Project description:We compare the transcriptome of gnotobiotic Ae. aegypti generated by contaminating axenic (bacteria-free) larvae with bacterial isolates found in natural mosquito breeding sites. We focused on four bacterial isolates (Lysobacter, Flavobacterium, Paenibacillus and Enterobacteriaceae) and found that different gnotobiotic treatments resulted in massive transcriptomic changes throughout the mosquito development.
Project description:Sequencing of the Aedes aegypti genome has enabled genome-wide studies of gene expression in this mosquito. The large quantities of data produced from such studies require efficient cataloguing in order for new insight to be made into gene expression patterns and the underlying molecular mechanisms for producing these patterns. Our study provides a comprehensive catalogue of genes whose transcription products increase or decrease in abundance in adult females following blood feeding. We developed a publicly-accessible database and data-mining tool, aeGEPUCI, that integrates 1) stage-specific microarray analyses of gene expression in Ae. aegypti, 2) functional gene annotation, 3) genomic sequence data, and 4) computational sequence analysis tools. The database is accessible from the address http://www.aegep.bio.uci.edu.
Project description:Aedes aegypti mosquitoes are important vectors of viral diseases. Mosquito host factors play important roles in virus control and it has been suggested that Dengue virus replication is regulated by Dnmt2-mediated DNA methylation. However, recent studies have shown that Dnmt2 is a tRNA methyltransferase and that Dnmt2-dependent methylomes lack defined DNA methylation patterns, thus necessitating a systematic re-evaluation of the mosquito genome methylation status. We have now searched the A. aegypti genome for candidate DNA modification enzymes. This failed to reveal any known (cytosine-5) DNA methyltransferases, but identified homologues for the Dnmt2 tRNA methyltransferase, the Mettl4 (adenine-6) DNA methyltransferase, and the Tet DNA demethylase. All genes were expressed at variable levels throughout mosquito development. Mass spectrometry demonstrated that DNA methylation levels were several orders of magnitude below the levels that are usually detected in organisms with DNA methylation-dependent epigenetic regulation. Furthermore, whole-genome bisulfite sequencing failed to reveal any evidence of defined DNA methylation patterns. These results suggest that the A. aegypti genome is unmethylated. Interestingly, additional RNA bisulfite sequencing provided evidence for Dnmt2-mediated tRNA methylation in mosquitoes. These findings have important implications for understanding the mechanism of Dnmt2-dependent virus control.
Project description:Transcriptomic analysis of mosquito larvae exposed to LC50 of Cry11Aa toxin from Bti at different times was conducted to determine defense response pathways in order to better understand the toxin's mode of action and identify possible cellular targets to enhance toxin activity.
Project description:Sequencing of the Aedes aegypti genome has enabled genome-wide studies of gene expression in this mosquito. The large quantities of data produced from such studies require efficient cataloguing in order for new insight to be made into gene expression patterns and the underlying molecular mechanisms for producing these patterns. Our study provides a comprehensive catalogue of genes whose transcription products increase or decrease in abundance in adult females following blood feeding. We developed a publicly-accessible database and data-mining tool, aeGEPUCI, that integrates 1) stage-specific microarray analyses of gene expression in Ae. aegypti, 2) functional gene annotation, 3) genomic sequence data, and 4) computational sequence analysis tools. The database is accessible from the address http://www.aegep.bio.uci.edu. 8 conditions, 3 replicates for each condition
Project description:As is the case in vertebrates, successful mosquito immune responses depend on a complex and carefully orchestrated network of processes in order to defend the host from a foreign invader and to simultaneously maintain a homeostatic environment. Although commonalities exist between immune response mechanisms across phyla, invertebrates (e.g., mosquitoes) lack anticipatory, adaptive immune responses. However, a number of immune response elements, both cellular and humoral, exist to combat infection. Organisms that successfully invade the mosquito body cavity (hemocoel) potentially are subjected to a variety of mosquito defenses including: melanotic encapsulation, the production of antimicrobial peptides (AMPs) or reactive oxygen and nitrogen intermediates, or to engulfment by phagocytic cells. These arrays are used to temporally compare and contrast transcriptome profiles associated with these responses, as manifest in the cells (hemocytes) that circulate in the mosquito body cavity. Hemocytes have documented association with phagocytosis and melanization reactions in mosquitoes, and these responses, to a variety of Gram positive and Gram negative bacteria, have been characterized. Inoculated Escherichia coli elicit a strong phagocytic response, and Micrococcus luteus are rapidly melanized. For transcriptome profiling, inoculated moquitoes are compared to naïve (uninoculated) mosquitoes. Data obtained will begin to clarify the interactive role of hemocyte genes and gene products in orchestrating phagocytic, melanization, and AMP responses against invading bacteria. Keywords: time course