Lecture 16: Fertilization

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Lecture 16: Fertilization Animal Science 434 John J. Parrish

Capacitation 1951 Austin - rat Chang - rabbit Fertilization does not occur until several hours after sperm are placed in the uterus or oviduct

Chang Experiment - 1951 Ovulation After Before Percent Fertilization

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

Effect of Capacitation Epididymal Ejaculated Seminal Plasma + Capacitated Female Tract +

Model of Capacitation D F Acrosome Nucleus Cytoplasm Plasma Membrane

D F Chol-Acceptor D F GAG D F D F D F Ca2+ Cholesterol Acrosome Ca2+ pH cAMP Nucleus Activity of Enzymes Cytoplasm Plasma Membrane

Capacitation No change in sperm morphology Motility change hyperactivation cAMP Power output

The Acrosome Reaction Fused Plasma Plasma Membrane Membrane and Outer Acrosomal Membrane Plasma Membrane Acrosin Hyaluronidase Acrosome Nucleus

Modification of the Equitorial Region

The Zona Pellucida ZP1 ZP2 ZP3

Sperm Interaction with the Zona Attachment Zona Pellucida Binding ZP3 Acrosome Reaction Induction Acrosome Reaction

Secondary Binding Zona Pellucida ZP2 ZP2 Zona Penetration ZP2 ZP2 Motility

Zona Pellucida

Sperm tract through zona pellucida

18-24 hr 6-12 hr

Maturation Promoting Factor (MPF) LH Surge Metaphase Promoting Factor Primary Oocyte GVBD Metaphase I MPF MPF GV-Intact MPF PB-1 MPF Ovulation Metaphase II Secondary Oocyte

Zona Pellucida Oocyte Ca2+ Ca2+ Perivitelline Space

Calcium Transients During Bovine Fertilization

Initiation of Calcium Pulse by Soluble Sperm Component

Calcium Pulse Initiated by IP3 ER

X Zona Pellucida Oocyte Pronuclei Ca2+ AII TII MPF Perivitelline MII PBII MPF MII Perivitelline Space

X Zygote Zona Pellucida Oocyte Decondensation Pronuclei Pronuclei Ca2+ MII AII MPF X Ca2+ Pronuclei TII PBII Perivitelline Space

Zona Block to Polyspermy Zona Pellucida Oocyte Ca2+ Cortical Granules Zona Block to Polyspermy Perivitelline Space

Blocks to Polyspermy Species 1. Mammals 3. Invertebrates Amphibians Fish Rabbits 4. Amphibians Avians 1 2 3 4

Perivitelline Space 1st Polar Body Vitelline Membrane Zona Pellucida Sperm acrosomal region binds to sperm receptor on zona Perivitelline Space Initiates completion of Meiosis II and release of 2nd polar body 1st Polar Body Vitelline Membrane Zona Pellucida Initiates - Zona & Vitelline Reaction Cortical Granule ZP3 Binding Fusion Cortical Granule Release Zona Reaction Initiation of Acrosome Reaction Excluded Sperm Continuation of Reaction Acrosome-Reacted Sperm Penetration Zona and Vitelline Reactions block polyspermy Sperm penetrates zona Redrawn from Wasserman, P.M. 1988 Scientific American. p78