title: Investigating the Mechanism by Which Gain-of-function Mutations to the α1 Glycine Receptor Cause Hyperekplexia.
abstract: Hyperekplexia is a rare human neuromotor disorder caused by mutations that impair the efficacy of glycinergic inhibitory neurotransmission. Loss-of-function mutations in the GLRA1 or GLRB genes, which encode the α1 and β glycine receptor (GlyR) subunits, are the major cause. Paradoxically, gain-of-function GLRA1 mutations also cause hyperekplexia, although the mechanism is unknown. Here we identify two new gain-of-function mutations (I43F and W170S) and characterize these along with known gain-of-function mutations (Q226E, V280M, and R414H) to identify how they cause hyperekplexia. Using artificial synapses, we show that all mutations prolong the decay of inhibitory postsynaptic currents (IPSCs) and induce spontaneous GlyR activation. As these effects may deplete the chloride electrochemical gradient, hyperekplexia could potentially result from reduced glycinergic inhibitory efficacy. However, we consider this unlikely as the depleted chloride gradient should also lead to pain sensitization and to a hyperekplexia phenotype that correlates with mutation severity, neither of which is observed in patients with GLRA1 hyperekplexia mutations. We also rule out small increases in IPSC decay times (as caused by W170S and R414H) as a possible mechanism given that the clinically important drug, tropisetron, significantly increases glycinergic IPSC decay times without causing motor side effects. A recent study on cultured spinal neurons concluded that an elevated intracellular chloride concentration late during development ablates α1β glycinergic synapses but spares GABAergic synapses. As this mechanism satisfies all our considerations, we propose it is primarily responsible for the hyperekplexia phenotype.
authors.affiliation: From the Queensland Brain Institute and.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: From the Queensland Brain Institute and.
authors.forename: Anna
authors.initials: A
authors.lastname: Bode
authors.affiliation: From the Queensland Brain Institute and.
authors.forename: Bindi
authors.initials: B
authors.lastname: Nguyen
authors.affiliation: From the Queensland Brain Institute and.
authors.forename: Angelo
authors.initials: A
authors.lastname: Keramidas
authors.affiliation: From the Queensland Brain Institute and School of Biomedical Sciences, University of Queensland, Brisbane, Queensland, Australia 4072 j.lynch@uq.edu.au.
authors.forename: Joseph W
authors.identifier: 0000-0002-1227-2252
authors.initials: JW
authors.lastname: Lynch
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D059330:GABAergic Neurons / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D000071017:Hyperekplexia / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D020125:Mutation, Missense; D051381:Rats; D018009:Receptors, Glycine / Q000235:genetics / Q000378:metabolism; D013569:Synapses / Q000235:genetics / Q000378:metabolism; D009435:Synaptic Transmission / Q000235:genetics*
chemical_list: C000657604:GLRA1 protein, human; C000657606:Glra1 protein, rat; D018009:Receptors, Glycine
keywords: glycine receptor; glycinergic; inhibition mechanism; patch clamp; site-directed mutagenesis; startle disease; synapse; tropisetron