Effects of Sex Steroid Receptor Agonists and Antagonists on the Expression of the FOXL2 Transcription Factor and its Target Genes AMH and CYP19A1 in the Neonatal Porcine Ovary

Abstract

Abstract Recently, we have demonstrated that neonatal exposure to androgen and estrogen agonists or antagonists influenced the number of ovarian follicles in piglets. Since the FOXL2 transcription factor is required for proper ovarian follicle formation and activation, the objective of the study was to examine effects of exposure of the neonatal porcine ovary to testosterone propionate (TP; an androgen), flutamide (FLU; an antiandrogen), 4-tert-octylphenol (OP; compound with estrogenic activity), ICI 182,780 (ICI; an antiestrogen), and methoxychlor (MXC; compound with estrogenic, antiestrogenic and antiandrogenic properties) on FOXL2 expression and expression of its target genes, AMH and CYP19A1. Piglets were injected subcutaneously with TP, FLU, OP, ICI, MXC, or corn oil (control) between postnatal days 1 and 10 (n = 4/each group). Ovaries were excised from the 11-day-old piglets and the expression of FOXL2, AMH and CYP19A1 was examined using immunohistochemistry and/or real-time PCR and Western blot. FOXL2 was localized in stroma cells surrounding egg nests and in granulosa cells. TP, OP and MXC increased both FOXL2 and AMH mRNAs, while FLU and ICI decreased CYP19A1 mRNA. The increased FOXL2 protein abundance was found in all examined groups. In addition, TP, OP, ICI and MXC increased AMH protein abundance, while TP, FLU and OP decreased CYP19A1 protein abundance. In conclusion, neonatal exposure to sex steroid receptor agonists and antagonists increased FOXL2 expression at mRNA and/or protein levels and affected FOXL2 target genes in the ovaries of 11-day-old piglets. Therefore, it seems that impaired ovarian folliculogenesis induced by altered steroid milieu during the neonatal development period in pigs may, at least in part, involve FOXL2.