Project description:Microarray analysis on days 1, 2 and 7 post-infection with dengue, yellow fever and West Nile virus in Aedes aegypti Rockefeller strain mosquitoes RNA was purified and hybridized with Nimblegen X4 microarray chips using 81-mer probes designed from 18,000 open reading frames (ORF) found in the Ae. aegypti genome, with 2 different probes per ORF
Project description:Wolbachia pipientis is an intracellular symbiotic bacterium found in insects and arthropods. Wolbachia can decrease the vectorial capacity for various pathogens, such as the dengue virus, in Aedes aegypti. The purpose of this study was to determine the effect of Wolbachia (wMel strain) on the vectorial capacity of Ae. aegypti for Dirofilaria immitis. We analyzed gene expression patterns by RNA-seq in addition to the D. immitis infection phenotype in Ae. aegypti infected with and without wMel. Four Ae. aegypti strains, MGYP2.tet, MGYP2, Liverpol (LVP)-Obihiro (OB), and LVP-OB-wMel (OB-wMel) were analyzed for transcriptome comparison in Malpighian tubule at 2 days post infection. The correlation between Wolbachia infection, D. immitis infection phenotype and immune-related genes expression in Ae. aegypti was investigated.
Project description:The ability of many viruses to manipulate the host antiviral immune response often results in complex host-pathogen interactions. In order to study the interaction of dengue virus (DENV) with the Aedes aegypti immune response, we have characterized the DENV infection-responsive transcriptome of the immune-competent A. aegypti cell line Aag2. As in mosquitoes, DENV infection transcriptionally activated the cell line Toll pathway and a variety of cellular physiological systems. Most notably, however, DENV infection down-regulated the expression levels of numerous immune signaling molecules and antimicrobial peptides (AMPs). Functional assays showed that transcriptional induction of AMPs from the Toll and IMD pathways in response to bacterial challenge is impaired in DENV-infected cells. In addition, Escherichia coli, a gram-negative bacteria species, grew better when co-cultured with DENV-infected cells than with uninfected cells, suggesting a decreased production of AMPs from the IMD pathway in virus-infected cells. Pre-stimulation of the cell line with gram-positive bacteria prior to DENV infection had no effect on DENV titers, while pre-stimulation with gram-negative bacteria resulted in an increase in DENV titers. These results indicate that DENV is capable of actively suppressing immune responses in the cells it infects, a phenomenon that may have important consequences for virus transmission and insect physiology. Infected (dengue virus or heat-inactivated dengue virus) vs. naive cells. 3 replicates each.
Project description:Microarray analysis on days 1, 2 and 7 post-infection with dengue, yellow fever and West Nile virus in Aedes aegypti Rockefeller strain mosquitoes RNA was purified and hybridized with Nimblegen X4 microarray chips using 81-mer probes designed from 18,000 open reading frames (ORF) found in the Ae. aegypti genome, with 2 different probes per ORF Nimblegen X4 array format, 81mer probes, RNA samples taken mostly in triplicate, microarray analysis done in triplicate. Thirty seven samples in total. Fold change data with infection/mock on the Series record.
Project description:Wolbachia is a vertically transmitted intracellular bacteria that infect most than 60% of insect species. The strains wMelPop and wMel were introduced in the dengue virus vector Aedes aegypti, naturally not infected by Wolbachia. Recently, it was shown that those two strains inhibit dengue virus replication into their new host, A. aegypti (Moreira et al. 2009 and Walker et al. in preparation). The aim of this project is to look at the transcriptional response of Aedes aegypti to infection with wMel and wMelPop and try to find some genes or pathway potentially involved in the viral interference.Four laboratory lines of A. aegypti were used throughout this study. The PGYP1 and Mel2 lines were generated by transinfection with wMelPop and wMel strains respectively. PGYP1.tet and Mel2tet lines were treated with the antibiotic tetracycline and cured from Wolbachia infection (McMeniman et al., 2009 and Walker et al in preparation). The Mosquitoes were reared under standard laboratory conditions (26 ± 2 °C, 12:12 light/dark cycle, 75% relative humidity). Mosquito larvae were fed 0.1mg/larvae of TetraMin Tropical Tablets once a day. Adults were transferred to cages (measuring 30 x 30 x 30 cm) at emergence at 400 individuals per cage. Adults were supplied with a basic diet of 10% sucrose solution (Turley et al., 2009).
Project description:Custom microarrays were used to examine global differences in female vs. male gene expression in the developing pupal head of the dengue vector mosquito Aedes aegypti.
Project description:The study provides a comparative of transcript levels in uninfected and CHIKV-infected Aedes aegypti derived Aag2 cells using RNA Seq
Project description:We used RNA-sequencing to identify differentially expressed genes in the midgut of Aedes aegypti that contribute to the field derivied dengue susceptible (Cali-S) and dengue refractory (Cali-R) phenotypes
Project description:Aedes aegypti mosquitoes infect hundreds of millions of people each year with dangerous viral pathogens including dengue, yellow fever, Zika, and chikungunya. Progress in understanding the biology of this insect, and developing tools to fight it, depends on the availablity of a high-quality genome assembly. Here we use DNA proximity ligaton (Hi-C) and Pacific Biosciences long reads to create AaegL5 - a highly contiguous A. aegypti reference.
Project description:Wolbachia are intracellular maternally inherited bacteria that can spread through insect populations and block virus transmission by mosquitoes, providing an important new dengue control strategy. To better understand the mechanisms of virus inhibition, proteomic quantification of the effects of Wolbachia in mosquito (Aedes aegypti) midguts was performed.