Study On The Regulation Of FSH On The Expression Of GDNF Of Piglet Testis Sertoli Cell

Normal and matured sperm are requirement for the mammalian reproduction. FSH is one of the most important endocrine hormone in regulating the spermatogenesis. By controling the sertoli cell's function, FSH stimulated spermatogenesis indirectly. It has been proved that Glial cell line-derived neurotrophic factor, which is produced by Sertoli cells and is regulated by FSH (Tadokoro Y, 2002), is a most important paracrine facor in regulation the stem cell proliferation and/or renewal (Meng X, 2000). By using primary immature boar sertoli cell, this study was conducted to analyze the effects of FSH on GDNF expression in sertoli cell.Firstly, we using semi-quantity RT-PCR and immunhistochemistry to assay the expression of Glial cell line-derived neurotrophic factor (GDNF) in piglet testes and to investigate the significance of GDNF in the regulation of testes development and spermatogenesis. In the immunostaining slides, GDNF was found positively expressed in the Leydig cell area from 2 to 4 weeks, and was found positively expressed in Leydig cells and in the seminiferous tubules after 2 months old. The expression of GDNF enhanced with age increased,. Results from RT-PCR indicated that, the level of GDNF mRNA elevated as age increased, it could be detected in 2 weeks piglet testis and was significant different in 2 month(P<0.05) and 6 month and 18 month old (P<0.01). These results showed that GDNF played a direct role in the regulation of testicular development and spermatogenesis.To study the mechanism of FSH on regulation of GDNF expression in sertoli cell, we used primary piglet SC to analyze the effects of FSH on the proliferative potential and expression of GNDF of Sertoli cell. Our experiment indicated that: (1) FSH can promoted Sertoli cell development and proliferation, prolong the living time of sertoli cell in vitro. The appropriated concentration and stimulated time were 50 ng/ml and 48 hour. It approved that FSH is the necessary component in primary piglet sertoli cell in vitro culture. (2) FSH increased the expression of proliferating cell nuclear antigen (PCNA) of sertoli cell: PCNA protein was detected when stimulated with FSH 15 min and significantly intensed in 30min, with a...