurs from the anterior pituitary.
• Ovulation approximately occurs 16–24 hours after the LH surge.
• LH peak persists for about 24 hours.
• The LH surge stimulates completion of reduction division of the oocyte
and initiates luteinization of the granulosa cells, synthesis of
progesterone and prostaglandins.
INTRODUCTION
• FSH rise: Preovulatory rise of progesterone facilitates the positive
feedback action of estrogen to induce FSH surge → increase in
plasminogen activator → plasminogen → plasmin → helps lysis of the
wall of the follicle.
2. Stretching factor
3. Contraction of the micromuscles in the theca externa and ovarian
stroma due to increased prostaglandin secretion.
• Following ovulation, the follicle is changed into corpus luteum.
• The ovum is picked up into the Fallopian tube and undergoes either
degeneration or further maturation
FERTILIZATION
• Fertilization is the process of fusion of the spermatozoon with the
mature ovum.
• It begins with sperm egg collision and ends with production of a
mononucleated single cell called the zygote.
Goal:
1. To initiate the embryonic development of the egg
2. To restore the chromosome number of the species.
APPROXIMATION OF THE GAMETES
• The ovum, after ovulation is picked up by the tubal fimbriae.
• This action is thought to be muscular or suction or ciliary action or a
positive chemotaxis exerted by the tubal secretion.
• The ovum is rapidly transported to the ampullary part.
• Fertilizable life span of oocyte ranges from 12 to 24 hours whereas
that of sperm is 48 to 72 hours.
• Out of hundreds of millions of sperms deposited in the vagina at
single ejaculation, only thousands enter the uterine tube while only
300–500 reach the ovum.
CONTACT AND FUSION OF THE
GAMETES
• Complete dissolution of the cells of the corona radiata occurs by the
chemical action of the hyaluronidase liberated from the acrosomal
cap of the hundreds of sperm present at the site
• Penetration of the zona pellucida is facilitated by the release of
hyaluronidase from the acrosomal cap.
• More than one sperm may penetrate the zona pellucida.
• One touches the oolemma.
CONTACT AND FUSION OF THE
GAMETES
• After the sperm fusion, penetration of other sperm is prevented by
zona reaction (hardening) and oolemma block.
• This is due to release of cortical granules by exocytosis from the
oocyte.
• Alteration of its electrical potential.
CONTACT AND FUSION OF THE
GAMETES
• Completion of the second meiotic division of the oocyte immediately
follows, each containing haploid number of chromosomes (23, X)
• The female pronucleus and the second polar body which is pushed to
the perivitelline space.
• The head and tail of the spermatozoon enter the cytoplasm of the
oocyte but the plasma membrane is left behind on the oocyte
surface.
• Head and the neck of the spermatozoon become male pronucleus
containing haploid number of chromosomes (23, X) or (23, Y).
CONTACT AND FUSION OF THE
GAMETES
• The male and the female pronuclei unite at the center with
restoration of the diploid number of chromosomes (46) which is
constant for the species.
• The zygote formed contains both the paternal and maternal genetic
materials.
Immunological reaction (chemotaxis).
• An antigen called fertilizin present on the cortex and its coat of the
ovum, reacts with the antibody called antifertilizin liberated at the
plasma membrane of the sperm head.
CONTACT AND FUSION OF THE
GAMETES
Sex Determination:
• Sex of the child is determined by the pattern of the sex chromosome
supplied by the spermatozoon.
• The mature ovum carries 22 autosomes and one X chromosome,
while the mature sperm carries 22 autosomes and either an X or Y
chromosome.
• If the fertilizing sperm is carrying X chromosome the baby will be a
female (46 XX), if it is carrying Y chromosome the baby will be a male
(46 XY).
MORULA
• After the zygote formation, typical mitotic division of the nucleus
occurs by producing two blastomeres.
• The two cell stage is reached approximately 30 hours after
fertilization.
• Each contains equal cytoplasmic volume and chromosome numbers.
• The blastomeres continue to divide by binary division through 4, 8,
16 cell stage until a cluster of cells is formed called morula,
resembling a mulberry.
MORULA
• In the 16-64 cell stage, after spending about 3 days in the uterine
tube, the morula enters the uterine cavity through the narrow uterine
ostium (1 mm) on the 4th day
• The transport is a slow process and is controlled by muscular
contraction and movement of the cilia.
• The central cell of the morula is known as inner cell mass which forms
the embryo proper and the peripheral cells are called outer cell mass
which will form protective and nutritive membranes of the embryo.
BLASTOCYST
• The morula remains free in the uterine cavity on the 4th and 5th day,
covered by a film of mucus.
• A cavity appears within the morula converting it into a cystic structure
called blastocyst.
• Due to blastocyst enlargement the zona pellucida becomes stretched,
thinned and gradually disappears.
• Lysis of zona and escape of embryo is called zona hatching.
BLASTOCYST
• The cells on the outer side of the morula (polar) become trophectoderm
and the inner cells (apolar) become inner cell mass by the mediation of
epithelial cadherin (E-cadherin) (protein).
• Trophectoderm differentiates into chorion (placenta) and the inner cell
mass into the embryo.
• Completely undifferentiated cells are called the pluripotent embryonic
stem (ES) cells.
• ES cells are able to produce mature somatic cells of any germ layer
(ectoderm, mesoderm and endoderm).
• The blastocyst remains free in the uterine cavity for 3-4 days, during
which it is nourished by the secretion of the endometrium (uterine milk).
IMPLANTATION (nidation)
• Implantation occurs in the endometrium of the anterior or posterior
wall of the body near the fundus on the 6th day which corresponds to
the 20th day of a regular menstrual cycle.
• Implantation occurs through four stages
• Apposition
• Adhesion
• Penetration
• Invasion
CHANGES IN THE BLASTOCYST
• The polar trophoblast cells adjacent to the inner cell mass are
primarily involved in adhesion to the endometrial cells.
• The factors responsible for blastocyst attachment are:
• P. selectin, heparin sulfate, proteoglycan, EGF (epidermal growth
factor), integrins, trophinin and others.
• The signals for trophoblast multiplication arise from the inner cell
mass.
ENDOMETRIUM AT THE
IMPLANTATION SITE
• The endometrium is in the secretory phase corresponding to 20–21
days of cycle.
• The microvilli on the surface of the trophectoderm interdigitate with
the decidual cells to form the junctional complexes.
• Endometrial receptivity and molecular signaling during implantation is
induced by progesterone, LIF (leukemia inhibitory factor),
prostaglandins and COX-2 (cyclooxygenase).
APPOSITION
• Occurs through pinopod formation.
• Pinopods are long finger like projections (microvilli) from the
endometrial cell surface.
• These pinopods absorb the endometrial fluid which is secreted by the
endometrial gland cells.
• This fluid, rich in glycogen and mucin provides nutrition to the
blastocyst initially.
• Unless this fluid is absorbed, adhesion phase cannot occur.
ADHESION
• Adhesion of blastocyst to the endometrium occurs through the
adhesion molecules like integrin, selectin and cadherin
(glycoproteins).
PENETRATION AND INVASION
• Actual penetration and invasion occur through the stromal cells in
between the glands and is facilitated by the histolytic action of the
blastocyst.
• With increasing lysis of the stromal cells, the blastocyst is burrowed
more and more inside the stratum compactum of the decidua.
• Vacuoles appear in the advancing syncytium which fuse to form large
lacunae.
• Concurrently, the syncytial cells penetrate deeper into the stroma and
erode the endothelium of the maternal capillaries
PENETRATION
• The syncytium by penetrating the vessels becomes continuous with the
endothelial lining and permits the maternal blood to enter into the lacunar
system.
• Erosion of few maternal arteries with formation of blood space (lacunae) occurs.
• Nutrition is now obtained by aerobic metabolic pathway from the maternal
blood.
• Further penetration is stopped probably by the maternal immunological factor
and the original point of entry is sealed by fibrin clot and later by epithelium.
• The process is completed by 10th or 11th day which corresponds to D 24-25
from LMP.
PENETRATION
• This type of deeper penetration of the human blastocyst is called
interstitial implantation and the blastocyst is covered on all sides by
the endometrium (decidua).
• Occasionally, there may be increased blood flow into the lacunar
spaces at the abembryonic pole.
• This results in disruption of the lacunae and extravasation of blood
into the endometrial cavity.
• This corresponds approximately to 13th day after fertilization the
expected day of the following period.
PENETRATION
• This may produce confusion in determination of the expected date of
delivery.
• The process of implantation is controlled by the immunomodulatory
role of various cytokines (interleukins 3, 4, 5, 6, 10, 13), many local
peptides like epidermal growth factor (EGF), insulin like growth factor
(IGF) and prostaglandins.
• Both the decidua and the embryo synthesize these molecules.
TROPHOBLAST
• The cells of the blastocyst differentiate into an outer trophectoderm and
an inner cell mass.
• Just before implantation, the trophectoderm is further differentiated
into an inner mononuclear cellular layer called cytotrophoblast or
Langhans’ layer and an outer layer of multinucleated syncytium called
syncytiotrophoblast.
• The cytotrophoblasts that line the villous stems are the villous
cytotrophoblasts.
• The cytotrophoblast cells that invade the decidua are known as
‘interstitial extravillous cytotrophoblast’ and those that invade the
lumens of the maternal spiral arteries are known as ‘intravascular
extravillous cytotrophoblast’
TROPHOBLAST
• Throughout pregnancy, syncytiotrophoblast is derived from the
cytotrophoblast.
• Placenta and the fetal membranes are developed from the
trophoblast.
• It is involved in most of the functions ascribed to the placenta as a
whole.
• Thus, it serves at least 3 important functions — invasion, nutrition and
production of hormones for the maintenance of pregnancy.
• Local cytokines regulate the invasion of the cytotrophoblasts in the
decidua.
THE DECIDUA
• It is the thickened vascular endometrium of the pregnant uterus.
• It is called so because it casts of after parturition.
• The glands become enlarged, tortuous and filled with secretion.
• The stroma cells become large with small nuclei and clear cytoplasm,
these are called decidual cells.
• The decidua, like secretory endometrium, consists of three layers:
• the superficial compact layer
• the intermediate spongy layer
• the thin basal layer
STRUCTURE OF DECIDUA
THE DECIDUA
• The separation of placenta occurs through the spongy layer while the
endometrium regenerates again from the basal layer.
• The trophoblast of the blastocyst invades the decidua to be
implanted in:
• the posterior surface of the upper uterine segment in about 2/3
of cases
• the anterior surface of the upper uterine segment in about 1/3
of cases.
THE DECIDUA
• After implantation the decidua becomes differentiated into:
• Decidua basalis (serotine) under the site of implantation (the portion of
the decidua in contact with the base of the blastocyst).
• Decidua capsularis (reflexa) covering the ovum (the thin superficial
compact layer covering the blastocyst).
• Decidua parietalis (vera) lining the rest of the uterine cavity.
• As the conceptus enlarges and fills the uterine cavity the decidua
capsularis fuses with the decidua parietalis. (decidual space)
• This occurs nearly at the end of 12 weeks.
THE DECIDUA
• At term, they become atrophied due to pressure and the two cannot be
defined as a double layer.
• The decidua basalis retains its characteristic appearance till term and
becomes the maternal portion of the placenta
• The decidua has the following functions:
1. It is the site of implantation.
2. It resists more invasion of the trophoblast.
3. It nourishes the early implanted ovum by its glycogen and lipid contents.
4. It shares in the formation of the placenta.
CHORION
• The chorion is the outermost layer of the two fetal membranes
(chorion and amnion).
• After implantation, the trophoblast differentiates into 2 layers:
a. an outer one called syncytium (syncytiotrophoblast) which is
multinucleated cells without cell boundaries,
b. an inner one called Langhan’s layer (Cytotrophoblast) which is
cuboidal cells with simple cytoplasm.
CHORION
• A third layer of mesoderm appears inner to the cytotrophoblast.
• The trophoblast and the lining mesoderm together form the
chorion.
• Mesodermal tissue (connecting stalk) connects the inner cell mass to
the chorion and will form the umbilical cord later on.
• Spaces (lacunae) appear in the syncytium, increase in size and fuse
together to form the " chorio-decidual space" or " intervillus space".
• Erosion of the decidual blood vessels by the trophoblast allows blood
to circulate in this space.
CHORION
• The outer syncytium and inner Langhan’s cells form finger-like buds
surrounding the developing ovum called primary villi.(9th day)
• When the mesoderm invades the center of the primary villi they are
called secondary villi. (16th day)
• When blood vessels (branches from the umbilical vessels) develop
inside the mesodermal core, they are called tertiary villi. (21st day)
CHORION
• At first, the chorionic villi surround the developing ovum.
• After the 12th week, the villi opposite the decidua capsularis atrophy
leaving the chorion laeve which forms the outer layer of the foetal
membrane and is attached to the margin of the placenta.
• The villi opposite the decidua basalis grow and branch to form the
chorion frondosum and together with the decidua basalis will form
the placenta.
• Some of these villi attach to the decidua basalis ( the basal plate)
called the "anchoring villi", other hang freely in the intervillous
spaces called "absorbing villi"
AMNION
• On the 8th day, the embryoblast differentiates into bilaminar germ disc
which consists of dorsal ectodermal layer and ventral endodermal layer.
• The bilaminar germ disc is connected with the trophoblast by
mesenchymal condensation (connecting stalk)
• Two cavities appear one on each side of the germ disc, the amniotic
cavity and yolk sac.
• The layer of cells at the floor of the amniotic cavity will give the
ectodermal structures of the foetus and the layer of cells at the roof of
the yolk sac will give the endodermal structures of the foetus and the
mesoderm in between will give the mesodermal structure.
AMNION
Phases of conceptus development:
1. The ovum: the products of conception in the first 2 weeks after
fertilization.
2. The embryo: from 3 to 5 weeks.
3. The foetus: the developing infant (6-40 wks).
THE PLACENTA
• The placenta develops from the chorion frondosum (foetal origin)
and decidua basalis (maternal origin).
• It begins at 6th week and is completed by 12th week
Anatomy At Term:
• Shape discoid. Diameter: 15-20 cm. Weight : 500 gm.
• Thickness 2.5 cm at its center and gradually tapers towards the
periphery.
• Position in the upper uterine segment (99.5%), either in the
posterior surface (2/3) or the anterior surface (1/3).
THE PLACENTA
Surfaces:
A. Foetal surface: smooth, glistening and is covered by the amnion which is
reflected on the cord.
• The umbilical cord is inserted near or at the center of this surface and its
radiating branches can be seen beneath the amnion.
• At term, about four-fifths of the placenta is of fetal origin.
B. Maternal surface: dull greyish red in colour and is divided into 15-20
cotyledons.
• Each cotyledon is formed of the branches of one main villus stem covered
by decidua basalis.
THE PLACENTA