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Fertilisation
Kersti Lundin
Reproductive Medicine
Sahlgrenska University Hospital
Learning objectives
Overview
Concepts of fertilisation
The sperm/oocyte contribution
The first ~20 hours
Sexual reproduction
Combination of genes from two
individuals (maternal / paternal)
Oocyte
Cytoplasmic maturation
1st meiosis
Binding to ZP
Acrosome reaction
Penetration
Capacitation
A post-ejaculatory event, <1h
Ca2+ increase, cAMP increase
Sperm plasma membrane increase
fluidity
y ((cholesterol loss))
Can be seen as a hyperactivated
movement pattern
Reversible
In the capacitated state, the sperm cell
can undergo the acrosome reaction in
response to appropriate stimulus (not
reversible)
K+
hyperpolarisation
+
Cholesterol
HCO3- and Ca2+
cAMP PTK
capacitation
Secretion of hyaluronidase
Penetration of cumulus matrix
Secretion of hyaluronic acid
Expansion of cumulus cells
Zona pellucida
The human zona pellucida is made up as an 3-D
matrix, by four major glycoproteins; ZP1, ZP2, ZP3
and ZP4, secreted by the oocyte
ZP1, ZP3 and ZP4 can bind capacitated human
t
d induce
i d
ti
spermatozoa
and
an acrosome reaction
Human ZP2 only binds to acrosome-reacted
spermatozoa and thus acts as a secondary sperm
receptor.
Plasma
membrane
Outer acrosomal
membrane
Inner
acrosomal
membrane
Nuclear
membrane
Centrioles
Acrosomal enzymes
+ vigorous
movements of the
sperm tail drives the
sperm trough the
zona pellucida into
the perivitelline
space
Sperm-oocyte fusion
During the acrosome reaction, the
equatorial segment of the sperm head
acquires the capacity to recognise and
fuse with the plasma membrane of the
oocyte
Specific recognition
and adhesion
Binding
Fusion
Decondensation
1. sperm
penetration
of expanded
cumulus
cells
6. Formation of
pronuclei
5. Sperm
decondensation
4. oocyte activation
(polyspermy block,
2nd meiosis
completion, 2nd PB)
The Author 2008. Published by Oxford University Press on behalf of the European Society of
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Polyspermy block / 1.
Membrane block
Decreased receptiveness of the oolemma
The oolemma gradually becomes resistant to
fusion with additional spermatozoa
Polyspermy block / 2.
The cortical reaction (z.p. block)
Release of cortical granules
Induced by the first calcium waves
Secretory vesicles derived from the Golgi complex
Translocated by microfilaments to the cortex during
oocyte maturation
Shown to contain e.g. trypsin-like proteinases,
peroxidases.
Prevents polyspermy by inducing changes in
the zona pellucida:
receptor inactivation (ZP3)
zona hardening (modification of ZP2)
Liu, 2011
Spermatozoon
1st + 2nd meiosis
Metamorphosis
Plasma maturation
Cytoplasmic maturation
1st meiosis
Binding
Acrosome reaction
Penetration
Fusion
Decondensation
SNDF
((nucleoplasmin)
l
l
i )
SAOAF
(oscillin, phospholipase Cz)
Dozortsev et al 1998
Oocyte activation
Modifications to allow
development to proceed
Triggered by sperm entry
Release of cortical granules
Resumption of meiosis
Formation of male and female pronuclei
Starts with:
Increase in internal free calcium from the
endoplasmic reticulum (ER)
Generation of a series of calcium waves
Ca2+ oscillations
SUZI
*
Initiated by PLCz => IP3
Begin a few minutes after
sperm entry, and cease
around PN formation
ICSI
intact
Regulation by mitochondria
/ATP production?
ICSI
manipulated
*
* release of
sperm factors
Metaphase II arrest
Pronuclear formation
Transcriptional
competence of the
gametes restored
Number of pronuclei
No sperm activation
Routine IVF
Sperm
Ability to swim, bind, acrosome react,
penetrate, fuse
Oocyte
Similar to in vivo situation
vs. In vivo
Abundance of sperm
Environment
ICSI
Sperm
Ability to decondense and activate oocyte
Selection, Competence?
Communication with oocyte?
Oocyte
Invasive
Contamination?
vs. In vivo
Environment