title: The modulatory role of transforming growth factor beta1 and androstenedione on follicle-stimulating hormone-induced gelatinase secretion and steroidogenesis in rat granulosa cells.
abstract: To investigate the potential roles of matrix metalloproteinases (MMPs) in ovarian granulosa cell differentiation, we studied the interactive effects of FSH and local ovarian factors, transforming growth factor beta1 (TGFbeta1) and androstenedione, on gelatinase secretion and progesterone production in rat ovarian granulosa cells. Granulosa cells of eCG-primed immature rats were treated once with various doses of FSH and TGFbeta1 and androstenedione alone or in combinations for 2 days. Conditioned media were analyzed for gelatinase activity using gelatin-zymography/densitometry and progesterone levels using enzyme immunoassay. Cell lysates were analyzed for steroidogenic acute regulatory (StAR) and cholesterol side-chain-cleavage (P450scc) enzyme protein levels. This study demonstrates for the first time that FSH dose-dependently increased the secretion of a major 63-kDa gelatinase and minor 92- and 67-kDa gelatinases. TGFbeta1 also dose-dependently increased the secretion of 63-kDa gelatinase, while androstenedione alone had no effect. The 92-kDa gelatinase was identified as the pro-MMP9 that could be cleaved by aminophenylmercuric acetate into the 83-kDa active form. Importantly, we show that TGFbeta1 and androgen act in an additive manner to enhance FSH stimulatory effects both on the secretion of gelatinases and the production of progesterone. We further show by immunoblotting that the enhancing effect of TGFbeta1 and androstenedione on FSH-stimulated steroidogenesis is partly mediated through the increased level of StAR protein and/or P450scc enzyme. In conclusion, this study indicates that, during antral follicle development, TGFbeta1 and androgen act to enhance FSH promotion of granulosa cell differentiation and that the process may involve the interplay of modulating cell- to-matrix/cell-to-cell interaction and steroidogenic activity.
authors.affiliation: Institute of Molecular and Cellular Biology, National Taiwan University, Taipei 106, Taiwan.
authors.forename: Ferng-Chun
authors.initials: FC
authors.lastname: Ke
authors.forename: Li-Chung
authors.initials: LC
authors.lastname: Chuang
authors.forename: Ming-Ting
authors.initials: MT
authors.lastname: Lee
authors.forename: Yun Ju
authors.initials: YJ
authors.lastname: Chen
authors.forename: Sui-Wen
authors.initials: SW
authors.lastname: Lin
authors.forename: Paulus S
authors.initials: PS
authors.lastname: Wang
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Stocco
authors.forename: Jiuan-Jiuan
authors.initials: JJ
authors.lastname: Hwang
mesh_terms: D000735:Androstenedione / Q000494:pharmacology*; D000818:Animals; D002478:Cells, Cultured; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D005260:Female; D005640:Follicle Stimulating Hormone / Q000008:administration & dosage / Q000494:pharmacology*; D018093:Gelatinases / Q000378:metabolism*; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D006728:Hormones / Q000008:administration & dosage / Q000494:pharmacology*; D011374:Progesterone / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D013256:Steroids / Q000096:biosynthesis*; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D053773:Transforming Growth Factor beta1
chemical_list: D006728:Hormones; D013256:Steroids; C508962:Tgfb1 protein, rat; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D000735:Androstenedione; D011374:Progesterone; D005640:Follicle Stimulating Hormone; D002786:Cholesterol Side-Chain Cleavage Enzyme; D018093:Gelatinases