Oxytocin receptor expression in stallion testes and epididymides

Endocrine, paracrine, and autocrine factors orchestrate the development and physiology of the stallion reproductive system. Oxytocin (OXT) is one of the critical endocrine, paracrine, and autocrine factors for the male reproductive system. Previous studies have investigated OXT receptor (OXTR) expression in testes and epididymides, including humans, marmosets, macaques, swine, and sheep. This study aimed to explore (1) OXTR localization in the testes and epididymides and (2) the seasonal modification of OXTR expression in the testes. Adult stallion testis and epididymis samples were prepared using routine castration procedures. Reverse-transcription PCR was performed to detect the presence of OXTR messenger RNA (mRNA) in the testes. Western blot procedure was performed to confirm the cross-reactivity of OXTR antibody to horse OXTR. Immunohistochemistry was performed to detect OXTR protein expression in the testes and epididymides. Oxytocin receptor mRNA was detected in the stallion testes. The OXTR protein band was observed at 55 kDa. Interestingly, the relative intensity of the OXTR protein band varied between nonbreeding and breeding season. The OXTR protein level in the testes collected during the breeding season was higher than that during the nonbreeding season. Oxytocin receptor localization was observed in the cytoplasm of Type A spermatogonia and spermatid. Oxytocin receptor protein expression was also observed in the cytoplasmic area of Leydig cells and the membrane of the seminiferous tubules. The cytoplasm of principal and basal cells in the caput, corpus, and cauda was also immunolabeled with OXTR antibody. In conclusion, based on the expression of OXTR in tissues of testes and epididymides, OXT-OXTR system may be a critical factor for stallion testicular and epididymal function. In addition, according to the seasonal alteration of intensity, the OXT-OXTR system may be associated with seasonal changes in the reproductive system in stallions.