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Protein expression in female salivary glands of pyrethroid-susceptible and resistant strains of Aedes aegypti mosquitoes.

ABSTRACT: BACKGROUND:A group of insecticides called pyrethroids has been used extensively worldwide and development of pyrethroid resistance within mosquito populations, especially in Aedes aegypti, has rapidly spread through populations. In this study, SDS-PAGE, 2-DE coupled with NanoLC-MS, and bioinformatics were used to analyze the female salivary gland proteins of pyrethroid-susceptible (PMD) and pyrethroid-resistant (PMD-R and UPK-R) strains of Ae. aegypti mosquitoes for the first time. RESULTS:SDS-PAGE analysis revealed that among the three strains at least nine major proteins were detected but one protein band (20 kDa) was found only in the PMD strain. Two-dimensional gel electrophoresis analysis revealed 19 similarly expressed proteins in the salivary glands of the three strains involved in blood-feeding process, stress response, immunogenic response, and metabolic process and five additional major protein spots differentially expressed in the susceptible and resistant strains. Comparative analysis of the expression volume of each protein spot between the PMD and the PMD-R strains showed three downregulated proteins of the PMD-R mosquitoes. For UPK-R strains, six major proteins were downregulated when compared to the PMD strain. Additionally, four downregulated proteins were found in the UPK-R when compared to the PMD-R strain. These results suggest that pyrethroids might induce alteration of salivary gland proteins in resistant mosquitoes. Network analysis by STITCH database 5.0 showed that SRPN23 interacted with sodium and calcium ions, suggesting that SRPN23 might be involved in insecticide resistance. CONCLUSIONS:Information obtained from this study will be useful for further studies on the roles of differentially expressed salivary gland proteins in resistance to insecticides and viral transmission.

SUBMITTER: Mano C

PROVIDER: S-EPMC6419353 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

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Protein expression in female salivary glands of pyrethroid-susceptible and resistant strains of Aedes aegypti mosquitoes.

Mano Chonlada C Jariyapan Narissara N Sor-Suwan Sriwatapron S Roytrakul Sittiruk S Kittisenachai Suthathip S Tippawangkosol Pongsri P Somboon Pradya P

Parasites & vectors 20190314 1

<h4>Background</h4>A group of insecticides called pyrethroids has been used extensively worldwide and development of pyrethroid resistance within mosquito populations, especially in Aedes aegypti, has rapidly spread through populations. In this study, SDS-PAGE, 2-DE coupled with NanoLC-MS, and bioinformatics were used to analyze the female salivary gland proteins of pyrethroid-susceptible (PMD) and pyrethroid-resistant (PMD-R and UPK-R) strains of Ae. aegypti mosquitoes for the first time.<h4>Re ...[more]

PMID: 30871604

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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.

Dataset Information

Protein expression in female salivary glands of pyrethroid-susceptible and resistant strains of Aedes aegypti mosquitoes.

Publications

Protein expression in female salivary glands of pyrethroid-susceptible and resistant strains of Aedes aegypti mosquitoes.

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