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Rabbit Epididymal Secretory Proteins. II. Immunolocalization and Sperm Association of REP38¹

^a Pest Animal Control Cooperative Research Centre, CSIRO Sustainable Ecosystems, Canberra, ACT 2601, Australia ^b Department of Biological Sciences, University of Newcastle, Callaghan, NSW 2308, Australia

Polyclonal antibody was used to partially characterize REP38, a major rabbit epididymal secretory protein. Western blot analyses and immunohistochemistry indicated that REP38 is only expressed in regions 5 and 6 of the epididymis (corpus epididy-midis) and is localized in the supranuclear region and microvilli of the principal cells in these regions. It was not expressed in other tissues of the body. In region 8 (cauda epididymidis), REP38 was detected in the luminal border and cytoplasm of scattered principal cells, indicating that it may be reabsorbed in this region. This protein accumulated on the sperm plasma membrane downstream of region 5 and was localized predominantly over the acrosomal and postacrosomal regions of the head and the middle piece. Although tightly bound to epididymal sperm, REP38 migrated to the equatorial segment under conditions in vivo that would promote capacitation. When tested in vitro, anti-REP38 IgG reduced the percentage of ova fertilized in a concentration-dependent manner, apparently by blocking sperm-egg fusion.

First decision: 24 May 1999.

¹ This work was supported by a grant from the Australian Research Council, the Research Management Committee, University of Newcastle, and the Vertebrate Biocontrol Cooperative Research Centre, Canberra, Australia. B.N. was supported by an Australian Postgraduate Award and a Pest Animal Control Cooperative Research Centre Student Award.

² Correspondence: Michael Holland, Pest Animal Control Cooperative Research Centre, CSIRO Sustainable Ecosystems, P.O. Box 284, Canberra, ACT 2601, Australia. FAX: 61 2 6242 1511; michael.holland{at}csiro.au

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