A point mutation in a Drosophila GABA receptor confers insecticide resistance

ffrench-Constant, Richard H.; Rocheleau, Thomas A.; Steichen, J.C.; Chalmers, Alison E.

doi:10.1038/363449a0

Cited by 509 publications

(326 citation statements)

References 26 publications

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“…Resistance to dieldrin involves a subunit of the insect gamma aminobutyric acid (GABA) receptor, The encoded Rdl subunit assembles with other GABA receptor subunits to form the target site of the cyclodiene insecticides [23]. Dieldrin resistance is associated with the replacement of a single amino acid (alanine at position 302) in the Drosophila melanogaster Rdl allele [24]. A homologous mutation has also been indicated to confer dieldrin resistance in a wide variety of insect species such as A. aegypti , Drosophila simulans, Musca demestica, Lucilia cuprina, Blattella germanica , Tribolium castaneum , Hypothenemus hampei, Bemisia , and Myzus persicae .…”

Section: Introductionmentioning

confidence: 99%

Existence of the rdl mutant alleles among the anopheles malaria vector in Indonesia

Asih

Syahrani

Rozi

et al. 2012

Malar J

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BackgroundThe gamma-aminobutyric acid (GABA) receptor-chloride channel complex is known to be the target site of dieldrin, a cyclodiene insecticide. GABA-receptors, with a naturally occurring amino acid substitution, A302S/G in the putative ion-channel lining region, confer resistance to cyclodiene insecticides that includes aldrin, chlordane, dieldrin, heptachlor, endrin and endosulphan.MethodsA total of 154 mosquito samples from 10 provinces of malaria-endemic areas across Indonesia (Aceh, North Sumatra, Bangka Belitung, Lampung, Central Java, East Nusa Tenggara, West Nusa Tenggara, West Sulawesi, Molucca and North Molucca) were obtained and identified by species, using morphological characteristic. The DNA was individually extracted using chelex-ion exchanger and the DNA obtained was used for analyses using sequencing method.ResultsMolecular analysis indicated 11% of the total 154 Anopheles samples examined, carried Rdl mutant alleles. All of the alleles were found in homozygous form. Rdl 302S allele was observed in Anopheles vagus (from Central Java, Lampung, and West Nusa Tenggara), Anopheles aconitus (from Central Java), Anopheles barbirostris (from Central Java and Lampung), Anopheles sundaicus (from North Sumatra and Lampung), Anopheles nigerrimus (from North Sumatra), whereas the 302 G allele was only found in Anopheles farauti from Molucca.ConclusionThe existence of the Rdl mutant allele indicates that, either insecticide pressure on the Anopheles population in these areas might still be ongoing (though not directly associated with the malaria control programme) or that the mutant form of the Rdl allele is relatively stable in the absence of insecticide. Nonetheless, the finding suggests that integrated pest management is warranted in malaria-endemic areas where insecticides are widely used for other purposes.

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Section: Introductionmentioning

confidence: 99%

Existence of the rdl mutant alleles among the anopheles malaria vector in Indonesia

Asih

Syahrani

Rozi

et al. 2012

Malar J

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“…The first GABA A receptor mutant was isolated from pesticide-resistant Drosophila and the locus was named Resistant to dieldrin (Rdl) 19,20 . Similar to mammalian GABA A receptors, RDL channels mediate fast inhibitory neurotransmission 21 and are expressed in the CNS 22 .…”

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Modulation of GABAA receptor desensitization uncouples sleep onset and maintenance in Drosophila

et al. 2008

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Many lines of evidence indicate that GABA and GABA A receptors make important contributions to human sleep regulation. Pharmacological manipulation of these receptors has differential effects on sleep onset and sleep maintenance insomnia. Here we show that sleep is regulated by GABA in Drosophila and that a mutant GABA A receptor, Rdl A302S , specifically decreases sleep latency. The drug carbamazepine (CBZ) has the opposite effect on sleep; it increases sleep latency as well as decreasing sleep. Behavioral and physiological experiments indicated that Rdl A302S mutant flies are resistant to the effects of CBZ on sleep latency and that mutant RDL A302S channels are resistant to the effects of CBZ on desensitization, respectively. These results suggest that this biophysical property of the channel, specifically channel desensitization, underlies the regulation of sleep latency in flies. These experiments uncouple the regulation of sleep latency from that of sleep duration and suggest that the kinetics of GABA A receptor signaling dictate sleep latency.Insomnia is the most common sleep problem and affects approximately one third of the adult American population 1 . Patients with insomnia are generally subdivided into three categories: sleep onset insomnia, sleep maintenance insomnia and terminal insomnia (early-morning awakening coupled with an inability to return to sleep) 2 . The biological basis for these insomnia classifications remains unknown. Nonetheless, a single class of drugs, agonistic modulators of GABA A receptors, effectively ameliorates these diverse symptoms 2,3 . GABA A receptors are a family of pentameric ligand-gated Cl -channels 4 and are a major source of inhibitory currents throughout the CNS 5,6 . These receptors are also an important target for pharmacologic treatment of many other neurological disorders in addition to sleep 7 .The fruit fly Drosophila melanogaster is an ideal model for dissecting the relationships between molecules and behaviors, as well as between different sleep states 8,9 . As in mammals, it has been shown that the sleep-like state of Drosophila is associated with reduced sensory responsiveness and reduced brain activity 10,11 , and is subject to both circadian and homeostatic regulation 12,13 . Researchers have also identified a number of genes 14,15 , Notably, the mutation that causes the insecticide resistance phenotype (A302S) 20 specifically decreases the rate of RDL desensitization with little or no effect on other channel properties 23 . As a consequence, the mutant receptor has a longer single channel open duration and, therefore, increased channel current, at least under certain conditions (see below). Because of these characteristics, and because this mutation does not have obvious effects on health or viability, we decided to establish the importance of GABAergic transmission to sleep in flies and to examine the effects of the Rdl mutation.Interestingly, flies with this mutant GABA A receptor subunit slept more, primarily because of decreased sleep lat...

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“…A point mutation in the chloride channel pore-lining domain, replacing alanine 301 with serine, was present in all resistant D. melanogaster strains (5). This mutation provided 4,000-fold resistance when homozygous and lower levels of resistance in heterozygotes (2,6). The homologous mutation was subsequently found in a large number of cyclodiene-resistant species from many insect orders (7)(8)(9), as well as a glycine replacement at the homologous site in some resistant strains of Drosophila simulans and other species (5,10,11).…”

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confidence: 99%

“…The role of Ser 301 in resistance is well established (5,6). However, to distinguish it as the causal mutation for the phenotypes assayed, transgenic lines containing a 55.7-kb Rdl-only duplication were generated.…”

mentioning

confidence: 99%

Gene duplication in the major insecticide target site,Rdl, inDrosophila melanogaster

Remnant

Good

Schmidt

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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The Resistance to Dieldrin gene, Rdl, encodes a GABA-gated chloride channel subunit that is targeted by cyclodiene and phenylpyrazole insecticides. The gene was first characterized in Drosophila melanogaster by genetic mapping of resistance to the cyclodiene dieldrin. The 4,000-fold resistance observed was due to a single amino acid replacement, Ala 301 to Ser. The equivalent change was subsequently identified in Rdl orthologs of a large range of resistant insect species. Here, we report identification of a duplication at the Rdl locus in D. melanogaster. The 113-kb duplication contains one WT copy of Rdl and a second copy with two point mutations: an Ala 301 to Ser resistance mutation and Met 360 to Ile replacement. Individuals with this duplication exhibit intermediate dieldrin resistance compared with single copy Ser 301 homozygotes, reduced temperature sensitivity, and altered RNA editing associated with the resistant allele. Ectopic recombination between Roo transposable elements is involved in generating this genomic rearrangement. The duplication phenotypes were confirmed by construction of a transgenic, artificial duplication integrating the 55.7-kb Rdl locus with a Ser 301 change into an Ala 301 background. Gene duplications can contribute significantly to the evolution of insecticide resistance, most commonly by increasing the amount of gene product produced. Here however, duplication of the Rdl target site creates permanent heterozygosity, providing unique potential for adaptive mutations to accrue in one copy, without abolishing the endogenous role of an essential gene.T he single point mutation in the Resistance to dieldrin (Rdl) gene represents one of the most significant cases of target site resistance to an insecticide yet observed. Cyclodiene resistance was reported in 62% of insecticide resistant species in the 1980s, following widespread use of cyclodiene insecticides, including dieldrin, which started in the 1950s (1). The nature of the genetic target, Rdl, was discovered after dieldrin was discontinued because of the widespread evolution of resistance in many species. Rdl was first discovered in Drosophila melanogaster using a positional cloning approach. High homology to human GABA receptors confirmed it was the first insect ligand-gated chloride channel subunit identified (2-4). A point mutation in the chloride channel pore-lining domain, replacing alanine 301 with serine, was present in all resistant D. melanogaster strains (5). This mutation provided 4,000-fold resistance when homozygous and lower levels of resistance in heterozygotes (2, 6). The homologous mutation was subsequently found in a large number of cyclodiene-resistant species from many insect orders (7-9), as well as a glycine replacement at the homologous site in some resistant strains of Drosophila simulans and other species (5, 10, 11).Characterization of deficiency lines and inversions in D. melanogaster showed that Rdl is an essential gene (3). Thus, the Ala 301 to Ser or Gly mutation in Rdl exhibits unique properties, p...

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A point mutation in a Drosophila GABA receptor confers insecticide resistance

Cited by 509 publications

References 26 publications

Existence of the rdl mutant alleles among the anopheles malaria vector in Indonesia

Existence of the rdl mutant alleles among the anopheles malaria vector in Indonesia

Modulation of GABAA receptor desensitization uncouples sleep onset and maintenance in Drosophila

Gene duplication in the major insecticide target site,Rdl, inDrosophila melanogaster

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