Frontiers in Genetics (Feb 2021)

Alteration of mRNA 5-Methylcytosine Modification in Neurons After OGD/R and Potential Roles in Cell Stress Response and Apoptosis

  • Huan Jian,
  • Huan Jian,
  • Chi Zhang,
  • Chi Zhang,
  • ZhangYang Qi,
  • ZhangYang Qi,
  • Xueying Li,
  • Xueying Li,
  • Xueying Li,
  • Yongfu Lou,
  • Yongfu Lou,
  • Yi Kang,
  • Yi Kang,
  • Weimin Deng,
  • Yigang Lv,
  • Yigang Lv,
  • Chaoyu Wang,
  • Chaoyu Wang,
  • Wei Wang,
  • Wei Wang,
  • Shenghui Shang,
  • Shenghui Shang,
  • Mengfan Hou,
  • Mengfan Hou,
  • Hengxing Zhou,
  • Hengxing Zhou,
  • Hengxing Zhou,
  • Hengxing Zhou,
  • Shiqing Feng,
  • Shiqing Feng,
  • Shiqing Feng,
  • Shiqing Feng

DOI
https://doi.org/10.3389/fgene.2021.633681
Journal volume & issue
Vol. 12

Abstract

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Epigenetic modifications play an important role in central nervous system disorders. As a widespread posttranscriptional RNA modification, the role of the m5C modification in cerebral ischemia-reperfusion injury (IRI) remains poorly defined. Here, we successfully constructed a neuronal oxygen-glucose deprivation/reoxygenation (OGD/R) model and obtained an overview of the transcriptome-wide m5C profiles using RNA-BS-seq. We discovered that the distribution of neuronal m5C modifications was highly conserved, significantly enriched in CG-rich regions and concentrated in the mRNA translation initiation regions. After OGD/R, modification level of m5C increased, whereas the number of methylated mRNA genes decreased. The amount of overlap of m5C sites with the binding sites of most RNA-binding proteins increased significantly, except for that of the RBM3-binding protein. Moreover, hypermethylated genes in neurons were significantly enriched in pathological processes, and the hub hypermethylated genes RPL8 and RPS9 identified by the protein-protein interaction network were significantly related to cerebral injury. Furthermore, the upregulated transcripts with hypermethylated modification were enriched in the processes involved in response to stress and regulation of apoptosis, and these processes were not identified in hypomethylated transcripts. In final, we verified that OGD/R induced neuronal apoptosis in vitro using TUNEL and western blot assays. Our study identified novel m5C mRNAs associated with ischemia-reperfusion in neurons, providing valuable perspectives for future studies on the role of the RNA methylation in cerebral IRI.

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