Abstract
Melatonin (MEL) is involved in homeostasis of the epididymis lumen environment. Dihydrotestosterone (DHT) partakes in the development of gonads and organs in male animals. However, whether MEL secretion, the expression of its receptors, MT1 and MT2, and sheep epididymal epithelial cell apoptosis is regulated by DHT remains unclear. In this study, we used immunohistochemical staining to detect the distribution patterns of DHT synthetases [5α-reductase (5α-red)] and its androgen receptor (AR) in sheep epididymides. 5α-red1, 5α-red2 and AR were positively expressed in sperm, epididymal epithelial cells, and the smooth muscle cells of the caput, corpus and cauda regions of the epididymis. DHT concentration and the expression levels of 5α-red and AR in the caput, corpus and cauda regions were measured by enzyme-linked immunosorbent assay, liquid chromatography-mass spectrometry, real-time quantitative polymerase chain reaction and western blot analysis. DHT concentration in the caput was significantly higher than those in corpus and cauda, probably because of the high expression of 5α-red2 in the caput and secretion and transport of DHT by the testicles. DHT inhibited MEL secretion, the expression of its membrane receptors and MEL synthetases in cultured sheep epididymal epithelial cells in vitro. In addition, the Bax/Bcl-2 ratio, ACT CASP3 and caspase-3 mRNA expression were also decreased. The decreasing effect was partially reversed after flutamide treatment. Therefore, DHT regulates sheep epididymal function by influencing MEL expression and apoptosis-related factors. This study provides basic data for further research on the reproductive physiology of male animals.