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

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

Chicago

Congreso; 100th Annual Meeting of the Endocrine Society and Expo (ENDO2018); 2018

Endocrine Society

Testosteroneacts through the androgen receptor (AR) in Sertoli cells to induce theirmaturation and support spermatogenesis, but the molecular and cellularmechanisms are still poorly understood. We have previously demonstrated that,in the pre-pubertal Sertoli cell line SMAT1, androgens exert a direct effectthrough the AR on two genes associated with Sertoli cell maturation: Amh- by inhibiting its expression - and Cyp26b1 - by upregulating itsexpression. Since the AR can act through classical (genomic) or non-classical(non-genomic) pathways, we sought to elucidate whether the effect of androgenson Amh and Cyp26b1 gene expression occurs through the classical,the non-classical or both pathways in Sertoli cells. SMAT1 cells wereco-transfected with expression vectors for mutant AR proteins that selectivelyactivate either the classical (ARÄ372-385) or the non-classical (ARC562G)testosterone signaling pathway together with Amh or Cyp26b1 promoterluciferase constructs. SMAT1 cells were incubated with DHT (10-7M)or vehicle for 24 h. Results, expressed as percentage (mean±SEM), are comparedagainst basal activity (theoretical value: 100%) using a one sample t-test. Amhexpression was inhibited when the ARC562G construct was present (62.9±7.864, P=0.042,n=3) but not after transfection of ARÄ372-385 (82.58±11.63, P=0.273, n=3). Cyp26b1expression was upregulated by the ARC562G construct (224.2±18.59, P=0.001,n=6) but not by ARÄ372-385(103.6±7.451, P=0.647, n=6). We conclude that the non-classical pathwayparticipates in the regulation of both Amh and Cyp26b1 in SMAT1cells, and could be involved in the maturation of Sertoli cells around theonset of puberty.