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Zygote formation, Cleavage & Blastula formation

Click to edit Master subtitle style R. Mareeswaran, (09BC011)

Second year M.Sc., Biochemistry

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Early Embryonic Development

Cleavage starts Fertilization of ovum Ovary Oviduct

Secondary oocyte Blastocyst (implanted) Ovulation Endometrium Uterus

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Fertilization.....

Fertilization is the process whereby two sex cells (gametes) fuse together to create a new individual with genetic potentials derived from both parents. known as Zygote formation a single event occurs in a series of steps

Also Not

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Events
1. 2.

Contact and recognition

Regulation of sperm entry into the egg. Fusion of the genetic material of sperm and egg. Activation of egg metabolism

3.

4.

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Sperm Capacitation..
Freshly Must

ejaculated sperm are unable or poorly able to fertilize. undergo a series of changes collectively known as Capacitation. is associated with removal of adherent seminal plasma proteins, reorganization of plasma membrane lipids and proteins.

Capacitation

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Capacitation

occurs while sperm reside in the female reproductive tract for a period of time, as they normally do during gamete transport. appears to destabilize the sperm's membrane to prepare it for the acrosome reaction

Capacitation

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Phases.
Phase Phase Phase

1 - Penetration of the corona radiata; 2 - Penetration of the zona pellucida

3 - Fusion of the oocyte and sperm cell membranes.

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PHASE 1: PENETRATION OF THE CORONA RADIATA

Of

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200 to 300 million spermatozoa only 300 to 500 reach the site of fertilization. one of these fertilizes the egg. sperm pass freely through corona

Only

Capacitated

cells

PHASE 2: PENETRATION OF THE ZONA PELLUCIDA

Zona

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- glycoprotein shell

Facilitates

and maintains sperm binding and induces the acrosome reaction. binding and the acrosome reaction are mediated by the ligand ZP3, a zona protein. of acrosomal enzymes (acrosin) allows sperm to penetrate the zona, of the zona pellucida changes when the head of the sperm comes in contact with the oocyte surface. in release of lysosomal enzymes from cortical granules lining the plasma membrane

Both

Release

Permeability

Results

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Acrosomal Reaction
The

acrosome reaction provides the sperm with an enzymatic drill to get throught the zona pellucida. same zona pellucida protein that serves as a sperm receptor also stimulates a series of events that lead to many areas of fusion between the plasma membrane and outer acrosomal membrane. fusion (actually an exocytosis) and vesiculation expose the acrosomal contents, leading to leakage of acrosomal enzymes from the sperm's head.

The

Membrane

 As

the acrosome reaction progresses and the sperm passes through the zona pellucida, more and more of the plasma membrane and acrosomal contents are lost. the time the sperm traverses the zona pellucida, the entire anterior surface of its head, down to the inner acrosomal membrane, is denuded.

By

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The

constant propulsive force from the sperm's flagellating tail, in combination with acrosomal enzymes, allow the sperm to create a tract through the zona pellucida. a sperm penetrates the zona pellucida, it binds to and fuses with the plasma membrane of the oocyte.

Once

PHASE 3: FUSION OF THE OOCYTE AND SPERM CELL MEMBRANES

Initial

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adhesion of sperm to the oocyte - by the integrins & disintegrins penetration egg responds in three ways: Cortical and zona reactions.

After 1. 2.

Resumption of the second meiotic division Metabolic activation of the egg

3.

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The Zona Reaction

The

cortical reaction refers to a massive exocytosis of cortical granules seen shortly after sperm-oocyte fusion. granules contain a mixture of enzymes, including several proteases alter the structure of the zona pellucida, inducing what is known as the zona reaction. of cortical granules may also interact with the oocyte plasma membrane.

Cortical

Proteases

Components

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The critical importance of the zona reaction is that it represents the major block to polyspermy in most mammals. This effect is the result of two measurable changes induced in the zona pellucida:
1. 2.

The zona pellucida hardens. Sperm receptors in the zona pellucida are destroyed

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The Zona Reaction Animation

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Egg Activation
Prior

to fertilization, the egg is in a quiescent state, arrested in metaphase of the second meiotic division. binding of a sperm, the egg rapidly undergoes a number of metabolic and physical changes that collectively are called egg activation. effects include a rise in the intracellular concentration of calcium, completion of the second meiotic division and the so-called cortical reaction.

Upon

Prominent

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Fusion Of Genetic Material

Pronuclear Male

migration takes about 12 hours

and female pronuclei are indistinguishable

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Summary.
1 The sperm

acrosomal enzymes digest the 3 Proteins on the eggs jelly SPERM sperm head bind coat to egg receptors 4 The plasma membranes of sperm and egg fuse Sperm head 5 The sperm nucleus enters Nucleus the egg cytoplasm Acrosome Acrosomal Plasma 6 A membrane enzymes fertilization envelope forms Receptor protein molecules Plasma Sperm membrane nucleus Vitelline Cytoplasm layer Egg Jelly nucleus coat 7 The nuclei EGG CELL of sperm Zygote and egg fuse nucleus

approaches the egg

2 The sperms

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Results Of Fertilization
Restoration

of the diploid number of chromosomes of the sex of cleavage

Determination Initiation

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Blocking Of Polyspermy
2

Types
Fast block Changing electric potential block Zona reaction

Slow

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Cleavage.
Mitosis

and cytokinesis of the zygote produces an increasing number of smaller cells (Blastomeres), each with an exact copy of the genome present in the zygote. Tight junction & Gap junction 16 cell ends with the formation of a blastula.

Compaction Morula

Cleavage

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Cleavage Pattern

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Blastocyst Formation
Morula

enters the uterine cavity Embryoblast & Tropoblast

Blastocele Blastocyst

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ENDOMETRIUM Inner cell mass

Cavity

Trophoblast

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Th an Yo k u!