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Compartmentalization of Prion Isoforms Within the Reproductive Tract of the Ram¹

Gamètes Males et Fertilité,³ UMR 6175 INRA-CNRS-Université de Tours-Haras Nationaux, Station de Physiologie de la Reproduction et des Comportements, 37380 Nouzilly, France Génétique et Physiopathologie des Encéphalopathies spongiformes transmissibles,⁴ Station de Pathologie Infectieuse et Immunologie, Institut National de la Recherche Agronomique, 37380 Nouzilly, France

Cellular prion protein (Prp^C) is a glycoprotein usually associated with membranes via its glycosylphosphatidylinositol (GPI) anchor. The trans-conformational form of this protein (Prp^SC) is the suggested agent responsible for transmissible neurodegenerative spongiform encephalopathies. This protein has been shown on sperm and in the reproductive fluids of males. Antibodies directed against the C-terminal sequence near the GPI-anchor site, an N-terminal sequence, and against the whole protein showed that the Prp isoforms were compartmentalized within the reproductive tract of the ram. Immunoblotting with the three antibodies showed that the complete protein and both N- and C-terminally truncated and glycosylated isoforms are present within cauda epididymal fluid and seminal plasma. Moreover, we demonstrate that in these fluids, the Prp^C isoforms are both in a soluble state as well as associated with small membranous vesicles (epididymosomes). We also report that only one major glycosylated 25 kDa C-terminally truncated Prp^C isoform is associated with sperm from the testis, cauda epididymis, and semen, and this form is also present in the sperm cytoplasmic droplets that are released during maturation. In sperm, this C-terminal truncated form was found to be associated with membrane lipid rafts present in the mature sperm, suggesting a role for it in the terminal stages of sperm maturation.

¹ Supported by the Commité Interministériel Français de Recherche sur les Prions (Gis Prion), the AIP Prion INRA, and by the Région Centre. H.E. is the holder of a postdoctoral grant cofinanced by the Région Centre and INRA.

² Correspondence. FAX: 33 247 427 743; gatti{at}tours.inra.fr

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