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Functional Expression in Xenopus Oocytes of Invertebrate Ligand-Gated Ion Channels

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Neurotox ’91

Abstract

Ligand-gated ion channels are hetero- or homo-oligomeric proteins exhibiting pseudosymmetry, with polypeptide subunits arranged around a central hydrophilic pore.1 The channels directly operated by neurotransmitters belong to this class of membrane proteins in which the channel is an integral part of the receptor protein.2 Ligand-gated ion channels have been particularly well studied in mammals, and targets for several chemically distinct classes of medicinal drugs have been recognised. Investigations of invertebrate ligand-gated ion channels have revealed target sites for insecticides and anthelmintics.3 DNA cloning of polypeptide subunits of ligand-gated ion channel molecules has been achieved in the case of skeletal muscle, electric tissue4 and nervous system nicotinic acetylcholine receptors5 of vertebrates. Subunits of vertebrate GABAA,6 glycine7 and L-glutamate receptors8 have also been cloned recently, and the cells most widely used to test for functional expression of putative subunits are the oocytes of Xenopus laevis. 9 These cells permit expression in the plasma membrane of functional neurotransmitter-operated ion channels following cytoplasmic injection of crude RNA, poly-adenylated (messenger) RNA (poly (A)+ mRNA) and cDNA-derived mRNA. Functional channels of this type can also be expressed as a result of nuclear injection of cDNA and an appropriate promoter.10

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Authors and Affiliations

  1. AFRC Laboratory of Molecular Signalling, Department of Zoology, University of Cambridge, Downing Street, Cambridge, CB2 3EJ, UK

    David B. Sattelle, Howard A. Riina & Nicola M. Anthony

  2. Department of Zoology, University of Cambridge, Downing Street, Cambridge, CB2 3EJ, UK

    Sarah C. R. Lummis

  3. Laboratory of Molecular Biology, MRC Centre, Hills Road, Cambridge, CB2 2QH, UK

    John T. Fleming & John Marshall

  4. Department of Pharmacology, Yale University, New Haven, Connecticut, 06510-8066, USA

    John Marshall

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  1. David B. Sattelle
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  1. Department of Life Science, University of Nottingham, University Park, NG7 2RD, Nottingham, UK

    I. R. Duce

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Sattelle, D.B., Lummis, S.C.R., Riina, H.A., Fleming, J.T., Anthony, N.M., Marshall, J. (1992). Functional Expression in Xenopus Oocytes of Invertebrate Ligand-Gated Ion Channels. In: Duce, I.R. (eds) Neurotox ’91. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-2898-8_15

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  • DOI: https://doi.org/10.1007/978-94-011-2898-8_15

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