This chapter will describe the major events of

fertilization accomplished in mammals.

Fertilization

sex

reproduction
 STRUCTURE OF THE GAMETES

Sperm
• discovered in the 1670s
• role in fertilization was not discovered until the mid-1800s

1840s - Albert von Kölliker described the formation of sperm from

cells in the adult testes
• sperm excited the egg to develop in much the same way a magnet
communicates its presence to iron
 STRUCTURE OF THE GAMETES

Sperm
1847 - Karl Ernst von Baer showed the union of sperm and egg in
sea urchins and tunicates

1870 - Oscar Hertwig and Herman Fol

repeated this work and detailed the union of the two cells’
nuclei.
MODIFICATION OF A GERM CELL TO
FORM A MAMMALIAN SPERM
SPERM ANATOMY
SPERM ANATOMY
 SPERM ANATOMY

Acrosomal vesicle, or Acrosome

• derived from the cell’s Golgi apparatus
• contains enzymes that digest proteins and
complex sugars
• Enzymes stored in the acrosome can digest a
path through the outer coverings of the egg
SPERM ANATOMY
SPERM ANATOMY

Flagellu
m
SPERM ANATOMY

major motor portion

of the flagellum
SPERM ANATOMY
SPERM ANATOMY

made exclusively of the dimeric

protein tubulin
SPERM ANATOMY
SPERM ANATOMY
SPERM CAPACITATION IN MAMMALS

Capacitation - final stages of sperm maturation

 THE EGG OR OVUM
Egg, or Ovum
female gamete capable of binding sperm and being fertilized
Oocyte
developing egg that cannot yet bind sperm or be fertilized

The human egg is in second meiotic metaphase when it binds sperm,

whereas the sea urchin egg has completed all of its meiotic divisions
when it binds sperm.
 THE EGG OR OVUM
Cytoplasm and Nucleus
 
The egg accumulates a remarkable cytoplasmic storehouse during
its maturation

Nutritive proteins
 supply of energy and amino acids
 THE EGG OR OVUM
The egg accumulates a remarkable cytoplasmic storehouse during
its maturation

Ribosomes and tRNA

 developing egg has special mechanisms for
synthesizing ribosomes
 THE EGG OR OVUM
The egg accumulates a remarkable cytoplasmic storehouse during
its maturation

Messenger RNAs
 encode proteins for the early stages of
development
 THE EGG OR OVUM
The egg accumulates a remarkable cytoplasmic storehouse during
its maturation

Morphogenetic factors
 These include transcription factors and
paracrine factors.
 THE EGG OR OVUM
The egg accumulates a remarkable cytoplasmic storehouse during
its maturation
Protective chemicals
 Many eggs contain ultraviolet filters and DNA
repair enzymes
 protect them from sunlight, and some eggs
contain molecules that potential predators find
distasteful
 THE EGG OR OVUM

The Cell membrane and Extracellular

Envelope
 regulates the flow of specific ions during
fertilization
 often involved in sperm-egg recognition
Invertebrates
 regulates the flow of specific ions
 THE EGG OR OVUM

vitelline envelope
 Essential for the species-specific binding of
sperm
 THE EGG OR OVUM

Cortex
 gel-like cytoplasm
 contains high concentrations of globular actin
molecules which polymerize to form long cables of
actin microfilaments
 extend the egg surface into small
projections called microvilli
aid sperm entry into the
cell
 THE EGG OR OVUM

cortical granules
 contain proteolytic enzymes
 homologous to the acrosomal vesicle of the
sperm
 contain mucopolysaccharides, adhesive
glycoproteins, and hyalin protein
 enzymes and mucopolysaccharides help prevent
polyspermy
 THE EGG OR OVUM

Mammalian egg
 extracellular envelope
is a separate, thick
matrix called the zona
pellucida
 THE EGG OR OVUM

Mammalian egg
 surrounded by a layer of
cells called the cumulus
 Mammalian sperm have to
get past these cells to
fertilize the egg.
 innermost layer of
cumulus cells is called the
corona radiata.
QUICK-QUIZ
and constitute the
sperm head
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acrosome and nucleus
major motor portion of the
flagellum
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axoneme
final stages of sperm
maturation
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Capacitation
essential for the species-
specific binding of sperm
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vitelline envelope
 additional sperm from
entering the egg after the
first sperm has entered
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polyspermy
 INTERNAL FERTILIZATION IN MAMMALS

 mammalian fertilization occurs inside the oviducts of

the female
 sperm population ejaculated into the female is
probably heterogeneous, containing spermatozoa at
different stages of maturation
 there may be multiple mechanisms by which
mammalian sperm can undergo the acrosome
reaction and bind to the zona pellucida
GETTING THE GAMETES INTO THE OVIDUCT:
TRANSLOCATION AND CAPACITATION

 Both the male and female gametes use a combination

of small-scale biochemical interactions and large-
scale physical propulsion to get to the ampulla
 region of the oviduct where fertilization takes place
 TRANSLOCATION

 A mammalian oocyte just released from the ovary is

surrounded by a matrix containing cumulus cells
 If this matrix is experimentally removed or
significantly altered, the fimbriae of the oviduct will
not “pick up” the oocyte-cumulus complex
 TRANSLOCATION

 Once it is picked up, a combination of ciliary beating

and muscle contractions transport the oocyte-
cumulus complex to the appropriate position for its
fertilization in the oviduct.
 significantly altered, the fimbriae of the oviduct will
not “pick up” the oocyte-cumulus complex
 TRANSLOCATION

In humans, about 300 million

sperm are ejaculated into the
vagina, but only one in a million
enters the Fallopian tubes
 TRANSLOCATION

Sperm motility
 in those mammals where the female is promiscuous,
sperm from the same male will often form “trains” or
aggregates where the combined propulsion of the flagella
make the sperm faster
 In those species without such female promiscuity, the
sperm usually remain individual
TRANSLOCATION

The “sperm train” of the

wood mouse Apodemus sylvaticus
 TRANSLOCATION

Uterine muscle contractions

 sperm appear to be transported to the
oviduct by the muscular activity of the
uterus
 TRANSLOCATION

Sperm rheotaxis
 migrate against the direction of the flow—using CatSper
calcium channels
 CAPACITATION
 newly ejaculated mammalian sperm are immature and
cannot fertilize the egg
 sperm must undergo a suite of sequential physiological
changes called capacitation
 “speedy” sperm may have little chance of fertilizing the
egg because they have not undergone capacitation
 The knowledge that recently ejaculated mammalian sperm
could not fertilize an egg was a critical breakthrough in the
development of successful in vitro fertilization techniques
 CAPACITATION
 A study found that nearly all human pregnancies result
from sexual intercourse during a 6-day period ending on
the day of ovulation.
 prepare the sperm for the acrosome reaction and enable the
sperm to become hyperactive
 CAPACITATION
Two sets of molecular changes are considered to be important:
1. Lipid changes
 sperm cell membrane is altered by the
removal of cholesterol by albumin
proteins
 CAPACITATION
Two sets of molecular changes are considered to be important:
2. Protein changes.
 proteins or carbohydrates on the sperm surface are
lost during capacitation
 membrane potential of the sperm cell becomes more
negative as potassium ions leave the sperm
 CAPACITATION

2. Protein changes.
 unmasking of these sites might be one of the effects
of cholesterol depletion
 membrane potential of the sperm cell becomes more
negative as potassium ions leave the sperm
 CAPACITATION
Two sets of molecular changes are considered to be important:
2. Protein changes.
 allow calcium channels to be opened
 activating cAMP production and in
facilitating the membrane fusion
events of the acrosome reaction
CAPACITATION
 CAPACITATION

2. Protein changes.
 activating cAMP production and facilitating the
membrane fusion events of the acrosome reaction
 membrane potential of the sperm cell becomes more
negative as potassium ions leave the sperm
 CAPACITATION

Izumo
 critical in sperm-egg fusion
 slowing the rate of sperm capacitation and extending
the active life of sperm may maximize the
probability that sperm will still be available to meet
the egg in the ampulla
 IN THE VICINITY OF THE OOCYTE:
HYPERACTIVATION,
THERMOTAXIS, AND CHEMOTAXIS
 sperm become hyperactivated
 detach and continue their journey to the egg
 Uncapacitated sperm move in a planar direction,
allowing more time for the sperm head to attach to
the oviduct epithelial cells.
 Capacitated sperm rotate around their long axis,
probably enhancing the detachment of the sperm
from the epithelia
 IN THE VICINITY OF THE OOCYTE:
HYPERACTIVATION,
THERMOTAXIS, AND CHEMOTAXIS

 Hyperactivation, along with a hyaluronidase enzyme

on the outside of the sperm cell membrane, enables
the sperm to digest a path through the extracellular
matrix of the cumulus cells
 IN THE VICINITY OF THE OOCYTE:
HYPERACTIVATION,
THERMOTAXIS, AND CHEMOTAXIS

What does provide the sperm with directions?

 Thermotaxis
ability to sense temperature difference and
preferentially swim from cooler to warmer sites
 IN THE VICINITY OF THE OOCYTE:
HYPERACTIVATION,
THERMOTAXIS, AND CHEMOTAXIS

Chemotaxis
 oocyte and its accompanying cumulus cells secrete molecules that
attract capacitated
Chemotactic compounds
 progesterone
 In humans, progesterone has been shown to bind to a receptor that
activates Ca2+ channels in the cell membrane of the sperm tail,
leading to sperm hyperactivity
 THREE CALCIUM-MEDIATED SENSING
PROCESSES GET THE MAMMALIAN SPERM
TO THE EGG:

Rheotaxis
Thermotaxis
chemotaxis
 THE ACROSOME REACTION AND
RECOGNITION AT THE ZONA PELLUCIDA

 The human zona pellucida has four major

glycoproteins—ZP1, ZP2, ZP3, and ZP4.
 most sperm undergo the acrosome reaction in
or around the cumulus
 GAMETE FUSION AND THE
PREVENTION OF POLYSPERMY
 The side of the sperm head makes contact with
the egg
 fusion between sperm and
egg begins
 Human sperm also contain
Izumo protein, and
antibodies directed against
Izumo prevent sperm-egg
fusion in humans as well.
 Izumo binds to an oocyte
protein called Juno
 eggs deficient in Juno cannot
bind or fuse with acrosome-
reacted sperm
 interaction of Izumo and
Juno recruits the egg
membrane protein CD9 to
the area of sperm egg
adhesion
 GAMETE FUSION AND THE
PREVENTION OF POLYSPERMY

 In mammals, blocks to polyspermy include modification

of the zona proteins by the contents of the cortical
granules so that sperm can no longer bind to the zona.
FUSION OF GENETIC MATERIAL

The mature unfertilized

oocyte completes the first
meiotic division, budding
off a polar body
FUSION OF GENETIC MATERIAL

As the sperm enters the

oocyte, microtubules
condense around it as the
oocyte completes its
second meiotic division at
the periphery.
FUSION OF GENETIC MATERIAL

By 15 hours after
fertilization, the two
pronuclei have come
together, and the centrosome
splits to organize a bipolar
microtubule array. The
sperm tail is still seen
FUSION OF GENETIC MATERIAL

At prometaphase, chromosomes
from the sperm and egg intermix
on the metaphase equator and a
mitotic spindle initiates the first
mitotic division.
The sperm tail can still be seen.
 FUSION OF GENETIC MATERIAL

 Each sperm brings into the egg not only its

nucleus but also its mitochondria, its centriole,
and a tiny amount of cytoplasm.
 The sperm mitochondria and their DNA are
degraded in the egg cytoplasm
 all of the new individual’s mitochondria are
derived from its mother
 FUSION OF GENETIC MATERIAL

 the sperm centriole not only survives but

appears to serve as the organizing agent for
making the new mitotic spindle
 sperm cytoplasm has recently been found to
contain enzymes that activate egg metabolism,
as well as RNA fragments that may alter gene
expression
 ACTIVATION OF THE MAMMALIAN
EGG
 The rise in intracellular free Ca2+ at fertilization
in amphibians and mammals causes the
degradation of cyclin and the inactivation of
MAP kinase, allowing the second meiotic
metaphase to be completed and the formation of
the haploid female pronucleus.
 ACTIVATION OF THE MAMMALIAN
EGG
 In mammals, DNA replication takes place as the
pronuclei are traveling toward each other. The
pronuclear membranes disintegrate as the
pronuclei approach each other, and their
chromosomes gather around a common
metaphase plate.
Why do think a “speedy” sperm
has a little chance of fertilizing
the egg?
Sometimes the egg and sperm fail to meet and
conception does not take place. What are the leading
causes of infertility in humans, and what procedures are
being used to circumvent these blocks?
One of the goals of modern pharmacology is to create a
male contraceptive. Reviewing the steps of fertilization,
what steps do you think it might be possible to block
pharmacologically in order to produce such a
contraceptive for males?
 CODA

Fertilization is not a moment or an event, but a

process of carefully orchestrated and coordinated
events including the contact and fusion of
gametes, the fusion of nuclei, and the activation of
development.
 CODA

It is a process whereby two cells, each at the verge

of death, unite to create a new organism that will
have numerous cell types and organs. It is just the
beginning of a series of cell-cell interactions that
characterize animal development.