Estrogens Are Involved in the Activation of Anti-Müllerian Hormone (AMH) Promoter through an Estrogen Response Element Hemi-Palindrome Present at -1772bp

SCHTEINGART HELENA F; VALERI CLARA; EDELSZTEIN NADIA; RIGGIO M; GIULIANELLI S; LANARI CLAUDIA; REY RODOLFO A.

San Diego , California

Congreso; Endocrine Society's 97th Annual Meeting; 2015

The Endocrine Society

AMH is expressed at high levels in prepubertal Sertoli cells of the testis. FSH increases, whereas androgens inhibit, testicular AMH production. In patients with Androgen Insensitivity Syndrome or with Peutz-Jeghers Syndrome, AMH testicular production is increased, concomitantly with estradiol. The AMH promoter contains an estrogen response element hemi-palindrome (½-ERE) at position -1772. In this work we assessed whether estrogens are involved in the increase of the AMH promoter activity in a prepubertal Sertoli cell line, SMAT1. Methods and Results: Reporter assays were performed in SMAT1 cells co-transfected with AMH promoter-luciferase plasmids and expression vectors for human estrogen receptor α (ERα) and/or β (ERβ), and incubated with 17β-estradiol (E2) at concentrations ranging from 10-12 to 10-6 M and/or the antiestrogen ICI 182,780 (10-6 M) for 24 h. Luciferase activity was used to estimate the AMH promoter activity, and results were expressed as % of the respective control (mean ± SEM). One sample t-tests were used to compare the results of the stimulated conditions to a 100% value corresponding to the basal activity. The expression of ERα and ERβ was ascertained in transfected SMAT1 cells by immunofluorescence and Western blotting. Maximum AMH promoter activity in ERα-transfected cells was observed with E2 between 10-10 and 10-8 M; subsequent experiments were performed with E2 10-9 M since it is a physiologic concentration within the testis. In ERα-transfected cells, E2 induced a significant increase of the AMH-3068-1 promoter activity (263.5 ± 12.5%, P