Abstract
Mosquito control based on chemical insecticides is considered as an important element of the current global strategies for the control of mosquito-borne diseases. Unfortunately, the development of insecticide resistance of important vector mosquito species jeopardizes the effectiveness of insecticide-based mosquito control. In contrast to target site resistance, other mechanisms are far from being fully understood. Global protein profiles among cypermethrin-resistant, propoxur-resistant, dimethyl-dichloro-vinyl-phosphate-resistant and susceptible strain of Culex pipiens pallens were obtained and proteomic differences were evaluated by using isobaric tags for relative and absolute quantification labeling coupled with liquid chromatography/tandem mass spectrometric analysis. A susceptible strain of Culex pipiens pallens showed elevated resistance levels after 25 generations of insecticide selection, through iTRAQ data analysis detected 2,502 proteins, of which 1,513 were differentially expressed in insecticide-selected strains compared to the susceptible strain. Finally, midgut differential protein expression profiles were analyzed, and 62 proteins were selected for verification of differential expression using iTRAQ and parallel reaction monitoring strategy, respectively. iTRAQ profiles of adaptation selection to three insecticide strains combined with midgut profiles revealed that multiple insecticide resistance mechanisms operate simultaneously in resistant insects of Culex pipiens pallens. Significant molecular resources were developed for Culex pipiens pallens, potential candidates were involved in metabolic resistance and reducing penetration or sequestering insecticide. Future research that is targeted towards RNA interference of the identified metabolic targets, such as cuticular proteins, cytochrome P450s, glutathione S-transferases and ribosomal proteins proteins and biological pathways (drug metabolism-cytochrome P450, metabolism of xenobiotics by cytochrome P450, oxidative phosphorylation, ribosome) could lay the foundation for a better understanding of the genetic basis of insecticide resistance in Culex pipiens pallens.
Highlights
Mosquitoes and mosquito-borne diseases continue to pose a huge public health burden worldwide; malaria, lymphatic filariasis, dengue, chikungunya, and West Nile virus cause significant medical and economic impacts that disproportionately affect developing countries[1,2,3].Since the 1950s, chemical insecticides have been used on a massive scale to control mosquito populations
Global protein profiles were compared among a susceptible strain of Cx. pipiens pallens and strains that were cypermethrin-resistant, propoxur-resistant, and dimethyl-dichlorovinyl-phosphate-resistant after 25 generations of selection by distinct chemical insecticide families, multiple mechanisms were found to operate simultaneously in resistant mosquitoes of Cx. pipiens pallens, including mechanisms to lower penetration of or sequester the insecticide or to increase biodegradation of the insecticide via subtle alterations in either the cuticular protein levels or the activities of detoxification enzymes (P450s and glutathione S-transferases)
Vector control of mosquitoes remains crucial to reduce disease transmission, and has long been a critical part of the global strategy to manage mosquito-associated diseases; the application of chemical insecticides is the most important component in mosquitoes control[10], and characterizing the molecular mechanisms underlying resistance is a key step for improving resistance management strategies
Summary
Mosquitoes and mosquito-borne diseases continue to pose a huge public health burden worldwide; malaria, lymphatic filariasis, dengue, chikungunya, and West Nile virus cause significant medical and economic impacts that disproportionately affect developing countries[1,2,3].Since the 1950s, chemical insecticides have been used on a massive scale to control mosquito populations. Long-term intensive and widespread overuse or misuse use of insecticides has applied intense selection pressure that has led to the development and subsequent intensification of various genetically modulated resistance mechanisms in mosquitoes [4,5,6] and has resulted in a rise in mosquito-borne diseases and outbreaks of mosquito-related diseases in recent years[7,8]. With the widespread development of resistance in mosquitoes to the most commonly used insecticides, insecticide resistance is regarded as the most serious threat to the control of mosquitoes and mosquito-borne diseases[9]. Vector control of mosquitoes remains crucial to reduce disease transmission, and has long been a critical part of the global strategy to manage mosquito-associated diseases; the application of chemical insecticides is the most important component in mosquitoes control[10], and characterizing the molecular mechanisms underlying resistance is a key step for improving resistance management strategies. A common phenomenon of insecticide resistance is that multiple mechanisms operate simultaneously in resistant insects such as the house fly[28,29,30], cockroach[31], mosquito[32,33,34,35], cotton bollworm[36], and bed bug[37]
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