Dataset Information

Identification of molecular response to blood meal in the fatbody of Anopheles stephensi using quantitative proteomics

ABSTRACT: Fat body is an important tissue in the context of vitellogenesis, vector immunity, vector physiology and vector-parasite interaction. However, the proteome of fatbody and impact of blood meal on the gene expression of this vital organ has not been investigated so far. Therefore, in this study, we made an attempt to identify proteins expressed in fatbody of An. stephensi and their altered expression in response to blood meal. In all, we identified 4,504 proteins in the fatbody using multiple fractionation strategies, which is by far the largest resource of fatbody proteome in any mosquito species. Further, comparative proteomic analysis of fatbody 24 and 48 hours post blood meal led to identification of over 300 differentially expressed proteins. Bioinformatics analysis of these proteins suggested their role in vitellogenesis, lipid transport, mosquito immunity and oxidation-reduction processes. Interestingly, we identified four novel genes,which were found to be differentially expressed upon blood meal. These proteins are potential target for vector control strategies and development of transmission blocking vaccines.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Anopheles Stephensi (asian Malaria Mosquito)

TISSUE(S): Fat Body

SUBMITTER: Keshava Prasad T. S.

LAB HEAD: T. S. Keshava Prasad

PROVIDER: PXD006462 | Pride | 2018-01-03

REPOSITORIES: Pride

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Publications

Response to Blood Meal in the Fat Body of Anopheles stephensi Using Quantitative Proteomics: Toward New Vector Control Strategies Against Malaria.

Kumar Manish M Mohanty Ajeet Kumar AK Sreenivasamurthy Sreelakshmi K SK Dey Gourav G Advani Jayshree J Pinto Sneha M SM Kumar Ashwani A Prasad Thottethodi Subrahmanya Keshava TSK

Omics : a journal of integrative biology 20170905 9

Malaria remains a grand challenge for disruptive innovation in global health therapeutics and diagnostics. Anopheles stephensi is one of the major vectors of malaria in Asia. Vector and transmission control are key focus areas in the fight against malaria, a field of postgenomics research where proteomics can play a substantive role. Moreover, to identify novel strategies to control the vector population, it is necessary to understand the vector life processes at a global and molecular scale. In ...[more]

PMID: 28873011

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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:Mosquitoes possess an innate immune system that is capable of limiting infection by a variety of pathogens, including the Plasmodium spp. parasites responsible for human malaria. The Anopheles immune deficiency (IMD) innate immune signaling pathway confers resistance to Plasmodium falciparum. While some previously identified Anopheles anti-Plasmodium effectors are regulated through signaling by Rel2, the transcription factor of the IMD pathway, many components of this defense system remain uncharacterized. To begin to better understand the regulation of immune effector proteins by the IMD pathway, we used oligonucleotide microarrays and iTRAQ to analyze differences in mRNA and protein expression, respectively, between transgenic An. stephensi mosquitoes exhibiting blood meal-inducible overexpression of an active recombinant Rel2 and their wild-type conspecifics. Numerous genes were differentially regulated at both the mRNA and protein levels following induction of Rel2. While multiple immune genes were up-regulated, a majority of the differentially expressed genes have no known immune function in mosquitoes. Identified sequences were assigned putative functions and gene ontology (GO) terms based on homology to previously annotated A. gambiae gene sequences. Selected up-regulated genes from multiple GO categories were tested for both anti-Plasmodium and anti-bacterial action using RNA interference (RNAi). Based on our experimental findings, we conclude that increased expression of the IMD immune pathway-controlled transcription factor Rel2 affects the expression of numerous genes with diverse functions, suggesting a broader physiological impact of immune activation and possible functional versatility of Rel2. Our study has identified multiple novel anti-Plasmodium effectors. Midguts from midgut-specific transgenic A. stephensi at 6 and 12 hours post-blood meal and fat bodies from fat body-specific transgenic A. stephensi at 12 and 18 hours post-blood meal were compared to wild-type A. stephensi at the same time points. 3 biological replicates and 1 pseudo-replicate per array.

Project description:With their genome sequenced, Anopheles gambiae mosquitoes now serve as a powerful tool for basic research in comparative, evolutionary and developmental biology. The knowledge generated by these studies is expected to reveal molecular targets for novel vector control and pathogen transmission blocking strategies. Comparisons of gene-expression profiles between adult male and nonblood-fed female Anopheles gambiae mosquitoes revealed that roughly 22% of the genes showed sex-dependent regulation. Blood-fed females switch the majority of their metabolism to blood digestion and egg formation within 3 h after the meal is ingested, in detriment to other activities such as flight and response to environment stimuli. Changes in gene expression are most evident during the first, second and third days after a blood meal, when as many as 50% of all genes showed significant variation in transcript accumulation. After laying the first cluster of eggs (between 72 and 96 h after the blood meal), mosquitoes return to a nongonotrophic stage, similar but not identical to that of 3-dayold nonblood-fed females. Ageing and/or the nutritional state of mosquitoes at 15 days after a blood meal is reflected by the down-regulation of 5% of all genes. A full description of the large number of genes regulated at each analysed time point and each biochemical pathway or biological processes in which they are involved is not possible within the scope of this contribution. Therefore, we present descriptions of groups of genes displaying major differences in transcript accumulation during the adult mosquito life. However, a publicly available searchable database (Anopheles gambiae Gene Expression Database at UC Irvine) has been made available so that detailed analyses of specific groups of genes based on their descriptions, functions or levels of gene expression variation can be performed by interested investigators according to their needs. Experiment Overall Design: 12 Samples analyzed, 3 replicates per sample, Affymetrix internal controls

Dataset Information

Identification of molecular response to blood meal in the fatbody of Anopheles stephensi using quantitative proteomics

Publications

Response to Blood Meal in the Fat Body of Anopheles stephensi Using Quantitative Proteomics: Toward New Vector Control Strategies Against Malaria.

Similar Datasets

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