REGULATION OF FOLLISTATIN MESSENGER-RIBONUCLEIC-ACID IN STEROIDOGENIC RAT GRANULOSA-CELL LINES

被引:35
作者
SHUKOVSKI, L
KERENTAL, I
DANTES, A
AMSTERDAM, A
机构
关键词
D O I
10.1210/en.136.7.2889
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Follistatin is a 35-kilodalton monomer isolated from follicular fluid that acts on pituitary gonadotropes to suppress the production of FSH. Transfection of rat granulosa cells with specific oncogenes, such as simian virus 40 (SV40) DNA and Ha-ras oncogene, leads to their immortalization concomitant with preservation of their capacity for inducible steroidogenesis. Experiments were designed to investigate the regulation of follistatin messenger RNA (mRNA) accumulation upon stimulation with forskolin, 2-O-tetradecanol-phorbol-13-acetate (TPA), FSH, and hCG in four different granulosa cell lines. Granulosa cells were transfected with SV40 DNA alone (POGS5); with SV40 DNA and Ha-ras oncogene (POGRS1); with SV40 DNA, Ha-ras oncogene, and LH/CG receptor (GLHR15); or with FSH receptor (GFSHR17) expression plasmid. Cells were cultured to reach confluence and then stimulated for 6 or 24 h with ovine FSH (oFSH; 0.004-4 nM), hCG (9 nM), forskolin (50 mu M), and TPA (50 nM), alone or in combination. In the POGS5 cell line, forskolin caused a 15-fold stimulation of follistatin mRNA after 24-h incubation. The POGRS1 cell line showed a time-dependent stimulation of follistatin gene expression induced by both forskolin (5.7-fold) and TPA (9.4-fold). In the GFSHR17 cells, forskolin, oFSH, and TPA induced an increase in follistatin mRNA. When oFSH(1.6 nM) was added to cells treated with forskolin (50 mu M) or TPA (50 nM), no additional stimulation was observed. The GLHR15 cell line treated with hCG showed a 2.7-fold increase in follistatin mRNA accumulation within 6 h. Our data demonstrate that 1) follistatin mRNA is detectable and induced by forskolin and TPA in transformed granulosa cell lines that do not express the FSH or LH receptors; 2) in the GFSHR17 cell line, FSH, forskolin, and TPA caused a time- and dose-dependent regulation of the gene; and 3) follistatin gene expression is up-regulated by hCG in the GLHR15; cell line. We conclude that these transformed steroidogenic cell lines can serve as a useful model to study the regulation of follistatin gene expression, a peptide known to regulate pituitary and ovarian hormone secretion and differentiation of granulosa cells by its activin-binding action. Moreover, this gene can be regulated in im mortalized granulosa cells by both the protein kinase A and protein kinase C pathways, although these cells express the large T antigen and the Ha-ras oncogenic proteins.
引用
收藏
页码:2889 / 2895
页数:7
相关论文
共 40 条
[31]   REGULATED EXPRESSION OF STEROL CARRIER PROTEIN-2 IN THE OVARY - A KEY ROLE FOR CYCLIC-AMP [J].
RENNERT, H ;
AMSTERDAM, A ;
BILLHEIMER, JT ;
STRAUSS, JF .
BIOCHEMISTRY, 1991, 30 (47) :11280-11285
[32]   THE ISOLATION OF POLYPEPTIDES WITH FSH SUPPRESSING ACTIVITY FROM BOVINE FOLLICULAR-FLUID WHICH ARE STRUCTURALLY DIFFERENT TO INHIBIN [J].
ROBERTSON, DM ;
KLEIN, R ;
DEVOS, FL ;
MCLACHLAN, RI ;
WETTENHALL, REH ;
HEARN, MTW ;
BURGER, HG ;
DEKRETSER, DM .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1987, 149 (02) :744-749
[33]   PRODUCTION OF ACTIVIN-BINDING PROTEIN BY RAT GRANULOSA-CELLS INVITRO [J].
SAITO, S ;
NAKAMURA, T ;
TITANI, K ;
SUGINO, H .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 176 (01) :413-422
[34]   CHARACTERIZATION OF GONADAL SEX CORD-STROMAL TUMOR-CELL LINES FROM INHIBIN-ALPHA AND P53-DEFICIENT MICE - THE ROLE OF ACTIVIN AS AN AUTOCRINE GROWTH-FACTOR [J].
SHIKONE, T ;
MATZUK, MM ;
PERLAS, E ;
FINEGOLD, MJ ;
LEWIS, KA ;
VALE, W ;
BRADLEY, A ;
HSUEH, AJW .
MOLECULAR ENDOCRINOLOGY, 1994, 8 (08) :983-995
[35]   FOLLISTATIN GENE-EXPRESSION IN THE OVARY AND EXTRAGONADAL TISSUES [J].
SHIMASAKI, S ;
KOGA, M ;
BUSCAGLIA, ML ;
SIMMONS, DM ;
BICSAK, TA ;
LING, N .
MOLECULAR ENDOCRINOLOGY, 1989, 3 (04) :651-659
[36]  
SHUKOVSKI L, 1992, J REPROD FERTIL, V95, P861, DOI 10.1530/jrf.0.0950861
[37]   INTRODUCTION OF A GONADOTROPIN RECEPTOR EXPRESSION PLASMID INTO IMMORTALIZED GRANULOSA-CELLS LEADS TO RECONSTITUTION OF HORMONE-DEPENDENT STEROIDOGENESIS [J].
SUH, BS ;
SPRENGEL, R ;
KERENTAL, I ;
HIMMELHOCH, S ;
AMSTERDAM, A .
JOURNAL OF CELL BIOLOGY, 1992, 119 (02) :439-450
[38]  
SUH BS, 1990, ENDOCRINOLOGY, V127, P2489
[39]   ISOLATION AND PARTIAL CHARACTERIZATION OF FOLLISTATIN - A SINGLE-CHAIN MR 35,000 MONOMERIC PROTEIN THAT INHIBITS THE RELEASE OF FOLLICLE-STIMULATING-HORMONE [J].
UENO, N ;
LING, N ;
YING, SY ;
ESCH, F ;
SHIMASAKI, S ;
GUILLEMIN, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1987, 84 (23) :8282-8286
[40]   THE EFFECT OF BOVINE ACTIVIN AND FOLLICLE-STIMULATING-HORMONE (FSH) SUPPRESSING PROTEIN FOLLISTATIN ON FSH-INDUCED DIFFERENTIATION OF RAT GRANULOSA-CELLS INVITRO [J].
XIAO, S ;
FINDLAY, JK ;
ROBERTSON, DM .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 1990, 69 (01) :1-8