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2 September 2009 Progesterone Receptor-Induced Gene Expression in Primary Mouse Granulosa Cell Cultures
Venkataraman Sriraman, Mala Sinha, JoAnne S. Richards
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The progesterone receptor (PGR) is induced by luteinizing hormone (LH) in granulosa cells of preovulatory follicles, and the PGR-A isoform is essential for ovulation based on the phenotypes of Pgr isoform-specific knockout mice. Although several genes regulated by PGR-A in vivo have been identified, whether these genes are primary targets of PGR-A or if their expression also depends on other signaling molecules that are induced by the LH surge has not been resolved. Therefore, to identify genes that are either induced or repressed by PGR in the absence of LH-mediated signaling cascades, we infected primary cultures of mouse granulosa cells with either PGR-A or PGR-B adenoviral vectors without or with R-5020 as a PGR ligand. Total RNA was extracted from infected cells at 16 h and analyzed by Affymetrix Mouse 430 2.0 microarrays. PGR-A in the presence or absence of ligand significantly induced approximately 50 genes 2-fold or more (local pooled error test at P ≤ 0.01). Fewer and different genes were induced by PGR-B in the absence of ligand. Edn1, Apoa1, and Cited1 were primarily regulated by PGR-A as verified by additional RT-PCR analyses, suppression by the PGR antagonist RU486, and the lack of induction by protein kinase A, protein kinase C, or epidermal growth factor (EGF)-like factors pathways. PGR regulation of these genes was confirmed further by gene expression analyses in hormonally primed Pgr mutant mouse ovaries. Because Edn1, Apoa1, and Cited1 are known to regulate angiogenesis, PGR may affect the neovascularization of follicles that is initiated with ovulation.

Venkataraman Sriraman, Mala Sinha, and JoAnne S. Richards "Progesterone Receptor-Induced Gene Expression in Primary Mouse Granulosa Cell Cultures," Biology of Reproduction 82(2), 402-412, (2 September 2009).
Received: 19 March 2009; Accepted: 1 August 2009; Published: 2 September 2009

granulosa cells
progesterone receptor
steroid hormone receptors
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