ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of c-jun on hCG-induced testosterone secretion in isolated rat Leydig cells by antisense oligodeoxynucleotides(ASODNs).</p><p><b>METHODS</b>c-jun ASODNs were used to antagonise the effects of c-jun, hCG was used to induce the testosterone secretion of LC cultured in vitro and testosterone was measured by radioimmunoassay.</p><p><b>RESULTS</b>The testosterone secretion of LC in vitro could be induced by hCG, which was a good model for the functional study of LC. c-jun ASODNs decreased the hCG-induced testosterone secretion of LC in a dose-dependent manner(P < 0.05).</p><p><b>CONCLUSION</b>It is suggested that c-jun proto-oncogene enhances the testosterone secretion of LC.</p>
Subject(s)
Animals , Male , Rats , Cells, Cultured , Chorionic Gonadotropin , Pharmacology , Dose-Response Relationship, Drug , Leydig Cells , Bodily Secretions , Oligonucleotides, Antisense , Pharmacology , Proto-Oncogene Proteins c-jun , Physiology , Rats, Sprague-Dawley , Testosterone , Bodily SecretionsABSTRACT
Objective To study isolation and identification and culture of rat type A spermatogonial cells in vitro.Methods Percoll discontinue density gradient centrifugation combined with different speeds of different cells adhering to dish was used to purify the type A spermatogonial cells.The c-kit and TERT special antibodies were used to identify the type A spermatogonial cells.The purified cells were cultured in vitro. Results 0.614?10~6 cells per testis finally were obtained and the percentage of viable cells was 92.1% by trypan blue dye exclusion test.The percentage of type A spermatogonial cells expressing c-kit and TERT were 91.7?1.2% and 90.8?1.0% respectively.Type A spermatogonial cells could proliferate and self-renew in the DMEM containing 10% NBS.Conclusion Percoll discontinue density gradient centrifugation combined with different speeds of different cells adhering to dish is an efficiency method for isolation of rat type A spermatogonial cells.The purified cells are type A spermatogonial cells by identification of the immunohistochemistry of c-kit and TERT antibodies.Type A spermatogonial cells can proliferate and self-renew in vitro.