Dataset Information

Proteomics studies confirm CYP450 core involvement in multiple resistance strains of Aedes aegypti from French Guiana

ABSTRACT: Insecticide resistance is a worldwide threat for vector control around the world, and Aedes aegypti , the main vector of several arboviruses, is a particular concern. To better understand the mechanisms of resistance, four isofemale strains originally from French Guiana were isolated and analysed using combined approaches. The activity of detoxification enzymes involved in insecticide resistance was assayed, and mutations located at positions 1016 and 1534 of the sodium voltage-gated channel gene, which have been associated with pyrethroid resistance in Aedes aegypti populations in Latin America, were monitored. Resistance to other insecticide families (organophosphates and carbamates) was evaluated. A large-scale proteomic analysis was performed to identify proteins involved in insecticide resistance. Our results revealed a metabolic resistance and resistance associated with a mutation of the sodium voltage-gated channel gene at position 1016. Metabolic resistance was mediated through an increase of esterase activity in most strains but also through the shifts in the abundance of several cytochrome P450 (CYP450s). Overall, resistance to deltamethrin was linked in the isofemale strains to resistance to other class of insecticides, suggesting that cross- and multiple resistance occur through selection of mechanisms of metabolic resistance. These results give some insights into resistance to deltamethrin and into multiple resistance phenomena in populations of Ae. aegypti

INSTRUMENT(S): Q Exactive

ORGANISM(S): Aedes Aegypti

TISSUE(S): Midgut

SUBMITTER: MARIETTE MATONDO

LAB HEAD: Isabelle Dusfour

PROVIDER: PXD020408 | Pride | 2020-12-09

REPOSITORIES: pride

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Dataset's files

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			Action	DRS
	20180618_Q1_TC_colQ1_58_Epelboin_IR03-NI-1.raw	Raw
	20180618_Q1_TC_colQ1_58_Epelboin_IR03-NI-2.raw	Raw
	20180618_Q1_TC_colQ1_58_Epelboin_IR03-NI-3.raw	Raw
	20180618_Q1_TC_colQ1_58_Epelboin_IR05-NI-1.raw	Raw
	20180618_Q1_TC_colQ1_58_Epelboin_IR05-NI-2.raw	Raw

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CYP450 core involvement in multiple resistance strains of Aedes aegypti from French Guiana highlighted by proteomics, molecular and biochemical studies.

Epelboin Yanouk Y Wang Lanjiao L Giai Gianetto Quentin Q Choumet Valérie V Gaborit Pascal P Issaly Jean J Guidez Amandine A Douché Thibaut T Chaze Thibault T Matondo Mariette M Dusfour Isabelle I

PloS one 20210111 1

Insecticide resistance is a worldwide threat for vector control around the world, and Aedes aegypti, the main vector of several arboviruses, is a particular concern. To better understand the mechanisms of resistance, four isofemale strains originally from French Guiana were isolated and analysed using combined approaches. The activity of detoxification enzymes involved in insecticide resistance was assayed, and mutations located at positions 1016 and 1534 of the sodium voltage-gated channel gene ...[more]

PMID: 33428654

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Project description:Aedes aegypti SP strain vs. SMK strain. Aedes aegypti is the major vector of yellow fever and dengue/dengue hemorrhagic fever. Starting with a population collected from Singapore, we established a pyrethroid-resistant A. aegypti strain (SP) and investigated three major possible mechanisms of insecticide resistance. After 10 generations of adult selection, an A. aegypti strain developed 1650-fold resistance to permethrin, which is one of the most widely used pyrethroid insecticides for mosquito control. SP larvae also developed 8790-fold resistance following selection of the adults. Prior to the selections, the frequencies of V1016G and F1534C mutations in domains II and III, respectively, of voltage-sensitive sodium channel genes (Vssc) were 0.44 and 0.56, respectively. In contrast, only G1016 alleles were present after two permethrin selections, indicating that G1016 can contribute more to the insensitivity of Vssc than C1534. In vivo metabolism studies showed that the SP strain excreted permethrin metabolites more rapidly than the susceptible SMK strain. Pretreatment with piperonyl butoxide caused strong inhibition of excretion of permethrin metabolites, suggesting that cytochrome P450 monooxygenases (P450s) play an important role in resistance development. In vitro metabolism studies also indicated an association of P450s with resistance. Microarray analysis showed that multiple P450 genes were over-expressed during the larval and adult stages in the SP strain. Following quantitative real time PCR, we focused on two P450 isoforms, CYP9M6 and CYP6BB2, and confirmed that they were capable of detoxifying permethrin to 4'-HO-permethrin. Over-expression of CYP9M6 was partially due to gene amplification. Association analysis demonstrated that CYP9M6 and CYP6BB2 complementarily conferred permethrin resistance. Two other P450s (CYP9J26 and CYP9J28), which are capable of metabolizing permethrin, were also over-expressed in the SP strain, indicating that at least four P450 isoforms are likely involved in resistance development. Our data show that it is unlikely that reduced cuticle penetration of permethrin contributes to resistance. One-color experiment with two strains (SP, SMK) and 3 developmental stages/genders (larvae, adult males, and adult females), 4 biological replicates each.