title: Anxiogenic and Stressor Effects of the Hypothalamic Neuropeptide RFRP-3 Are Overcome by the NPFFR Antagonist GJ14.
abstract: RFamide-related peptide-3 (RFRP-3) is a recently discovered neuropeptide that has been proposed to play a role in the stress response. We aimed to elucidate the role of RFRP-3 and its receptor, neuropeptide FF (NPFF1R), in modulation of stress and anxiety responses. To achieve this, we characterized a new NPFF1R antagonist because our results showed that the only commercially available putative antagonist, RF9, is in fact an agonist at both NPFF1R and the kisspeptin receptor (KISS1R). We report here the identification and pharmacological characterization of GJ14, a true NPFFR antagonist. In in vivo tests of hypothalamic-pituitary-adrenal (HPA) axis function, GJ14 completely blocked RFRP-3-induced corticosterone release and neuronal activation in CRH neurons. Furthermore, chronic infusion of GJ14 led to anxiolytic-like behavior, whereas RFRP-3 infusion had anxiogenic effects. Mice receiving chronic RFRP-3 infusion also had higher basal circulating corticosterone levels. These results indicate a stimulatory action of RFRP-3 on the HPA axis, consistent with the dense expression of NPFF1R in the vicinity of CRH neurons. Importantly, coinfusion of RFRP-3 and GJ14 completely reversed the anxiogenic and HPA axis-stimulatory effects of RFRP-3. Here we have established the role of RFRP-3 as a regulator of stress and anxiety. We also show that GJ14 can reverse the effects of RFRP-3 both in vitro and in vivo. Infusion of GJ14 causes anxiolysis, revealing a novel potential target for treating anxiety disorders.
authors.affiliation: Centre for Neuroendocrinology and Department of Anatomy (J.S.K., P.W.B., G.M.A.) and National School of Pharmacy (J.D.A.T.), University of Otago, Dunedin 9054, New Zealand; Glycosyn (B.S.D.), Callaghan Innovation (B.S.D.), and The Ferrier Research Institute (G.F.P.), University of Wellington, Lower Hutt 5010, Victoria, New Zealand; Institut National de la Recherche Agronomique (M.B.), Nouzilly 37380, France; and School of Biological Sciences (C.S.W., D.L.H.), University of Auckland, Auckland 1142, New Zealand.
authors.forename: Joon S
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Centre for Neuroendocrinology and Department of Anatomy (J.S.K., P.W.B., G.M.A.) and National School of Pharmacy (J.D.A.T.), University of Otago, Dunedin 9054, New Zealand; Glycosyn (B.S.D.), Callaghan Innovation (B.S.D.), and The Ferrier Research Institute (G.F.P.), University of Wellington, Lower Hutt 5010, Victoria, New Zealand; Institut National de la Recherche Agronomique (M.B.), Nouzilly 37380, France; and School of Biological Sciences (C.S.W., D.L.H.), University of Auckland, Auckland 1142, New Zealand.
authors.forename: Phil W
authors.initials: PW
authors.lastname: Brownjohn
authors.affiliation: Centre for Neuroendocrinology and Department of Anatomy (J.S.K., P.W.B., G.M.A.) and National School of Pharmacy (J.D.A.T.), University of Otago, Dunedin 9054, New Zealand; Glycosyn (B.S.D.), Callaghan Innovation (B.S.D.), and The Ferrier Research Institute (G.F.P.), University of Wellington, Lower Hutt 5010, Victoria, New Zealand; Institut National de la Recherche Agronomique (M.B.), Nouzilly 37380, France; and School of Biological Sciences (C.S.W., D.L.H.), University of Auckland, Auckland 1142, New Zealand.
authors.forename: Blake S
authors.initials: BS
authors.lastname: Dyer
authors.affiliation: Centre for Neuroendocrinology and Department of Anatomy (J.S.K., P.W.B., G.M.A.) and National School of Pharmacy (J.D.A.T.), University of Otago, Dunedin 9054, New Zealand; Glycosyn (B.S.D.), Callaghan Innovation (B.S.D.), and The Ferrier Research Institute (G.F.P.), University of Wellington, Lower Hutt 5010, Victoria, New Zealand; Institut National de la Recherche Agronomique (M.B.), Nouzilly 37380, France; and School of Biological Sciences (C.S.W., D.L.H.), University of Auckland, Auckland 1142, New Zealand.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Beltramo
authors.affiliation: Centre for Neuroendocrinology and Department of Anatomy (J.S.K., P.W.B., G.M.A.) and National School of Pharmacy (J.D.A.T.), University of Otago, Dunedin 9054, New Zealand; Glycosyn (B.S.D.), Callaghan Innovation (B.S.D.), and The Ferrier Research Institute (G.F.P.), University of Wellington, Lower Hutt 5010, Victoria, New Zealand; Institut National de la Recherche Agronomique (M.B.), Nouzilly 37380, France; and School of Biological Sciences (C.S.W., D.L.H.), University of Auckland, Auckland 1142, New Zealand.
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Walker
authors.affiliation: Centre for Neuroendocrinology and Department of Anatomy (J.S.K., P.W.B., G.M.A.) and National School of Pharmacy (J.D.A.T.), University of Otago, Dunedin 9054, New Zealand; Glycosyn (B.S.D.), Callaghan Innovation (B.S.D.), and The Ferrier Research Institute (G.F.P.), University of Wellington, Lower Hutt 5010, Victoria, New Zealand; Institut National de la Recherche Agronomique (M.B.), Nouzilly 37380, France; and School of Biological Sciences (C.S.W., D.L.H.), University of Auckland, Auckland 1142, New Zealand.
authors.forename: Debbie L
authors.initials: DL
authors.lastname: Hay
authors.affiliation: Centre for Neuroendocrinology and Department of Anatomy (J.S.K., P.W.B., G.M.A.) and National School of Pharmacy (J.D.A.T.), University of Otago, Dunedin 9054, New Zealand; Glycosyn (B.S.D.), Callaghan Innovation (B.S.D.), and The Ferrier Research Institute (G.F.P.), University of Wellington, Lower Hutt 5010, Victoria, New Zealand; Institut National de la Recherche Agronomique (M.B.), Nouzilly 37380, France; and School of Biological Sciences (C.S.W., D.L.H.), University of Auckland, Auckland 1142, New Zealand.
authors.forename: Gavin F
authors.initials: GF
authors.lastname: Painter
authors.affiliation: Centre for Neuroendocrinology and Department of Anatomy (J.S.K., P.W.B., G.M.A.) and National School of Pharmacy (J.D.A.T.), University of Otago, Dunedin 9054, New Zealand; Glycosyn (B.S.D.), Callaghan Innovation (B.S.D.), and The Ferrier Research Institute (G.F.P.), University of Wellington, Lower Hutt 5010, Victoria, New Zealand; Institut National de la Recherche Agronomique (M.B.), Nouzilly 37380, France; and School of Biological Sciences (C.S.W., D.L.H.), University of Auckland, Auckland 1142, New Zealand.
authors.forename: Joel D A
authors.initials: JD
authors.lastname: Tyndall
authors.affiliation: Centre for Neuroendocrinology and Department of Anatomy (J.S.K., P.W.B., G.M.A.) and National School of Pharmacy (J.D.A.T.), University of Otago, Dunedin 9054, New Zealand; Glycosyn (B.S.D.), Callaghan Innovation (B.S.D.), and The Ferrier Research Institute (G.F.P.), University of Wellington, Lower Hutt 5010, Victoria, New Zealand; Institut National de la Recherche Agronomique (M.B.), Nouzilly 37380, France; and School of Biological Sciences (C.S.W., D.L.H.), University of Auckland, Auckland 1142, New Zealand.
authors.forename: Greg M
authors.initials: GM
authors.lastname: Anderson
mesh_terms: D000818:Animals; D001007:Anxiety / Q000139:chemically induced / Q000188:drug therapy* / Q000378:metabolism; D001522:Behavior, Animal / Q000187:drug effects* / Q000502:physiology; D016466:CHO Cells; D003345:Corticosterone / Q000097:blood; D003412:Cricetulus; D007030:Hypothalamo-Hypophyseal System / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D009479:Neuropeptides / Q000494:pharmacology*; D010913:Pituitary-Adrenal System / Q000187:drug effects / Q000378:metabolism; D018013:Receptors, Neuropeptide / Q000037:antagonists & inhibitors*; D013315:Stress, Psychological / Q000139:chemically induced / Q000188:drug therapy* / Q000378:metabolism
chemical_list: D009479:Neuropeptides; C416987:RFamide peptide; D018013:Receptors, Neuropeptide; C083929:neuropeptide FF receptor; D003345:Corticosterone