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Gonadotropin regulation of the rat proopiomelanocortin promoter: characterization by transfection of primary ovarian granulosa cells.

Publication ,  Journal Article
Young, SL; Nielsen, CP; Lundblad, JR; Roberts, JL; Melner, MH
Published in: Mol Endocrinol
January 1989

To characterize the transcriptional effects of human (h)FSH and hCG on the POMC gene, primary rat granulosa cells were transiently transfected with a chloramphenicol acetyltransferase (CAT) reporter plasmid under the control of the POMC promoter and 5' region. POMC-CAT contains a fragment of the rat POMC gene, extending from nucleotide -704 to nucleotide +63, fused to the CAT gene. Treatment of POMC-CAT-transfected cells with either hFSH (20 ng/ml) or hCG (10 ng/ml) significantly increased CAT enzyme activity; however, neither hCG nor hFSH increased CAT enzyme activity in cells transfected with pSV2-CAT, a reporter plasmid under the control of the SV40 virus promoter and 5' region. The phosphodiesterase inhibitor isobutylmethylxanthine or the nonhydrolyzable cAMP analog cAMP-chlorothiophenyl significantly increased CAT activity in POMC-CAT-transfected granulosa cells. Human FSH stimulated transcription 10, 20, and 40 h after treatment, but FSH stimulation at the two earlier time points was 2.5- to 5.5-fold greater than that at 40 h. Gonadotropin-stimulated steroidogenesis was equivalent in POMC-CAT-transfected granulosa cells, untransfected, and mock-transfected cells. This indicates that transfection left the physiological hormone response intact. These data demonstrate the following. 1) 767 basepairs of the rat POMC gene are enough to confer gonadotropin stimulation on the CAT marker gene in granulosa cells. 2) Although the POMC promotor lacks a well conserved cAMP response element, either of two different pharmacological manipulations of granulosa cells that raise intracellular cAMP can also stimulate POMC-driven CAT expression. 3) Transfected primary cultures of granulosa cells provide a nontransformed, physiologically relevant model with which to study hormone-regulated gene expression.(ABSTRACT TRUNCATED AT 250 WORDS)

Duke Scholars

Published In

Mol Endocrinol

DOI

ISSN

0888-8809

Publication Date

January 1989

Volume

3

Issue

1

Start / End Page

15 / 21

Location

United States

Related Subject Headings

  • Transfection
  • Transcription, Genetic
  • Thionucleotides
  • Rats
  • Promoter Regions, Genetic
  • Progesterone
  • Pro-Opiomelanocortin
  • Plasmids
  • Granulosa Cells
  • Gene Expression Regulation
 

Citation

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ICMJE
MLA
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Young, S. L., Nielsen, C. P., Lundblad, J. R., Roberts, J. L., & Melner, M. H. (1989). Gonadotropin regulation of the rat proopiomelanocortin promoter: characterization by transfection of primary ovarian granulosa cells. Mol Endocrinol, 3(1), 15–21. https://doi.org/10.1210/mend-3-1-15
Young, S. L., C. P. Nielsen, J. R. Lundblad, J. L. Roberts, and M. H. Melner. “Gonadotropin regulation of the rat proopiomelanocortin promoter: characterization by transfection of primary ovarian granulosa cells.Mol Endocrinol 3, no. 1 (January 1989): 15–21. https://doi.org/10.1210/mend-3-1-15.
Young SL, Nielsen CP, Lundblad JR, Roberts JL, Melner MH. Gonadotropin regulation of the rat proopiomelanocortin promoter: characterization by transfection of primary ovarian granulosa cells. Mol Endocrinol. 1989 Jan;3(1):15–21.
Young, S. L., et al. “Gonadotropin regulation of the rat proopiomelanocortin promoter: characterization by transfection of primary ovarian granulosa cells.Mol Endocrinol, vol. 3, no. 1, Jan. 1989, pp. 15–21. Pubmed, doi:10.1210/mend-3-1-15.
Young SL, Nielsen CP, Lundblad JR, Roberts JL, Melner MH. Gonadotropin regulation of the rat proopiomelanocortin promoter: characterization by transfection of primary ovarian granulosa cells. Mol Endocrinol. 1989 Jan;3(1):15–21.

Published In

Mol Endocrinol

DOI

ISSN

0888-8809

Publication Date

January 1989

Volume

3

Issue

1

Start / End Page

15 / 21

Location

United States

Related Subject Headings

  • Transfection
  • Transcription, Genetic
  • Thionucleotides
  • Rats
  • Promoter Regions, Genetic
  • Progesterone
  • Pro-Opiomelanocortin
  • Plasmids
  • Granulosa Cells
  • Gene Expression Regulation