CHAPTER 5 FERTILIZATION

Fertilization occurs through sexual reproduction. Where two haploid gametes fused to produce an
genetically unique individual.

AMPULLARY REGION OF THE OVIDUCT - place where the egg and spermatozoon unite.

Mammalian Egg Complex (ovulated) enters the oviduct via the Infundibulum, consist of three
components:

1. The oocyte, arrested at metaphase of meiosis II in most domestic mammals.

2. The zona pellucida and extracellular matrix surrounding the oocyte.

3. The cumulus cells, consisting of several layers of cells from the cumulus oophorus embedded in an
extra cellular matrix composed mainly of hyaluronic acid.

Ovulated egg complex is describe as the Cumulus-oocyte Complex

- Sperm Transport in the Female Genitall Tract -

The duration of copulation varies among different domestic animals.

In several mammalian species the semen is deposited in to the cranial vagina.

In others the semen is either ejaculated directly into the cervix or via the processus urethralis into both
the cervix and uterus.

In ruminants the ejaculate is small in volume but with great concentration

In boar the volume of ejaculation is large but low in concentration

In stallion they ejaculate in a series of jets.

In the dog the first three fractions of the ejaculate originates from the prostrate. The volume of this
clear and acellular so called Presperm Fraction.

In cat the ejaculate volume is small and it is unclear whether it compromises multitask fractions.

Retograde Loss of Spermatozoa from the female tract.

A major barrier for spernatozoa transport is the CERVIX UTERI which can also serve as a reservoir of
spermatozoa.

The cervix act as filter for removing non-viable spermatozoa.

- Capacitation -

Spermatozoa are not able to fertilize the oocyte immediately after arrival in the female genital tract to
acquire fertility must reside there for a certain period of time. The changes that occur during this period
constitute CAPACITATION.

It begins in the uterus and ends in the isthmus of the oviduct.

Plasma membrane of the spermatozoon undergoes marked changes during capacitation.

Important processes during capacitation includes:

-removal of the glycoprotein coat and seminal plasma membrane

-the functional coupling of the signal-transducing pathways that regulate the initiation of the acrosome
reaction by the zona pellucida glycoproteins

-alterations in the flagellar motility that are necessary to penetrate so

-development of the capacity to fuse with the plasma membrane of the oocyte.

The reorganization of the sperm membrane after cholesterol depletion is considered an early step in
capacitation.

Spermatozoa express a bicarbonate-sensitive soluble form of adenyl cyclase that may control these
phosphorylation events.

- Interactions Between Spermatozoa and the Zona Pellucida -

ZONA PELLUCIDA - is an extracellular matrix surrounding the oocyte.

- it is composed of three glycoproteins:

ZPA = ZP2

ZPB = ZP1

ZPC = ZP3

Zona pellucida have been obtained from studies in the mouse.

It has been involved in several critical stages of fertilization: Adhesion, Binding, Capacitated
Spermatozoa to the ZP
*ADHESION OF SPERMATOZOA TO ZP

ADHESION - is the first contact between the spermatozoon and the zona pellucida it is followed by
relatively firm binding.

*ACROSOME REACTION

ACROSOME REACTION - is a result of which hydrolytic enzymes are released from the acrosome of the
sperm head. This permit the spermatozoon to penetrate the ZP.

When the plasma membrane forms multiple fusion sites with the outer acrosomal membrane resulting
in the formation of many small vesicles (vesiculation)

Acrosin and Hyaluronidase are enzymes released during the acrosomal reaction. Acrosin hydrolyzes ZP
proteins and also enhances the ability of spermatozoa to bind to these proteins.

Proacrosin is the inactive form of the enzyme acrosin and it has a strong affinity for the ZP. This,
proacrosin aids in binding to the ZO as the acrosomal reaction proceeds. As proacrosin is converted into
acrosin, the spermatozoon penetrates by using enzyme to digest a small hole in the zp and pass through
it.

- Adhesion and Fusion of the Spermatozoon and Oocyte -

After the penetration of the ZP, sperm adhere to and fuse with the plasma membrane of the oocyte.

The fertilizing spermatozoon including its tail is engulfed by the oocyte, this process is called SYNGAMY.

The membrane fusion of the male and female gametes involves;

Sperm Fertilin-a (ADAM1)

Fertilin-B (ADAM2)

Cyritestin (ADAM3)

CRISP1V (cystein-rich secretory protein 1)

- OOCYTE ACTIVATION -

After entry of the spermatozzon the oocyte undergoes OOCYTE ACTIVATION, which establishes a block
to polyspermic fertilization, resumption of meiosis and initiation of embryonic development.
In all animals investigated that activation involves and Increase In The Cytosoloc Calcium Ion
Concentration.

Oocyte activation responses include recruitment of maternal mRNAs for translation and changes in
protein synthesis.

Constitutes of the sperm nucleus presumably it's perinuclear theca, have been associated with oocyte
activating activities.

*BLOCK TO POLYSPERMIC FERTILIZATION

The block to polyspermic fertilization is established through exocytosis of a set of secretory granules the
Corticol Granules. This is referred to as the CORTICAL REACTION

Any further penetration of spermatozoa into the oocyte is prevented and the so called Zona-Block to
Polyspermy is establish.

*RESUMPTION OF MEIOSIS AND PRONUCLEUS FORMATION

As another consequence of oocyte activation meiosis is resumed and the second meiotic division is
completed.

The daughter cell that receives hardly any cytoplasm is called the Second Polar Body.

The other daughter cell is the definitive oocyte now referred to as the zygote.

The oocyte haploid chromosome set becomes surrounded by layers of smooth endoplasmic reticulum
that contribute to the formations of a nuclear envelope and a vesicular nucleus known as the Female or
Maternal Pronucleus. The sperm undergoes marked changes within the oocytes cytoplasm. It becomes
swollen, surrounded by smoothe endoplasmic reticulum contributing to a nuclear envelope and form
the Malebor Paternal Pronucleus.

The primary reducing agent is Glutathione from the oocyte cytoplasm.

Upon dismantling of the nuclear envelopes the male and female haploid genomes become United in the
center of the zygote. The mixing us referred to as KARYOGAMY OR SYNKARYOSIS.

At the dissolution of the nuclear envelopes of the pronuclei the chromatin condenses to form the
Prophase of the first mitotic Division.

The subsequent cleavage is normally completed within 24 hours after ovulation. If the oocyte is not
fertilized within this period of time it's loses its developmental potential.
OVERVIEW OF MAMMALIAN FERTILIZATION

A. Anaphase of the first meiotic division in the follicle

B. Penetration of the spermatozoon into the periviteline space; metaphase of the second meiotic
division and activation of the oocyte results in release of corticol granules.

C. First sperm is taken up into the oocyte by a phagocytic process: anaphase of the second meiotic
division

D. Formation of the male and female pronuclei, the tail of the spermatozoon degenerates.

E. Karyogamy

F. First mitotic division of the zygote.