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Characterization of a Novel Rat Epididymal Cell Line to Study Epididymal Function

Institut National de la Recherche Scientifique-Institut Armand-Frappier (J.D., N.S., D.G.C.), Université du Québec, Pointe-Claire, Québec, Canada H9R 1G6; Ontogeny-Reproduction Research Unit (R.S.V.), Centre Hospitalier Universitaire Laval Research Centre and Centre de Recherche en Biologie de la Reproduction, Department of Obstetrics and Gynecology, Université Laval, Sainte-Foy, Québec, Canada G1K 7P4; and Department of Anatomy and Cell Biology (L.H., D.G.C.), McGill University, Montreal, Québec, Canada H3A 2B2

Address all correspondence and requests for reprints to: Dr. Daniel G. Cyr, Institut National de la Recherche Scientifique-Institut Armand Frappier, Université du Québec, 245 Hymus Boulevard, Pointe Claire, Quebec, Canada H9R 1G6. E-mail. daniel.cyr{at}iaf.inrs.ca.

The epididymis is an androgen-dependent organ that allows spermatozoa to become fully functional as they pass through this tissue. The specialized functions of the epididymis are mediated by interactions between epididymal epithelial cells and between epididymal cells and spermatozoa. Although the critical role of the epididymis in sperm maturation is well established, the mechanisms regulating cell-cell interactions remain poorly understood because of the lack of appropriate cell line models. We now report the characterization of a novel rat caput epididymal cell line (RCE) that was immortalized by transfecting primary cultures of rat epididymal cells with the simian virus 40 large T antigen. At the electron microscope level, the cell line was composed of epithelial principal cells with characteristics of in vivo cells; principal cells had well-developed Golgi apparatus, abundant endoplasmic reticulum cisternae, and few endosomes. RCE cells expressed the mRNAs coding for the androgen receptor, estrogen receptor , and 4-ene-steroid-5--reductase types 1 and 2 as well as epididymal-specific markers Crisp-1 and epididymal retinoic acid binding protein. Epididymal retinoic acid binding protein expression was significantly induced with dihydrotestosterone, although this effect was not blocked by flutamide, suggesting that RCE cells are not androgen responsive. Neighboring cells formed tight and gap junctions characteristic of epididymal cells in vivo and expressed tight (occludin and claudin-1, -3, and -4) and gap junctional proteins (connexin-26, -30.3, -32, and -43). The RCE cell line displays many characteristics of epithelial principal cells, thus providing a model for studying epididymal cell functions.

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