Characterization of progesterone biosynthesis in a transformed granulosa cell line1 1 A preliminary report was presented at the Serono Symposium 'Molecular Basis of Reproductive Endocrinology' held in Vancouver, B.C., Canada, July 1991.

I. M. Rao, P. F. Gadson, E. Anderson, Peter J Hornsby, V. B. Mahesh

Research output: Contribution to journalArticle

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Abstract

Original languageEnglish
Pages (from-to)121-128
Number of pages8
JournalMolecular and Cellular Endocrinology
Volume94
Issue number1
DOIs
StatePublished - 1993
Externally publishedYes

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Endocrinology
Granulosa Cells
Biosynthesis
Canada
Progesterone
Cells
Cell Line
Cholesterol Side-Chain Cleavage Enzyme
Cyclic AMP
Rats
Serum
Polyomavirus Transforming Antigens
Diethylstilbestrol
Gap Junctions
Gene expression
Population
Horses
Estradiol
Messenger RNA
Genes

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

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title = "Characterization of progesterone biosynthesis in a transformed granulosa cell line1 1 A preliminary report was presented at the Serono Symposium 'Molecular Basis of Reproductive Endocrinology' held in Vancouver, B.C., Canada, July 1991.",
abstract = "The study of regulation of steroidogenesis in primary cultures of rat granulosa cells is difficult because the cells do not undergo more than one cell doubling in culture. Furthermore, there is size and steroidogenic heterogeneity in granulosa cells and it is difficult to obtain pure, functionally defined populations. Hence, it is advantageous to develop a homogeneous population of granulosa cells. In this report we describe the characterization of one such cell line (Rao-gcl-29) developed from diethylstilbestrol treated immature rat granulosa cells by transformation with SV40 T antigen. In this cell line cyclic AMP analogs induce high levels of progesterone biosynthesis, though there was no effect on estradiol biosynthesis. Also, FSH and hCG have no effect on progesterone biosynthesis. In the presence of FBS medium (20{\%} fetal bovine serum in DMEM/F-12) and enriched medium (10{\%} fetal bovine serum, 10{\%} horse serum and 2{\%} UltraSer G in DMEM/F-12 medium), 1 mM cAMP analogs induce high levels of progesterone biosynthesis upto 96 h. Ultrastructural features of the cell line resemble those of primary granulosa cells, in addition to forming gap junctions. Cyclic AMP analogs also induced cytochrome P450scc mRNA in this cell line by 48 h, and this effect is apparent by 24 h. Thus, this cell line could be useful in understanding the molecular mechanisms of regulation of cytochrome P450scc gene regulation.",
keywords = "Cytochrome P450scc, Granulosa cell, Progesterone, SV40 T antigen, Transformation",
author = "Rao, {I. M.} and Gadson, {P. F.} and E. Anderson and Hornsby, {Peter J} and Mahesh, {V. B.}",
year = "1993",
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T1 - Characterization of progesterone biosynthesis in a transformed granulosa cell line1 1 A preliminary report was presented at the Serono Symposium 'Molecular Basis of Reproductive Endocrinology' held in Vancouver, B.C., Canada, July 1991.

AU - Rao, I. M.

AU - Gadson, P. F.

AU - Anderson, E.

AU - Hornsby, Peter J

AU - Mahesh, V. B.

PY - 1993

Y1 - 1993

N2 - The study of regulation of steroidogenesis in primary cultures of rat granulosa cells is difficult because the cells do not undergo more than one cell doubling in culture. Furthermore, there is size and steroidogenic heterogeneity in granulosa cells and it is difficult to obtain pure, functionally defined populations. Hence, it is advantageous to develop a homogeneous population of granulosa cells. In this report we describe the characterization of one such cell line (Rao-gcl-29) developed from diethylstilbestrol treated immature rat granulosa cells by transformation with SV40 T antigen. In this cell line cyclic AMP analogs induce high levels of progesterone biosynthesis, though there was no effect on estradiol biosynthesis. Also, FSH and hCG have no effect on progesterone biosynthesis. In the presence of FBS medium (20% fetal bovine serum in DMEM/F-12) and enriched medium (10% fetal bovine serum, 10% horse serum and 2% UltraSer G in DMEM/F-12 medium), 1 mM cAMP analogs induce high levels of progesterone biosynthesis upto 96 h. Ultrastructural features of the cell line resemble those of primary granulosa cells, in addition to forming gap junctions. Cyclic AMP analogs also induced cytochrome P450scc mRNA in this cell line by 48 h, and this effect is apparent by 24 h. Thus, this cell line could be useful in understanding the molecular mechanisms of regulation of cytochrome P450scc gene regulation.

AB - The study of regulation of steroidogenesis in primary cultures of rat granulosa cells is difficult because the cells do not undergo more than one cell doubling in culture. Furthermore, there is size and steroidogenic heterogeneity in granulosa cells and it is difficult to obtain pure, functionally defined populations. Hence, it is advantageous to develop a homogeneous population of granulosa cells. In this report we describe the characterization of one such cell line (Rao-gcl-29) developed from diethylstilbestrol treated immature rat granulosa cells by transformation with SV40 T antigen. In this cell line cyclic AMP analogs induce high levels of progesterone biosynthesis, though there was no effect on estradiol biosynthesis. Also, FSH and hCG have no effect on progesterone biosynthesis. In the presence of FBS medium (20% fetal bovine serum in DMEM/F-12) and enriched medium (10% fetal bovine serum, 10% horse serum and 2% UltraSer G in DMEM/F-12 medium), 1 mM cAMP analogs induce high levels of progesterone biosynthesis upto 96 h. Ultrastructural features of the cell line resemble those of primary granulosa cells, in addition to forming gap junctions. Cyclic AMP analogs also induced cytochrome P450scc mRNA in this cell line by 48 h, and this effect is apparent by 24 h. Thus, this cell line could be useful in understanding the molecular mechanisms of regulation of cytochrome P450scc gene regulation.

KW - Cytochrome P450scc

KW - Granulosa cell

KW - Progesterone

KW - SV40 T antigen

KW - Transformation

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