Linkage of pyrethroid insecticide resistance to a sodium channel locus in the tobacco budworm

doi:10.1016/0965-1748(93)90064-Y

Insect Biochemistry and Molecular Biology

Volume 23, Issue 7, October 1993, Pages 763-775

https://doi.org/10.1016/0965-1748(93)90064-Y Get rights and content

Abstract

Pyrethroids, with their lack of persistence and low mammalian toxicity, have been important insecticides since the early 1970s. However, heavy use has selected for resistance to pyrethroids in populations of many insects, in particular the tobacco budworm Heliothis virescens, a major cotton pest in the Americas. Several studies have identified the voltage-gated sodium channel as the principal target of pyrethroid action, and the sodium channel has been implicated in pyrethroid resistance in Musca domestica and Drosophila melanogaster. We present molecular genetic evidence that pyrethroid resistance is linked to a sodium channel locus in a strain of H. virescens. This is the first such evidence for any major agricultural pest, and is an important step towards understanding the molecular basis of resistance. This in turn will facilitate assessment, modeling, and control of resistance in pest populations, and increase our understanding of sodium channel function.

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The effects of knock-down resistance mutations and alternative splicing on voltage-gated sodium channels in Musca domestica and Drosophila melanogaster
2020, Insect Biochemistry and Molecular Biology
Citation Excerpt :
Later, ‘super-kdr (s-kdr)’ was also identified as an allelic form of kdr which can provide up to 500-fold resistance to Type-II pyrethroids such as deltamethrin (Sawicki, 1978). A number of studies associated the kdr and s-kdr phenotype in houseflies (Williamson et al., 1993; Knipple et al., 1994) and similar resistance mechanisms in other insect species (Taylor et al., 1993; Dong and Scott, 1994) with the para-type VGSC. The para-type sodium channel in housefly was fully sequenced by Williamson et al. (1996) and single nucleotide polymorphisms were identified in resistant insects.
Voltage-gated sodium channels (VGSCs) are a major target site for the action of pyrethroid insecticides and resistance to pyrethroids has been ascribed to mutations in the VGSC gene. VGSCs in insects are encoded by only one gene and their structural and functional diversity results from posttranscriptional modification, particularly, alternative splicing. Using whole cell patch clamping of neurons from pyrethroid susceptible (wild-type) and resistant strains (s-kdr) of housefly, Musca domestica, we have shown that the V₅₀ for activation and steady state inactivation of sodium currents (I_Na+) is significantly depolarised in s-kdr neurons compared with wild-type and that 10 nM deltamethrin significantly hyperpolarised both of these parameters in the neurons from susceptible but not s-kdr houseflies. Similarly, tail currents were more sensitive to deltamethrin in wild-type neurons (EC₁₅ 14.5 nM) than s-kdr (EC₁₅ 133 nM). We also found that in both strains, I_Na+ are of two types: a strongly inactivating (to 6.8% of peak) current, and a more persistent (to 17.1% of peak) current. Analysis of tail currents showed that the persistent current in both strains (wild-type EC₁₅ 5.84 nM) was more sensitive to deltamethrin than was the inactivating type (wild-type EC₁₅ 35.1 nM). It has been shown previously, that the presence of exon l in the Drosophila melanogaster VGSC gives rise to a more persistent I_Na+ than does the alternative splice variant containing exon k and we used PCR with housefly head cDNA to confirm the presence of the housefly orthologues of splice variants k and l. Their effect on deltamethrin sensitivity was determined by examining I_Na+ in Xenopus oocytes expressing either the k or l variants of the Drosophila para VGSC. Analysis of tail currents, in the presence of various concentrations of deltamethrin, showed that the l splice variant was significantly more sensitive (EC₅₀ 42 nM) than the k splice variant (EC₅₀ 866 nM). We conclude that in addition to the presence of point mutations, target site resistance to pyrethroids may involve the differential expression of splice variants.
The investigation of the pyrethroid insecticide lambda-cyhalothrin (LCT)-affected Ca 2+ homeostasis and -activated Ca 2+ -associated mitochondrial apoptotic pathway in normal human astrocytes: The evaluation of protective effects of BAPTA-AM (a selective Ca 2+ chelator)
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Citation Excerpt :
Furthermore, this study evaluated the protection of BAPTA-AM on LCT-treated cells. Several studies have identified voltage-gated sodium channel (Nav) as the principal target of pyrethroid action (Taylor et al., 1993; Liu et al., 2006). Pyrethroids have been shown to exert their toxicity by acting on Nav and causing hyperexcitability in the nervous system (Magby and Richardson, 2017).
Exposure to insecticides has been found to have deleterious effects on human health. Lambda-cyhalothrin (LCT), a mixture of isomers of cyhalothrin, is a pyrethroid insecticide routinely used in pest control programs. LCT was reported to cause neurotoxic effects in various models. However, the mechanism of underlying effect of LCT on cytotoxicity in normal human brain cells is still elusive. This study examined whether LCT affected Ca²⁺ homeostasis and Ca²⁺-related physiology in Gibco® Human Astrocytes (GHA cells), and explored whether BAPTA-AM (1,2-bis(2-aminophenoxy)ethane-N,N,N'N'-tetraacetic acid), a selective Ca²⁺ chelator, has protective effects on LCT-treated GHA cells. The data show that LCT (10–15 μM) concentration-dependently induced cytotoxicity in both GHA cells and DI TNC1 normal rat astrocytes but only induced intracellular Ca²⁺ concentration ([Ca²⁺]_i) rises in GHA cells. In terms of Ca²⁺ signaling in GHA cells, LCT-induced [Ca²⁺]_i rises were reduced by removing extracellular Ca²⁺ and were inhibited by store-operated Ca²⁺ channel modulators (2-APB, econazole or SKF96365). In Ca²⁺-free medium, pretreatment with the endoplasmic reticulum Ca²⁺ pump inhibitor thapsigargin abolished LCT-induced [Ca²⁺]_i rises. Conversely, incubation with LCT abolished thapsigargin-induced [Ca²⁺]_i rises. Regarding cytotoxicity, LCT evoked apoptosis by regulating apoptotic protein expressions (Bax, BCl-2, cleaved caspase-9/-3). This apoptotic response was significantly inhibited by prechelating cytosolic Ca²⁺ with BAPTA-AM. Together, in GHA cells, LCT induced [Ca²⁺]_i rises by inducing Ca²⁺ entry via store-operated Ca²⁺ channels and Ca²⁺ release from the endoplasmic reticulum. Moreover, BAPTA-AM has a protective effect on inhibiting LCT-activated mitochondrial apoptotic pathway. This study provided new insights into the molecular protective mechanism of LCT-induced cytotoxicity in normal human astrocytes.
Molecular characterization of a sodium channel gene in the rice leaffolder, Cnaphalocrocis medinalis (Guenée)
2013, Pesticide Biochemistry and Physiology
Citation Excerpt :
Sodium channels are targeted by pyrethroids and sodium channel blocking insecticides including indoxacarb and metaflumizone. Since the reports of tight linkage between kdr resistance and the para-like sodium channel gene in the house fly, Musca domestica, the tobacco budworm, Heliothis virescens, and the German cockroach, B. germanica [27–29], ten sodium channel mutations have been confirmed to be responsible for kdr resistance [1]. On the other hand, while development of indoxacarb resistance has been reported in several insects including M. domestica, P. xylostella, and Spodoptera litura [30–33], little is known about the relationship between sodium channel mutations and indoxacarb resistance.
Voltage-gated sodium channels are the targets of several important classes of synthetic insecticides including pyrethroids, indoxacarb and metaflumizone. The rice leaffolder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae), is an important rice insect pest throughout Southeast Asia. In this study, we report the cDNA sequence (named as CmNav) of C. medinalis sodium channel α-subunit homologous to the para gene from Drosophila melanogaster. The 5643-bp contiguous CmNav gene sequence encoded 1877 amino acid residues, which spans the 5′ UTR and covers the open reading frame from the start codon to beyond the last transmembrane segment of transmembrane domain IV (IV S6). The deduced amino acid sequence of CmNav shares 79% identity with Drosophila para. All hallmarks of sodium channel proteins are conserved in the CmNav including an overall organization of four large hydrophobic domains and the sodium channel’s DEKA selectivity filter motif. In addition, two conserved alternative splicing sites were identified in the CmNav gene, which form the optional exon B and the mutually exclusive exon K/L, respectively. Diagnostic PCR showed that the inclusion frequencies of alternative exons differed between developmental stages or adult anatomical regions.
Kdr allelic variation in a sodium channel gene from a population of South Carolina Heliothis virescens (Fabricius)
2008, Journal of Asia-Pacific Entomology
Mutations at V421M and L1029H in the hscp sodium channel gene are known to contribute to knockdown resistance (kdr) in the Woodrow, Dalzell, and PTJ strains of H. virescens (tobacco budworm) from the cotton fields of South Carolina, USA. In the IS6 region of the sodium channel gene, the frequencies of the mutant allele methionine in the Woodrow and Dalzell strains were 0.07 and 0.1, respectively. For the IIS6 region, the frequencies of the mutant allele histidine in Woodrow and Dalzell strains were 0.175 and 0.263, respectively. In the PTJ strain, the frequencies of methionine and histidine alleles were 0 and 0.1, respectively. The Hpy3 allele, which is strongly linked to the histidine mutant allele, was also found in Woodrow and Dalzell strains. In addition, we found a new allele, which is one nucleotide different from Hpy3, called Hpy3-1, and found that it is also linked to histidine.
Three mutations identified in the voltage-sensitive sodium channel α-subunit gene of permethrin-resistant human head lice reduce the permethrin sensitivity of house fly Vssc1 sodium channels expressed in Xenopus oocytes
2008, Insect Biochemistry and Molecular Biology
Point mutations in the para-orthologous sodium channel α-subunit of the head louse (M815I, T917I, and L920F) are associated with permethrin resistance and DDT resistance. These mutations were inserted in all combinations using site-directed mutagenesis at the corresponding amino acid sequence positions (M827I, T929I, and L932F) of the house fly para-orthologous voltage-sensitive sodium channel α-subunit (Vssc1^WT) gene and heterologously co-expressed with the sodium channel auxiliary subunit of house fly (Vsscβ) in Xenopus oocytes. The double mutant possessing M827I and T929I (Vssc1^MITI/Vsscβ) caused a ∼4.0 mV hyperpolarizing shift and the triple mutant, Vssc1^MITILF/Vsscβ, caused a ∼3.2 mV depolarizing shift in the voltage dependence of activation curves. Vssc1^MITI/Vsscβ, Vssc1^TILF/Vsscβ, and Vssc1^MITILF/Vsscβ caused depolarizing shifts (∼6.6, ∼7.6, and ∼8.8 mV, respectively) in the voltage dependence of steady-state inactivation curves. The M827I and L932F mutations reduced permethrin sensitivity when expressed alone but the T929I mutation, either alone or in combination, virtually abolished permethrin sensitivity. Thus, the T929I mutation is the principal cause of permethrin resistance in head lice. Comparison of the expression rates of channels containing single, double and triple mutations with that of Vssc1^WT/Vsscβ channels indicates that the M827I mutation may play a role in rescuing the decreased expression of channels containing T929I.
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2005, Comprehensive Molecular Insect Science

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^†: Present address for correspondence: Department of Entomology, University of Arizona, Tucson, AZ 85721, U.S.A.

^¶: Present address: Department of Ecology and Evolution, University of Chicago, 1103 East 57th Street, Chicago, IL 60637, U.S.A.

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Linkage of pyrethroid insecticide resistance to a sodium channel locus in the tobacco budworm

Abstract

Pestic. Biochem. Physiol.

Pestic. Biochem. Physiol.

Pestic. Biochem. Physiol.

Insect Biochem. Molec. Biol.

Pestic. Biochem. Physiol.

Pestic. Biochem. Physiol.

FEBS Lett.

Pestic. Biochem. Physiol.

Pestic. Biochem. Physiol.

Pest. Biochem. Physiol.

Cell

Trends Ecol. Evol.

Biochim. biophys. Acta

Pestic. Biochem. Physiol.

Pestic. Biochem. Physiol.

Pestic. Biochem. Physiol.

Pestic. Biochem. Physiol.

Biochem. biophys. Res. Commun.

Neuron

Biochem. biophys. Res. Commun.

Current Protocols in Molecular Biology

Probing the molecular structure of the voltage-dependent sodium channel

Ann. rev. Neurosci.

Knockdown resistance to dichlorodiphenyl-trichloroethane and pyrethroid insecticides in the napts mutant of Drosophila melanogaster is correlated with reduced neuronal sensitivity

Arch. Insect Biochem. Physiol.

Insecticide resistance: challenge to pest management and basic research

Science

Biochemical and genetic mechanisms of insecticide resistance

Improved detection of insecticide resistance through conventional and molecular techniques

Ann. rev. Ent.

Target site and enzyme changes associated with selection of subcolonies of a multiresistant house fly strain with methyl parathion or permethrin

Pestic. Biochem. Physiol.

Knockdown resistance to dichlorodiphenyl-trichloroethane and pyrethroid insecticides in the nap^ts mutant of Drosophila melanogaster is correlated with reduced neuronal sensitivity