Identification of proteins associated with pyrethroid resistance by iTRAQ-based quantitative proteomic analysis in Culex pipiens pallens

Abstract

BackgroundMosquito control based on chemical insecticides is considered as an important element in the current global strategies for the control of mosquito-borne diseases. Unfortunately, the development of pyrethroid resistance in important vector mosquito species jeopardizes the effectiveness of insecticide-based mosquito control. To date, the mechanisms of pyrethroid resistance are still unclear. Recent advances in proteomic techniques can facilitate to identify pyrethroid resistance-associated proteins at a large-scale for improving our understanding of resistance mechanisms, and more importantly, for seeking some genetic markers used for monitoring and predicting the development of resistance.MethodsWe performed a quantitative proteomic analysis between a deltamethrin-susceptible strain and a deltamethrin-resistant strain of laboratory population of Culex pipiens pallens using isobaric tags for relative and absolute quantitation (iTRAQ) analysis. Gene Ontology (GO) analysis was used to find the relative processes that these differentially expressed proteins were involved in. One differentially expressed protein was chosen to confirm by Western blot in the laboratory and field populations of Cx. pipiens pallens.ResultsWe identified 30 differentially expressed proteins assigned into 10 different categories, including oxidoreductase activity, transporter activity, catalytic activity, structural constituent of cuticle and hypothetical proteins. GO analysis revealed that 25 proteins were sub-categorized into 35 hierarchically-structured GO classifications. Western blot results showed that CYP6AA9 as one of the up-regulated proteins was confirmed to be overexpressed in the deltamethrin-resistant strains compared with the deltamethrin-susceptible strains both in the laboratory and field populations.ConclusionsThis is the first study to use modern proteomic tools for identifying pyrethroid resistance-related proteins in Cx. pipiens. The present study brought to light many proteins that were not previously thought to be associated with pyrethroid resistance, which further expands our understanding of pyrethroid resistance mechanisms. CYP6AA9 was overexpressed in the deltamethrin-resistant strains, indicating that CYP6AA9 may be involved in pyrethroid resistance and may be used as a potential genetic marker to monitor and predict the pyrethroid resistance level of field populations.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-015-0709-5) contains supplementary material, which is available to authorized users.

Proteomic analysis of Bayvarol® resistance mechanisms in the honey bee parasite Varroa destructor

“…This protein is possibly a cuticular protein as it shows homology to gasp from Drosophila melanogaster, a chitin binding cuticle constituent. Previous research has demonstrated that a down-regulation of some cuticle proteins alongside an up-regulation of others does occur, suggesting that an alteration in cuticle composition may occur, leading to decreased rates of entry of the chemical Wang et al, 2015). However, no cuticle proteins were observed here to be of higher abundance in resistant mites which may indicate that the cuticular changes in resistant mites involve the removal of certain proteins without their replacement.…”

“…One possible means by which organisms are thought to become resistant is by alteration of the route of entry of the chemical such as a thickening or hardening of the cuticle or other epithelial barriers (Hemingway et al, 2004). A number of cuticular proteins were present in lower amounts in resistant mites (Figure 2(b)), which partly contradicts the trend observed in previous reports (Puinean et al, 2010;Silva, Jander, Samaniego, Ramsey, & Figueroa, 2012;Vontas et al, 2005;Wang et al, 2015). The protein that showed the lowest abundance in the resistant mites, exclusively expressed in the profile of sensitive mites, was a conserved hypothetical protein (Table 3).…”

“…The cross resistance of V. destructor mites to unrelated chemicals supports the theory that metabolic resistance could play a role in the development of resistant populations (Martin, 2004;Vontas et al, 2005). Resistance due to changes in the rate or efficacy of metabolism is thought to be mainly attributed to up-regulation of metabolic enzymes such as esterases, gluthathione-S transferases, and P450 mono-oxygenases (Puinean et al, 2010;Wang et al, 2015). Notes: Relative fold changes in expression of proteins in the profile of Bayvarol ® resistant mites, and the number of matched peptides, sequence coverage, PEP and mean LFQ intensities.…”

Preliminary characterization of putative structural cuticular proteins in the malaria vector Anopheles sinensis

“…aegypti , C. quinquefasciatus and An. gambiae , but fewer than in D. melanogaster . The phenomenon of many genes physically clustered on the same scaffold was not observed in the An.…”

Caste‐ and pesticide‐specific effects of neonicotinoid pesticide exposure on gene expression in bumblebees

“…Some of the genes affected by clothianidin exposure in our study have also been affected by neonicotinoids or other pesticides in other studies and species. These include muscular genes such as troponin and calponin (Kimura‐Kuroda et al, ; Lewis, Szilagyi, Gehman, Dennis, & Jackson, ; Wang et al, ) and metabolic enzymes such as glucose dehydrogenase (Christen et al, ) and hexosaminidase D (Qi et al, ; Yang, Liu, Liu, Qu, & Qian, ). At a different level, cellular transport genes such as the ABC transport family (Dermauw & Van Leeuwen, ), one member of which was differentially expressed in our study, have been suggested to provide tolerance of neonicotinoids, such as imidacloprid, acetamiprid and thiacloprid, with greater mortality identified for neonicotinoid‐exposed honeybee larvae treated with an ABC inhibitor (Hawthorne & Dively, ).…”