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Volume 16, Number 9, Issue of May 1, 1996 pp. 2869-2880
Copyright ©1996 Society for NeuroscienceStructure and Pharmacological Properties of a Molluscan Glutamate-Gated Cation Channel and its Likely Role in Feeding Behavior
Received Oct. 11, 1995; revised Jan. 29, 1996; accepted Feb. 2, 1996.
Thorsten Stühmer1, Muriel Amar1, 2, Robert J. Harvey1, Isabel Bermudez2, Jan van Minnen3, and Mark G. Darlison1 1 Institut für Zellbiochemie und Klinische Neurobiologie, Universitäts-Krankenhaus Eppendorf, Universität Hamburg, 20246 Hamburg, Germany, 2 School of Biological and Molecular Sciences, Oxford Brookes University, Headington, Oxford OX3 0BP, United Kingdom, and 3 Department of Organismic Zoology, Vrije Universiteit, 1081 HV Amsterdam, The Netherlands
We describe the isolation of a molluscan (Lymnaea stagnalis) full-length complementary DNA that encodes a mature polypeptide (which we have named Lym-eGluR2) with a predicted molecular weight of 105 kDa that exhibits 44-48% identity to the mammalian kainate-selective glutamate receptor GluR5, GluR6, and GluR7 subunits. Injection of in vitro-transcribed RNA from this clone into Xenopus laevis oocytes results in the robust expression of homo-oligomeric cation channels that can be gated by L-glutamate (EC50 = 1.2 ± 0.3 µM) and several other glutamate receptor agonists; rank order of potency: glutamate
kainate > ibotenate > AMPA. These currents can be blocked by the mammalian non-NMDA receptor antagonists 6,7-dinitroquinoxaline-2,3-dione, 6-cyano-7-nitroquinoxaline-2,3-dione, and 1-(4-chlorobenzoyl)piperazine-2,3-dicarboxylic acid. Ionic-replacement experiments have shown that the agonist-induced current is carried entirely by sodium and potassium ions. In situ hybridization has revealed that the Lym-eGluR2 transcript is present in all 11 ganglia of the Lymnaea CNS, including the 4-cluster motorneurons within the paired buccal ganglia. The pharmacological properties and deduced location of Lym-eGluR2 are entirely consistent with it being (a component of) the receptor, which has been identified previously on buccal motorneurons, that mediates the excitatory effects of glutamate released from neurons within the feeding central pattern generator.
Key words: buccal ganglion; complementary DNA cloning; feeding behavior; in situ hybridization; ion channel; ionotropic glutamate receptor; kainate receptor; mollusc (Lymnaea stagnalis); Xenopus oocyte expression; 4-cluster motorneurons
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