Proteomics reveals localization of cuticular proteins in Anopheles gambiae

Zhou, Yihong; Badgett, Majors J.; Orlando, Ron; Willis, Judith H.

doi:10.1016/j.ibmb.2018.09.011

Cited by 16 publications

(15 citation statements)

References 35 publications

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“…Furthermore, the role of certain enzymes ( CYP4G16 , CYP4G17 ) in enhancing the biosynthesis of epicuticular hydrocarbons and the upregulation of cuticular genes in resistant strains has been demonstrated. So far, 293 cuticle proteins (CPR) have been characterized in Anopheles gambiae [ 46 ], and at least 300 are present in the recently annotated Ae . aegypti genome assembly.…”

Section: Discussionmentioning

confidence: 99%

Permethrin resistance in Aedes aegypti: Genomic variants that confer knockdown resistance, recovery, and death

et al. 2021

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Pyrethroids are one of the few classes of insecticides available to control Aedes aegypti, the major vector of dengue, chikungunya, and Zika viruses. Unfortunately, evolving mechanisms of pyrethroid resistance in mosquito populations threaten our ability to control disease outbreaks. Two common pyrethroid resistance mechanisms occur in Ae. aegypti: 1) knockdown resistance, which involves amino acid substitutions at the pyrethroid target site—the voltage-gated sodium channel (VGSC)—and 2) enhanced metabolism by detoxification enzymes. When a heterogeneous population of mosquitoes is exposed to pyrethroids, different responses occur. During exposure, a proportion of mosquitoes exhibit immediate knockdown, whereas others are not knocked-down and are designated knockdown resistant (kdr). When these individuals are removed from the source of insecticide, the knocked-down mosquitoes can either remain in this status and lead to dead or recover within a few hours. The proportion of these phenotypic responses is dependent on the pyrethroid concentration and the genetic background of the population tested. In this study, we sequenced and performed pairwise genome comparisons between kdr, recovered, and dead phenotypes in a pyrethroid-resistant colony from Tapachula, Mexico. We identified single-nucleotide polymorphisms (SNPs) associated with each phenotype and identified genes that are likely associated with the mechanisms of pyrethroid resistance, including detoxification, the cuticle, and insecticide target sites. We identified high association between kdr and mutations at VGSC and moderate association with additional insecticide target site, detoxification, and cuticle protein coding genes. Recovery was associated with cuticle proteins, the voltage-dependent calcium channel, and a different group of detoxification genes. We provide a list of detoxification genes under directional selection in this field-resistant population. Their functional roles in pyrethroid metabolism and their potential uses as genomic markers of resistance require validation.

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Section: Discussionmentioning

confidence: 99%

Permethrin resistance in Aedes aegypti: Genomic variants that confer knockdown resistance, recovery, and death

et al. 2021

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“…Some RR-1 and RR-2 genes however had transcripts both immediately prior to and immediately after ecdysis and in different larval stages [ 54 ]. Besides chitin-binding, and possibly predominant localization within the endocuticle or exocuticle, defined functions have not yet been ascribed to an individual cuticle protein or cuticle protein family in mosquitoes [ 55 – 59 ]. The marked increase in transcripts for some cuticle proteins in all structures in BW L4 is consistent with observations in anopheline L4 subject to a short-term salinity stress [ 44 ].…”

Section: Discussionmentioning

confidence: 99%

Transcriptomic, proteomic and ultrastructural studies on salinity-tolerant Aedes aegypti in the context of rising sea levels and arboviral disease epidemiology

et al. 2021

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Background Aedes aegypti mosquito, the principal global vector of arboviral diseases, lays eggs and undergoes larval and pupal development to become adult mosquitoes in fresh water (FW). It has recently been observed to develop in coastal brackish water (BW) habitats of up to 50% sea water, and such salinity tolerance shown to be an inheritable trait. Genomics of salinity tolerance in Ae. aegypti has not been previously studied, but it is of fundamental biological interest and important for controlling arboviral diseases in the context of rising sea levels increasing coastal ground water salinity. Results BW- and FW-Ae. aegypti were compared by RNA-seq analysis on the gut, anal papillae and rest of the carcass in fourth instar larvae (L4), proteomics of cuticles shed when L4 metamorphose into pupae, and transmission electron microscopy of cuticles in L4 and adults. Genes for specific cuticle proteins, signalling proteins, moulting hormone-related proteins, membrane transporters, enzymes involved in cuticle metabolism, and cytochrome P450 showed different mRNA levels in BW and FW L4 tissues. The salinity-tolerant Ae. aegypti were also characterized by altered L4 cuticle proteomics and changes in cuticle ultrastructure of L4 and adults. Conclusions The findings provide new information on molecular and ultrastructural changes associated with salinity adaptation in FW mosquitoes. Changes in cuticles of larvae and adults of salinity-tolerant Ae. aegypti are expected to reduce the efficacy of insecticides used for controlling arboviral diseases. Expansion of coastal BW habitats and their neglect for control measures facilitates the spread of salinity-tolerant Ae. aegypti and genes for salinity tolerance. The transmission of arboviral diseases can therefore be amplified in multiple ways by salinity-tolerant Ae. aegypti and requires appropriate mitigating measures. The findings in Ae. aegypti have attendant implications for the development of salinity tolerance in other fresh water mosquito vectors and the diseases they transmit.

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“…LC-MS/MS studies to examine CPs of adult An. gambiae, including legs, have identified very few CPs restricted to only one structure [62][63][64]. Specifically for legs peptides for CPR139 (not present in our dataset) were restricted to this tissue, while CPR73 , present in our leg-specific dataset was identified in legs and eye lens protein extracts [62].…”

Section: Cuticular Proteinsmentioning

confidence: 65%

“…gambiae, including legs, have identified very few CPs restricted to only one structure [62][63][64]. Specifically for legs peptides for CPR139 (not present in our dataset) were restricted to this tissue, while CPR73 , present in our leg-specific dataset was identified in legs and eye lens protein extracts [62]. In the same work peptides for CPR115, CPR118, CPR119, and CPR146 were identified in legs but also in different adult tissues and developmental stages while CPR1, CPR116 and CPR143 were identified in adult structures other than legs [62].…”

Section: Cuticular Proteinsmentioning

confidence: 99%

Transcriptomic analysis of resistance and short-term induction response to pyrethroids, in Anopheles coluzzii legs

Kefi¹,

Charamis²,

Balabanidou³

et al. 2021

Preprint

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BackgroundInsecticide-treated bed nets and indoor residual spraying comprise the major control measures against Anopheles gambiae sl, the dominant vector in sub-Saharan Africa. The primary site of contact with insecticide is through the mosquitoes’ legs, which represents the first barrier insecticides have to bypass to reach their neuronal targets. Proteomic changes and leg cuticle modifications have been associated with insecticide resistance that may reduce the rate of penetration of insecticides. Here, we performed a multiple transcriptomic analyses focusing on An. coluzzii legs. ResultsFirstly, leg-specific enrichment analysis identified 359 genes including the pyrethroid-binder SAP2 and 2 other chemosensory proteins, along with 4 ABCG transporters previously shown to be leg enriched. Enrichment gene families included those involved in detecting chemical stimuli including gustatory and ionotropic receptors and genes implicated in hydrocarbon-synthesis. Subsequently, we compared transcript expression in the legs of a highly resistant strain (VK7-HR) to both a strain with very similar genetic background which has reverted to susceptibility after several generations without insecticide pressure (VK7-LR) and a lab susceptible population. 232 differentially expressed genes (73 up-regulated and 159 down-regulated) were identified in the resistant strain when compared to the two susceptible counterparts, indicating an over-expression of phase I detoxification enzymes and cuticular proteins, with decrease in hormone-related metabolic processes in legs from the insecticide resistant population. Finally, we analysed the short-term effect of pyrethroid exposure on An. coluzzii legs, comparing 1hour-deltamethrin-exposed (VK7-IN) to unexposed (VK7-HR) leg transcriptomes and identified 348 up-regulated genes including those encoding for GPCRs, ABC transporters, odorant-binding proteins and members of the divergent salivary gland protein family. ConclusionsThe data on An. coluzzii leg-specific transcriptome provides valuable insights into the first line of defense in pyrethroid resistant and short-term deltamethrin-exposed mosquitoes. Our results suggest that xenobiotic detoxification is likely occurring in legs, while the enrichment of sensory proteins, ABCG transporters and cuticular genes is also evident. Constitutive resistance is primarily associated with elevated levels of detoxification and cuticular genes, while short-term insecticide-induced tolerance is linked with overexpression of transporters, GPCRs and GPCR-related genes, sensory/binding and salivary gland proteins.

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Proteomics reveals localization of cuticular proteins in Anopheles gambiae

Cited by 16 publications

References 35 publications

Permethrin resistance in Aedes aegypti: Genomic variants that confer knockdown resistance, recovery, and death

Permethrin resistance in Aedes aegypti: Genomic variants that confer knockdown resistance, recovery, and death

Transcriptomic, proteomic and ultrastructural studies on salinity-tolerant Aedes aegypti in the context of rising sea levels and arboviral disease epidemiology

Transcriptomic analysis of resistance and short-term induction response to pyrethroids, in Anopheles coluzzii legs

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