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MicroRNA analysis in mouse neuro-2a cells after pseudorabies virus infection
Yongtao Li1, Guanmin Zheng1, Yujuan Zhang1
1College of Animal Husbandry and Veterinary Science, Henan Agricultural University, Zhengzhou, 450002, China.
Abstract:
Pseudorabies virus (PRV), an alpha herpesvirus can enter the mammalian nervous system, causing Aujezsky's disease. Previous studies have reported an alteration of microRNA (miRNA) expression levels during PRV infections. However, knowledge regarding miRNA response in nervous cells to PRV infection is still unknown. To address this issue, small RNA libraries from infected and uninfected mouse neuroblastoma cells were assessed after Illumina deep sequencing. A total of eight viral miRNA were identified, and ten host miRNAs showed significantly different expression upon PRV infection. Among these, five were analyzed by stem-loop RT-qPCR, which confirmed the above data. Interestingly, these viral miRNAs were mainly found in the large latency transcript region of PRV, and predicted to target a variety of genes, forming a complicated regulatory network. Moreover, ten cellular miRNAs were expressed differently upon PRV infection, including nine upregulated and one downregulated miRNAs. Host targets of these miRNAs obtained by bioinformatics analysis belonged to large signaling networks, mainly encompassing calcium signaling pathway, cAMP signaling pathway, MAPK signaling pathway, and other nervous-associated pathways. These findings further highlighted miRNA features in nervous cells after PRV infection and contributed to unveil the underlying mechanisms of neurotropism as well as the neuropathogenesis of PRV.
Insights
This study reveals how pseudorabies virus (PRV) affects microRNA (miRNA) expression in mouse nervous cells. PRV infection alters host miRNA levels, impacting nervous system pathways and contributing to disease mechanisms.
Area of Science:
- Virology
- Neuroscience
- Molecular Biology
Background:
- Pseudorabies virus (PRV), an alphaherpesvirus, causes Aujeszky's disease by infecting the mammalian nervous system.
- Previous research indicates altered microRNA (miRNA) expression during PRV infections, but the response in nervous cells remains unclear.
Purpose of the Study:
- To investigate the role of miRNAs in nervous cells during PRV infection.
- To identify viral and host miRNAs affected by PRV and elucidate their potential functions.
Main Methods:
- Small RNA sequencing of infected and uninfected mouse neuroblastoma cells.
- Stem-loop RT-qPCR validation of selected miRNA expression.
- Bioinformatic analysis to predict miRNA targets and associated signaling pathways.
Main Results:
- Identified eight viral miRNAs, primarily from the PRV latency region, and ten differentially expressed host miRNAs (nine upregulated, one downregulated).
- Confirmed altered miRNA expression using stem-loop RT-qPCR.
- Predicted viral miRNAs target various genes, forming a complex regulatory network.
- Host miRNA targets are involved in crucial nervous system signaling pathways, including calcium, cAMP, and MAPK signaling.
Conclusions:
- PRV infection significantly alters miRNA expression profiles in nervous cells.
- Viral miRNAs may play a role in PRV pathogenesis and neurotropism.
- Host miRNAs are implicated in regulating nervous system pathways affected by PRV infection, offering insights into neuropathogenesis.
