Abstract
We have isolated a cDNA clone from the nematode Caenorhabditis elegans that encodes a protein of greatest sequence similarity to muscarinic acetylcholine receptors. This gene codes for a polypeptide of 682 amino acids containing seven putative transmembrane domains. The amino acid identities, excluding a highly variable middle portion of the third intracellular loop, to the human m1-m5 receptors are 28-34%. When this cloned receptor was coexpressed with a G protein-gated inwardly rectifying K+ channel (GIRK1) in Xenopus oocyte, acetylcholine was able to elicit the GIRK current. This acetylcholine-induced current was substantially inhibited by the muscarinic antagonist atropine in a reversible manner. However, another muscarinic agonist oxotremorine and antagonists scopolamine and pirenzepine had little or negligible effects on this receptor. Taken together, these results suggest that the cloned gene encodes a G protein-linked acetylcholine receptor that is most similar to but pharmacologically distinct from muscarinic acetylcholine receptors.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acetylcholine / pharmacology
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Animals
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Atropine / pharmacology
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Base Sequence
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Blotting, Southern
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Caenorhabditis elegans / genetics*
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Cloning, Molecular
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DNA, Complementary
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Electrophysiology
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Free Radical Scavengers / pharmacology
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G Protein-Coupled Inwardly-Rectifying Potassium Channels
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GTP-Binding Proteins / metabolism*
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Ion Channel Gating / drug effects
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Ion Channel Gating / physiology*
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Isoproterenol / pharmacology
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Membrane Potentials / drug effects
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Membrane Potentials / physiology
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Molecular Sequence Data
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Muscarinic Agonists / pharmacology
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Muscarinic Antagonists / pharmacology
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Oocytes / physiology
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Oxotremorine / pharmacology
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Pirenzepine / pharmacology
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Potassium Channels / physiology
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Potassium Channels, Inwardly Rectifying*
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Receptors, Muscarinic / genetics*
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Receptors, Muscarinic / metabolism*
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Scopolamine / pharmacology
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Sequence Homology, Amino Acid
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Serotonin / pharmacology
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Sympathomimetics / pharmacology
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Vasodilator Agents / pharmacology
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Xenopus
Substances
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DNA, Complementary
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Free Radical Scavengers
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G Protein-Coupled Inwardly-Rectifying Potassium Channels
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Muscarinic Agonists
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Muscarinic Antagonists
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Potassium Channels
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Potassium Channels, Inwardly Rectifying
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Receptors, Muscarinic
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Sympathomimetics
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Vasodilator Agents
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Serotonin
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Pirenzepine
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Oxotremorine
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Atropine
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Scopolamine
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GTP-Binding Proteins
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Isoproterenol
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Acetylcholine