title: Identification of suitable reference genes for BDV-infected primary rat hippocampal neurons.
abstract: Borna disease virus (BDV) is a neurotropic RNA virus that infects the limbic system of mammals and results in behavioral disorders. The hippocampus is a core region in the limbic system, which contributes to memory and learning and is important in the regulation of emotion. However, no validated microRNA housekeeping genes have yet been identified in BDV‑infected rat primary hippocampal neurons. Proper normalization is key in accurate miRNA expression analysis. The present study used reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) to evaluate the expression stability of 10 commonly used reference genes [miR‑92a, 5S, U6, miR‑103, miR‑101a, miR-let-7a, miR‑16, E2 small nucleolar RNA (snoRNA), U87 and miR‑191] in BDV‑infected rat hippocampal neurons and non‑infected controls across 12 days post‑infection. The data was analyzed by four statistical algorithms: geNorm, NormFinder, BestKeeper, and the comparative Δ‑Ct method. Subsequently, the most suitable reference genes (miR‑101a and U87) and the least suitable (snoRNA) were determined by the RankAggreg package. miR‑155 was selected as a standard by which to evaluate the most and least suitable reference genes. When normalized to the most stable reference gene there were significant differences between the two groups. However, when the data were normalized to the less stably expressed gene, the results were not significant. miR‑101a was recommended as a suitable reference gene for BDV-infected rat primary hippocampal neurons.
authors.affiliation: Department of Neurology, Yongchuan Hospital, Chongqing Medical University, Chongqing 402460, P.R. China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Mao
authors.affiliation: Department of Neurology, Yongchuan Hospital, Chongqing Medical University, Chongqing 402460, P.R. China.
authors.forename: Lujun
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Neurology, Yongchuan Hospital, Chongqing Medical University, Chongqing 402460, P.R. China.
authors.forename: Yujie
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Institute of Neuroscience and The Collaborative Innovation Center for Brain Science, Chongqing Medical University, Chongqing 400016, P.R. China.
authors.forename: Lu
authors.initials: L
authors.lastname: Sun
authors.affiliation: Institute of Neuroscience and The Collaborative Innovation Center for Brain Science, Chongqing Medical University, Chongqing 400016, P.R. China.
authors.forename: Siwen
authors.initials: S
authors.lastname: Liu
authors.affiliation: Institute of Neuroscience and The Collaborative Innovation Center for Brain Science, Chongqing Medical University, Chongqing 400016, P.R. China.
authors.forename: Peng
authors.initials: P
authors.lastname: He
authors.affiliation: Department of Rehabilitation, The Second Affiliated Hospital, Chongqing Medical University, Chongqing 400016, P.R. China.
authors.forename: Rongzhong
authors.initials: R
authors.lastname: Huang
authors.affiliation: Chongqing Key Laboratory of Neurobiology, Chongqing Medical University, Chongqing 400016, P.R. China.
authors.forename: Lin
authors.initials: L
authors.lastname: Sun
authors.affiliation: Institute of Neuroscience and The Collaborative Innovation Center for Brain Science, Chongqing Medical University, Chongqing 400016, P.R. China.
authors.forename: Shigang
authors.initials: S
authors.lastname: Chen
authors.affiliation: Institute of Neuroscience and The Collaborative Innovation Center for Brain Science, Chongqing Medical University, Chongqing 400016, P.R. China.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Neurology, Yongchuan Hospital, Chongqing Medical University, Chongqing 402460, P.R. China.
authors.forename: Peng
authors.initials: P
authors.lastname: Xie
mesh_terms: D000818:Animals; D001890:Borna Disease / Q000235:genetics / Q000821:virology; D001891:Borna disease virus / Q000502:physiology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D008297:Male; D035683:MicroRNAs / Q000235:genetics; D017966:Pyramidal Cells / Q000378:metabolism* / Q000821:virology*; D020871:RNA Stability; D051381:Rats
chemical_list: D035683:MicroRNAs