Project description:Proteomic analysis of Anopheles gambiae brain tissue after in-gel trypsin digestion. To gain insights into neurobiology of the Anopheles gambiae mosquito, we carried out a proteomic analysis of its brain using a comprehensive proteomic approach.
Project description:The goal of this study is to analyze the gene expression profile of Anopheles gambiae Ag55 cells and compare it with gene expression profile of blood fed female Anopheles gambiae adult to establish Ag55 cells as a potential model to study mosquito-pathogen interactions Methods: The transcript profiles of Ag55 cells were generated by RNA-sequencing, in triplicate, using Illumina HiSeq 2000 v3 platform. The 7 samples (paired-end reads, including replicates ( 3 samples from Ag55 cells and 4 samples from blood-fed female adults downloaded from NCBI GEO (Series GSE55453, SRA id : SRP039058))) were independently mapped on to A. gambiae genome (Anopheles-gambiae-PEST_CHROMOSOMES_AgamP4.fa.gz) downloaded from vectorbase by using TopHat followed by Cufflinks to estimate the expression values of the transcripts in FPKM (Fragments Per Kilobase per Million mapped reads) with the cuffdiff2 default geometric normalization. Differentially expressed genes (FDR<0.05 after Benjamini-Hochberg correction for multiple-testing) were identified for Ag55 cells relative to Blood fed female adults Results: We provide detailed gene expression and proteome profiles of Ag55 cells. We further compare the gene expression profiles of Ag55 cells and blood fed female adults. Gene Ontology enrichment analysis of our transcriptomic data suggested that Ag55 cells have phagocytic properties, a hypothesis which we confirmed using confocal imaging. Transcriptomic data further backed by proteomic data suggest that Ag55 cells express hemocyte like properties, and are immune competent. Conclusion: As Ag55 cells are immune competent and express hemocyte like properties they can be used as a model to study vector-pathogen immune response. Furthermore, the availability of transcriptomic and proteomic data of Ag55 cells will help researchers use and engineer the Ag55 cell line in an efficient way, for example by developing strategies to make it more suitable for studies of interactions with Plasmodium and other microbes.
Project description:We custom-built a bioinformatics pipeline to search for 20E-modifying enzymes in the accessory glands of Anopheles gambiae males, searching for ecdysteroid kinases (EcK), ecdysone oxidases (EO), and ecdysteroid-phosphate phosphatases (EPP). To this end, we generated RNAseq datasets of different An. gambiae tissues dissected from virgin and mated females and males, and produced similar datasets for Anopheles albimanus, a South American species that does not synthetize and transfer ecdysteroids during mating. These analyses led to the identification of one candidate EPP and two potential EcKs (EcK1 and EcK2), which we demonstrated are involved in the activity of a male-specific oxidized ecdysteroid (3D20E). We further determined that 3D20E is specifically produced by the An. gambiae male accessory glands and is transferred to females during copulation, where it triggers a series of post-mating responses.
Project description:we report the RNA-seq based analyses of the transcriptional changes in the Anopheles gambiae mosquitoes from East Africa classified as deltamethrin-resistant or -suscpetible accordign the WHO test