MECHANISM OF FERILIZATION

Fertilization, union of a sperm nucleus, of paternal origin, with an egg nucleus, of

maternal origin, to form the primary nucleus of an embryo. In all organisms the essence
of fertilization is, in fact, the fusion of the hereditary material of two different sex cells,
or gametes, each of which carries half the number of chromosomes typical of
the species. The most primitive form of fertilization, found in microorganisms and
protozoans, consists of an exchange of genetic material between two cells.

The first significant event in fertilization is the fusion of the membranes of the two
gametes, resulting in the formation of a channel that allows the passage of material from
one cell to the other. Fusion in advanced animals is usually followed by penetration of
the egg by a single spermatozoon. The result of fertilization is a cell (zygote) capable of
undergoing cell division to form a new individual

The fusion of two gametes initiates several reactions in the egg. One of these
causes a change in the egg membrane(s), so that the attachment of and
penetration by more than one spermatozoon cannot occur. In species in which
more than one spermatozoon normally enters an egg (polyspermy), only one
spermatozoal nucleus actually merges with the egg nucleus. The most important
result of fertilization is egg activation, which allows the egg to undergo cell
division. Activation, however, does not necessarily require the intervention of a
spermatozoon; during parthenogenesis, in which fertilization does not occur,
activation of an egg may be accomplished through the intervention of physical
and chemical agents. Invertebrates such as aphids, bees, and rotifers normally
reproduce by parthenogenesis.

Capacitation of sperm
Capacitation, generally speaking, is the change sperm undergo in the female reproductive
tract that enables them to penetrate and fertilize an egg.

This step is a biochemical event; the sperm moves normally and looks mature prior to capacitation.
It is important to note that once the sperm reaches the egg, it does not mean that it is capable
of fertilizing it immediately. In order to fertilize the egg, the sperm must undergo the process of
capacitation in the reproductive tract where a number of enzymes and signaling molecules
are involved. This process can take around 10 hours, which means that the fertilization time
is approximately 24 hours.
 Capacitation causes a series of changes in sperm’s biochemical composition and structure
such that they develop increased motility and are prepared for interactions with the egg (or oocyte)
at fertilization. Essentially, the sperm becomes able to break through the membrane (known as
the zona pellucida) that surrounds the egg. Once sperm is able to traverse the zona pellucida,
it can begin the process of fertilizing the egg.

It is also important to consider the two distinct types of fertilization when thinking about
capacitation and how it occurs:

• For in vivo fertilization, or fertilization happening in living organisms, capacitation occurs

after ejaculation when sperm leaves the vagina and enters later sections of female
reproductive tract. The uterus aids in the steps of capacitation by secreting sterol-
binding albumin, lipoproteins, and certain enzymes.
• For in vitro fertilization, or assistive reproductive technologies, capacitation occurs by
incubating spermatozoa that have either undergone ejaculation or have been extracted
from the epididymis and incubated outside the body. In this process, an ovum or ova
is also removed from the woman's ovaries and co-incubated with sperm for fertilization
before being implanted into the female body.

What happens during capacitation?

• Membrane remodeling: The sperm's membrane becomes more fluid, allowing it to fuse
with the egg
• Motility changes: Sperm move in a more hyperactive, asymmetrical pattern
• Protein phosphorylation: Protein kinases are activated, and protein phosphatases are
inactivated
• Removal of seminal plasma proteins: The sperm's membrane is destabilized to prepare
for the acrosome reaction
• Cholesterol removal: Cholesterol is removed from the sperm
• Ion permeability changes: The sperm's ion permeability changes
 When does capacitation occur?
• Capacitation begins after ejaculation when sperm come into contact with fluids in the female
reproductive tract
• It continues as the sperm move through the female reproductive tract
What is the result of capacitation?
• Sperm are able to respond to zona pellucida ligands and undergo the acrosome reaction

Maturation of the egg

Maturation is the final step in the production of functional eggs (oogenesis) that can
associate with a spermatozoon and develop a reaction that prevents the entry of more
than one spermatozoon. In addition, the cytoplasm of a mature egg can support the
changes that lead to fusion of spermatozoal and egg nuclei and initiate embryonic
development
Egg surface
Certain components of an egg’s surface, especially the cortical granules, are associated
with a mature condition. Cortical granules of sea urchin eggs, aligned beneath
the plasma membrane (thin, soft, pliable layer) of mature eggs, have a diameter of 0.8–
1.0 micron (0.0008–0.001 millimetre) and are surrounded by a membrane similar in
structure to the plasma membrane surrounding the egg. Cortical granules are formed in
a cell component known as a Golgi complex, from which they migrate to the surface of
the maturing egg.

Egg coats
The surfaces of most animal eggs are surrounded by envelopes, which may be soft
gelatinous coats (as in echinoderms and some amphibians) or thick membranes (as
in fishes, insects, and mammals). In order to reach the egg surface, therefore,
spermatozoa must penetrate these envelopes; indeed, spermatozoa
contain enzymes (organic catalysts) that break them down. In some cases (e.g., fishes
and insects) there is a channel, or micropyle, in the envelope, through which a
spermatozoon can reach the egg.

The jelly coats of echinoderm and amphibian eggs consist of

complex carbohydrates called sulfated mucopolysaccharides. The envelope of a
mammalian egg is more complex. The egg is surrounded by a thick coat composed of a
carbohydrate protein complex called zona pellucida. The zona is surrounded by an outer
envelope, the corona radiata, which is many cell layers thick and formed by follicle cells
adhering to the oocyte before it leaves the ovarian follicle.

Although it once was postulated that the jelly coat of an echinoderm egg contains a
substance (fertilizin) thought to have an important role not only in the establishment of
sperm-egg interaction but also in egg activation, fertilizin now has been shown identical
with jelly-coat material, rather than a substance continuously secreted from it. Yet there
is evidence that the egg envelopes do play a role in fertilization; i.e., contact with the egg
coat elicits the acrosome reaction (described below) in spermatozoa.
Events of fertilization
Sperm-egg association
fertilization

Specificity of sperm-egg interaction

Fertilization is strictly species-specific, and the egg’s coating, the zona

pellucida, plays an important role in the binding process between sperm
and egg. In general, the biochemistry of the zona pellucida of one species
differs from that of another, and thus it only matches up and binds with
sperm of the appropriate species. For example, among the echinoderms,
solutions of the jelly coat clump, or agglutinate, only spermatozoa of their
own species.
 In both echinoderms and amphibians, however, slight damage to an egg
surface makes fertilization possible with spermatozoa of different species
(heterologous fertilization); this procedure has been used to obtain certain
hybrid larvae. In addition, binding between sperm and egg of different
species may occur when the zona pellucida of the egg is removed.

The eggs of ascidians, or sea squirts, members of

the chordate subphylum Tunicata, are covered with a thick membrane
called a chorion. The space between the chorion and the egg is filled with
cells called test cells. The gametes of ascidians, which have both male and
female reproductive organs in one animal, mature at the same
time, yet self-fertilization does not occur. If the chorion and the test cells
are removed, however, not only is fertilization with spermatozoa of
different species possible, but self-fertilization also can occur.
The steps in fertilization (Fig. 40.4) are:

I. Approach of the spermatozoon to the egg:

a. By chemotactic movement the sperm swims towards the egg.

b. The jelly coat of the egg produces fertilizin and the sperms produce antifertilizin.

c. The fertilizin and the antifertilizin combine to form an initial bond to facilitate
penetration of spermatozoon into the egg.
The zona pellucida (ZP) proteins are ZP1, ZP2, ZP3, and ZP4, which are glycosylated
proteins that make up the zona pellucida. The ZP is a transparent, porous coat that
surrounds mammalian eggs and plays a vital role in fertilization.
Function
• ZP proteins are involved in oogenesis, fertilization, and preimplantation development
• ZP proteins bind to spermatozoa
• ZP proteins induce the acrosome reaction
• ZP proteins establish the ZP block to polyspermy
II. Penetration of the sperm:

a. The bound or agglutinated spermatozoon produces an enzyme, lysine, which

dissolves egg membranes in the local area and clears the path for spermatozoon to
reach the egg surface.
b. The acrosomal filament is pushed through the jelly and vitelline membrane, and
touches the surface membrane of the egg cytoplasm.

III. Reaction in egg during sperm penetration:

a. Coming in contact with the acrosomal filament the egg cytoplasm bulges forward
to produce a conical projection, the fertilization cone.

b. It gradually engulfs the spermatozoon and carries it inward.

c. Simultaneously profound cortical reaction occurs in the cytoplasm.

d. Immediately after penetration of the spermatozoon, the vitelline membrane sepa-

rates from the plasma membrane. The vitelline membrane thickens and forms the
fertilization membrane which prevents entry of other sperms and thus ensures en-
trance of only one sperm into the egg.

e. The cortical granules swell, become liquefied and the fluid is released in the space
between egg cytoplasm and fertilization membrane.

IV. Behaviour of sperm within the ovum:

a. The head and middle piece enter the egg cytoplasm but the tail is left outside.

b. The egg completes its meiosis immediately and loses the centriole.

c. The sperm rotates 180° and its middle part comes to the front end. The path along
which the sperm moves within the egg is known as fertilization path.

d. The egg pro-nucleus moves to the periphery

e. The sperm pro-nucleus swells up. The centrosome and the centriole in the middle
piece form aster around the two pronuclei.

f. The sperm centriole divides into two, move to the poles, being connected by fibres.

g. The nuclear membranes disappear, the chromosomes of the male and female
pronuclei are arranged in the equator of the spindle.

h. It is similar to metaphase plate of meiosis and immediately followed by the

cleavage of the zygote.

Importance of Fertilization:

a. The sperm entry activates the secondary oocyte to complete its maturation divi-
sion.

b. Restores diploid number of chromosomes and recombines the maternal and

paternal genetic materials.