Immunohistochemical and hybridocytochemical study on ghrelin signalling in the rat seminiferous epithelium

The results of presented study demonstrate expression of ghrelin, its functional receptor GHSR-1a and their genes in spermatogenic cells of rat testis suggesting their functioning within seminiferous epithelium. The immunohistochemical and hybrydocytochemical expression, of proteins and transcripts, was estimated taking into account the cycle of seminiferous epithelium and phases of spermatogenesis. Both transcripts and ghrelin was found to show nuclear expression and scarcely cytoplasmic. Expression of genes for ghrelin and GHSR-1a was shown in early spermatocytes and round spermatids representing transcriptional phases of meiosis and spermiogenesis. Ghrelin was evidenced to show nuclear expression in two stage-specific windows, in late spermatogonia, in spermatocytes up to early pachytenes, and again in spermatids of acrosome and early maturation phase of spermiogenesis. In late pachytenes, secondary spermatocytes, round spermatids, maturing spermatids and spermatozoa the reaction is lacking. With two types of antibodies against the GHSR-1a used the two different patterns of immunostaining was evidenced suggesting two isoforms of GHSR-1a. The first evidenced GHSR-1a in cytoplasm of spermatocytes, cell membrane and acrosomes of spermatids, Sertoli cell processes and heads of spermatozoa. With second type of antibodies the immunostaining marks all steps of evolution of acrosome in spermatids. It is believed that site of ghrelin expression in seminiferous epithelium may indicate its role in local regulations, not excepting the intracellular signalling. Immunostaining pattern for GHSR-1a seems to suggest both its participation in the cross-talk among the cells and also process of furnishing gametes with GHSR-1a for its response to ghrelin in seminal plasma or female reproductive tract.