title: Luteinizing hormone beta promoter stimulation by adenylyl cyclase and cooperation with gonadotropin-releasing hormone 1 in transgenic mice and LBetaT2 Cells.
abstract: Rat luteinizing hormone beta (Lhb) gene transcription is stimulated by hypothalamic gonadotropin-releasing hormone 1 (GnRH1), and this response may be modulated by other signaling pathways such as cAMP. Here we characterize the ability of cAMP, alone or with GnRH1, to stimulate Lhb gene transcription in mouse pituitary and clonal gonadotroph cells. Both cAMP and pituitary adenylyl cyclase-activating peptide increase GnRH1 stimulation of luciferase activity in pituitaries of mice expressing the rat Lhb-luciferase transgene, suggesting cAMP and GnRH1 pathways interact in vivo. cAMP stimulation of the Lhb-luciferase transgene was similar between females in metestrus and proestrus, but GnRH1 stimulation was greater at proestrus. Additive effects with combined treatments were observed at metestrus and proestrus. Elevated intracellular cAMP stimulated Lhb promoter activity in LbetaT2 clonal gonadotroph cells, alone and with GnRH1. In LbetaT2 cells, cAMP stimulation of the Lhb promoter was eliminated by inhibition of protein kinase A (PKA); GnRH1 stimulation was partially suppressed by either PKA or protein kinase C inhibitors. Only the proximal GnRH1-responsive region of the promoter was required for cAMP stimulation, and mutation of the 3' NR5A1 site diminished the response. Regulation of primary mRNA transcripts from the endogenous Lhb gene by cAMP and GnRH1 correlated with results from the Lhb-luciferase transgene or transfected promoter. Occupancy of the endogenous promoter by EGR1 was increased by GnRH1 with or without forskolin, but forskolin alone had little effect. Thus, cAMP stimulation of Lhb promoter activity, and enhancement of GnRH1 stimulation, occurs in multiple physiological states independent of steroid status, via a PKA-dependent mechanism.
authors: Department of Physiology, University of Virginia Medical School, Charlottesville, Virginia 22903, USA.
authors: Heather A
authors: HA
authors: Ferris
authors: Heidi E
authors: HE
authors: Walsh
authors: Jonathan
authors: J
authors: Stevens
authors: Patricia C
authors: PC
authors: Fallest
authors: Margaret A
authors: MA
authors: Shupnik
mesh_terms: D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D012739:Gonadal Steroid Hormones / Q000378:metabolism*; D052681:Gonadotrophs / Q000378:metabolism*; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism*; D037101:Luteinizing Hormone, beta Subunit / Q000378:metabolism*; D008686:Metestrus / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide / Q000378:metabolism; D011359:Proestrus / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism*; D054339:Steroidogenic Factor 1 / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: C495402:Adcyap1 protein, mouse; D012739:Gonadal Steroid Hormones; D037101:Luteinizing Hormone, beta Subunit; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D011498:Protein Precursors; D054339:Steroidogenic Factor 1; D014157:Transcription Factors; C050866:progonadoliberin I; D005576:Colforsin; D007987:Gonadotropin-Releasing Hormone; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C