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Keywords:

  • Nicotinic acetylcholine receptor;
  • α7 subunit;
  • negative subsite mutations;
  • neonicotinoids;
  • imidacloprid;
  • nitenpyram;
  • (+)-epibatidine
  • The nitroguanidine insecticide imidacloprid along with a second generation of related compounds including nitenpyram, all nicotinic acetylcholine (ACh) receptor ligands, are used increasingly in many countries. Site-directed mutagenesis and heterologous expression in Xenopus laevis oocytes have been deployed to investigate mutants (G189D and G189E) of the chicken α7 homomer-forming nicotinic receptor subunit which are predicted to enhance the negative charge at the negative subsite (loop D) of the ACh binding site.

  • Xenopus oocytes expressing wild-type α7 nicotinic receptors respond to imidacloprid with rapid inward currents. Imidacloprid and nitenpyram are partial agonists, whereas ACh, (−)-nicotine and (+)-epibatidine are full agonists.

  • Compared to wild-type α7, the mutant G189D and G189E receptors are much less sensitive to the insecticides, whereas their sensitivity to (−)-nicotine, ACh and (+)-epibatidine is only slightly reduced. In contrast, G189N and G189Q mutants are sensitive not only to ACh, (−)-nicotine and (+)-epibatidine, but also to the two insecticides. Thus reduction of the insecticide-sensitivity by the mutations G189D and G189E are attributed to an increase in negativity of loop D. Desnitro-imidacloprid (DN-IMI), an imidacloprid derivative lacking the nitro group is a potent agonist on the G189D and G189E mutants suggesting an important role of loop D in nicotinic receptor interactions with the nitro group of nitroguanidine insecticides.

British Journal of Pharmacology (2000) 130, 981–986; doi:10.1038/sj.bjp.0703374