Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Culex quinquefasciatus Jpal-per strain vs. Ogasawara strain

ABSTRACT: A pyrethroid-resistant strain of Culex quinquefasciatus, JPal-per, exhibits 2500-fold greater larval resistance to permethrin than the insecticide-susceptible strain Ogasawara. An increased microsome monooxygenase metabolism is involved in the resistance mechanism. Microarray analysis revealed altered expressions of cytochrome P450 genes in the fourth instar larvae of JPal-per compared to those in OGS. An oligo DNA array was designed for 62 different cDNA segments encoding unique P450 isoforms of the Cx. pipiens complex. Other probes for non-P450 Cx. pipiens complex genes were also mounted on the array. Two strains with two color experiment, total 10 replicates, 5 biological replicates (each strain), 5 dye-swap experiments.

ORGANISM(S): Culex quinquefasciatus

SUBMITTER: Osamu Komagata

PROVIDER: E-GEOD-17326 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Overexpression of cytochrome P450 genes in pyrethroid-resistant Culex quinquefasciatus.

Komagata Osamu O Kasai Shinji S Tomita Takashi T

Insect biochemistry and molecular biology 20100118 2

JPal-per strain of Culex quinquefasciatus exhibits extremely high resistance against pyrethroids in larvae, though the resistance is greatly lower in adults. Increased microsome monooxygenase metabolism is one of the major factors of the larval resistance in this strain. We cloned 46 novel cytochrome P450 cDNAs from JPal-per strain. An oligonucleotide microarray was designed for the novel 46 genes plus 16 previously reported P450 genes along with other non-P450 gene probes. Of these, five P450 g ...[more]

PMID: 20080182

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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.

Dataset Information

Culex quinquefasciatus Jpal-per strain vs. Ogasawara strain

Publications

Overexpression of cytochrome P450 genes in pyrethroid-resistant Culex quinquefasciatus.

Similar Datasets

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