Project description:Flaviviruses, particularly Japanese encephalitis virus (JEV) and West Nile virus (WNV), are important causes of virus-induced central nervous system (CNS) disease in humans. We used microarray analysis to identify cellular genes that are differentially regulated following infection of the brain with JEV (P3) or WNV (New York 99). Gene expression data for these flaviviruses was compared to that induced following infection of the brain with reovirus (Type 3 Dearing), an unrelated neurotropic virus. Although several studies have described gene expression changes following virus infection of the brain, this report is the first to directly compare large-scale gene expression data from different viruses. We found that a large number of genes were up-regulated in common to infections with all 3 viruses (fold change > 2, P < 0.001), including genes associated with interferon signaling, the immune system, inflammation and cell death/survival signaling. In addition, genes associated with glutamate signaling were down-regulated in common to infections with all 3 viruses (fold change > 2, P < 0.001). These genes may serve broad spectrum therapeutic targets for virus-induced CNS disease. A distinct set of genes were up-regulated following flavivirus-infection, but not following infection with reovirus. These genes were associated with tRNA charging and may serve as therapeutic targets for flavivirus-induce CNS disease. Gene expression in the brain following WNV or JEV infection. WNV- or JEV-infected (N=3) vs. mock-infected (N=3) mouse brain.
Project description:Neurotropic flavivirus Japanese encephalitis virus (JEV) and West Nile virus (WNV) are amongst the leading causes of encephalitis. Using label-free quantitative proteomics, we identified proteins differentially expressed upon JEV (gp-3, RP9) or WNV (IS98) infection of human neuroblastoma cells. Both viruses were associated with the up-regulation of immune response (IFIT1/3/5, ISG15, OAS, STAT1, IRF9) and the down-regulation of SSBP2, involved in gene expression, as well as PAM, involved in neuropeptide amidation. Proteins associated to membranes, involved in extracellular matrix organization and collagen metabolism represented major clusters down-regulated by JEV and WNV. Moreover, transcription regulation and mRNA processing clusters were also heavily regulated by both neurotropic flaviviruses. If the proteome of neuroblastoma cells infected by JEV or WNV was significantly modulated in the presence of mosquito saliva, both viruses showed distinct patterns. Mosquito saliva favored the modulation of proteins associated with gene regulation in JEV infected neuroblastoma cells while it was the modulation of proteins associated with protein maturation, signal transduction and ion transporters in the case of WNV infected neuroblastoma cells.
Project description:PURPOSE: To provide a detailed gene expression profile of the normal postnatal mouse cornea. METHODS: Serial analysis of gene expression (SAGE) was performed on postnatal day (PN)9 and adult mouse (6 week) total corneas. The expression of selected genes was analyzed by in situ hybridization. RESULTS: A total of 64,272 PN9 and 62,206 adult tags were sequenced. Mouse corneal transcriptomes are composed of at least 19,544 and 18,509 unique mRNAs, respectively. One third of the unique tags were expressed at both stages, whereas a third was identified exclusively in PN9 or adult corneas. Three hundred thirty-four PN9 and 339 adult tags were enriched more than fivefold over other published nonocular libraries. Abundant transcripts were associated with metabolic functions, redox activities, and barrier integrity. Three members of the Ly-6/uPAR family whose functions are unknown in the cornea constitute more than 1% of the total mRNA. Aquaporin 5, epithelial membrane protein and glutathione-S-transferase (GST) omega-1, and GST alpha-4 mRNAs were preferentially expressed in distinct corneal epithelial layers, providing new markers for stratification. More than 200 tags were differentially expressed, of which 25 mediate transcription. CONCLUSIONS: In addition to providing a detailed profile of expressed genes in the PN9 and mature mouse cornea, the present SAGE data demonstrate dynamic changes in gene expression after eye opening and provide new probes for exploring corneal epithelial cell stratification, development, and function and for exploring the intricate relationship between programmed and environmentally induced gene expression in the cornea. Keywords: other
Project description:JEV is a neurotropic pathogen that causes lethal encephalitis in humans. The high susceptibility and massive proliferation of JEV in neurons lead to neuronal damage and dramatical inflammation in central nerves system (CNS). However, the mechanisms by which JEV preferentially infects neurons and induces massive neuroinflammation are largely unknown. Here, we found that JEV mainly distributed in cerebral cortex, striatum and thalamus in brain of infected mice, indicating the tissue or cell type bias. Cells in these regions of JEV-infected mice showing different symptoms (mild, moderate and severe) were isolated and subjected to scRNA-seq. 88000 single cells were obtained based on top differentially expressed genes, and 34 clusters and 10 major cell types were identified. As demonstrated by scRNA-seq and flow-cytometry assay, activated microglia cells and infiltrating immune cells including monocyte & macrophage, T cells and NK cells which are responsible for inflammatory response were increased in JEV-infected brain in a symptom severity-dependent manner. Furthermore, the subclusters of these cells and the cell-cell interaction network were analyzed. More communications between individual cells and significant increase in ligand receptor pairs associated with tight junctions, chemokines and antigen-presenting molecules were shown in JEV-infected mouse brain, suggesting upregulation of cellular permeability, inflammation and antiviral response. To identify which subtype(s) of neurons is/are targeted by JEV, 27874 neuronal cells were reclustered to 15 subsets. By analyzing the correlation between the expression of JEV E gene and neuronal genes, we found that Baiap2high neurons are highly permissive to JEV.
Project description:Introgressed variants from other species can be an important source of genetic variation because they may arise rapidly, can include multiple mutations on a single haplotype, and have often been pretested by selection in the species of origin. Although introgressed alleles are generally deleterious, several studies have reported introgression as the source of adaptive alleles-including the rodenticide-resistant variant of Vkorc1 that introgressed from Mus spretus into European populations of Mus musculus domesticus. Here, we conducted bidirectional genome scans to characterize introgressed regions into one wild population of M. spretus from Spain and three wild populations of M. m. domesticus from France, Germany, and Iran. Despite the fact that these species show considerable intrinsic postzygotic reproductive isolation, introgression was observed in all individuals, including in the M. musculus reference genome (GRCm38). Mus spretus individuals had a greater proportion of introgression compared with M. m. domesticus, and within M. m. domesticus, the proportion of introgression decreased with geographic distance from the area of sympatry. Introgression was observed on all autosomes for both species, but not on the X-chromosome in M. m. domesticus, consistent with known X-linked hybrid sterility and inviability genes that have been mapped to the M. spretus X-chromosome. Tract lengths were generally short with a few outliers of up to 2.7 Mb. Interestingly, the longest introgressed tracts were in olfactory receptor regions, and introgressed tracts were significantly enriched for olfactory receptor genes in both species, suggesting that introgression may be a source of functional novelty even between species with high barriers to gene flow.
Project description:The purpose is to obtain samples for mRNA, miRNA, proteomics, lipidomics, metabolomics, and histopathology analysis in mouse cortex infected with wild-type West Nile virus (WNV; WNV-NY99 382), and mutant WNV-E218A (WNV-NY99 382 E218A 2 nt).
Project description:The purpose is to obtain samples for mRNA, miRNA, proteomics, lipidomics, metabolomics, and histopathology analysis in mouse cortex infected with wild-type West Nile virus (WNV; WNV-NY99 382), and mutant WNV-E218A (WNV-NY99 382 E218A 2 nt).
Project description:The purpose is to obtain samples for mRNA, miRNA, proteomics, lipidomics, metabolomics, and histopathology analysis in mouse cerebellum infected with wild-type West Nile virus (WNV; WNV-NY99 382), and mutant WNV-E218A (WNV-NY99 382 E218A 2 nt).
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from seven Mus musculus tissues (Heart, Brain, Liver, Lung, Kidney, Skeletal Muscle, Thymus)