Inhibition of capacitation-associated tyrosine phosphorylation signaling in rat sperm by epididymal protein Crisp-1.
Ejaculated sperm are unable to fertilize an egg until they undergo capacitation. Capacitation results in the acquisition of hyperactivated motility, changes in the properties of the plasma membrane, including changes in proteins and glycoproteins, and acquisition of the ability to undergo the acrosome reaction. In all mammalian species examined, capacitation requires removal of cholesterol from the plasma membrane and the presence of extracellular Ca2+ and HCO3-. We designed experiments to elucidate the conditions required for in vitro capacitation of rat spermatozoa and the effects of Crisp-1, an epididymal secretory protein, on capacitation. Protein tyrosine phosphorylation, a hallmark of capacitation in sperm of other species, occurs during 5 h of in vitro incubation, and this phosphorylation is dependent upon HCO3-, Ca2+, and the removal of cholesterol from the membrane. Crisp-1, which is added to the sperm surface in the epididymis in vivo, is lost during capacitation, and addition of exogenous Crisp-1 to the incubation medium inhibits tyrosine phosphorylation in a dose-dependent manner, thus inhibiting capacitation and ultimately the acrosome reaction. Inhibition of capacitation by Crisp-1 occurs upstream of the production of cAMP by the sperm.
Roberts
Kenneth P
KP
Department of Urologic Surgery, University of Minnesota, Minneapolis 55455, USA.
Wamstad
Joseph A
JA
Ensrud
Kathy M
KM
Hamilton
David W
DW
D020101:Acrosome Reaction / Q000502:physiology; D000818:Animals; D001639:Bicarbonates / Q000378:metabolism; D015153:Blotting, Western; D002118:Calcium / Q000502:physiology; D002784:Cholesterol / Q000378:metabolism / Q000502:physiology; D003470:Culture Media; D000242:Cyclic AMP / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004822:Epididymis / Q000187:drug effects / Q000502:physiology*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D008562:Membrane Glycoproteins / Q000494:pharmacology*; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013075:Sperm Capacitation / Q000187:drug effects / Q000502:physiology*; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*; D013107:Sphingolipids / Q000378:metabolism; D014443:Tyrosine / Q000502:physiology*
D001639:Bicarbonates; C486320:Crisp3 protein, rat; D003470:Culture Media; D008562:Membrane Glycoproteins; D013107:Sphingolipids; D014443:Tyrosine; D002784:Cholesterol; D000242:Cyclic AMP; D002118:Calcium