Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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Biology of Reproduction, Vol 14, 586-598, Copyright © 1976 by Society for the Study of Reproduction

Isolation of an "Immobilizing Antigen" from Rabbit Sperm Membranes

MICHAEL G. O'RAND 1, and CHARLES B. METZ 1

1 Institute for Molecular and Cellular Evolution, University of Miami, Coral Gables, Florida 33134


To understand more precisely how individual sperm antigens function during sperm immobilization and fertilization, we describe a method for their isolation from rabbit sperm membranes. Three fractions were prepared by centrifugation of a rabbit epididymal sperm homogenate, examined by SDS-PAGE, and by electron microscopy, and tested for hyaluronidase and succinic dehydrogenase activity. The membrane-rich fraction was extracted with lithium diiodosalicylate and a sperm surface membrane glycoprotein (MGP; "immobilizing antigen") isolated from it by ion-exchange chromatography. The MGP showed a single immunodiffusion precipitin band against goat anti-rabbit sperm globulin. Antibodies to MGP immobilized rabbit sperm in the presence of complement. This MGP-"immobiizing antigen" was localized on the head and tail plasma membrane of epididymal, ejaculate and capacitated sperm by exchange agglutination and immobilization in the presence of guinea pig complement.

Note:
ACKNOWLEDGMENT Contribution Number 281 from the Institute for Molecular and Cellular Evolution. This study was aided by Grant No. M 74-31 from the Population Council and Grant No. 1-R01-HD06663-01 from the National Institutes of Child Health and Human Development. Dr. M. O'Rand was supported by United States Public Health Service, Postdoctoral fellowship NIH 5 F02 HD53900-02. The authors would like to thank Mr. R. Bartram for help with the hyaluronidase assays, Ms. M. A. Hart for help with preparing the photographs and Dr. C. Cordle for reading the manuscript.

Submitted on September 25, 1975
Accepted on February 2, 1976






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Copyright © 1976 by the Society for the Study of Reproduction.