Suppression of the GnRH Pulse Generator by Neurokinin B Involves a κ-Opioid Receptor-Dependent Mechanism

• Published in: Endocrinology (Philadelphia) 2012-10, Vol.153 (10), p.4894-4904
• Main Authors: Grachev, P, Li, X. F, Kinsey-Jones, J. S, di Domenico, A. L, Millar, R. P, Lightman, S. L, O'Byrne, K. T
• Format: Article
• Language: English
• Subjects: 17β-Estradiol; Agonists; Animals; Antagonists; Arcuate nucleus; Arcuate Nucleus of Hypothalamus - drug effects; Arcuate Nucleus of Hypothalamus - metabolism; Biological and medical sciences; Dynorphin A; Estradiol - blood; Estrous Cycle - drug effects; Estrous Cycle - metabolism; Fundamental and applied biological sciences. Psychology; G protein-coupled receptors; Gene expression; Genes; Gonadotropin-releasing hormone; Gonadotropin-Releasing Hormone - genetics; Gonadotropin-Releasing Hormone - metabolism; Hypothalamus; In vivo methods and tests; Kiss1 protein; Kisspeptins - genetics; Kisspeptins - metabolism; Luteinizing Hormone - metabolism; Narcotics; Neurogenesis; Neurokinin; Neurokinin B; Neurons; Opioid receptors; Ovariectomy; Peptide Fragments - pharmacology; Preoptic Area - drug effects; Preoptic Area - metabolism; Pulse generators; Rats; Rats, Sprague-Dawley; Receptors; Receptors, G-Protein-Coupled - genetics; Receptors, G-Protein-Coupled - metabolism; Receptors, Kisspeptin-1; Receptors, Neurokinin-3 - agonists; Receptors, Opioid, kappa - metabolism; Sampling methods; Secretion; Senktide; Sex hormones; Signalling systems; Substance P - analogs & derivatives; Substance P - pharmacology; Vertebrates: endocrinology
• ISSN: 0013-7227; 1945-7170
• DOI: 10.1210/en.2012-1574

Neurokinin B (NKB) and its receptor (NK3R) are coexpressed with kisspeptin, Dynorphin A (Dyn), and their receptors [G-protein-coupled receptor-54 (GPR54)] and κ-opioid receptor (KOR), respectively] within kisspeptin/NKB/Dyn (KNDy) neurons in the hypothalamic arcuate nucleus (ARC), the proposed site of the GnRH pulse generator. Much previous research has employed intracerebroventricular (icv) administration of KNDy agonists and antagonists to address the functions of KNDy neurons. We performed a series of in vivo neuropharmacological experiments aiming to determine the role of NKB/NK3R signaling in modulating the GnRH pulse generator and elucidate the interaction between KNDy neuropeptide signaling systems, targeting our interventions to ARC KNDy neurons. First, we investigated the effect of intra-ARC administration of the selective NK3R agonist, senktide, on pulsatile LH secretion using a frequent automated serial sampling method to obtain blood samples from freely moving ovariectomized 17β-estradiol-replaced rats. Our results show that senktide suppresses LH pulses in a dose-dependent manner. Intra-ARC administration of U50488, a selective KOR agonist, also caused a dose-dependent, albeit more modest, decrease in LH pulse frequency. Thus we tested the hypothesis that Dyn/KOR signaling localized to the ARC mediates the senktide-induced suppression of the LH pulse by profiling pulsatile LH secretion in response to senktide in rats pretreated with nor-binaltorphimine, a selective KOR antagonist. We show that nor-binaltorphimine blocks the senktide-induced suppression of pulsatile LH secretion but does not affect LH pulse frequency per se. In order to address the effects of acute activation of ARC NK3R, we quantified (using quantitative RT-PCR) changes in mRNA levels of KNDy-associated genes in hypothalamic micropunches following intra-ARC administration of senktide. Senktide down-regulated expression of genes encoding GnRH and GPR54 (GNRH1 and Kiss1r, respectively), but did not affect the expression of Kiss1 (which encodes kisspeptin). We conclude that NKB suppresses the GnRH pulse generator in a KOR-dependent fashion and regulates gene expression in GnRH neurons.