Expression and localization of guanine nucleotide-binding protein alpha S in the testis and epididymis of rams at different developmental stages

•The Gαs subunit is expressed in ram testicles in an age-dependent manner.•Gαs levels are significantly upregulated in the epididymis compared to the testes.•Gαs expression is the highest in the cauda and corpus epididymis of rams.•Gαs is synthesized by Sertoli and Leydig cells, spermatozoa, and spermatocytes.

The guanine nucleotide-binding alpha S subunit (Gαs) is an important element of key signaling pathways, which is widely expressed in mammalian tissues; however, its role in the reproductive system is still unclear. In this study, we investigated the expression and localization of Gαs in the testes and epididymis of rams at different developmental stages using quantitative RT-PCR, immunohistochemistry, and western blotting. In 1-, 6-, and 12-month-old rams, the transcription of Gαs-encoding gene (Gnαs) was significantly upregulated in the corpus and cauda epididymis compared to the testes and caput epididymis (P < 0.05). At 12 months, the level of Gnαs mRNA was higher than that at 1 and 6 months for all tested tissues (P < 0.05). The Gαs protein was detected in the principal cells and interstitial epididymal cells, including Sertoli and Leydig cells, as well as in testicular cells, spermatogonial stem cells, and spermatocytes. Gαs expression was the highest in the cauda epididymis (P < 0.05), followed by the corpus epididymis, caput epididymis, and testes. The results indicate that in the reproductive organs of rams, Gαs is expressed in a tissue-specific and age-dependent manner. The high levels of Gαs observed in the epididymis suggest that Gαs may influence the composition of the epididymal lumen fluid and, consequently, the microenvironment for spermatozoa maturation. Thus, Gαs could play an important role in spermatogenesis and the development of the testes and epididymis in the reproductive system of rams.