Comparative proteomics reveals mechanisms that underlie insecticide resistance in Culex pipiens pallens Coquillett

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.

Quantitative proteomics analysis of permethrin and temephos-resistant Ae. aegypti revealed diverse differentially expressed proteins associated with insecticide resistance from Penang Island, Malaysia

“…pipiens pallens . Furthermore, Zhang et al [ 37 ] revealed the differentially expressed proteins in insecticide resistance Cx . pipiens pallens coquillett in Shandong Province, China, similar to differentially expressed proteins found in our study, i.e., cytochromes-related, motor-related, ribosomal, GST, cuticle, and HSPs ( S1 File ).…”

“…Furthermore, cuticle proteins and their assembly pattern influenced the integrity of the cuticle [ 37 , 51 ]. This study identified several insect cuticle proteins differentially expressed in ANOVA and t-test analyses, including pupal cuticle protein, putative (A0A1S4FZ80), cuticle protein 2 (Q17LN8), cuticle protein 5 (Q16I56) in the permethrin and temephos resistant adults and larvae Ae .…”

Insecticide Resistance in Vectors of Medically Important Parasitic Infections

“…To observe changes at the RNA level, molecular techniques such as real-time polymerase chain reaction (RT-PCR), differential display reverse transcription PCR, northern blot and microarrays may be used. For protein estimation enzyme assays, various techniques such as enzyme linked enzyme-linked immunosorbent assay (ELISA), western blot, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) may be used [65,66]. In addition to this, to study the mechanism of resistance at the molecular level through proteomics, the identification of resistance-related proteins with their expression profiling can provide knowledge of the activity of proteins related to insecticide resistance in insects [67].…”

“…quinquefasciatus from Zanzibar was determined [86]. These assays provided evidence for insecticide resistance caused due to target site mutations and metabolic alterations at the genetic level [87].…”

Isobaric tags for relative and absolute quantification-based proteomic analysis of host-pathogen protein interactions in the midgut of Aedes albopictus during dengue virus infection

“…Isobaric tags for relative and absolute quantification (iTRAQ), as a highly advanced proteomic platform, can identify differentially regulated proteins more accurately and provide reliable quantitative information. This method labels protein samples with unique isotopes to enable robust comparisons of different samples in a mixture and to study changes in protein levels under different experimental conditions ( De Mandal et al, 2020 ; Zhang et al, 2021 ). iTRAQ has been applied successfully to screen differentially expressed proteins (DEPs) after pathogen infection in mosquitos to understand host immune defense mechanisms ( Baldridge et al, 2017 ; Xin et al, 2017 ).…”