Spermatozoa modulate epididymal cell proliferation and protein secretion in vitro

Abstract Normal epididymal function, such as protein expression and secretion, is primarily regulated by testicular androgens and temperature. However, the role of spermatozoa in this critical process has never been studied. In order to determine whether sperm itself could regulate epididymal function, we have developed a cell culture system of bovine epididymal cells to study the interactions between spermatozoa and the epididymal epithelium. Primary cells from caput, corpus, and cauda epididymal tissues were cultured in the presence of androgens at 32°C (scrotal) and 37°C (abdominal). Newly synthesized proteins were metabolically labeled with 35 S‐methionine after sperm co‐incubation and the pattern of secreted proteins was analyzed by two‐dimensional polyacrylamide gel electrophoresis. Proliferation rate, protein secretion rate and electrophoretic patterns of secreted proteins were evaluated 48 hr post‐co‐incubation. Incubation at 32°C indicated that spermatozoa stimulation increases the level of protein secretion of cultured cells from all epididymal sections while it slightly decreases proliferation of corpus cells. At 37°C, spermatozoa co‐incubation significantly decreases the protein secretion rate of cultured cells from all epididymal sections. Independently of cell incubation temperature, spermatozoa stimulation induces both an increase in the intensity of radiolabeled proteins and the appearance of new secreted proteins of caput cells without affecting the protein pattern of corpus or cauda cells. Incubation at 37°C, however, greatly modifies the pattern of proteins expressed at 32°C by cauda cells. Taken together, these results support the hypothesis that spermatozoa themselves affect epididymal cell function, most importantly for caput epididymides. Mol. Reprod. Dev. 75: 512–520, 2008. © 2007 Wiley‐Liss, Inc.