CEDIE   05498
CENTRO DE INVESTIGACIONES ENDOCRINOLOGICAS "DR. CESAR BERGADA"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
FSH regulates anti-Müllerian hormone gene expression through PKA-mediated SOX9 activation and SF1 nuclear translocation
Autor/es:
LASALA, C.; SCHTEINGART, H.F.; AROUCHE, N.; BEDECARRAS, P; DI CLEMENTE, N; REY, R
Lugar:
New York, USA
Reunión:
Congreso; Lawson Wilkins Pediatric Endocrinology Society / European Society for Pediatric Endocrinology 2009 Meeting; 2009
Institución organizadora:
LWPES/ESPE
Resumen:
FSH increases anti-Müllerian hormone (AMH) gene expression in prepubertal Sertoli cells through the classical cyclic AMP (cAMP) pathway. No classical cAMP response element is present on the AMH promoter. SOX9, SF1, GATA4 and AP1 are activated by cAMP in diverse cellular types and have response elements on the AMH promoter. Our aim was to test whether AP1, GATA4, SF1 and/or SOX9 could be involved in cAMP-mediated upregulation of AMH expression in prepubertal Sertoli cells. A previously validated prepubertal Sertoli cell line (SMAT1) was incubated for 24 h with or without 1 mM dbcAMP in the presence or absence of inhibitors of PKA (H89), PI3-K (LY294002), p38-K (SB203580) or MAPK (PD98059). AMH promoter activity was assessed after 30 Hormone Research 2009 Free Communication transfection with luciferase vectors containing 3 kb of the AMH promoter, either wild-type or with mutated sites for GATA (-74, -168 and -408), AP1 (-203), SOX9 (-141) or SF1 (-92 and -218). Mutations of SOX9 and SF1 sites, but not those of GATA4 and AP1 sites, abolished AMH promoter response to dbcAMP. The PKA inhibitor H89, but not the other kinase inhibitors, impaired wild-type promoter response to cAMP. We next assessed whether an increased availability of SOX9 and SF1 in Sertoli cell nuclei was observed after cAMP incubation. In real time RT-PCR and Western blot analyses, SOX9 expression was upregulated in SMAT1 cells after cAMP incubation. H89 precluded SOX9 upregulation. SF1 expression was not modified by cAMP. By immunocytochemistry, we observed that SOX9 was always nuclear, whereas SF1 was translocated from the cytoplasm to the nucleus after cAMP incubation. In conclusion, cAMP activation of PKA results in an increased bioavailability of SF1 and SOX9 in prepubertal Sertoli cells, resulting from SF1 nuclear translocation and SOX9 upregulated expression. cAMP-activated SF1 and SOX9 are responsible for the increase in AMH promoter activity, after binding to specific response elements present in the proximal AMH promoter.