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Detection of MicroRNAs in Microglia by Real-time PCR in Normal CNS and During Neuroinflammation
Published on: July 23, 2012
Exploitation of microRNAs by Japanese Encephalitis virus in human microglial cells
Meghana Rastogi1, Neha Srivastava1, Sunit K Singh1
1Laboratory of Human Molecular Virology and Immunology, Molecular Biology Unit, Faculty of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.
Abstract:
JEV infection in CNS leads to the JE neuroinflammation. Children and old age individual have been reported to be more prone to JEV infection. MicroRNAs are endogenous, small non-coding RNAs, which regulate the gene expression. These are ∼22 nucleotide long, conserved RNA sequence that binds at the 3'UTR of a target mRNA and regulate the post-transcriptional gene expression. The role of microRNAs has been reported in several diseases like cancer, viral infection, neuro-degeneration, diabetes etc. In the present study, the human microglial cells were infected with JEV (JaOArS982). The control and infected samples were subject to microarray profiling for microRNA expression. The microarray profile yielded differentially expressed microRNAs from JEV infected samples. The microRNA gene targets, gene ontology, annotations, and pathways were identified through various bioinformatics tools. Additionally, the pathways were mostly found common to "ubiquitin mediated proteolysis," "cytokine signaling," "maintenance of barrier function/cell junctions," JAK/STAT pathway" "Toll-like receptor signaling," "Wnt-signaling," "adhesion molecules," "apoptosis," "endocytosis," "vesicle mediated transport" etc.
Insights
Japanese encephalitis virus (JEV) infection causes neuroinflammation in the central nervous system (CNS). This study identified differentially expressed microRNAs in human microglial cells following JEV infection, revealing key affected biological pathways.
Area of Science:
- Neuroscience
- Molecular Biology
- Virology
Background:
- Japanese encephalitis virus (JEV) infection causes neuroinflammation in the central nervous system (CNS), with higher susceptibility in children and the elderly.
- MicroRNAs (miRNAs) are small, non-coding RNAs regulating gene expression post-transcriptionally and are implicated in various diseases, including viral infections and neurodegeneration.
Purpose of the Study:
- To investigate the role of microRNAs in JEV-induced neuroinflammation.
- To identify differentially expressed microRNAs and their associated pathways in human microglial cells upon JEV infection.
Main Methods:
- Infection of human microglial cells with JEV (JaOArS982).
- Microarray profiling to assess microRNA expression in control and infected cells.
- Bioinformatics analysis to identify microRNA targets, gene ontology, annotations, and signaling pathways.
Main Results:
- Microarray analysis revealed differentially expressed microRNAs in JEV-infected microglial cells compared to controls.
- Identified pathways significantly associated with JEV infection include ubiquitin-mediated proteolysis, cytokine signaling, JAK/STAT, Toll-like receptor signaling, Wnt-signaling, and apoptosis.
Conclusions:
- MicroRNAs play a significant role in the host response to JEV infection in microglial cells.
- The identified pathways provide insights into the molecular mechanisms underlying JEV-induced neuroinflammation and potential therapeutic targets.
