Molecular mechanism underlying promotion of neuronal regeneration

本研究旨在探讨“促进神经元再生的分子机制”。主要研究两个主题：(1)与神经元再生相关的转录因子和表观遗传因子的鉴定；(2)促进神经元再生的蛋白修饰的鉴定。我们发现转录因子如ATF3， cJun和STAT3对促进神经再生至关重要，并进一步试图了解这些转录因子使用神经再生相关分子之一的启动子，损伤诱导的神经元内肽酶DINE的相互作用。这一分析表明，这些转录因子似乎并没有直接结合启动子区域，而是表明存在一种用于这些转录因子组装的平台分子。本研究发现的转录调控可能是神经再生的特异性调控，并将为促进神经再生提供一种新的、更多的转录机制。我们还试图确定促进神经再生的蛋白质修饰，如磷酸化、甲基化和乙酰化。为了鉴定对神经损伤反应的修饰分子，我们使用了2-DG电泳和抗体印迹，例如可以识别某个激酶特异性磷酸化位点。对表达上调的候选位点进行TOFMAS分析。我们已经确定pherpherin是Akt对神经损伤的磷酸化分子。虽然磷酸化的功能意义尚不清楚，但本实验建立的方法可以作为鉴定磷酸化、甲基化或乙酰化修饰的蛋白质的良好工具。本项目的结果为我们提供了一种新的神经再生的转录机制，也为鉴定磷酸化、甲基化和乙酰化的分子修饰提供了有用的工具。少

This study was carried out to investigate "Molecular mechanism underlying promotion of neuronal regeneration". Two major topics were investigated : (1) Identification of transcription factors and epigenetic factors, which are associated with neuronal regeneration, (2) Identification of protein modification, which promotes neuronal regeneration. We have revealed that transcription factors such as ATF3, cJun and STAT3 were crucial to promote nerve regeneration, and further tried to understand the interaction of those transcription factors using the promoter for one of nerve regeneration associated molecules, damage induced neuronal endopeptidase DINE. This analysis demonstrated that those transcription factors seemed not to bind the promoter region directly, and instead suggested the existence of a sort of platform molecule for the assembly of those transcription factors. The transcriptional control identified in this study may be specific to nerve regeneration and would provide a novel … More transcription mechanism for the promotion of nerve regeneration. We have also attempted to identify protein modifications such as phosphorylation, methylation, and acetylation for the promotion of nerve regeneration. To identify the modified molecules in response to nerve injury, we used 2-DG electrophoresis and blotted with antibodies, which recognized a certain kinase specific phosphorylation site for instance. The candidate spots, which showed up-regulation of the modification were further analyzed by TOFMAS. We have identified pheripherin as a phosphorylated molecule by Akt in response to nerve injury. Although the functional significance of the phosphorylation is not clear, the method established in this experiment could be a good tool for the identification of the proteins, which are modified by phosphorylation, methylation, or acetylation. The results obtained in the present project provide us with a novel transcription mechanism underlying the nerve regeneration, and also provide useful tool for the identification of molecular modification by phosphorylation, methylation, and acetylation. Less