Glycogen branching enzyme: a novel deltamethrin resistance-associated gene from Culex pipiens pallens

Xu, Yang; Yang, Mingxia; Sun, Jing; Qian, Jin; Zhang, Donghui; Sun, Yan; Ma, Lei; Zhu, Changliang

doi:10.1007/s00436-008-1003-7

Cited by 10 publications

(5 citation statements)

References 32 publications

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“…Pipiens pallens . As a result, several genes associated with novel DM resistance have been isolated [17-19]. One of the highly expressed genes in the DR strain was opsin, which belongs to G protein-coupled receptors (GPCRs) [20].…”

Section: Introductionmentioning

confidence: 99%

Functional characterization of an arrestin gene on insecticide resistance of Culex pipiens pallens

Sun

Zou

et al. 2012

Parasites Vectors

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BackgroundContinuous and excessive application of insecticides has resulted in the rapid development of insecticide resistance in several mosquito species, including Culex pipiens pallens. Previous studies in our laboratory found that arrestin gene expression was higher in the deltamethrin-resistant (DR) strain than in the deltamethrin-susceptible (DS) strain of Cx. pipiens pallens. Similarly, other studies reported that arrestin was highly expressed in permethrin-resistant Cx. quinquefasciatus and in dichlorodiphenyltrichloroethane (DDT)-resistant Drosophila melanogaster.MethodsFull-length cDNAs of an arrestin gene were cloned from Cx. pipiens pallens via polymerase chain reaction (PCR) and rapid amplification of cDNA end (RACE). The mRNA levels of the arrestin gene in the whole life cycle of DR and DS strains of Cx. pipiens pallens were investigated via quantitative real-time PCR. In addition, the relationship between arrestin and deltamethrin (DM) resistance were identified using genetic overexpression strategies and arrestin RNAi in mosquito cells. Cell viability was analyzed with cholecystokinin octapeptide after DM treatment. Moreover, the mRNA levels of cytochrome P450 6A1 (CYP6A1) and opsin in the transfected cells and controls were analyzed.ResultsComplete arrestin gene sequence was cloned and expressed throughout the life cycle of Cx. pipiens pallens. Moreover, arrestin was significantly upregulated in the DR strain, compared with that in the DS strain at the egg, pupae, and adult stages. Arrestin overexpression comparably increased the mosquito cell viability, whereas arrestin knockdown by siRNA decreased mosquito cell viability with deltamethrin (DM) treatment. Meanwhile, the mRNA levels of CYP6A1 and opsin were upregulated in mosquito cells transfected with arrestin and downregulated in mosquito cells with arrestin knockdown.ConclusionThis study presented the first evidence that arrestin might be associated with insecticide resistance in Cx. pipiens pallens.

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Section: Introductionmentioning

confidence: 99%

Functional characterization of an arrestin gene on insecticide resistance of Culex pipiens pallens

Sun

Zou

et al. 2012

Parasites Vectors

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“…Here, we report the characterization of one cypermethrin resistance associated gene of C. pipiens pallens. The writer reported insecticide-resistant ribosomal protein L39 gene of C. pipiens pallens , while other researchers in the same laboratory identified several genes differentially expressed in cypermethrin-resistant strain of the mosquito previously (Chen et al 2010;Gong et al 2005a,b;He et al 2009;Hu et al 2007;Shen et al 2002;Tian et al 2001;Xu et al 2008;Yang et al 2008;Zhang et al 2011;Zhu et al 1998Zhu et al , 2000Sun et al 2006). Some of the candidate genes encode for enzymes such as chymotrypsin (Gong et al 2005a), trypsin (Shen et al 2002), and CYP450 (Gong et al 2005b), which are likely involved in more direct functions, e.g., oxidation of cypermethrin, others are likely more upstream factors, such as ribosomal protein L22 (He et al 2009), ribosomal protein S4 , and ribosomal protein S29 .…”

Section: Discussionmentioning

confidence: 95%

Cloning and overexpression of transferrin gene from cypermethrin-resistant Culex pipiens pallens

et al. 2011

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The complete sequence of transferrin has been cloned from cypermethrin-resistant strain of Culex pipiens pallens(Cr-C strain). Quantitative reverse transcription PCR analysis indicated that the transferrin transcription level was 12.6 times higher in Cr-C strain than in susceptible strain at fourth instar larvae. The transferrin expression was also found to be consistently higher throughout the life cycle of Cr-C strain. A protein of predicted size 90.8 kDa has been detected by Western blotting in transferrin-transfected mosquito C6/36 cells. These transferrin-transfected cells also showed enhanced cypermethrin resistance compared to null-transfected or plasmid vector-transfected cells as determined by methyl tritiated thymidine((3)H-TdR) incorporation. These results indicate that transferrin is expressed at higher levels in Cr-C strain and may confer some insecticide resistance in C. pipiens pallens.

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“…Similarly, searches of the ten most highly expressed genes from these previous studies with our data found that none of these previously identified genes were overexpressed in our resistant strains. Candidate genes previously associated with pyrethroid resistance in other species of mosquitoes include transferrin [ 82 ], iron responsive element binding protein [ 83 ], glycogen branching enzyme [ 84 ], chymotrypsin [ 82 ] and G-protein coupled receptor [ 85 ]. None of these genes were overexpressed in our resistant strains, leading us to conclude that they do not play a role in pyrethroid resistance in CKR or SP.…”

Section: Discussionmentioning

confidence: 99%

Transcriptomic and proteomic analysis of pyrethroid resistance in the CKR strain of Aedes aegypti

et al. 2021

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Aedes aegypti is an important vector of human viral diseases. This mosquito is distributed globally and thrives in urban environments, making it a serious risk to human health. Pyrethroid insecticides have been the mainstay for control of adult A. aegypti for decades, but resistance has evolved, making control problematic in some areas. One major mechanism of pyrethroid resistance is detoxification by cytochrome P450 monooxygenases (CYPs), commonly associated with the overexpression of one or more CYPs. Unfortunately, the molecular basis underlying this mechanism remains unknown. We used a combination of RNA-seq and proteomic analysis to evaluate the molecular basis of pyrethroid resistance in the highly resistant CKR strain of A. aegypti. The CKR strain has the resistance mechanisms from the well-studied Singapore (SP) strain introgressed into the susceptible Rockefeller (ROCK) strain genome. The RNA-seq and proteomics data were complimentary; each offering insights that the other technique did not provide. However, transcriptomic results did not quantitatively mirror results of the proteomics. There were 10 CYPs which had increased expression of both transcripts and proteins. These CYPs appeared to be largely trans-regulated, except for some CYPs for which we could not rule out gene duplication. We identified 65 genes and lncRNAs as potentially being responsible for elevating the expression of CYPs in CKR. Resistance was associated with multiple loci on chromosome 1 and at least one locus on chromosome 3. We also identified five CYPs that were overexpressed only as proteins, suggesting that stabilization of CYP proteins could be a mechanism of resistance. Future studies to increase the resolution of the resistance loci, and to examine the candidate genes and lncRNAs identified here will greatly enhance our understanding of CYP-mediated resistance in A. aegypti.

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Glycogen branching enzyme: a novel deltamethrin resistance-associated gene from Culex pipiens pallens

Cited by 10 publications

References 32 publications

Functional characterization of an arrestin gene on insecticide resistance of Culex pipiens pallens

Functional characterization of an arrestin gene on insecticide resistance of Culex pipiens pallens

Cloning and overexpression of transferrin gene from cypermethrin-resistant Culex pipiens pallens

Transcriptomic and proteomic analysis of pyrethroid resistance in the CKR strain of Aedes aegypti

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