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Lecture 16: Fertilization
Animal Science 434John J. Parrish
Capacitation
1951Austin - ratChang - rabbit Fertilization does not occur until several
hours after sperm are placed in the uterus or oviduct
Chang Experiment - 1951
OvulationOvulation
Per
cen
t F
erti
liza
tio
n
AfterAfter BeforeBefore
Capacitation
The changes within sperm that confer upon the sperm the ability to acrosome react in response to the appropriate stimulus.
Zona Pellucida
Cumulus Cells
Follicular Fluid
Zona Pellucida
Cumulus Cells
Follicular Fluid
Effect of CapacitationEpididymal Ejaculated
SeminalPlasmaand/or
EpididymalSecretions
+
Capacitated
FemaleTract
+
DF (BSP in Bovine)
Plasma Membrane
Decapacitation Factor – Stabilizes plasma membrane and inhibits a premature acrosome reaction
CholesterolAcceptor
BSPHeparin
Plasma Membrane
CholesterolAcceptor
BSPHeparin
Plasma Membrane Raft MigrationZp bindingclustering
sAC
HCO3-
HCO3-
(+)
(-)
cAMP
PK-A
PTK Ptyr-Ptase
Protein Tyrosine Phosphorylation
(-)
(+)
(+)
(+)
pHi
(+)
H+
H+
CholesterolAcceptor
BSPHeparin
Raft MigrationZp bindingclustering
CO2 + H2O H2CO3 H+ + HCO3
-
Carbonic Anhydrase
Diffusion
Extracellular Space
sAC
HCO3-
HCO3-
(+)
(-)
cAMP
PK-A
PTK Ptyr-Ptase
Protein Tyrosine Phosphorylation
(-)
(+)
(+)
(+)
pHi
(+)
H+
H+
CholesterolAcceptor
BSPHeparin
Raft MigrationZp bindingclustering
ATP ADPCa2+
ATP ADP
Ca2+
Acrosome
Ca2+
CholesterolAcceptor
BSPHeparin
Raft Migration
ATP ADPCa2+
ATP ADP
Ca2+
Acrosome
Ca2+
Ca2+
sAC
Ca2+HCO3-
HCO3-
(+)
(-)
(+)
cAMP
PK-A
PTK Ptyr-Ptase
Protein Tyrosine Phosphorylation
(-)
(+)
(+)
(+)
pHi
(+)
H+
H+
CholesterolAcceptor
BSPHeparin
Ca2+
Acrosome
Ca2+
Capacitation
No change in sperm morphologyMotility change– hyperactivation
cAMP
Power output
cAMP
Power output
The Acrosome ReactionPlasma Membrane
Acrosome
Fused PlasmaMembrane and
Outer AcrosomalMembrane
Fused PlasmaMembrane and
Outer AcrosomalMembrane
Nucleus
AcrosinHyaluronidase
AcrosinHyaluronidase
Modificationof the
EquitorialRegion
Modificationof the
EquitorialRegion
The Zona PellucidaThe Zona Pellucida
Zona Pellucida
ZP3
Attachment
Binding
Acrosome ReactionInduction
Acrosome Reaction
Sperm Interaction with the Zona
Zona Pellucida
ZP2
ZP2
ZP2
ZP2
Secondary Binding
Zona Penetration
MotilityMotility
Zona Pellucida
Sperm tract through zona pellucidaSperm tract through zona pellucida
6-12 hr6-12 hr
18-24 hr18-24 hr
Maturation Promoting Factor(MPF)
Metaphase Promoting FactorMetaphase Promoting Factor
MPF MPF
MPFMPF
Primary Oocyte
GV-Intact
GVBD Metaphase I
PB-1
Secondary OocyteMetaphase II
Ovulation
LH Surge
Zona Pellucida
PerivitellineSpace
Oocyte
Ca2+
Ca2+
Ca2+
Ca2+
Zona Pellucida
PerivitellineSpace
Oocyte
Ca2+
Ca2+
Ca2+
Ca2+
Zona Pellucida
PerivitellineSpace
Oocyte
Ca2+
Ca2+
Ca2+
Ca2+
Zona Pellucida
PerivitellineSpace
Oocyte
Ca2+
Ca2+
Ca2+
Ca2+
Zona Pellucida
PerivitellineSpace
Oocyte
Ca2+
Ca2+
Ca2+
Ca2+
Calcium Transients During Bovine FertilizationCalcium Transients During Bovine Fertilization
Initiation of Calcium Pulse by Soluble Sperm Component
Endoplasmic Reticulum
Ca2+
PLCzetaPIP2
DAG
IP3
Ca2+
Oocyte
Zona Pellucida
PerivitellineSpace
Oocyte
MII
AIIMPF X
Ca2+
Pronuclei
TII
PBII
Zona Pellucida
PerivitellineSpace
Oocyte
MII
AIIMPF X
Ca2+Ca2+
Pronuclei
Decondensation
Pronuclei
Zygote
TII
PBII
Zona Pellucida
PerivitellineSpace
Oocyte
Ca2+
CorticalGranules
Zona Block to Polyspermy
Zona Pellucida
PerivitellineSpace
Oocyte
Ca2+
CorticalGranules
Zona Block to Polyspermy
3
Blocks to Polyspermy
1
2
4
Species
1.Mammals
3.InvertebratesAmphibiansFishRabbits
4.AmphibiansAvians
Gamete Aging
Oocyte life span approx. 6 – 12 hours after ovulation
Sperm life span approx. 24 – 48 hours after ejaculation