title: The influence of the antiandrogen 2-hydroxyflutamide on the androgen receptor expression in the porcine ovarian follicles - an in vitro study.
abstract: Androgens are one of the most important agents influencing ovarian follicles growth and development. The biological action of androgens is primarily exerted through transcriptional regulation by the androgen receptor (AR), a member of the steroid hormone receptor superfamily. The purpose of this study was to test the role of androgen receptor agonist testosterone (T) or antagonist 2-hydroxyflutamide (2-Hf) and in combination on AR expression in cultured porcine granulosa cells (GC) or whole follicles. Granulosa cells isolated from mature pig follicles were cultured for 48 h. During the last 12 and 24 h of culture, they were incubated in the presence of T (10(-7)  m/ml), 2-Hf (1.7 × 10(-4)  m) or both T and 2-Hf (T + 2-Hf, at the same concentrations as when added separately). To better imitate in vivo conditions, whole follicles (6-8 mm in diameter) isolated from porcine ovaries have been incubated (for 12 and 24 h) in an organ culture system with the addition of the same factors. Thereafter, cells or sections obtained from cultured follicles were processed for AR detection by immunocytochemistry or immunohistochemistry. Moreover, expression of AR mRNA and protein was determined by real-time PCR and Western blot analysis. It was shown that the addition of 2-Hf in the presence of T had a positive effect on AR mRNA and protein expression in porcine GC and ovarian follicles. Moreover, the addition of 2-Hf influenced AR distribution in GC cultures which is seen as change of its localization from nuclear to perinuclear. Our results suggest that androgens acting through AR could be involved in the control of AR expression in porcine GC in vitro and in vivo.
authors.affiliation: Department of Endocrinology, Institute of Zoology, Jagiellonian University, Krakow, Poland. duda.m@op.pl
authors.forename: M
authors.initials: M
authors.lastname: Duda
authors.forename: A
authors.initials: A
authors.lastname: Wolna
authors.forename: K
authors.initials: K
authors.lastname: Knapczyk-Stwora
authors.forename: M
authors.initials: M
authors.lastname: Durlej-Grzesiak
authors.forename: M
authors.initials: M
authors.lastname: Knet
authors.forename: Z
authors.initials: Z
authors.lastname: Tabarowski
authors.forename: M
authors.initials: M
authors.lastname: Slomczynska
mesh_terms: D000726:Androgen Antagonists / Q000494:pharmacology; D000818:Animals; D005260:Female; D005485:Flutamide / Q000031:analogs & derivatives* / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D006080:Ovarian Follicle / Q000187:drug effects*; D011944:Receptors, Androgen / Q000235:genetics / Q000378:metabolism*; D013552:Swine / Q000502:physiology*; D013739:Testosterone / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000726:Androgen Antagonists; D011944:Receptors, Androgen; C014290:hydroxyflutamide; D013739:Testosterone; D005485:Flutamide