title: Identification of proteins associated with pyrethroid resistance by iTRAQ-based quantitative proteomic analysis in Culex pipiens pallens.
abstract: BACKGROUND
  Mosquito 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.
  
  
  METHODS
  We 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.
  
  
  RESULTS
  We 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.
  
  
  CONCLUSIONS
  This 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.
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. wwjsix@163.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. wwjsix@163.com.|Department of Pathogen Biology, Hebei Medical University, Shijiazhuang, China. wwjsix@163.com.
authors.forename: Weijie
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. lvyuan19870212@163.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. lvyuan19870212@163.com.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lv
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. fangfujinnjmu@gmail.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. fangfujinnjmu@gmail.com.
authors.forename: Fujin
authors.initials: F
authors.lastname: Fang
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. hongshanchao1@163.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. hongshanchao1@163.com.
authors.forename: Shanchao
authors.initials: S
authors.lastname: Hong
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. guoqin2782@163.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. guoqin2782@163.com.
authors.forename: Qin
authors.initials: Q
authors.lastname: Guo
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. 1824361543@qq.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. 1824361543@qq.com.
authors.forename: Shengli
authors.initials: S
authors.lastname: Hu
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. feifei19870603@126.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. feifei19870603@126.com.
authors.forename: Feifei
authors.initials: F
authors.lastname: Zou
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. selina-007-520@163.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. selina-007-520@163.com.
authors.forename: Linna
authors.initials: L
authors.lastname: Shi
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. 463226459@qq.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. 463226459@qq.com.
authors.forename: Zhentao
authors.initials: Z
authors.lastname: Lei
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. 635398076@qq.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. 635398076@qq.com.
authors.forename: Kai
authors.initials: K
authors.lastname: Ma
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. zdd1209love@163.com.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. zdd1209love@163.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Zhou
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. iota@njmu.edu.cn.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. iota@njmu.edu.cn.
authors.forename: Donghui
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. sunyan@njmu.edu.cn.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. sunyan@njmu.edu.cn.
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. malei@njmu.edu.cn.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. malei@njmu.edu.cn.
authors.forename: Lei
authors.initials: L
authors.lastname: Ma
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. shenbo@njmu.edu.cn.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. shenbo@njmu.edu.cn.
authors.forename: Bo
authors.initials: B
authors.lastname: Shen
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, China. clzhu@njmu.edu.cn.|Jiangsu Province Key Laboratory of Modern Pathogen Biology, Nanjing Medical University, Nanjing, China. clzhu@njmu.edu.cn.
authors.forename: Changliang
authors.initials: C
authors.lastname: Zhu
mesh_terms: D000818:Animals; D003465:Culex / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007305:Insecticide Resistance; D007306:Insecticides / Q000494:pharmacology*; D009032:Mosquito Control; D040901:Proteomics; D011722:Pyrethrins / Q000494:pharmacology*
chemical_list: D019476:Insect Proteins; D007306:Insecticides; D011722:Pyrethrins