Comprehensive analysis of m6A methylation modification in chronic spinal cord injury in mice.

Chronic spinal cord injury (CSCI) is a catastrophic disease of the central nervous system (CNS), resulting in partial or complete loss of neurological function. N6‐methyladenosine (m6A) is the most common form of reversible posttranslational modification at the RNA level. However, the role of m6A modification in CSCI remains unknown. In this study, we established a CSCI model using a water‐absorbable polyurethane polymer, with behavioral assessment, electrophysiological analysis, and histochemical staining for validation. Methylated RNA immunoprecipitation sequencing (meRIP‐seq) and messenger RNA sequencing (mRNA‐seq) were jointly explored to compare the differences between CSCI spinal tissue and normal spinal tissue. Furthermore, real‐time quantitative reverse transcription pcr (qRT–PCR), western blot analysis, and immunofluorescence staining were used to analyze m6A modification‐related proteins. We found that water‐absorbable polyurethane polymer simulated well chronic spinal cord compression. Basso mouse scale scores and electrophysiological analysis showed continuous neurological function decline after chronic compression of the spinal cord. meRIP‐seq identified 642 differentially modified m6A genes, among which 263 genes were downregulated and 379 genes were upregulated. mRNA‐seq showed that 1544 genes were upregulated and 290 genes were downregulated after CSCI. Gene Ontology terms and enriched Kyoto Encyclopedia of Genes and Genomes pathways were also identified. qRT–PCR, western blotting, and immunofluorescence staining showed that Mettl14, Ythdf1, and Ythdf3 were significantly upregulated after CSCI. Our study revealed a comprehensive profile of m6A modifications in CSCI which may act as a valuable key for future research on CSCI. [ABSTRACT FROM AUTHOR]