Fertilization Capacitation Chang Experiment

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Fertilization Capacitation Chang Experiment

Lecture 16: Fertilization Animal Science 434 John J. Parrish Capacitation Chang Experiment - 1951 1951 n 100 o i Ovulation t Austin - rat a 80 z i l i t Chang - rabbit r 60 After Before e F Fertilization does not occur until several t 40 n e hours after sperm are placed in the c r 20 e uterus or oviduct P 0 -2 -1 0 2 4 6 8 Time of Insemination (hr relative to ovulation) Effect of Capacitation Capacitation Epididymal Ejaculated Capacitated The changes within sperm that confer upon the sperm the ability to acrosome react in response to the appropriate Seminal Plasma stimulus. Female + + and/or Tract Zona Pellucida Epididymal Secretions Cumulus Cells Follicular Fluid 1 Model of Capacitation GAG D F D F Acrosome Acrosome Nucleus Nucleus Cytoplasm Cytoplasm Plasma Membrane Plasma Membrane D F Chol-Acceptor GAG Capacitation Ca2+ Cholesterol No change in sperm morphology Motility change – hyperactivation Acrosome Ca2+ Nucleus pH Activity of cAMP Enzymes cAMP Cytoplasm Power output Plasma Membrane The Acrosome Reaction Plasma Membrane Fused Plasma Membrane and Outer Acrosomal Membrane Acrosin Acrosome Hyaluronidase Nucleus Modiﬁcation of the Equitorial Region 2 The Zona Pellucida Sperm Interaction with the Zona Attachment Z o n a Binding P e ZP3 l Acrosome Reaction l Induction u c Acrosome Reaction i d a Z Secondary Binding Zona Pellucida o ZP n 2 a ZP2 P Zona Penetration e ZP l 2 l u ZP2 c Motility i d a 6-12 hr 18-24 hr Sperm tract through zona pellucida 3 Maturation Promoting Factor Zona Pellucida LH Surge (MPF) Oocyte Metaphase Promoting Factor 2+ Primary Oocyte GVBD Ca Metaphase I Ca2+ MPF MPF Ca2+ GV-Intact Ca2+ PB-1 Ovulation MPF MPF Perivitelline Space Metaphase II Secondary Oocyte Zona Pellucida Zona Pellucida Oocyte Oocyte Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Perivitelline Perivitelline Space Space Zona Pellucida Zona Pellucida Oocyte Oocyte Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Ca2+ Perivitelline Perivitelline Space Space 4 Calcium Transients During Bovine Fertilization Initiation of Calcium Pulse by 500 500 450 450 Soluble Sperm Component 400 400 ) ) 350 350 M M 300 300 (n (n 250 250 um i 200 200 c l a 150 150 Calcium C 100 100 50 50 DAG 0 0 0 10 20 30 40 50 60 70 80 90 0 10 20 30 40 50 60 70 80 90 Oocyte PIP Time (m in) Time (min ) PLCzeta 2 500 500 IP 450 450 3 400 400 ) 2+ 350 ) 350 Ca nM 300 nM 300 ( 250 ( 250 ium cium 200 200 l alc Ca 150 150 C 100 100 50 50 2+ 0 0 Ca 0 10 20 30 40 50 60 70 80 90 0 10 20 30 40 50 60 70 80 90 Time (min) Time (m in) Endoplasmic Reticulum Sperm Receptor - Calcium Zona Pellucida Oocyte Pulse Initiation PLC Ca++ IP3 ER Pronuclei Ca2+ AII TII MXPF Perivitelline MII Space PBII Zona Pellucida Zona Pellucida Oocyte Oocyte Decondensation Ca2+ Pronuclei Zona Block to Cortical Zygote Granules Polyspermy Pronuclei Ca2+ AII TII MXPF Perivitelline MII Perivitelline Space PBII Space 5 Zona Pellucida Blocks to Polyspermy Oocyte Species Ca2+ 1. Mammals 1 3. Invertebrates Zona Block to Amphibians Cortical Fish 2 Granules Polyspermy Rabbits 4. Amphibians 3 Avians 4 Perivitelline Space 6.

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What Can We Learn About Fertilization from Cystic Fibrosis?
COMMENTARY What can we learn about fertilization from cystic fibrosis? Harvey M. Florman*, Melissa K. Jungnickel, and Keith A. Sutton Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01655 t is a curious and intriguing situa- tion that mammalian sperm are introduced into the female repro- ductive tract in an infertile state Iand must be educated there before they are able to fertilize oocytes. The recog- nition that mammalian sperm must first be switched into a competent state, or capacitated, was exploited in the devel- opment of in vitro fertilization methods and has proved essential for the depen- dent technologies of clinical assisted re- production and infertility treatments. Yet many aspects of the mechanisms of capacitation remain unclear (for recent reviews, see refs. 1 and 2). In a recent issue of PNAS, Xu et al. (3) advanced our understanding of capacitation by showing that CFTR, the cystic fibrosis transmembrane regulator, plays an es- sential role in some aspects of this process. During capacitation there is a com- plex alteration of the biochemical, bio- physical, and cell biological properties Fig. 1. Capacitation and fertilization. (A) As a result of capacitation, sperm develop hyperactivated motility, the ability to respond to chemotactic signals and acrosome reaction-inducing signals. Capaci- of sperm. These include (but are not tated sperm penetrate the cumulus and reach the zona pellucida. Contact with the zona pellucida triggers restricted to) alterations in membrane acrosome reactions, permitting sperm to penetrate to the oocyte surface and fuse with the oocyte. potential and membrane sterol content, Multiple steps in this process are controlled by capacitation.
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11/5/12! Lecture 16: Fertilization! Animal Science 434! John J. Parrish! Capacitation! Chang Experiment - 1951! 1951! Ovulation! Austin - rat! Chang - rabbit ! After! Before! Fertilization does not occur until several hours after sperm are placed in the uterus or oviduct! Percent Fertilization! Effect of Capacitation! Capacitation! Epididymal! Ejaculated! Capacitated! The changes within sperm that confer upon the sperm the ability to acrosome react in response to the appropriate Seminal! stimulus.! Plasma! Female! +! +! and/or ! Tract! Zona Pellucida! Epididymal! Secretions! Cumulus Cells! Follicular Fluid! 1! 11/5/12! DF (BSP in Bovine) Cholesterol BSP Heparin Acceptor Plasma Membrane! Plasma Membrane! Decapacitation Factor – Stabilizes plasma membrane and inhibits a premature acrosome reaction! BSP + - BSP Cholesterol H HCO3 Cholesterol Heparin Heparin Acceptor Acceptor Zp binding Zp binding Plasma Membrane! Ra: Migra=on clustering Ra: Migra=on clustering (+) sAC + H - HCO3 (+) cAMP (+) pH i PK-A (+) (-) PTK Ptyr-Ptase (+) (-) Protein Tyrosine Phosphorylaon + - BSP H HCO3 Cholesterol Heparin Extracellular Space! Acceptor Zp binding Ra: Migra=on clustering Diffusion! (+) sAC + H - + - HCO3 CO2 + H2O H2CO3 H + HCO3 (+) cAMP (+) pH i PK-A Carbonic Anhydrase! (+) (-) PTK Ptyr-Ptase (+) (-) Protein Tyrosine Phosphorylaon 2! 11/5/12! BSP 2+ Ca2+ Cholesterol Ca Heparin Acceptor Ra: Migra=on ATP ADP ATP ADP Ca2+ Ca2+ Ca2+ ATP ADP ATP ADP Ca2+ Ca2+ Acrosome Acrosome + - BSP H HCO3 Cholesterol 2+ Heparin Ca Acceptor Capacitation ! No
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Cysteine-Rich Secretory Protein 4 Is an Inhibitor of Transient Receptor Potential M8 with a Role in Establishing Sperm Function
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