title: Direct regulation of gonadotrophin-releasing hormone (GnRH) transcription by RF-amide-related peptide-3 and kisspeptin in a novel GnRH-secreting cell line, mHypoA-GnRH/GFP.
abstract: RF-amide-related peptide-3 [RFRP-3; also often referred to as the mammalian orthologue of the avian gonadotrophin-inhibitory hormone (GnIH)] and kisspeptin have emerged as potent modulators of neuroendocrine function via direct regulation of the reproductive axis in the hypothalamus and pituitary. There are few studies focusing on the direct regulatory effects of RFRP-3 and kisspeptin on gonadotrophin-releasing hormones (GnRH) neurones. We report their effect on GnRH mRNA expression and release in a novel GnRH neuronal cell model, mHypoA-GnRH/GFP, generated from adult-derived GnRH-GFP neurones. The neurones express receptors for both RFRP-3 and kisspeptin, Gpr147 and Gpr54, respectively. Incubation with 100 nm RFRP-3 results in attenuation of GnRH mRNA expression by approximately 60%. Conversely, incubation with 10 nm of Kiss-10 induced GnRH mRNA expression, whereas the combined effect was an overall repression of GnRH mRNA levels. With transcription inhibitors, the repression of GnRH mRNA levels was linked to a transcriptional mechanism but not mRNA stability. No significant changes in GnRH secretion were observed upon RFRP-3 exposure in these neurones. Our findings suggest that the suppressive signalling of RFRP-3 on GnRH transcription may dominate over kisspeptin induction in the mHypoA-GnRH/GFP GnRH neuronal cell model.
authors.affiliation: Department of Physiology, University of Toronto, Toronto, ON, Canada.
authors.forename: N M
authors.initials: NM
authors.lastname: Gojska
authors.forename: Z
authors.initials: Z
authors.lastname: Friedman
authors.forename: D D
authors.initials: DD
authors.lastname: Belsham
mesh_terms: D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology; D002460:Cell Line; D003609:Dactinomycin / Q000494:pharmacology; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics* / Q000378:metabolism; D059648:Kisspeptins / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology*; D051379:Mice; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D000074502:Receptors, Kisspeptin-1; D018013:Receptors, Neuropeptide / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*
chemical_list: C110962:5,6-dichlorobenzimidazole; D001562:Benzimidazoles; C492248:Kiss1r protein, mouse; D059648:Kisspeptins; D009479:Neuropeptides; C416987:RFamide peptide; D012333:RNA, Messenger; D043562:Receptors, G-Protein-Coupled; D000074502:Receptors, Kisspeptin-1; D018013:Receptors, Neuropeptide; C083929:neuropeptide FF receptor; D003609:Dactinomycin; D007987:Gonadotropin-Releasing Hormone
keywords: RF-amide-related peptide; gonadotrophin-inhibitory hormone; gonadotrophin-releasing hormone; hypothalamus; kisspeptin; neuronal cell biology; secretion; transcription