Dataset Information

Proteomic changes occurring in Anopheles gambiae and Anopheles stephensi during aging

ABSTRACT: The age of mosquitoes is a crucial determinant of their susceptibility to infection, probability of survival to transmit pathogens and tolerance to insecticides. We investigated changes to the abundance of proteins found in heads and thoraces of Anopheles gambiae and Anopheles stephensi as they aged. Protein expression changes were assessed using two-dimensional difference gel electrophoresis and the identity of differentially expressed proteins was determined by using either matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry or capillary high-pressure liquid chromatography coupled with a linear ion-trap (LTQ)-Orbitrap XL hybrid mass spectrometer. Protein biomarkers were validated by quantitative Western blot analysis.

INSTRUMENT(S): LTQ Orbitrap, ultraflex

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

DISEASE(S): Plasmodium Falciparum Malaria

SUBMITTER: Keyur Dave

LAB HEAD: Jeffrey J. Gorman

PROVIDER: PXD002153 | Pride | 2015-08-26

REPOSITORIES: Pride

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Proteomic changes occurring in the malaria mosquitoes Anopheles gambiae and Anopheles stephensi during aging.

Sikulu Maggy T MT Monkman James J Dave Keyur A KA Hastie Marcus L ML Dale Patricia E PE Kitching Roger L RL Killeen Gerry F GF Kay Brian H BH Gorman Jeffery J JJ Hugo Leon E LE

Journal of proteomics 20150620

The age of mosquitoes is a crucial determinant of their ability to transmit pathogens and their resistance to insecticides. We investigated changes to the abundance of proteins found in heads and thoraces of the malaria mosquitoes Anopheles gambiae and Anopheles stephensi as they aged. Protein expression changes were assessed using two-dimensional difference gel electrophoresis and the identity of differentially expressed proteins was determined by using either matrix-assisted laser desorption i ...[more]

PMID: 26100052

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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.

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	MALDI.zip	Other
	Spot1073.dat	Other
	Spot1073.dat-pride.pride.mgf.gz	Mgf
	Spot1073.dat-pride.pride.mztab.gz	Mztab
	Spot1073.dat-pride.xml.gz	Xml