The Alteration of M6A-tagged Transcript Profiles in The Retina of Rats After Traumatic Optic Neuropathy

Messager RNA (mRNA) can be modified in a variety of ways, among which the modification of N6-methyladenosine (m6A) is one of the most common ones. Recent studies have found that the m6A modification in mRNA could functionally regulate the splicing, localization, translation and stability of mRNA, which might be closely related to multiple diseases. However, the roles of m6A modification in traumatic optic neuropathy (TON) are unknown. Herein, we detected the expression of m6A-related genes via quantitative real-time PCR (qRT-PCR) and performed methylated RNA immunoprecipitation sequencing (MeRIP-seq) as well as RNA-sequencing to analyze the alteration profiles of m6A modification after TON. The results showed that the expression of m6A-related genes (METTL3, WTAP, FTO and ALKBH5) were all upregulated after TON. In all, 2810 m6A peaks were differentially upregulated and 689 m6A peaks were downregulated. In addition, the hypermethylated and hypomethylated profiles of mRNA transcripts were also identified. To sum up, our study revealed the differentially expressed m6A modification in the early stage of TON, which may provide novel insights into the mechanism and treatment of TON.信使RNA（Messenger RNA, mRNA）可发生多种修饰，其中N6-甲基腺嘌呤（N6-methyladenosine, m6A）是最为常见的修饰类型之一。近期研究表明，mRNA上的m6A修饰可在功能上调控mRNA的剪接、定位、翻译及稳定性，且该修饰可能与多种疾病密切相关。然而，m6A修饰在创伤性视神经病变（traumatic optic neuropathy, TON）中的作用尚不明确。本研究通过定量实时聚合酶链反应（quantitative real-time PCR, qRT-PCR）检测m6A相关基因的表达水平，并联合甲基化RNA免疫沉淀测序（methylated RNA immunoprecipitation sequencing, MeRIP-seq）与RNA测序技术，分析创伤性视神经病变发生后m6A修饰的表达谱变化。结果显示，创伤性视神经病变发生后，m6A相关基因（METTL3、WTAP、FTO及ALKBH5）的表达均显著上调。共计2810个m6A峰呈现差异上调，689个m6A峰呈现差异下调。此外，本研究还鉴定出了mRNA转录本的高甲基化与低甲基化表达谱。综上，本研究揭示了创伤性视神经病变早期阶段的差异表达m6A修饰模式，可为该病的发病机制研究与治疗策略开发提供全新的视角。