The non-classical androgen signaling pathway is involved in the regulation of Amh and Cyp26b1 in SMAT1 pre-pubertal Sertoli cells.

SCHTEINGART, HELENA F.; EDELSZTEIN, NADIA Y.; REY, RODOLFO A.

Buenos Aires

Congreso; II Reunión Conjunta de Sociedades de Biociencia; 2017

Sociedad Argentina de Investigación Clínica (SAIC)

Testosterone acts through the androgen receptor (AR) in Sertoli cells to induce their maturation and support spermatogenesis, but the molecular and cellular mechanisms are still poorly understood. We have previously demonstrated that, in the pre-pubertal Sertoli cell line SMAT1, androgens exert a direct effect through the AR on two genes associated with Sertoli cell maturation: Amh - by inhibiting its expression ? and Cyp26b1 ? by upregulating its expression - (Edelsztein et al., SAIC 2015, 2016). Since the AR can act through classical (genomic) or non-classical (non-genomic) pathways, we sought to elucidate whether the effect of androgens on Amh and Cyp26b1 gene expression occur through the classical, the non-classical or both pathways in Sertoli cells. SMAT1 cells were co-transfected with expression vectors for mutant AR proteins that selectively activate either the classical (ARΔ372-385) or the non-classical (ARC562G) testosterone signaling pathway together with Amh or Cyp26b1 promoter luciferase constructs. SMAT1 cells were incubated with DHT (10-7M) or vehicle for 24 h. Results, expressed as percentage (mean±SEM), are compared against basal activity (theoretical value: 100%) using a one sample t-test. Amh expression was inhibited when the ARC562G construct was present (62.9±7.864, P=0.042, n=3) but not after transfection of ARΔ372-385 (82.58±11.63, P=0.273, n=3). Cyp26b1 expression was upregulated by the ARC562G construct (224.2±18.59, P=0.001, n=6) but not by ARΔ372-385 (103.6±7.451, P=0.647, n=6). We conclude that the non-classical pathway participates in the regulation of both Amh and Cyp26b1 in SMAT1 cells, and could be involved in the maturation of Sertoli cells around the onset of puberty.