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The Journal of Neuroscience, November 24, 2004, 24(47):10628-10635; doi:10.1523/JNEUROSCI.5540-03.2004

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Cellular/Molecular
Estrogen Receptor (ER) Isoforms Rather Than ER Regulate Corticotropin-Releasing Hormone Promoter Activity through an Alternate Pathway

William J. Schouler Miller,^1,2 Shotaro Suzuki,³ Lydia K. Miller,^1,2 Robert Handa,³ and Rosalie M. Uht¹

¹Departments of Pathology and Biochemistry and Molecular Genetics and ²University of Virginia College, University of Virginia, Charlottesville, Virginia 22908-0904, and ³Colorado State University, Fort Collins, Colorado 80523

The hypothalamic-pituitary-adrenal axis regulates mammalian stress responses by secreting glucocorticoids. The magnitude of the response is in part determined by gender, for in response to a given stressor, circulating glucocorticoids reach higher levels in female rats than in males. This gender difference could result from estrogen regulation of the corticotropin-releasing hormone (CRH) promoter via either of its receptors: estrogen receptor (ER) or ER. Immunocytochemistry revealed that a subset (12%) of medial parvocellular CRH neurons in the rat hypothalamus contain ER but not ER. To determine whether ERs could regulate CRH promoter activity, we cotransfected cells with a CRH promoter construct and either ER or individual ER isoforms. ER weakly stimulated CRH promoter transcriptional activity in a ligand-independent manner. Conversely, all ER isoforms tested stimulated CRH promoter activity with different ligand profiles. ER1 and ER23 displayed constitutive activity (ER1 more than ER23). Ligand-dependent activity of isoforms 1 and 2 was altered by an Exon3 splice variant (3) or by the additional 18 amino acids in the ligand-binding domain of ER2 isoforms. Lastly, we suggest that ER regulation of CRH takes place through an alternate pathway, one that requires protein-protein interactions with other transcription factors or their associated complexes. However, a pure ER-activator protein-1 alternate pathway does not appear to be involved.

Key words: estrogen receptor ; estrogen receptor ; corticotropin-releasing hormone; estradiol; tamoxifen; stress

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Received June 4, 2003; revised September 16, 2004; accepted October 7, 2004.

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