Power Point 2.o: Fertilization.

Learning Goals-
Topic 2: Development
Understand key features of fertilization and the stages of development
by comparing and contrasting these processes in model organisms
Sea Urchin, Frog, Chick, Drosophila, Human
Describe important developmental mechanisms
Types of cell movement
Cell migration
Cell fate and differentiation
 Organization during development (eg., axis, pattern formation, germ
layers)
Key genes
Apoptosis
 Fertilization
Formation of a diploid zygote from a haploid egg & sperm
Key Features:
Recognition at a distance

Contact Recognition and binding

Egg and Sperm Fusion

Blocks to Polyspermy

Egg Activation
 Recognition at a distance
Sea Urchin

Resact - soluble glycoprotein derived from the jelly layer

of the egg.

Resact is released from the jelly layer into the

surrounding seawater.

Sperm recognize and bind resact (species- specific) and

swim in the direction of higher resact concentration.

Chemotaxis- The migration of cells towards a soluble

concentration gradient of a stimulant.
 Contact Recognition
Sea urchin
Facilitated by a carbohydrate molecule
(fucose sulfate) in egg jelly layer binding to a
receptor on sperm plasma membrane
Overview of Fertilization
 Sea Urchin Fertilization

Sperm contacts the jelly coat

Acrosomal reaction: The acrosomal vesicle fuses with the plasma membrane
causing the extracellular release of digestive enzymes that penetrate the jelly
coat.

The acrosomal process forms due the polymerization of actin monomers

to form an actin filament.

Proteins on the surface of the acrosomal process bind to receptors

on the egg membrane
 Egg and Sperm Binding
Sea urchin

Bindin - protein molecule on the acrosomal

membrane that binds with species specificity to
the vitelline layer of the egg
Fast Block to Polyspermy: fusion of sperm and egg membranes
triggers depolarization (Na+ channels open, Na+ enters, inside
becomes more positive=depolarizes) of the membrane 1-3 sec after
sperm binds to egg; prevents additional sperm from fusing
(depolarization only last for about 1 minute)
 Blocks to Polyspermy
Fast transient block - membrane depolarization
Rapid change in the egg plasma membrane potential caused by
increase in Na+ concentration.

Unfertilized sea urchin eggs have a net negative charge inside,

but the charge becomes more positive upon fertilization.

When sea urchin sperm encounter an egg with a positive charge,

sperm-egg fusion is blocked
Cortical Reaction/Slow Block to Polyspermy: triggered by intracellular
calcium release which causes cortical granules (vesicles) to fuse with
membrane. Enzymes released from granules clip receptors, lifting
vitelline layer, which then hardens to form the fertilization envelope.
Sperm nucleus enters.
Slow permanent block - the cortical reaction
Increase in intracellular calcium levels triggers cortical granule
fusion .

Cortical Granules fuse with egg plasma membrane resulting in the

release of the following molecules into the extracellular space:

Proteinases and Glycosidases – separate vitelline layer

from plasma membrane
Mucopolysaccharides – osmotic gradient

Peroxidases –crosslinks macromolecules of the vitelline membrane

Hyalin protein- modifies the extracellular matrix of the egg to

help block sperm entry; coats outer surface of egg.
Putting it all together-Sea Urchin Fertilization
 Egg Activation

Early Events - increase in cell metabolism

Late Events - initiation of protein and DNA

synthesis in preparation of first cleavage (division)
Timeline for the fertilization of sea urchin eggs
 Fertilization in Mammals
Internal fertilization (unlike Sea Urchin)

Capacitation: process by which the glycoprotein coat and the

seminal proteins are removed from the surface of the sperm's
acrosome by substances secreted by the uterus or fallopian tubes
– Increases sperm metabolism and motility
– Necessary for future sperm and egg binding
– Triggered by bicarbonate ions (HCO –) in the vagina
3
– Requires about 5–6 hours in humans.

Zona pellucida: extracellular matrix of the egg

 Sperm Entry
http://www.sciencedirect.com/science/article/pii/S0014579307001792

Microvilli in
egg plasma
membrane
help facilitate
fusion
 Mammalian Fertilization

The acrosomal reaction occurs when sperm comes into contact with the
oocyte's zona pellucida (zp). Acrosomal enzymes begin to dissolve zp; actin
filament comes into contact with the zp. Then- calcium influx occurs, causing
cortical granules inside the oocyte then fuse to the outer membrane.
Contact Recognition and Binding
in Mammals
• The Zona Pellucida is
composed of three
proteins, ZP1, ZP2
and ZP3.
• Sperm plasma
membrane receptors
bind to ZP3
The cortical reaction in a
mouse prevents additional
sperm from entering the egg

The released contents of the

cortical granules both
remove carbohydrate from
ZP3 so it no longer can bind to
the sperm plasma
membrane and partly cleave
ZP2, hardening the zona
pellucida.

Together these changes

provide a block to polyspermy.
 Fertilization in mammals
Sperm binding and fusion of membranes triggers enzymes
release from cortical granules.

This cause changes in the zona pellucida resulting in the

cortical reaction and slow block to polyspermy: prevents
other sperm from fertilizing the same egg.

The oocyte now undergoes its second meiotic division

producing the haploid ovum and releasing a polar body.

The sperm and egg genetic material fuse

Timing
First cell division in sea urchins occur ~ 90 minutes after
fertilization

In humans: 12-36hrs

Recap
Understand the process of fertilization (and
steps involved)

Compare and contrast the unique features of

fertilization in various organisms