The ımmunohistochemistry, light and electron microscopic study of the effects of kisspeptin on spermatogenesis and apoptosis in the adult male rats

In this study, it is intended to investigate the effects of acute and chronic administration of Kisspeptin on testis tissue by biochemical methods, light and electron microscopy.Kisspeptin is a very strong neuropeptide inducing the LH and FSH secretion from the hypophysis and shows this effect by increasing the level of GnRH. In the recent studies, Kisspeptin known as a ?chemical kiss? has been reported to be effective about the reaching puberty.In this research, seven groups were formed as 1 day saline (SF), 5 days SF, 13 days SF, 1 day Kisspeptin-10(Kiss-10), 5 days Kiss-10, 13 days Kiss-10, 7 days waiting period after 13 days. The different SF groups were assessed as a control group, both 1 and 5 days adminstration as acute group, 13 days as chronic group. 50 nmol Kiss-10 was treated intraperitoneal (i.p.) one time in a day. Expression of Kisspeptin and GPR54 in the testis tissue was examined with immunfluerosans microscope. Hematoxylene- Eosine, PAS (+), Masson Trichrome staining, Ki-67 immunohistochemistry staining for defining proliferation, TUNEL immunreaction for apoptosis staining were performed for light microscopical evaluations. The level of testosterone was measured in serum by ELISA. The ultrastructure of testis tissues were evaluated.In the Kisspeptin group, the level of testosterone increased in acute period, decreased in chronic period compared to controls. Although the spermatogenetic cells stained with Ki-67 increased according to control group, these decreased in (13+7) group. The number of apoptotic cells defined with TUNEL decreased in acute group according to control, increased in chronic period. There was no differences about testis weight and diameter of tubules. In electron microscopic observation, seminifer tubule epithelium was normal appearance in control groups. In acute groups (1-5 days), apoptotic cells on the basement membran and epithelium of the seminifer tubule and also mitochondrial crystalisis in Sertoli cells and spermatogonium cytoplasm were determined. In 5 days Kiss group, the spaces among the spermatogenetic cells caused by degeneration of the connection units of spermatogenetic cord cell line were observed. In 13 days Kiss group, apoptotic cells were determined at the basal region of the seminifer tubule epithelium. In (13+7) group, as the tubule cells were observed in normal structure, vacuolization was determined in the cytoplasm of the dark spermatogonium placed in basal. In all experiment groups, active Leydig cells were situated in interstitial tissue.As a result, Kisspeptin-10 used in this study increased proliferation in seminifer tubule in acute and chronic adminstration groups. When it gave up, testosteron level and proliferation decreased, apoptosis increased.