Lesson 4:

Fertilization
 Oocyte Preparation for Fertilization
• Watch the informative video Life's Greatest Miracle at the PBS Nova Web
site:.
 Development of an Oocyte
1. Inside the primordial (“first”) follicle,
meiosis I stops in the primary oocyte.
2. The zona pellucida begins to form around
the oocyte.
3. The antrum, a fluid-filled cavity, begins to
form in the follicle.
4. When the follicle is mature, meiosis I is
complete. The secondary oocyte and first
polar body form.
5. During ovulation, the mature follicle bursts
and releases the secondary oocyte and first
polar body.
6. The remains of the follicle form the corpus
luteum.
7. If conception does not occur, the corpus
luteum breaks down.
 Oocyte in Preovulatory Follicle
• Oocyte (O) possesses a
plasma membrane (vitelline
membrane).
• Just outside of cell membrane
is acellular coat: the zona
pellucida (Z).
• Cells [Granulosal (corona
radiata and cumulus
oophorus) cells] (CR)
surround the zona pellucida.
 Oocyte in Preovulatory Follicle
The whole structure
(attached at one pole) sits
inside fluid-filled cavity
(antrum).

The antrum is
surrounded by layers of
cells that make up the
follicle wall.

The oocyte is ~280 um in diameter (follicle is 25 mm)

 Preparation for Fertilization-Oocyte
• An ovulated oocyte possesses one
polar body; the polar body contains
excess DNA.
• The oocyte is arrested in meiosis.

Micrograph by
ekem
 The Oocyte and the Oviduct
• The upper-most portion of the
oviduct [infundibulum (If)]
sweeps over the surface of the
ovary (O).
• It collects oocyte into oviduct
(OD).
• Oocyte is propelled down toward
site of fertilization.
 Sperm Anatomy and Size
• Three components:
• Head [nucleus &
acrosome (giant secretory
vesicle)]
• Midpiece (mitochondria)
• Tail
• Size:
• Length = 55 um
• Width = 4 um

(Note: A sperm travels over

30,000 times its length as it
moves through the female tract.
That is the equivalent of a 6 ft Diagram by Mariana Ruiz
man traveling over 300 miles!)
 Sperm Transport

Diagram of the female reproductive tract by Miraceti.

How does the sperm navigate the female reproductive

tract?
 Human Semen
• Semen (seminal plasma) consists of:
• Sperm from testes.
• Seminal fluid from accessory glands.
• Semen characteristics:
• General properties:
• Creamy texture.
• Gray to yellow in color.
• Volume: 2.5-3.5 ml (3 days abstinence) (Range= 2-6 ml).
• pH: 7.35-7.5 (slightly basic).
• Fertility index:
• 20 million sperm/ml.
• >40% of sperm have progressive linear motility.
• >60% are normal shape and size.
 Semen in the Vagina
• Within a minute after deposition in vagina, semen coagulates.
• Enzymes convert fibrinogen to fibrin.
• Within 20 minutes, semen liquefies again.
• Vaginal environment is usually acidic (pH=~4.2).
• Semen causes vaginal pH to elevate to ~7.2.
• Retrograde flow: Fluid and sperm go in wrong direction-exit out of
vulva.
 Sperm in the Vagina
• Phagocytosis: neutrophils
(immune cells) are present in large
numbers (estrogen-induced)
throughout tract.
• Neutrophils recognize sperm as
foreign and actively
phagocytize.
• One neutrophil can engulf
several sperm at once.

Neutrophil engulfing anthrax bacillus. Micrograph byVolker

Brinkmann.
 Sperm in the Cervix
• Cervical mucus is semi-impenetrable.
• Only mucus produced in an estrogen-dominated environment will
allow sperm transport through cervix.
• Sperm entering cervical crypts (established reservoirs) survive and enter
into uterus.
• Establish “privileged pathways.”
 Sperm in the Uterus
• Phagocytosis!!!
• Lost in the endometrium—most never find the oviduct.
• Uterine “waves” transport sperm to oviduct:
• Estrogen plays role.
• Prostaglandins in semen may also help.
 Sperm in the Oviduct
• Junction between oviduct & uterus can serve as barrier in some species,
including humans.
• Transport in oviduct is dependent on smooth muscle contractions, cilia
motion, and sperm motility.
 Preparation for Fertilization
Sperm cell Capacitation:
• Changes that a sperm must go through to
become capable of fertilization.
• Involves addition and removal of surface
molecules.
• Occurs in female reproductive tract.

Capacitated sperm become

hyperactivated in oviductal
environment.

Is the prepared (“ripe”) sperm attracted

to the oocyte by chemical factors?
Micrograph magnification is X 14.
 Fertilization: Acrosome Reaction
• Sperm undergo acrosome reaction.
• This involves fusion of plasma membrane with acrosomal membrane,
thereby releasing acrosome contents (proacrosin & other enzymes).
• See the schematic representation of sperm after acrosome reaction. Note
the presence of the equatorial segment of the sperm plasma membrane.
Fertilization: Sequence
1. Penetration of oocyte investments:
Hyperactivated sperm penetrate the surrounding
cells.

2. Sperm contact & bind to zona pellucida.

a. Involves hypothesized species-specific
receptors on both sperm plasma membrane
& zona pellucida (ZP).
b. Receptor on ZP has been isolated & is a
glycoprotein.
Fertilization: Sequence
3. Sperm penetrate the zona pellucida: Involves both
mechanical & chemical forces.

4. Sperm cell makes contact with oocyte plasma

membrane.
a. Equatorial segment of sperm cell interacts with
(binds?) to specific area of the oocyte’s plasma
membrane.
b. This triggers critical reactions in the oocyte.
 Fertilization
Blocks to Polyspermy:
• Rapid: Resting membrane potential of the oocyte plasma membrane goes
from -65 mV to +10 mV within 1-5 seconds of sperm-oocyte fusion.
• Slow: Involves chemical changes in the zona pellucida and, perhaps, plasma
membrane.
 Fertilization: Cortical Reaction
• Dependent on presence of cortical granules in cytoplasm of oocyte.
• Electron-dense vesicles located at periphery of oocyte.
• 1-3 layers in human oocyte.
• Some 15,000 total present in xenopus oocyte.
• Fusion of sperm & oocyte stimulates release of contents of the cortical
granules (exocytosis).
• Induces changes in the zona pellucida (zona block).
 Fertilization: Zona Block or Hardening
• Enzymes released from
cortical granules alter the
structure of the zona
pellucida.
• Result: Sperm cannot
penetrate the zona.
Fertilization: Vitelline or Plasma Membrane
Block
Evidence in some species suggests that cortical granules also alter the plasma
membrane of the oocyte so that sperm can no longer associate with or bind to
it.
 Fertilization: Several Simultaneous Events

• Completion of meiosis within oocyte.

• Formation & ejection of 2nd polar body.
• Engulfment of entire sperm cell.
Fertilization: Additional Events
1. Male & 2. Male & female pronuclei
female migrate closer together.
pronuclei
form.

3. Syngamy:
the fusion of
male and
female
pronuclei
occurs. Note
that the second
polar body is
now visible.
 Fertilization: Completed
The zygote (one-cell
embryo) is formed.

With its DNA replicated, it is

ready to undergo specialized
cell division, called
cleavage.
 Sequence of Fertilization Events
• Oocyte in oviduct, one polar body present.
• Sperm in vagina undergo capacitation.
• Sperm in oviduct: hyperactive sperm motility.
• Penetration of cell investments around oocyte.
• Binding to zona pellucida.
• Sperm acrosome reaction.
• Penetration of zona pellucida.
• Sperm-oocyte fusion.
 Nearly Simultaneous Events following
Sperm-Oocyte Fusion
• Erection of barriers to polyspermy:
• Cortical reaction
• Zona hardening
• Vitelline block
• Resumption and completion of meiosis; two polar bodies present outside
oocyte plasma membrane.
• Engulfment of sperm.
 Fertilization: Additional Events

• Male and female pronuclei form.

• The pronuclei migrate to close proximity of one another.
• DNA replication, in preparation for cell division, occurs.
• Male and female pronuclei fuse: syngamy.
• A zygote is formed.
• Fertilization is complete!
 The Zygote
• The zygote is one of the largest cells in the body.
• This cell is totipotent, capable of forming every cell type present in the
fully formed individual.
• The cell, formed from two “haploid” cells, has been restored to a diploid
state. Note, however, that DNA division has occurred.
 Fertilization: The End Result

A new potential individual has been formed!

• This new potential individual is formed from the union of two highly
specialized, genetically unique cells that were able to come together
through the sacred union of two bodies!
• By successfully completing a series of mitoses, the resulting offspring
will be genetically distinct from every other individual of its species to
live before and all that will come after, including its own offspring!
 Probability of Fertilization
• Probability is a function
of the timing of
intercourse.
• A female’s fertile period
begins 6 days prior to
ovulation and ends
within 24 hours after
ovulation.

From: Wilcox et al. 1995 NEJM 333:1517

 Contraceptives
Sexual practices, devices, and/or medications that reduce the chance of
pregnancy.

The only 100% reliable method of preventing pregnancy is abstinence!!

Added benefit for some forms of contraceptive: Reduction of spread of sexually

transmitted diseases (STDs).

The only 100% reliable method of preventing STD is life-long monogamy.

Forms of Contraception Contraceptive Devices

• Barrier • Male condom
• Chemical • Female condom
• Vaginal sponge
• Intrauterine device
• Spermicidal jelly, cream, or depository
Note: Be sure you know the • Diaphragm
effectiveness and failure rates of • Cervical cap
common birth-control methods.