BIOL124 Lecture 7

Gametes and fertilisation

Dr Jess Hammond
[email protected]
 Learning objectives
Today’s lecture should enable to you:
• draw a sketch of a sperm cell and describe its structure
• describe what is meant by the ‘fast block to polyspermy’
• describe the ‘acrosomal reaction’ in sea urchin fertilisation
• describe the ‘cortical reaction’ and discuss what is meant by
the ‘slow block to polyspermy’
• describe egg-sperm interactions at the molecular level for
sea urchin and human fertilisation
• describe events occurring in the human egg from the point
at which the sperm makes contact with the corona radiata,
to the first cleavage division.

Further reading
• Campbell and Reece Biology 11th Edition p1074-77,
• Tortora and Derrickson Principles of Anatomy and
th
 Spermatozoon (sperm cell)

Flagellum

ATP

p1058 Campbell
 Fertilisation

Egg activation Zygote formation

SEA URCHIN
• External fertilisation
• Large eggs
• Transparent embryos
 The steps of sea urchin fertilisation

3. Contact and fusion of

2. Acrosomal reaction sperm/egg membranes 4. Cortical reaction

1. Contact 5. Entry of sperm

nucleus

p1075 Campbell
The acrosomal reaction

p1075 Campbell
 Conspecific fertilisation
(egg fertilised by sperm from same species)

Bindin receptor on surface

of vitelline
membrane

Bindin on surface
of acrosomal
process
More enzymes
break down vitelline
layer and plasma
membrane

p1075 Campbell
 An illustration of the acrosomal reaction

http://worms.zoology.wisc.edu/dd2/echino/fert/acrosome/acrosome.html
Fusion leads to the fast block to polyspermy

p1075 Campbell
Fast block to polyspermy
• Membrane potential
present across sea urchin
egg plasma membrane
• Fusion of plasma
membranes causes
– Ion channels in plasma
membrane open
– Na+ influx from sea
water
= Membrane depolarisation
• Change in plasma
membrane  no more
sperm fusion
• Transient effect
Slow block to polyspermy

p1075 Campbell
The calcium wave of the cortical reaction
Binding of a sperm to the egg activates a signal transduction
pathway triggering release of Ca2+ into the cytosol from the ER

p1076 Campbell
 The Ca2+ wave correlates with
appearance of fertilisation envelope
 The Ca2+ wave correlates with
appearance of fertilisation envelope

Note formation of fertilisation envelope

Slow block to polyspermy
• Ca2+ wave increases cytosolic
conc of Ca2+ where sperm
fuses
• Cortical granules fuse with
plasma membrane and
release enzymes:
• break down adhesion
between vitelline layer
and membrane
• increase osmotic pressure
causing water influx
• snip off sperm receptors
• harden fertilisation
envelope

• Also stimulates egg activation

p1075 Campbell
 Egg activation: an increase in
protein synthesis and metabolic rate

Protein synthesis
 Pronuclei fusion (@45sec)

• Migration of female pronucleus guided by microtubules from

male pronucleus centrosome
• Directly fuse to form zygote
• DNA synthesis begins
Brain Break
Human fertilisation
Oogenesis
(revision)

Human egg
arrested at
metaphase
of Meiosis II

p1059 Campbell
 Ovarian cycle
(revision)

(Graafian follicle)
 Oocyte at Ovulation
• Ovulation releases a secondary oocyte and the first polar body
• Surrounded by the zona pellucida (glycoprotein layer) and corona
radiata (multicellular)
• Both layers must be penetrated by the sperm
Corona First polar
radiata body

Zona
pellucida

Martini, Fundamentals of Anatomy and Physiology 9th Edition, p1079

 Human fertilisation requires multiple
sperm to interact with the egg
(Sperm capacitation required)

1. Sperm contact
corona radiata

Hyaluronidase on
surface of sperm

ZP3
Granulosa cells
of corona radiata
embedded in
hyaluronic acid

p1077 Campbell
 Human fertilisation requires multiple
sperm to interact with the egg
2. Acrosomal
reaction to digest
zona pellicuda

3. Fusion of
membranes and
sperm enters

4. Fast and slow

blocks to ZP3
polyspermy
Cortical reaction

p1077 Campbell
 Mammalian sperm-egg molecular
interactions
• zona pellucida glycoprotein ZP3 binds to  1-4-
galactosyltransferase on sperm
• triggers acrosomal contents release
• acrosomal enzymes acrosin and -N-acetylglucosaminidase digest
zona pellucida glycoproteins
• access cell membrane
• fertilin on sperm head binds integrin-like protein and CD9 in
secondary oocyte plasma membrane
• allows membrane fusion
• to block polyspermy:
• depolarisation of oocyte cell membrane (within a few seconds)
• release of intracellular Ca2+  exocytosis of oocyte secretory
vesicles to harden zona pellucida
 Fertilisation
1 and oocyte activation
by sperm entry
• Acrosomal enzymes and strong flagella movements from multiple
sperm create gaps in the corona radiata and zona pellucida
• Only one sperm cell then makes contact with the oocyte
membrane  membrane fusion occurs
Fertilizing spermatozoon Second polar body

Martini, Fundamentals of Anatomy and Physiology 9th Edition, p1079

 Pronucleus formation
• The entire sperm enters and is absorbed into the cytoplasm
• Meiosis II resumes
– Secondary oocyte splits in two haploid cells of unequal size – the
ovum and the second polar body
– The female pronucleus develops

Nucleus of fertilising Female pronucleus (haploid)

Spermatozoon (haploid)

Martini, Fundamentals of Anatomy and Physiology 9th Edition, p1079

 Spindle formation and cleavage
preparation
• The male pronucleus develops
• Spindle fibres from centrosome appear in preparation for
the first cleavage division
Male Female
pronucleus pronucleus

Martini, Fundamentals of Anatomy and Physiology 9th Edition, p1079

 Preparation for first mitotic division

4h 8h 12h
chromosomes
• Each pronucleus enters mitosis separately
• Nuclear membranes break down and pronuclei fuse  zygote
• Chromosomes attach to common spindle
 Cleavage Begins… to be continued

• Chromosomes line up along metaphase plate and prepare

to divide
Metaphase of first cleavage division

Martini, Fundamentals of Anatomy and Physiology 9th Edition, p1079

Before the next lecture, please read

pp 1074-1094 Campbell (11th

edition)

Early embryogenesis
 Any questions?
• Ask me at the front
• Email me: [email protected]