title: Ligand-independent effects of estrogen receptor beta on mouse gonadotropin-releasing hormone promoter activity.
abstract: GnRH is the most upstream regulator of reproduction in vertebrates, and its synthesis and release are regulated by gonadal steroid hormones. The proposed sites of hormone action were historically thought to be upstream from GnRH neurons; however, the discovery of ERbeta in a subset of GnRH neurons suggests that this hypothesis should be reevaluated. To determine a functional role for ERbeta in GnRH neurons, we examined ERbeta's regulation of GnRH promoter activity. The GnRH-producing cell line, GT1-7, was cotransfected with expression vectors containing one of three ERbeta splice variants and a luciferase-reporter construct containing the full-length mouse GnRH promoter sequence or one of two deletions upstream of the transcription start site (-225/-201; -184/-150). Transfected cells were treated with 100 nm 17beta-estradiol (E(2)), diarylpropionitrile, raloxifene, or vehicle. There was a robust increase in GnRH-luciferase activity by all ERbeta splice variants in the absence of hormone. Furthermore, E(2) treatment abolished this response for ER-beta1 and ER-beta2, but not ER-beta1delta3. The -225/-201 and -184/-150 regions were critical for ERbeta-induced promoter activity because deletion of these regions eliminated the ligand-independent effects of ERbeta. ER-beta1 binds directly to these promoter regions and because there are no classical estrogen response elements in the mouse GnRH promoter, these data raise the possibility that this region contains a novel estrogen response element specific for ERbeta. Overall, our data suggest that ERbeta functions as a basic transcription factor in GnRH neurons and demonstrate a potential molecular mechanism for the negative feedback effects of E(2) on GnRH.
authors.affiliation: Department of Biomedical Sciences, College of Veterinary Medicine, Colorado State University, Fort Collins, 80523, USA. Toni.Pak@colostate.edu
authors.forename: Toni R
authors.initials: TR
authors.lastname: Pak
authors.forename: Wilson C J
authors.initials: WC
authors.lastname: Chung
authors.forename: James L
authors.initials: JL
authors.lastname: Roberts
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Handa
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D000242:Cyclic AMP / Q000096:biosynthesis; D047629:Estrogen Receptor beta / Q000235:genetics / Q000502:physiology*; D025461:Feedback, Physiological; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics*; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic
chemical_list: D047629:Estrogen Receptor beta; D008024:Ligands; D007987:Gonadotropin-Releasing Hormone; D000242:Cyclic AMP