title: Structure-activity relationships of acetylcholine derivatives with Lucilia cuprina nicotinic acetylcholine receptor α1 and α2 subunits in chicken β2 subunit hybrid receptors in comparison with chicken nicotinic acetylcholine receptor α4/β2.
abstract: Insect nicotinic acetylcholine (ACh) receptors (nAChRs) are the targets of several insecticide classes. In the present study, we report the gene identification and cloning of nAChR α1 and α2 subunits (Lcα1 and Lcα2) from the sheep blowfly Lucilia cuprina. Xenopus oocytes voltage clamp experiments as hybrids with the chicken β2 nAChR (Ggβ2) subunit resulted in ACh-gated ion channels with distinct dose-response curves for Lcα1/Ggβ2 (effective concentration 50% [EC50 ] = 80 nM; nH  = 1.05), and Lcα2/Ggβ2 (EC50  = 5.37 μM, nH  = 1.46). The neonicotinoid imidacloprid was a potent agonist for the α-bungarotoxin-sensitive Lcα1/Ggβ2 (EC50 ∼ 20 nM), while the α-bungarotoxin-resistant Lcα2/Ggβ2 showed a 30-fold lower sensitivity to this insecticide (EC50  = 0.62 μM). Thirteen close derivatives of ACh were analysed in EC50 , Hill coefficient and maximum current (relative to ACh) determinations for Lcα1/Ggβ2 and Lcα2/Ggβ2 and the chicken Ggα4/Ggβ2 nAChRs, and comparisons relative to ACh allowed the definition of novel structure-activity and structure-selectivity relationships. In the case of N-ethyl-acetylcholine, the EC50 of the chicken Ggα4/Ggβ2 rose by a factor of 1000, while for both Lcα1/Ggβ2 and Lcα2/Ggβ2, potency remained unchanged. Further derivatives with insect nAChR selectivity potential were acetyl-α-methylcholine and trimethyl-(3-methoxy-3-oxopropyl)ammonium, followed by acetylhomocholine and trimethyl-(4-oxopentyl) ammonium. Our results may provide guidance for the identification or design of insect-specific nAChR agonists using structure-based or in silico methods.
authors: MSD Animal Health Innovation GmbH, Schwabenheim, Germany.
authors: H
authors: H
authors: Dederer
authors: M
authors: M
authors: Berger
authors: T
authors: T
authors: Meyer
authors: M
authors: M
authors: Werr
authors: T
authors: T
authors: Ilg
mesh_terms: D000109:Acetylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002038:Bungarotoxins / Q000494:pharmacology; D002645:Chickens; D003001:Cloning, Molecular; D004175:Diptera; D004305:Dose-Response Relationship, Drug; D005260:Female; D007093:Imidazoles / Q000494:pharmacology; D019476:Insect Proteins / Q000235:genetics; D007306:Insecticides / Q000737:chemistry / Q000494:pharmacology; D008969:Molecular Sequence Data; D000073943:Neonicotinoids; D018722:Nicotinic Agonists / Q000737:chemistry* / Q000494:pharmacology*; D009574:Nitro Compounds / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects; D010802:Phylogeny; D021122:Protein Subunits; D011978:Receptors, Nicotinic / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics; D013329:Structure-Activity Relationship
chemical_list: D000109:Acetylcholine; D002038:Bungarotoxins; D007093:Imidazoles; D019476:Insect Proteins; D007306:Insecticides; D000073943:Neonicotinoids; D018722:Nicotinic Agonists; D009574:Nitro Compounds; D021122:Protein Subunits; D011978:Receptors, Nicotinic; D011994:Recombinant Proteins; C403531:nicotinic receptor alpha4beta2; C023993:acetylhomocholine; C082359:imidacloprid