Developmental Programming: Gestational Exposure To Excess Testosterone Alters The Ovarian Expression Of SF1 And DAX1 Proteins, Mediators Of AMH Expression, In The Sheep

MATILLER V; PADMANABHAN V; SALVETTI NR; PUTTABYATAPPA M ; ORTEGA HH

Chicago

Congreso; Endocrine Society's annual meeting, Chicago, USA,; 2018

Endocrine Society

Prenatalexposure to excess testosterone (T) induces reproductive defects that includemultifollicular ovarian phenotype, features seen in women with polycysticovarian syndrome (PCOS). Multifollicular phenotype appear to be the result ofenhanced recruitment and follicular persistence. Decreased anti-Mullerianhormone (AMH) in early growing follicles and its increased expression in antralfollicles underlie, in part, the increased recruitment and follicular arrest/persistence,respectively. The mechanisms by which disruption in AMH is mediated is unclear.Considering that AMH expression is regulated by transcription factors, SplicingFactor 1 (SF1) and dosage-sensitive sex reversal, adrenal hypoplasia criticalregion, on chromosome X, gene 1 (DAX1), with SF1 enhancing AMH expression andDAX1 antagonizing SF1?s action, the goal of this study was to determine if developmentalchanges in SF1 and DAX1 underlie prenatal T excess-induced changes in AMHexpression. Ovaries were collected from control and prenatal T-treated (100 mgtwice weekly from days 30-90 of gestation (term 147 days)) animals at fetal day90 and 140 and 1 and 2 years-of-age with sample size of 5-8/ treatmentgroup/developmental time point. Changes in SF1 and DAX1 levels in the granulosacells of the primordial, primary, and small pre-antral (SPA) follicles, granulosa/thecacells of the large pre-antral (LPA) and antral (LA) follicles (5-10 folliclesper follicular type) were assessed by immunohistochemistry, quantified usingImagePro Plus software, and data analyzed by ANOVA. SF1 and DAX1 weredetectable in granulosa cells of all follicle classes at all ages and thecacells of LPA and LA follicles. Prenatal T-treatment (1) had no effect on SF1and DAX1 at fetal day 90; (2) reduced SF1 in the theca cells of LPA folliclesat fetal day 140; (3) increased DAX1 in granulosa cells of primordial throughLPA and theca cells of LPA follicles at fetal day 140; (4) increased SF1 proteincontent in the granulosa cells of SPA, LPA and LA and theca cells of LPAfollicles and DAX1 in the granulosa cells of LPA at 1 year-of-age and (5) hadno effect on SF1 or DAX1 at year 2. These data indicate age-specific changes inSF1 and DAX1 protein. Increased expression of DAX1 in early follicular stages atday 140, increased expression of SF1 in LA follicles at 1 year and lack ofchanges at 2 years-of-age are consistent with decrease in AMH expression inearly stages, increased expression in later stages of follicular development andno changes at 2 years-of-age (Fertil Steril 97:748, 2012). The changes in SF1/DAX1 may therefore contribute to the enhanced follicular recruitment,follicular persistence, and multifollicular ovarian phenotype of prenatalT-treated females and thus may be of translational relevance to the ovarianphenotype of women with PCOS.