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Distinct Regulation by Steroids of Messenger RNAs for FSHR and CYP19A1 in Bovine Granulosa Cells

Endocrinology-Reproductive Physiology Program and Department of Dairy Science, University of Wisconsin, Madison, Wisconsin 53706

ABSTRACT

Steroidalregulation of gene expression in follicular cells is not completely defined. Granulosa cells from 5 mm bovine follicles were cultured and treated and steady-state mRNA levels determined forFSHR(follicle-stimulating hormone receptor) andCYP19A1(aromatase). Cells were treated for 5 days with (0.1–300 ng/ml) 17beta-estradiol (E₂), testosterone (T), or 5alpha-dihydrotestosterone (DHT).FSHRmRNA was increased by T and DHT but not E₂. In contrast,CYP19A1mRNA was induced by all doses of E₂ but only high doses of T and DHT. Similarly, varying treatment duration (1–5 days) showed thatFSHRwas increased by T and DHT andCYP19A1mRNA increased by E₂ and T at all times. Synergism between steroid hormones and FSH or forskolin was also evaluated. FSH or E₂ did not alterFSHRmRNA and did not enhance DHT stimulation ofFSHRmRNA. In contrast, DHT alone had no effect onCYP19A1mRNA but synergized with FSH plus E₂ to increaseCYP19A1mRNA, probably due to induction ofFSHRby DHT. Effects of E₂ and T onCYP19A1were blocked by ICI 182,780, indicating mediation by estrogen receptors. However, the specific androgen receptor antagonist bicalutamide did not block E₂ or T effects onCYP19A1but did block T and DHT stimulation ofFSHR. Thus,FSHRis specifically regulated through androgen receptor, whereasCYP19A1is regulated by multiple pathways, including estrogen receptors and cAMP/protein kinase A induced by FSHR activation in granulosa cells. These inter- and intracellular regulatory mechanisms may be critical for normal follicle growth and dominant follicle selection.

androgen receptor, estradiol, estradiol receptor, follicle-stimulating hormone receptor, granulosa cells

¹ Correspondence: Milo C. Wiltbank, 1675 Observatory Dr., Madison, WI 53706. FAX: 608 262 9412; e-mail:Wiltbank{at}wisc.edu

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