title: Glycine and GABA(A) ultra-sensitive ethanol receptors as novel tools for alcohol and brain research.
abstract: A critical obstacle to developing effective medications to prevent and/or treat alcohol use disorders is the lack of specific knowledge regarding the plethora of molecular targets and mechanisms underlying alcohol (ethanol) action in the brain. To identify the role of individual receptor subunits in ethanol-induced behaviors, we developed a novel class of ultra-sensitive ethanol receptors (USERs) that allow activation of a single receptor subunit population sensitized to extremely low ethanol concentrations. USERs were created by mutating as few as four residues in the extracellular loop 2 region of glycine receptors (GlyRs) or γ-aminobutyric acid type A receptors (GABA(A)Rs), which are implicated in causing many behavioral effects linked to ethanol abuse. USERs, expressed in Xenopus oocytes and tested using two-electrode voltage clamp, demonstrated an increase in ethanol sensitivity of 100-fold over wild-type receptors by significantly decreasing the threshold and increasing the magnitude of ethanol response, without altering general receptor properties including sensitivity to the neurosteroid, allopregnanolone. These profound changes in ethanol sensitivity were observed across multiple subunits of GlyRs and GABA(A)Rs. Collectively, our studies set the stage for using USER technology in genetically engineered animals as a unique tool to increase understanding of the neurobiological basis of the behavioral effects of ethanol.
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences (A.N., K.H.M., R.L.A.) and Titus Family Department of Clinical Pharmacy and Pharmaceutical Economics and Policy (L.A., D.L.D.), University of Southern California School of Pharmacy, Los Angeles, California; Department of Anesthesia, Beckman Program for Molecular and Genetic Medicine, Stanford University, Stanford University Medical Center, Stanford, California (J.R.T.); Departments of Anesthesiology and Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (G.E.H.); and Amgen Inc., Thousand Oaks, California (D.I.P.).
authors.forename: Anna
authors.initials: A
authors.lastname: Naito
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences (A.N., K.H.M., R.L.A.) and Titus Family Department of Clinical Pharmacy and Pharmaceutical Economics and Policy (L.A., D.L.D.), University of Southern California School of Pharmacy, Los Angeles, California; Department of Anesthesia, Beckman Program for Molecular and Genetic Medicine, Stanford University, Stanford University Medical Center, Stanford, California (J.R.T.); Departments of Anesthesiology and Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (G.E.H.); and Amgen Inc., Thousand Oaks, California (D.I.P.).
authors.forename: Karan H
authors.initials: KH
authors.lastname: Muchhala
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences (A.N., K.H.M., R.L.A.) and Titus Family Department of Clinical Pharmacy and Pharmaceutical Economics and Policy (L.A., D.L.D.), University of Southern California School of Pharmacy, Los Angeles, California; Department of Anesthesia, Beckman Program for Molecular and Genetic Medicine, Stanford University, Stanford University Medical Center, Stanford, California (J.R.T.); Departments of Anesthesiology and Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (G.E.H.); and Amgen Inc., Thousand Oaks, California (D.I.P.).
authors.forename: Liana
authors.initials: L
authors.lastname: Asatryan
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences (A.N., K.H.M., R.L.A.) and Titus Family Department of Clinical Pharmacy and Pharmaceutical Economics and Policy (L.A., D.L.D.), University of Southern California School of Pharmacy, Los Angeles, California; Department of Anesthesia, Beckman Program for Molecular and Genetic Medicine, Stanford University, Stanford University Medical Center, Stanford, California (J.R.T.); Departments of Anesthesiology and Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (G.E.H.); and Amgen Inc., Thousand Oaks, California (D.I.P.).
authors.forename: James R
authors.initials: JR
authors.lastname: Trudell
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences (A.N., K.H.M., R.L.A.) and Titus Family Department of Clinical Pharmacy and Pharmaceutical Economics and Policy (L.A., D.L.D.), University of Southern California School of Pharmacy, Los Angeles, California; Department of Anesthesia, Beckman Program for Molecular and Genetic Medicine, Stanford University, Stanford University Medical Center, Stanford, California (J.R.T.); Departments of Anesthesiology and Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (G.E.H.); and Amgen Inc., Thousand Oaks, California (D.I.P.).
authors.forename: Gregg E
authors.identifier: http://orcid.org/0000-0003-3641-8153
authors.initials: GE
authors.lastname: Homanics
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences (A.N., K.H.M., R.L.A.) and Titus Family Department of Clinical Pharmacy and Pharmaceutical Economics and Policy (L.A., D.L.D.), University of Southern California School of Pharmacy, Los Angeles, California; Department of Anesthesia, Beckman Program for Molecular and Genetic Medicine, Stanford University, Stanford University Medical Center, Stanford, California (J.R.T.); Departments of Anesthesiology and Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (G.E.H.); and Amgen Inc., Thousand Oaks, California (D.I.P.).
authors.forename: Daya I
authors.initials: DI
authors.lastname: Perkins
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences (A.N., K.H.M., R.L.A.) and Titus Family Department of Clinical Pharmacy and Pharmaceutical Economics and Policy (L.A., D.L.D.), University of Southern California School of Pharmacy, Los Angeles, California; Department of Anesthesia, Beckman Program for Molecular and Genetic Medicine, Stanford University, Stanford University Medical Center, Stanford, California (J.R.T.); Departments of Anesthesiology and Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (G.E.H.); and Amgen Inc., Thousand Oaks, California (D.I.P.) ddavies@usc.edu.
authors.forename: Daryl L
authors.initials: DL
authors.lastname: Davies
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences (A.N., K.H.M., R.L.A.) and Titus Family Department of Clinical Pharmacy and Pharmaceutical Economics and Policy (L.A., D.L.D.), University of Southern California School of Pharmacy, Los Angeles, California; Department of Anesthesia, Beckman Program for Molecular and Genetic Medicine, Stanford University, Stanford University Medical Center, Stanford, California (J.R.T.); Departments of Anesthesiology and Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania (G.E.H.); and Amgen Inc., Thousand Oaks, California (D.I.P.).
authors.forename: Ronald L
authors.initials: RL
authors.lastname: Alkana
mesh_terms: D000818:Animals; D001921:Brain / Q000187:drug effects*; D000431:Ethanol / Q000494:pharmacology*; D005260:Female; D008958:Models, Molecular; D011280:Pregnanolone / Q000494:pharmacology; D011963:Receptors, GABA-A / Q000737:chemistry / Q000187:drug effects*; D018009:Receptors, Glycine / Q000737:chemistry / Q000187:drug effects*; D013329:Structure-Activity Relationship; D014982:Xenopus laevis; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
chemical_list: D011963:Receptors, GABA-A; D018009:Receptors, Glycine; D000431:Ethanol; D005680:gamma-Aminobutyric Acid; D011280:Pregnanolone