Up-Regulation of SOX9 in Sertoli Cells from Testiculopathic Patients Accounts for Increasing Anti-Mullerian Hormone Expression via Impaired Androgen Receptor Signaling
Figure 2
Immunohistochemical analysis of mouse testes.
(A) AR, SOX9 and AMH immunostaining of testes obtained from wild-type and AR knockout (bottom panel) mice. Note that all Sertoli (S) cell nuclei, peritubular myoid (M) cell nuclei and Leydig (L) cells were AR-positive in wild-type mouse testes. Sertoli (S) cell nuclei showing SOX9 expression and Sertoli (S) cell cytoplasm with weaker AMH immunostaining for a wild-type mouse are shown in the upper panel. Sertoli (S) cell nuclei showing SOX9 distribution in an AR knockout mouse. Specific and robust AMH immunostaining was detected exclusively in the cytoplasm of Sertoli (S) cells in AR-knockout mouse testes with deficient spermatogenesis. Magnification ×400; bar = 20 µm. (B) Quantitative evaluation of AR ,SOX9 AMH in testicular tissues from wild-type and AR-knockout mice by real time RT-PCR. Each bar represents the mean ± SD; results were normalized with respect to 18S/GAPDH mRNA expression. SOX9 and AMH mRNA expression was significantly higher in testes from AR-knockout mice compared to controls. (C) SOX9 and AMH protein expression was significantly higher in testes from AR-knockout mice compared to controls by Western blot. (D) Co-immunostaining for AR and SOX9 Sertoli (S) cell nuclei of wild-type mouse testis. Magnification ×400. bar = 20 µm.