Lees K , Musgaard M , Suwanmanee S , Buckingham SD , Biggin P , Sattelle D .

084655 Wellcome Trust , WT084655 Wellcome Trust

	Figure 2. Binding models for the different agonists.(A) Overall structure of the homology model of RDLbd. Chains A and E are shown in cartoon representation, A in orange and E in cyan. Chains B, C and D are shown in spacefill representation in three different shades of grey. The agonist-binding site is indicated by the sidechains of R111 in chain A (light orange spacefill) and D204 in chain E (light cyan spacefill). (B–G): binding models for the docked agonists. The agonists are shown with grey carbon atoms, and relevant protein sidechains from chain A and chain E are shown with light orange and light blue carbon atoms, respectively. Only polar hydrogen atoms on agonists and sidechains are included. Potential hydrogen bonds (<3 Å) between ligand and protein are marked with black lines. (H) Superposition of the binding models in (B–F). The conformations of R111 and E204 shown are from the GABA binding model.
	Figure 3. Concentration-inhibition curves showing responses elicited by PTX to 1 mM GABA in RDL, RDLbdIV and RDLbdIV/A301S.Each data point was normalised to the amplitude of the response to 1 mM GABA and represents the mean amplitude of the responses recorded in 7–13 oocytes from at least 3 separate batches. Structures of the two compounds found in picrotoxin (PTX), picrotoxinin and picrotin, are shown.
	Figure 4. Concentration-inhibition curves showing responses elicited by the phenylpyrazoles (A) fipronil and (B) pyrafluprole to 1 mM GABA in wild-type, RDLbdIV and RDLbdIV/A301S.Each point represents the mean amplitude of the responses recorded in 2–13 oocytes from at least 3 separate batches. Structures of the two phenylpyrazoles are inserted.
	Figure 5. Effect of ivermectin on RDLbd.Concentration inhibition curves showing responses elicited by IVM to 100 µM GABA in wild-type, RDLbdIV and RDLbdIV/A301S. Each point represents the mean amplitude of the responses recorded in 2–6 oocytes from at least 3 separate batches. The structure of ivermectin is inserted.
	Figure 1. Concentration response curves for GABA and analogues mediated by variants of the RDLbd splice variant.(A) Concentration response curves for the three RDL subunits: RDLbd, RDLbdIV and RDLbdIVA301S to GABA (structure inserted). Data were normalised to 1 mM GABA. Each data point is the mean ± S.E.M from four to six oocytes from at least three separate frogs. Representative responses to 100 µM GABA are shown for each variant. Scale bars indicate 0.5 µA, 1 s. (B) Concentration response curves showing responses elicited by GABA and analogues at RDLbdIV receptors in Xenopus oocytes. GABA is the most potent agonist, and the rank order is GABA> muscimol> TACA>>β-alanine. Isoguvacine did not reach saturation. Data were normalised to the response to 1 mM GABA. Each data point represents mean±S.E.M from five to eight oocytes. (C) The chemical structures of the agonists tested in the present study: GABA (γ-amino butyric acid), TACA (trans-4-aminocrotonic acid), β-alanine, muscimol and isoguvacine.

Accardi, Nematode cys-loop GABA receptors: biological function, pharmacology and sites of action for anthelmintics. 2012, Pubmed

Accardi, Nematode cys-loop GABA receptors: biological function, pharmacology and sites of action for anthelmintics. 2012, Pubmed
Adelsberger, Activation of rat recombinant alpha(1)beta(2)gamma(2S) GABA(A) receptor by the insecticide ivermectin. 2000, Pubmed
Andreev, Multiple origins of cyclodiene insecticide resistance in Tribolium castaneum (Coleoptera: Tenebrionidae). 1999, Pubmed
Anthony, Duplication of the Rdl GABA receptor subunit gene in an insecticide-resistant aphid, Myzus persicae. 1998, Pubmed
Bass, Identification of the Rdl mutation in laboratory and field strains of the cat flea, Ctenocephalides felis (Siphonaptera: Pulicidae). 2004, Pubmed
Bergmann, A unified model of the GABA(A) receptor comprising agonist and benzodiazepine binding sites. 2013, Pubmed
Boulanger, 3- and 5-isoxazolol zwitterions: an ab initio molecular orbital study relating to GABA agonism and antagonism. 1987, Pubmed
Brehm, The structure of muscimol, a GABA analogue of restricted conformation. 1972, Pubmed
Buckingham, Insect GABA receptors: splicing, editing, and targeting by antiparasitics and insecticides. 2005, Pubmed
Eguchi, Functional characterization of Musca glutamate- and GABA-gated chloride channels expressed independently and coexpressed in Xenopus oocytes. 2006, Pubmed , Xenbase
Es-Salah, RNA editing regulates insect gamma-aminobutyric acid receptor function and insecticide sensitivity. 2008, Pubmed , Xenbase
Ffrench-Constant, Isolation of dieldrin resistance from field populations of Drosophila melanogaster (Diptera: Drosophilidae). 1990, Pubmed
Ffrench-Constant, Molecular cloning and transformation of cyclodiene resistance in Drosophila: an invertebrate gamma-aminobutyric acid subtype A receptor locus. 1991, Pubmed
Ffrench-Constant, A point mutation in a Drosophila GABA receptor confers insecticide resistance. 1993, Pubmed , Xenbase
Filippova, Evidence that the TM1-TM2 loop contributes to the rho1 GABA receptor pore. 2004, Pubmed , Xenbase
Fisher, The alpha 1 and alpha 6 subunit subtypes of the mammalian GABA(A) receptor confer distinct channel gating kinetics. 2004, Pubmed
Gielen, Benzodiazepines modulate GABAA receptors by regulating the preactivation step after GABA binding. 2012, Pubmed , Xenbase
Goujon, A new bioinformatics analysis tools framework at EMBL-EBI. 2010, Pubmed
Hibbs, Principles of activation and permeation in an anion-selective Cys-loop receptor. 2011, Pubmed
Hoopengardner, Nervous system targets of RNA editing identified by comparative genomics. 2003, Pubmed
Hosie, Alternative splicing of a Drosophila GABA receptor subunit gene identifies determinants of agonist potency. 2001, Pubmed , Xenbase
Hosie, Agonist pharmacology of two Drosophila GABA receptor splice variants. 1996, Pubmed , Xenbase
Hosie, Molecular biology of insect neuronal GABA receptors. 1997, Pubmed
Hosie, Replacement of asparagine with arginine at the extracellular end of the second transmembrane (M2) region of insect GABA receptors increases sensitivity to penicillin G. 2006, Pubmed
Jones, Splice-variant- and stage-specific RNA editing of the Drosophila GABA receptor modulates agonist potency. 2009, Pubmed , Xenbase
Kane, Drug-resistant Drosophila indicate glutamate-gated chloride channels are targets for the antiparasitics nodulisporic acid and ivermectin. 2000, Pubmed , Xenbase
King, Thermodynamics of the reduction of NADP with 2-propanol catalyzed by an NADP-dependent alcohol dehydrogenase. 2003, Pubmed
Krause, Ivermectin: a positive allosteric effector of the alpha7 neuronal nicotinic acetylcholine receptor. 1998, Pubmed , Xenbase
Krogsgaard-Larsen, GABA agonists. Development and interactions with the GABA receptor complex. 1981, Pubmed
Larkin, Clustal W and Clustal X version 2.0. 2007, Pubmed
Le Goff, Resistance to fipronil in Drosophila simulans: influence of two point mutations in the RDL GABA receptor subunit. 2005, Pubmed , Xenbase
Ludmerer, Ivermectin and nodulisporic acid receptors in Drosophila melanogaster contain both gamma-aminobutyric acid-gated Rdl and glutamate-gated GluCl alpha chloride channel subunits. 2002, Pubmed
Lummis, Two amino acid residues contribute to a cation-π binding interaction in the binding site of an insect GABA receptor. 2011, Pubmed , Xenbase
Lynagh, An improved ivermectin-activated chloride channel receptor for inhibiting electrical activity in defined neuronal populations. 2010, Pubmed
Martin, Electrophysiological effects of piperazine and diethylcarbamazine on Ascaris suum somatic muscle. 1982, Pubmed
McGonigle, Molecular characterization of agonists that bind to an insect GABA receptor. 2010, Pubmed , Xenbase
Melis, Molecular dynamics simulations of GABA binding to the GABAC receptor: the role of Arg104. 2008, Pubmed
Miller, Binding, activation and modulation of Cys-loop receptors. 2010, Pubmed
Miyazaki, DNA sequence and site of mutation of the GABA receptor of cyclodiene-resistant red flour beetle, Tribolium castaneum. 1995, Pubmed
Morris, AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility. 2009, Pubmed
Osolodkin, Molecular modeling of ligand-receptor interactions in GABA C receptor. 2009, Pubmed
Sattelle, Comparative pharmacology and computational modelling yield insights into allosteric modulation of human alpha7 nicotinic acetylcholine receptors. 2009, Pubmed , Xenbase
Shan, Ivermectin, an unconventional agonist of the glycine receptor chloride channel. 2001, Pubmed
Silver, Mechanism of action of sodium channel blocker insecticides (SCBIs) on insect sodium channels. 2010, Pubmed
Smart, Do motor-nerve terminals have gamma-aminobutyric acid receptors? 1980, Pubmed
Trapani, Propofol analogues. Synthesis, relationships between structure and affinity at GABAA receptor in rat brain, and differential electrophysiological profile at recombinant human GABAA receptors. 1998, Pubmed , Xenbase
Trott, AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. 2010, Pubmed
Wolstenholme, Recent progress in understanding the interaction between avermectins and ligand-gated ion channels: putting the pests to sleep. 2010, Pubmed
Wolstenholme, Glutamate-gated chloride channels. 2012, Pubmed