Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong 226001, China.
Department of Orthopaedics, Affiliated Hospital of Nantong University, Nantong 226001, China.
Exp Neurol. 2018 Oct;308:80-89. doi: 10.1016/j.expneurol.2018.07.003. Epub 2018 Jul 4.
The intrinsic axon regeneration capacity is crucial for peripheral nerve regeneration after injury. Identifying key molecules involved in this process makes great contribution to the investigation of peripheral nerve injury repair. Alternative splicing (AS) is an important regulation mode of eukaryotic gene expression, which has been widely studied both in physiological and pathological processes. However, less is known about the role of AS in peripheral nerve regeneration. In this work, to identify the AS events associated with axon regeneration capacity, we analyzed the AS events during sciatic nerve injury repair by RNA sequencing (RNA-Seq) and replicate multivariate analysis of transcript splicing (rMATS). The differential AS events were underwent gene ontology enrichment and pathway analyses. Moreover, we identified a significantly increased AS event of neuronal cell adhesion molecule Nrcam (Nrcam-S), and demonstrated down-regulation of Nrcam-S by specific siRNAs inhibited axon regeneration of Dorsal Root Ganglion (DRG) neurons after sciatic nerve injury in vitro and in vivo. Additionally, we found expression levels of RNA binding proteins (RBPs) CUGBP Elav-like family member 3 (CELF3) and RNA binding protein fox-1 homolog 2 (Rbfox2) were markedly increased after sciatic nerve injury. Our data may serve as a resource useful for further understanding how AS contributes to molecular regulations in DRG during sciatic nerve regeneration.
轴突内在的再生能力对于损伤后外周神经的再生至关重要。鉴定参与这一过程的关键分子,对于研究外周神经损伤修复具有重要意义。可变剪接(AS)是真核基因表达的一种重要调控模式,在生理和病理过程中都得到了广泛的研究。然而,对于 AS 在周围神经再生中的作用知之甚少。在这项工作中,为了鉴定与轴突再生能力相关的 AS 事件,我们通过 RNA 测序(RNA-Seq)和重复转录剪接多元分析(rMATS)分析了坐骨神经损伤修复过程中的 AS 事件。差异 AS 事件进行了基因本体富集和通路分析。此外,我们鉴定了神经元细胞黏附分子 Nrcam(Nrcam-S)的一个明显增加的 AS 事件,并证明了特异性 siRNAs 下调 Nrcam-S 抑制了体外和体内坐骨神经损伤后背根神经节(DRG)神经元的轴突再生。此外,我们发现 RNA 结合蛋白(RBPs)CUGBP 埃拉夫样家族成员 3(CELF3)和 RNA 结合蛋白 FOX-1 同源物 2(Rbfox2)的表达水平在坐骨神经损伤后明显增加。我们的数据可以作为进一步了解 AS 如何在外周神经再生过程中对 DRG 中的分子调控做出贡献的有用资源。