FERTILIZATION

Prof Dr Shaista Ali

 LEARNING OBJECTIVES
• Enlist the factors involved in transport of
oocyte and sperm
• Know the process and importance of
capacitation
• Define fertilization
• Discuss the phases of fertilization
• Enlist the results of fertilization
OOCYTE TRANSPORT
• Fimbriae of the uterine
tube sweep over the
surface of the ovary
• The fluid produced by
the mucosal cells of
uterine tube and their
cilia, sweep the
secondary oocyte into
the tube
• The muscles of the tube
contract rhythmically
and this peristalsis takes
the oocyte to the
ampulla
 SPERM TRANSPORT
• The VOLUME of ejaculate: averages 3.5 ml
range of 2 to 6 ml (200million – 600 million sperms)
• MOVEMENT SPEED 2 to 3 mm per minute
• SPEED varies with the pH of the environment, non motile during
storage in the epididymis, become motile in the ejaculate, move
slowly in the acid environment of the vagina, but move more
rapidly in the alkaline environment of the uterus
• Out of about 300 million, approximately 200-500 sperms reach the
fertilization site. Most sperms degenerate and are resorbed
• TIME OF TRANSPORT : recovered from the ampulla in 5 minutes
after their deposition. May take as long as 30 minutes - 6 days
•
 FACTORS INVOLVED IN SPERM TRANSPORT

• Travel towards the uterine tube with the help of

muscular contraction of the uterus and tube
• Chemo attractants produced by oocyte and
surrounding cells causes sperm chemotaxis
• Carbohydrate binding molecules and gamete specific
proteins on sperm surface are involved in sperm-egg
recognition
 VIABILITY OF GAMETES
• Oocytes are usually fertilized within 12 hours after ovulation
• They cannot be fertilized after 24 hours and get degenerated
• Sperms usually do not survive for more than 48 hours in the
female genital tract
• Some are stored in folds of the mucosa of the cervix and are
gradually released
• The short-term storage of sperms in the cervix and a gradual
release of sperms increases the chances of fertilization
 CAPACITATION
THE PERIOD OF CONDITIONING OF SPERM IN THE
FEMALE GENITAL TRACT
• Newly ejaculated sperms cannot
fertilize the oocytes
• The process of capacitation takes
5-7 hours
• Capacitated sperms are more
active
• Location: in the uterus but mostly
in oviducts, facilitated by
substances of the female genital
tract
• It is the removal of glycoprotein
and a seminal plasma protein
from the acrosomal region of the
spermatozoa
• The acrosomal reaction cannot
occur until capacitation has
occured
FERTILIZATION
DEFINITION :
Fertilization is a complex
sequence of coordinated
molecular events that begins
with contact between a sperm
and an oocyte and ends with
the intermingling of maternal
and paternal chromosomes at
metaphase of the first mitotic
division of the zygote, a
unicellular embryo
SITE:
Occurs in the ampullary region
of the uterine tube
PHASES
OF
FERTILIZATION
 PENETRATION OF THECORONA RADIATA
• Capacitated sperm disperse
the follicular cells of the
corona radiata, mainly by the
action of the enzyme
hyaluronidase released from
the acrosome of the sperm
• Tubal mucosal enzymes also
appear to assist the dispersal
• Movements of the tail of
the sperm are also important
in its penetration of the
corona radiata
STRUCTURE OF ZONA PALLUCIDA
• zona pellucida is
composed of 3 glycoproteins
• ZP1, ZP2, ZP3
• Repeating subunits of ZP2
and ZP3 form filaments
that are bound together by
ZP1
• The sperm binds to ZP3 and
this association, the zona
proteins causes a change in
ionic composition across the
sperm plasmalemma that
results in the acrosome
reaction, resulting in the
development of perforations in
acrosome
PENETRATION OF THE ZONA PELLUCIDA
• Results from the action of
enzymes released from the
acrosome
• The most important of
these enzymes is acrosin, a
proteolytic enzyme
• The enzymes esterases,
and neuraminidase are
also released, causing lysis
of the zona pellucida,
forming a path for the
sperm to reach the oocyte
SUMMARY ACROSOME REACTION
• Occurs after binding of
sperm with the zona
pellucida
• Reaction results in
acrosomal perforation
and the release of
enzymes needed to
penetrate the zona
pellucida
• Enzymes
❑ Acrosine
❑ Trypsin like substance
FUSION OF PLASMA MEMBRANES OF THE OOCYTE AND SPERM
• After penetration of the zona
pellucida, zona reaction, a change
in the composition of the zona
pellucida, that makes it
impermeable to other sperms
• The zona reaction result from the
action of lysosomal enzymes
released by cortical granules near
the plasma membrane of the
oocyte
• The contents of these granules are
released into the perivitelline
space, also cause changes in the
plasma membrane of oocyte,
making it impermeable to other
sperms, prevent polyspermy
FUSION OF PLASMA MEMBRANES OF THE OOCYTE AND
SPERM
• The plasma/cell membranes
of the oocyte and sperm fuse
and break down at the area
of fusion
• The head and tail of the
sperm enter the cytoplasm of
the oocyte, but the sperm's
plasma membrane remains
behind
COMPLETION OF THE SECOND MEIOTIC DIVISION OF
OOCYTE AND FORMATION OF FEMALE PRONUCLEUS

• Penetration of the oocyte by a

sperm activates the oocyte into
completing the second meiotic
division forming a definite
oocyte and a second polar body

• Following decondensation and

DNA replication of the maternal
chromosomes, the nucleus of
the definite oocyte becomes
the female pronucleus
FORMATION OF THE MALE PRONUCLEUS AND OOTID
• Within the cytoplasm of the oocyte, the
nucleus of the sperm enlarges, form
the male pronucleus
• The tail of the sperm degenerates so
the embryo contains only maternal
mitochondria
• Morphologically, the male and female
pronuclei are indistinguishable

• During growth of the pronuclei, they

replicate their DNA

• The oocyte containing two haploid

pronuclei is called an ootid
THE FUSION OF THE PRONUCLEI
• As the pronuclei fuse into a
single diploid aggregation of
chromosomes, the ootid
becomes a zygote
• The chromosomes in
the zygote become
arranged on a cleavage
spindle in preparation
for cleavage of the
zygote
 THE RESULTS OF FERTILIZATION
• Stimulates the secondary oocyte to complete
meiosis
• Restores the normal diploid number of
chromosomes (46)
• Results in variation of human species as maternal
and paternal chromosomes intermingle
• Determines the sex of the embryo
The sex chromosome (Y or X) carried by the successful sperm
determines embryonic sex
• Causes metabolic activation of ootid and initiates
cleavage
 TRIPLOIDY (IN VITRO)
• There are 3 pronuclei
within one zygote
• In the laboratory, such
embryos are discarded
• In vivo, such embryos
almost always abort
spontaneously
Advanced Fertility Center of Chicago
[Link]