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Intracellular mechanisms of gonadotropin-stimulated gene expression in granulosa cells.

Publication ,  Journal Article
Melner, MH; Young, SL
Published in: Steroids
May 1991

Previous studies have shown that the gonadotropins follicle-stimulating hormone and luteinizing hormone stimulate proopiomelanocortin (POMC) promoter activity and mRNA levels in ovarian granulosa cells. The objective of these studies was to determine the role of cAMP-dependent protein kinases (pKA) in gonadotropin-stimulated gene expression. Primary cultures of rat granulosa cells were transfected with a gene construct consisting of the POMC promoter (-150 to +63; designated pOMC-CAT) fused to the chloramphenicol acetyltransferase (CAT) reporter gene either alone or cotransfected with an expression plasmid (designated mutant RI), which overexpresses a mutant form of the murine RI subunit incapable of binding cAMP and serving as an irreversible inhibitor of the catalytic subunit of pKA. Follicle-stimulating hormone or isoproterenol caused a significant stimulation of pOMC-CAT activity in transfected cells. Cotransfection of pOMC-CAT with mutant RI caused a significant inhibition of basal pOMC-CAT activity and abolished the gonadotropin stimulation. As a control, transfection of the SV-40 viral enhancer-promoter fused to CAT (pSV2-CAT) was unresponsive to follicle-stimulating hormone stimulation and cotransfection with mutant RI had no significant effect on pSV2-CAT activity. These studies suggest that gonadotropin regulation of the POMC promoter is mediated by pKA and that promoter activity is stringently controlled by pKA.

Duke Scholars

Published In

Steroids

DOI

ISSN

0039-128X

Publication Date

May 1991

Volume

56

Issue

5

Start / End Page

232 / 236

Location

United States

Related Subject Headings

  • Transfection
  • Rats
  • Protein Kinases
  • Protein Kinase Inhibitors
  • Plasmids
  • Granulosa Cells
  • Gonadotropins, Equine
  • Gene Expression Regulation
  • Follicle Stimulating Hormone
  • Female
 

Citation

APA
Chicago
ICMJE
MLA
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Melner, M. H., & Young, S. L. (1991). Intracellular mechanisms of gonadotropin-stimulated gene expression in granulosa cells. Steroids, 56(5), 232–236. https://doi.org/10.1016/0039-128x(91)90039-x
Melner, M. H., and S. L. Young. “Intracellular mechanisms of gonadotropin-stimulated gene expression in granulosa cells.Steroids 56, no. 5 (May 1991): 232–36. https://doi.org/10.1016/0039-128x(91)90039-x.
Melner, M. H., and S. L. Young. “Intracellular mechanisms of gonadotropin-stimulated gene expression in granulosa cells.Steroids, vol. 56, no. 5, May 1991, pp. 232–36. Pubmed, doi:10.1016/0039-128x(91)90039-x.
Journal cover image

Published In

Steroids

DOI

ISSN

0039-128X

Publication Date

May 1991

Volume

56

Issue

5

Start / End Page

232 / 236

Location

United States

Related Subject Headings

  • Transfection
  • Rats
  • Protein Kinases
  • Protein Kinase Inhibitors
  • Plasmids
  • Granulosa Cells
  • Gonadotropins, Equine
  • Gene Expression Regulation
  • Follicle Stimulating Hormone
  • Female