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BOR - Papers in Press, published online ahead of print April 16, 2003.
Biol Reprod 2003, 10.1095/biolreprod.102.013771
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BIOLOGY OF REPRODUCTION 69, 572–581 (2003)
DOI: 10.1095/biolreprod.102.013771
© 2003 by the Society for the Study of Reproduction, Inc.

Gamete Biology

Inhibition of Capacitation-Associated Tyrosine Phosphorylation Signaling in Rat Sperm by Epididymal Protein Crisp-11

Kenneth P. Roberts3, Joseph A. Wamstad4, Kathy M. Ensrud4, and David W. Hamilton2,4

Department of Urologic Surgery,3 the Department of Genetics, Cell Biology, and Development,4 University of Minnesota, Minneapolis, Minnesota 55455

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.

1 This work supported by NIH grant HD-11962.

2 Correspondence: David W. Hamilton, Department of Genetics, Cell Biology, and Development, University of Minnesota, 420 Johnston Hall, 101 Pleasant St. SE, Minneapolis, MN 55455. FAX: 612 626 7431; dwh{at}umn.edu






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Copyright © 2003 by the Society for the Study of Reproduction.