Dataset Information

Anopheles culicifacies species A and species B Midgut Proteome

ABSTRACT: Background: Mosquito midgut is an important target for the host parasite interaction studies as it plays a major role in parasite growth and maturation and vector susceptibility. Proteomic approaches coupled with bioinformatics analysis have been used to study expression of functional proteins/enzymes of An. culicifacies susceptible and refractory species midgut in order to understand the mechanism of refractoriness that may help in contributing to unravel the host pathogen interactions. Methodology/Principle findings: In the present study proteomics approaches namely in solution and in gel digestion strategies followed by LC/MS/MS analysis were used. Further bioinformatics analysis were carried out to find out the functional annotated proteins, biological process, molecular function and their sub-cellular location using Gene ontology, SMART analysis, CELLO etc. In solution and in gel approach coupled with LC-MS/MS identified a total of 91 proteins in susceptible species and 69 proteins in refractory species. Comparative analysis between susceptible and refractory species of An. culicifacies indicated that mainly proteins involved in proteolysis mechanism, catalytic activity, peptidases activity and immune related proteins were found to be dominating in refractory as compared to susceptible species. Conclusion/Significance: Based on the present data a significant increase in number of proteins in midgut of refractory An. culicifacies species B were found that may conclude that these proteins may be responsible for the inhibiting parasite growth and linked to the melanization of oocysts or parasite lysis mechanisms in natural populations of refractory mosquito. Hence the progress of our studies at protein level suggests that these identified annotated putative proteins/enzymes may help to explore natural vector-parasite systems and reveal valuable insights into the mechanism of refractoriness which in turn further useful for bringing of novel strategies for control of malaria.

INSTRUMENT(S): Thermo Scientific instrument model

ORGANISM(S): Anopheles Culicifacies A Anopheles Culicifacies B

TISSUE(S): Midgut

SUBMITTER: sonam vijay

LAB HEAD: Dr Arun Sharma

PROVIDER: PXD002828 | Pride | 2018-10-24

REPOSITORIES: Pride

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Proteome-wide analysis of Anopheles culicifacies mosquito midgut: new insights into the mechanism of refractoriness.

Vijay Sonam S Rawal Ritu R Kadian Kavita K Singh Jagbir J Adak Tridibesh T Sharma Arun A

BMC genomics 20180508 1

<h4>Background</h4>Midgut invasion, a major bottleneck for malaria parasites transmission is considered as a potential target for vector-parasite interaction studies. New intervention strategies are required to explore the midgut proteins and their potential role in refractoriness for malaria control in Anopheles mosquitoes. To better understand the midgut functional proteins of An. culicifacies susceptible and refractory species, proteomic approaches coupled with bioinformatics analysis is an e ...[more]

PMID: 29739330

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Project description:Background: Mosquito midgut is an important target for the host parasite interaction studies as this major immune organ plays a role in parasite growth, maturation and vector competence. Most important rural malaria vector of India is Proteomic approaches coupled with bioinformatics analysis have been used to study expression of functional proteins/enzymes of An. culicifacies susceptible and refractory species midgut in order to understand the mechanism of refractoriness that may help in contributing to unravel the host pathogen interactions Methodology/Principle findings: Basic shot gun proteomics approaches along with iTRAQ based differential studies followed by LC/MS/MS were employed to study the proteome of sugar fed An. culicifacies midgut. Further bioinformatics analysis was carried out using Gene ontology, SMART analysis, CELLO algorithm etc. Here proteins identified with iTRAQ Ratio more than 1.5 was considered up regulated and less than 0.67 were considered down regulated. Ratio from 1.5 – 0.67 were considered as moderate to no change. Using these criteria we observed total 80 proteins in An. culicifacies midgut and when compared susceptible species against refractory species, total 30 proteins were found to be downregulated. 3 proteins were found to be upregulated in refractory species Conclusion/Significance: Based on the present data a significant increase in number of proteins in midgut of refractory An. culicifacies species B were found that may conclude that these proteins may be responsible for the inhibiting parasite growth and linked to the melanization of oocysts or parasite lysis mechanisms in natural populations of refractory mosquito. Hence the progress of our studies at protein level suggests that these identified annotated putative proteins/enzymes may help to explore natural vector-parasite systems and reveal valuable insights into the mechanism of refractoriness which in turn further useful for bringing of novel strategies for control of malaria.

Project description:Background: Anopheles culicifacies is a rural vector of malaria in tropical and sub tropical South East Asian region. The salivary gland of the mosquito is the target for sporozoite interaction, blood feeding behavior, haemostasis and vector-parasite interactions. Malaria parasite matures inside the salivary gland, gain competence and transmitted to the host along with the saliva during biting. The importance of the proteins expressed in salivary gland is the first step in understanding the physiology of blood feeding and may provide insights into vector- parasite interactions. Since, no genomic or transcriptomics information is available of Anopheles culicifacies, therefore locally expressed functional proteins in salivary glands are of much importance. . Method: In this study, 1DE protein and in solution digestion was combined with tandem mass spectrometry (nano LC-MS/MS) and computational bioinformatics for data mining was employed to study the proteome profile of salivary glands of sugar fed An. culicifacies mosquito species. Functional annotation of all the identified proteins was carried out using gene ontology tools, CELLO and SMART analysis software. Results: Total 102 proteins were identified and analysed by SEQUEST algorithm against mosquito protein database from Uniprot/NCBI. Out of which 81 proteins were identified using gel free approach and 21 proteins using in-gel approach and 15 were common among these two approaches. All the identified proteins were categorized in to 23 groups of biological processes using GO tool. 7 proteins were depicted to be secretary in nature by investigating the signal peptide present. Potential proteins with unknown function were predicted by analyzing their functional association with other characterized proteins by STRING algorithm and were categorized in cell adhesion, cytoskeleton and membrane trafficking networks. Conclusion: Our study elucidates the first proteomic dataset of An. culicifacies salivary gland proteins. Functional annotation of salivary proteins and complementary gene ontology assignments in An. culicifacies species may contribute towards understanding the complex physiology of the tissues in this species. This proteome baseline data may facilitate the discernment of salivary glands and parasite correlation during blood feeding. Furthermore, this mass spectrometry based proteomic data may also provide insights into the elucidation of role of differential functional proteins present in refractory An. culicifacies mosquito and may be useful for development of effective malaria control strategies.

Project description:A deeper understanding of malaria parasite development inside the Anopheles mosquito may lead to the identification of processes that can be targeted by transmission-blocking interventions. Paraquat (1,1'-dimethyl-4,4'-bipyridylium dichloride) is a potent superoxide-inducing agent that impacts Plasmodium ookinete development, especially at higher concentrations. Compounds like Paraquat can potentially induce an oxidative imbalance in the mosquito midgut during ookinete maturation, essentially super-stressing the parasite leading to the arrested development of ookinetes, the only stage that can invade through a mosquito midgut cell to establish an oocyst infection in the mosquito. The mosquito midgut has evolved to handle the natural production of reactive oxygen and nitrogen species (ROS and RNS, respectively) as a result of feeding on blood. The addition of Paraquat to a bloodmeal is expected to induce a cognate response in the midgut to handle the excess ROS/RNS, and high concentrations of this compound can potentially overwhelm the midgut response leading to mosquito death. While several studies have explored the effect of Paraquat on malaria parasites, a fundamental understanding of the mosquito response to this compound remains unknown. Here, we quantified the mosquito midgut proteomic response to a Paraquat-laced sugar meal to understand the intrinsic midgut response (in the absence of a bloodmeal). We then carried out transcriptomic analysis of the mosquito midgut for several antioxidants of the Trx and GSH pathways to compare concordance or discordance between protein and its transcripts during different oxidative stress conditions. Finally, we determined whether the same Trx and GSH pathways are upregulated following infection with either P. falciparum or P. berghei at 24 hrs post-blood feeding, coinciding with the time point for maximal ookinete traversal of the midgut. We discuss the potential selective action of Paraquat on the parasite and the intrinsic tolerance of the mosquito midgut to Paraquat-mediated oxidative stress.

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