PAPER 1 - PART B

Embryology - fertilization

References
BD Chaurasia's Dream Human Embryology, 2nd Edition
Inderbir Singh’s Human Embryology. 11th edition

Notes for first year BAMS students 2020-2021 batch

Prof (Dr K G Beena)

For internal circulation only

1
Fertilization

Fertilization is the process of fusion of two haploid germ cells, an ovum and a
spermatozoon resulting in the formation of a diploid, mono nucleated single cell,
the zygote. It takes place in the middle segment of the fallopian tube. The fertilized
ovum is a large cell which undergoes a series of divisions known as cleavage.

Out of a few hundred capacitated sperms, that surround the ovum, only one pierces
the zona pellucida and enters the ovum. As soon as one spermatozoon enters the
ovum, the second meiotic division is completed, and the second polar body is
extruded.

Fertilization involves the following steps

1. Approximation of gametes
2. Contact and fusion of gametes
3. Effects/Results of fertilization.

1. Approximation of gamete is by transport of male and female gametes towards

the fallopian tube of the uterus.

Transport of ovum and sperm towards the fallopian tube

At the time of ovulation the ovum is directly expelled into the peritoneal cavity.

2
The ovum that is shed from the ovary is closely embraced by the fimbriated end of
the uterine tube. The ovum is easily carried into the tube partly by the follicular
fluid discharged from the follicle and partly by the activity of ciliated cells lining the
tube. The ovum slowly travels through the tube toward the uterus, taking 3–4 days
to do so.
The fimbriae of uterine tube moves over the ovary at ovulation. The ciliary beats of
fimbriae sweep the ovulatory mass into the infundibulum. The ciliary beats of
uterine epithelium and muscular contractions of uterine tube are responsible for
the migration of ovum from the surface of ovary into the ampulla of uterine tube.
The ovum reaches ampulla, the site of fertilization in 25 minutes.

The ovum that is released at ovulation is viable for 24–48 hours. In the absence of
fertilization it degenerates.

Transport Spermatozoon

The prostaglandins present in semen stimulate peristaltic contractions of female

genital tract when the semen is deposited in the vagina . During their transport
there will be reduction in the number of spermatozoa due to the constrictions in
the female genital tract. Movements of their tails through uterus and tubes assisted
by muscular contraction are responsible for the movement of spermatozoa. Time
taken for transport to uterus is 5–45 minutes.

Around 200–500 million sperms are deposited in the female genital tract and about
300–500 spermatozoa only reach the site of fertilization. The life viability of
spermatozoa after ejaculation is 24–48 hours.

Capacitation is the final step in maturation of spermatozoon before fertilization and

it takes place in female genital tract. It starts in the uterus and continues into the
tubes. It involves number of changes including changes in the sperm cell membrane
and signal transduction (calcium influx, release of acrosomal enzymes, etc.). Certain
major changes in surface glycoproteins (removal of glycoprotein cut and exposing
zona binding proteins) is caused by secretions of the female genital tract thus
allowing sperm-ovum binding. Follicular fluid is thought to enhance the
capacitation.

Maturity and motility of sperms

During their passage through male genital tract the spermatozoa mature.
Movements of tail are important for their motility. Motility is important for
penetration of three barriers surrounding the ovum.
The motility of the spermatozoa depends on certain factors
3
 1. pH of the fluid medium
2. secretion of cervical mucus
3. fluid currents
4. temperature
5. hormones
Neutral and alkaline pH enhances the activity of spermatozoa . the vagina fluid
is acidic . The alkaline semen neutralizes the vaginal acidic fluid and enhances
the motility of spermatozoa
Cervical secretion acts as a mechanical barrier for spermatozoa. During the
time of ovulation cervical secretion becomes thin and watery which favours the
movement of spermatozoa. With rise of temperature the activity spermatozoa
increases.
Oxytocin causes propulsive movements of uterus which help to aspirate
seminal fluid from vagina into the fallopian tube. Oestrogen makes the cervical
secretion thin and watery and favours the movement of spermatozoa.
Prostaglandins in the semen also increase the movement female genital tract.
Progesterone released from follicle stimulates movements of spermatozoa.

2. Contact and fusion of gametes

Fusion of gametes involes the following steps
1. Chemo attraction
2. Penetration of spermatozoa through covering of ovum.

Chemo attraction of spermatozoa to ovum occurs by substance produced by

the ovum.
There are three barriers which the sperm has to penetrate before fusing with
the ovum. They are
1. Corona radiate
2. Zona pellucid
3. Vitelline membrane
Four processes are involved in the penetration of these barriers. They are acrosome
reaction, disintegration of barriers,depolarization in oocyte cytoplasm and nuclear
fusion

4
Coverings of sperm head - Acrosomal cap discharges chemical substances that
facilitate penetration of barriers around oocyte. This process is called acrosomal
reaction. The three acrosome enzymes that are released are hyaluronidase, the
protease enzyme acrosin and acid phosphatase. The glycoprotein of the zona
pellucida is responsible for induction of the acrosomal reaction.

Hyaluronidase from the acrosome of sperm, tubal mucosal enzymes and

movements of tail of spermatozoon aids in penetration of corona radiata. The
glycoproteins on the outer surface of sperm head binds with glycoproteins on the
zona pellucida of ovum. The reaction of zona is to prevent polyspermy. Alterations
in the plasma membrane of oocyte and zona pellucida ensure that no other
spermatozoon can enter the oocyte. The zona pellucida is altered due to release of
lysosomal enzymes by plasma membrane of the oocyte. This process is called zona
reaction.

When a spermatozoon comes in contact with the oocyte, plasma membranes of the
two cells fuse. The disintegrin peptide released from sperm head initiates fusion.
The vitelline membrane contains integrin peptides. This process takes 30 minutes.
The glycoproteins on the outer surface of sperm head binds with glycoproteins on
the zona pellucida of ovum.

The contact of sperm with vitelline membrane of oocyte triggers calcium wave
(depolarization) in oocyte cytoplasm and secondary oocyte resumes second meiotic
division

Entry of the sperm leads to metabolic changes within the ovum that facilitate its
development into an embryo.
Both head and tail of the spermatozoon (excluding plasma membrane) enters the
cytoplasm of oocyte. Approximation of pronuclei takes place near the middle of
cytoplasm of ovum. Immediately after the entry of sperm head into the cytoplasm
of the oocyte the latter completes its second meiotic division. Second polar body
extruded into perivitelline space

3. Results of fertilization

The results of fertilization are completion of second meiotic division of female

gamete, restoration of diploid number (46) of chromosomes, determination of sex,
initiation of cleavage (mitotic) division of zygote, determination of polarity and
bilateral symmetry of embryo and genetic diversity.