title: Molecular characterisation of nicotinic acetylcholine receptor subunits from the cat flea, Ctenocephalides felis (Siphonaptera: Pulicidae).
abstract: As part of a program to monitor the susceptibility of cat flea populations to the insecticide imidacloprid we have examined the cat flea nicotinic acetylcholine receptor, the target site protein of the neonicotinoid group of insecticides. Seven nAChR subunits (six alpha-type and one beta-type) were identified in cat flea using a degenerate PCR-based strategy. Five of these were expressed in vitro by creating chimeras containing the N-terminal ligand-binding domain of the cat flea subunits and the C-terminal region of the Drosophila Dalpha2 (SAD) subunit. Two of the five chimeric subunits, Cfalpha1/Dalpha2 and Cfalpha3/Dalpha2, when co-expressed with rat beta2 in Drosophila S2 cells, showed high-affinity binding of both epibatidine (Kd=1.6+/-0.6 and 0.13+/-0.06nM, respectively), and imidacloprid (Ki=142+/-34 and 28.7+/-2.4nM, respectively). It is likely therefore that Cfalpha1 and Cfalpha3 contribute to nAChR populations in vivo that are sensitive to imidacloprid. The identification of cat flea nAChR subunits that have a high affinity for imidacloprid presents candidate genes in which to look for resistance-associated mutations if target-site resistance to imidacloprid arises in domestic pet flea populations.
authors.affiliation: Department of Biological and Ecological Chemistry, Rothamsted Research, Harpenden, AL5 2JQ, UK. chris.bass@bbsrc.ac.uk
authors.forename: Chris
authors.initials: C
authors.lastname: Bass
authors.forename: Stuart J
authors.initials: SJ
authors.lastname: Lansdell
authors.forename: Neil S
authors.initials: NS
authors.lastname: Millar
authors.forename: Iris
authors.initials: I
authors.lastname: Schroeder
authors.forename: Andreas
authors.initials: A
authors.lastname: Turberg
authors.forename: Linda M
authors.initials: LM
authors.lastname: Field
authors.forename: Martin S
authors.initials: MS
authors.lastname: Williamson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002415:Cats; D003001:Cloning, Molecular; D007093:Imidazoles / Q000494:pharmacology; D007305:Insecticide Resistance / Q000235:genetics; D007306:Insecticides / Q000494:pharmacology; D008969:Molecular Sequence Data; D000073943:Neonicotinoids; D009574:Nitro Compounds; D010802:Phylogeny; D021122:Protein Subunits; D011978:Receptors, Nicotinic / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D005423:Siphonaptera / Q000235:genetics / Q000378:metabolism*
chemical_list: D007093:Imidazoles; D007306:Insecticides; D000073943:Neonicotinoids; D009574:Nitro Compounds; D021122:Protein Subunits; D011978:Receptors, Nicotinic; C082359:imidacloprid