FERTILIZATION

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. You
can simply say that it is the complex interaction between the male and female gametes to form
a zygote. It occurs in the ampulla of the uterine tube within 24 hours after ovulation.

The sperm pass through the cervix and the uterus and into the fallopian tubes by means of the
flagellar (whiplike) activity of their tails and can reach the fallopian tubes within 5 minutes after
coitus.

The time during which fertilization can occur is brief because of the short life span of mature
gametes. The ovum is estimated to survive for up to 24 hours after ovulation. The sperm
remains capable of fertilizing the ovum for up to 5 days after being ejaculated into the area of
the cervix.

Fig 1: Shows the movement of sperm and the optimal site of fertilization

Phases of Fertilization

1. Passage of sperm through the corona radiata: Dispersal of the follicular cells of the corona
radiata surrounding the oocyte and the zona pellucida appears to result mainly from 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.

2. Penetration of the zona pellucida: Passage of a sperm through the zona pellucida is the
important phase in the initiation of fertilization. The enzymes esterases, acrosin, and
neuraminidase appear to cause lysis of the zona pellucida, thereby forming a path for the
sperm to follow to the oocyte. The most important of these enzymes is acrosin, a proteolytic
enzyme. Once the sperm penetrates the zona pellucida, a zona reaction (a change in the
properties of the zona pellucida) occurs that makes it impermeable to other sperms. It is
believed that granules released from the secondary oocyte, which contain lysosomal enzymes,
produce this zona reaction.

3. Fusion of plasma membranes of the oocyte and sperm: The plasma or 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.

4. 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 and forming a mature oocyte and a second polar body. Following decondensation of
the maternal chromosomes, the nucleus of the mature oocyte becomes the female pronucleus.

5. Formation of the male pronucleus: Within the cytoplasm of the oocyte, the nucleus of the
sperm enlarges to form the male pronucleus and the tail of the sperm degenerates.

6. Fusion of the pronuclei and formation of a zygote: The male and female pronuclei fuse into
a single diploid aggregation of chromosomes to form a zygote. The chromosomes in the zygote
become arranged on a cleavage spindle in preparation for cleavage of the zygote.

Important Events of Fertilization

 Stimulation of the penetrated oocyte to complete the second meiotic division.

 Restoration of the normal diploid number of chromosomes (46) in the zygote.
 Variation of the human species through mingling of maternal and paternal
chromosomes.
 Determination of chromosomal sex of the embryo.
 Initiation of cleavage (cell division) of the zygote.

Clinical Correlates

1. Assisted Reproductive Technologies (ART)

a. In-vitro Fertilization (IVF) and Embryo Transfer

b. Cryopreservation of Embryos
c. Intracytoplasmic Sperm Injection
d. Surrogate Mothers

CLEAVAGES
Approximately 30 hours following fertilization, the zygote undergoes repeated mitotic divisions.
This is termed as Cleavage. This results in a rapid increase in the number of cells. These cells
become smaller with each cleavage division, known as blastomeres.

The zygote is the cell formed by the union of the sperm and the ovum, and it is transported
through the fallopian tube and into the uterus. During transport through the fallopian tube, the
zygote undergoes rapid mitotic division, or cleavage.
The first cleavage produces a 2-cell embryo, followed by 4-cell, 8-cell, 16-cell embryos, to form
the blastomere. The size of the zygote does not increase; rather, the individual cells become
smaller as they divide and eventually form a solid ball called the morula (about 16 cell or more)
3 days after fertilization.

At the 8 cell stage the human embryo undergoes a process called compaction. It results from
cells adhering together more tightly. The embryo becomes more compact, but the cells still
remain separate from each other. The cell adhesion protein, E-cadherin appears at the time of
compaction and causes the cells to adhere together more tightly than previously.

BLASTOCYST FORMATION

Shortly after the morula enters the uterus (approximately 4 days after fertilization), a fluid-filled
space called the blastocystic cavity appears inside the morula. This fluid passes from the
uterine cavity through the zona pellucida to form this space. As fluid increases in the
blastocystic cavity, it separates the blastomeres into two parts:

 A thin, outer cell later, the trophoblast, which gives rise to the embryonic part of the
placenta.
 A group of centrally located blastomeres, the inner cell mass, which gives rise to the
embryo; because it is the primordium of the embryo, the inner cell mass is called the
embryoblast.

During this stage of development - blastogenesis - the conceptus is called a blastocyst. After
the free blastocyst has floated in the uterine secretions for approximately 2 days, the zona
pellucida gradually degenerates and disappears. Shedding of the zona pellucida permits the
hatched blastocyst to increase rapidly in size.

Approximately 6 days after fertilization, the blastocyst attaches to the endometrial epithelium,
usually adjacent to the embryonic pole. As soon as it attaches to the endometrial epithelium,
the trophoblast starts to proliferate rapidly and gradually differentiates into two layers:

 An inner layer of cytotrophoblast

 An outer layer of syncytiotrophoblast
IMPLANTATION

Implantation of the blastocyst begins at the end of the first week and is completed during the
second week. It usually occurs in the endometrium of the uterus. Once implantation has taken
place, the trophoblast cells and other adjacent cells (from the blastocyst and the uterine
endometrium) proliferate rapidly, forming the placenta and the various membranes of
pregnancy.

Effects of hormones in Implantation of Embryo

Estrogen & progesterone hormones secreted from the ovary under the influence of FSH & LH,
help in the growth of the intra uterine endometrial layer and prepare it for implantation.

Clinical Correlates

1. Ectopic pregnancy

Fig 2: An overview of first week of development (fertilization to implantation).