Establishment of steroidogenic granulosa cell lines expressing follicle stimulating hormone receptors.
Follicle stimulating hormone (FSH) plays an important role in the regulation of oogenesis, spermatogenesis and production of steroid hormones. Receptors to FSH, which are uniquely expressed in ovarian granulosa and testicular Sertoli cells, are rapidly lost in tissue culture conditions and upon cell transformation. We have succeeded, by triple transfection of primary rat granulosa cells with SV40 DNA, Ha-ras oncogene and an FSH receptor expression plasmid, to establish stable steroidogenic cell lines expressing FSH receptors. The cell lines respond to rat, ovine and bovine FSH, which stimulate progesterone production at levels comparable to primary granulosa cells obtained from preovulatory follicles. No steroidogenic response is detected upon stimulation with ovine luteinizing hormone or human chorionic gonadotropin. The steroidogenic response is accompanied by de novo appearance of adrenodoxin which serves as a marker for the mitochondrial steroidogenic enzyme system. These cells express approximately 27,000 receptors per cell with a Kd of 100-115 pM. This Kd is close to the value calculated for the native receptor. The ED50 for the steroidogenic response to ovine FSH is 200 pM, suggesting a tight coupling between receptor activation and the steroidogenic response. FSH induces pronounced morphological changes in the established cell lines, which are also characteristic of primary granulosa cells. These FSH responsive cell lines can serve as a useful model for the study of the structure and function of the FSH receptor and the effect of oncogenes on its expression.
Keren-Tal
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Department of Hormone Research, Weizmann Institute of Science, Rehovot, Israel.
Dantes
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Sprengel
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Amsterdam
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A
D000818:Animals; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D005576:Colforsin / Q000494:pharmacology; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D011905:Genes, ras; D005822:Genetic Vectors; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D011374:Progesterone / Q000096:biosynthesis; D011401:Promoter Regions, Genetic; D051381:Rats; D011962:Receptors, FSH / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection
D004279:DNA, Viral; D011962:Receptors, FSH; D011993:Recombinant Fusion Proteins; D005576:Colforsin; D011374:Progesterone; D005640:Follicle Stimulating Hormone