Molecular characterization of Dalpha6 and Dalpha7 nicotinic acetylcholine receptor subunits from Drosophila: formation of a high-affinity alpha-bungarotoxin binding site revealed by expression of subunit chimeras

Journal of Neurochemistry
Stuart J Lansdell, Neil S Millar

Abstract

Nicotinic acetylcholine receptors (nAChRs) mediate fast synaptic transmission in the insect brain and are target sites for neonicotinoid insecticides. Seven nAChR subunits (four alpha-type and three beta-type) have been cloned previously from Drosophila melanogaster, the model insect system and characterized by heterologous expression. Recently, three further putative nAChR alpha subunits (Dalpha5, Dalpha6 and Dalpha7) with sequence similarity to the vertebrate alpha7 subunit have been identified from Drosophila genome sequence data but there have been no reports, as yet, of their characterization by heterologous expression. In the present study, we report the first isolation of a full-length Dalpha7 cDNA and the independent molecular cloning of Dalpha6. Binding of nicotinic radioligands was not detected to full-length Dalpha6 or Dalpha7 subunits when expressed alone or when or co-expressed with other nAChR subunits in Drosophila or mammalian cell lines, but specific cell-surface binding of [(125)I]alpha-bungarotoxin (K(d) = 0.68 +/- 0.22 nm) and [(3)H]methyllycaconitine (K(d) = 0.27 +/- 0.06 nm) was detected after expression of a subunit chimera containing the ligand-binding domains of Dalpha6 fused to the C-terminal domain of...Continue Reading

Citations

Feb 3, 2006·Invertebrate Neuroscience : in·James E C JepsonDavid B Sattelle
Jan 12, 2007·Invertebrate Neuroscience : in·Neil S Millar, Ian Denholm
Mar 23, 2005·The Journal of Biological Chemistry·Peipei WuThomas Schmidt-Glenewinkel
Mar 17, 2012·Insect Biochemistry and Molecular Biology·Monique Mathé-AllainmatSteeve H Thany
Dec 13, 2006·Trends in Pharmacological Sciences·Steeve H ThanyBruno Lapied
Jul 22, 2010·Journal of Veterinary Pharmacology and Therapeutics·D T VoR J Martin
Jun 4, 2011·Current Biology : CB·Susanna CampesanFlaviano Giorgini
Aug 21, 2010·Toxicon : Official Journal of the International Society on Toxinology·Monica MejiaTanja A Godenschwege
Aug 20, 2017·Journal of Pesticide Science·Andrew J CrossthwaitePeter Maienfisch
Aug 22, 2007·The Journal of Immunology : Official Journal of the American Association of Immunologists·Seddigheh Razani-BoroujerdiMohan L Sopori
May 28, 2020·Frontiers in Physiology·Alison CartereauSteeve H Thany
Jun 3, 2021·Proceedings of the National Academy of Sciences of the United States of America·Justin S RosenthalQuan Yuan

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