title: 17 beta-estradiol stimulates mouse neuropeptide Y-Y(1) receptor gene transcription by binding to estrogen receptor alpha in neuroblastoma cells.
abstract: Several studies have shown that neuropeptide Y (NPY) is involved in the stimulation of gonadotropin hormone releasing hormone (GnRH) and luteinizing hormone (LH) secretion and that these effects are modulated by gonadal steroid feedback. The NPY regulation of GnRH release is probably mediated by the activation of the Y(1) receptor subtype. In this study we examined the regulation of the Y(1) receptor gene transcription by estrogens in transiently transfected NG108-15 neuroblastoma glioma cells. A chimeric plasmid containing the murine Y(1) receptor promoter fused to the firefly luciferase reporter gene was induced by approximately 2-fold in response to 17 beta-estradiol treatment. The estrogen-mediated enhancement of luciferase activity was dose-dependent, blocked by the estrogen receptor (ER) antagonist ICI 182,780, and was strictly dependent on the presence of ER alpha, since it occurred only in NG108-15 cells cotransfected with an expression vector for the human ER. Mutational analysis was performed to investigate whether the hemipalindromic estrogen-responsive elements (EREs) flanking the Y(1) receptor gene are responsible for conferring estradiol inducibility to the Y(1) receptor gene promoter. Mutation of the ERE1 half site at position -932, or mutation of the ERE2 half site at position -809, relative to the ATG, failed to affect the 17 beta-estradiol-mediated enhancement of luciferase activity. Conversely, mutation of both ERE1 and ERE2 half sites completely abolished activation of luciferase activity induced by estrogen. We also examined whether 17 beta-estradiol stimulates the transcriptional activity of the Y(1) receptor gene by binding to ER beta. Results demonstrated that luciferase activity was not modulated by estrogens when cells were transfected with the expression plasmid bearing the human ER beta. Moreover coexpression of both ER alpha and ER beta completely abolished the estrogen-induced activation of luciferase activity observed in the presence of ER alpha. Our data suggest that estrogens activate Y(1) receptor gene transcription possibly via a direct interaction of ER alpha with the hemipalindromic EREs flanking the Y(1) receptor gene.
authors.affiliation: Section of Pharmacology, Department of Anatomy, Pharmacology and Forensic Medicine, Torino, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Musso
authors.forename: A
authors.initials: A
authors.lastname: Maggi
authors.forename: C
authors.initials: C
authors.lastname: Eva
mesh_terms: D000818:Animals; D017931:DNA Primers; D004958:Estradiol / Q000378:metabolism / Q000494:pharmacology*; D047628:Estrogen Receptor alpha; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D005910:Glioma; D008156:Luciferases / Q000235:genetics; D051379:Mice; D016296:Mutagenesis / Q000502:physiology; D009447:Neuroblastoma; D009478:Neuropeptide Y / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000187:drug effects; D051381:Rats; D011960:Receptors, Estrogen / Q000378:metabolism*; D017476:Receptors, Neuropeptide Y / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D017931:DNA Primers; D047628:Estrogen Receptor alpha; D009478:Neuropeptide Y; D011960:Receptors, Estrogen; D017476:Receptors, Neuropeptide Y; C097212:neuropeptide Y-Y1 receptor; D004958:Estradiol; D008156:Luciferases