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Comparison Between Epididymosomes Collected in the Intraluminal Compartment of the Bovine Caput and Cauda Epididymidis¹

Centre de Recherche en Biologie de la Reproduction and Département d'Obstétrique-Gynécologie, Faculté de Médecine, Université Laval, Ste-Foy, Quebec, Canada G1V 4G2

ABSTRACT

During their transit along the epididymidis, mammalian spermatozoa acquire new proteins involved in the acquisition of male gamete fertilizing ability. We previously described membranous vesicles called epididymosomes, which are secreted in an apocrine manner by the epididymal epithelium. Some selected proteins associated with epididymosomes are transferred to spermatozoa during epididymal transit. The present study compared epididymosomes collected from caput epididymal fluid with vesicles from the cauda epididymidis in the bull. Two-dimensional gel electrophoresis revealed major differences in protein composition of epididymosomes isolated from the caput and cauda epididymidis. LC-QToF analysis of major protein spots as well as Western blot analysis confirmed the differences in proteins associated with these two populations of epididymosomes. Biotinylated proteins associated with caput and cauda epididymosomes also revealed differences. When incubated with caput epididymal spermatozoa, epididymosomes prepared from these two segments transferred different protein patterns. By contrast, cauda epididymosomes transferred the same pattern of proteins to spermatozoa from the caput and cauda epididymidis. Transfer of biotinylated proteins from cauda epididymosomes to caput spermatozoa decreased in a dose-dependent manner when biotinylated epididymosomes were diluted with unbiotinylated vesicles. Caput epididymosomes added in excess were unable to inhibit transfer of biotinylated proteins from cauda epididymosomes to caput spermatozoa. Following transfer of biotinylated proteins from cauda epididymosomes to caput spermatozoa, addition of unbiotinylated cauda epididymosomes was unable to displace already transferred biotinylated proteins. These results established that epididymosomes from caput and cauda epididymidis have different protein composition and interact differently with maturing spermatozoa.

apocrine secretion, bovine spermatozoa, epididymis, epididymosomes, gamete biology, male reproductive tract, sperm, sperm maturation

¹Supported by Canadian Institutes for Health Research (CIHR) and Natural Sciences and Engineering Research Council of Canada (NSERC) grants to R.S. J.G. is supported by a PhD scholarship from the CIHR.

Correspondence: ² Robert Sullivan, Unité d'Ontogénie-Reproduction, Centre de Recherche, Centre Hospitalier de l'Université Laval, 2705 Blvd. Laurier, Ste-Foy, PQ, Canada, G1V 4G2. FAX: 418 654 2765; e-mail: robert.sullivan{at}crchul.ulaval.ca

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				V. Thimon, G. Frenette, F. Saez, M. Thabet, and R. Sullivan Protein composition of human epididymosomes collected during surgical vasectomy reversal: a proteomic and genomic approach Hum. Reprod., August 1, 2008; 23(8): 1698 - 1707. [Abstract] [Full Text] [PDF]