title: Effects of 2-hydroxyoestradiol, oestradiol and testosterone on FSH-induction of catecholamine- and gonadotrophin-responsive progesterone biosynthesis in rat granulosa cell cultures.
abstract: Catecholamine- and gonadotrophin-responsive progesterone biosynthesis increases during FSH-induced granulosa cell maturation. In this study, we compared in vitro effects of interrelated classes of follicular steroid on both endpoints: an androgen (testosterone), an oestrogen (oestradiol) and an oestradiol metabolite (2-hydroxyoestradiol). Granulosa cells from diethylstilboestrol-pretreated, immature rat ovaries were cultured for 48 h (pretreatment) in serum-free medium containing human FSH with or without steroid. The cell monolayers were then washed and reincubated for a further 24 h (test-treatment) in fresh medium with and without a catecholamine (isoproterenol or norepinephrine) or a gonadotrophin (FSH or hCG); test-treatment culture medium was collected and assayed for progesterone content. At concentrations up to 10(-6) M, each steroid enhanced dose-dependently basal and catecholamine-responsive progesterone production with a ranked potency-order of testosterone greater than 2-hydroxyoestradiol much greater than oestradiol. All three compounds also enhanced FSH and hCG responsiveness but this endpoint was affected most markedly by oestradiol. Pretreatment in the presence of specific antiandrogen (hydroxyflutamide; SCH16423) blocked the stimulatory effects of testosterone and 2-hydroxyoestradiol but did not inhibit stimulation by oestradiol, highlighting similarities between the actions of testosterone and the catechol oestrogen distinct from that of oestradiol. Test-treatment in the presence of beta-adrenergic antagonist (propranolol) blocked the stimulatory action of isoproterenol and reduced the response to FSH, suggesting the involvement of beta-adrenergic receptors in FSH as well as catecholamine action on steroidogenesis. We conclude that testosterone and catechol oestradiol have major effects on FSH-induced development of catecholamine responsive steroidogenesis whereas oestradiol mainly affects gonadotrophin responsiveness in this granulosa cell culture system. All three steroid types could interact with gonadotrophins and locally produced catecholamines to influence granulosa cell function in vivo.
authors.affiliation: Institute of Obstetrics and Gynaecology, Hammersmith Hospital, London, England.
authors.forename: K E
authors.initials: KE
authors.lastname: Hudson
authors.forename: E J
authors.initials: EJ
authors.lastname: Wickings
authors.forename: S G
authors.initials: SG
authors.lastname: Hillier
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology*; D002393:Estrogens, Catechol / Q000494:pharmacology*; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D007545:Isoproterenol / Q000494:pharmacology; D007700:Kinetics; D009638:Norepinephrine / Q000494:pharmacology*; D011374:Progesterone / Q000096:biosynthesis*; D051381:Rats; D013739:Testosterone / Q000494:pharmacology*
chemical_list: D006063:Chorionic Gonadotropin; D002393:Estrogens, Catechol; D013739:Testosterone; D011374:Progesterone; D004958:Estradiol; D005640:Follicle Stimulating Hormone; C001390:2-hydroxyestradiol; D007545:Isoproterenol; D009638:Norepinephrine