eIF3f与olig1结合及其在少突胶质前体细胞分化成熟中的作用研究

Myelin is crucial for neural system functioning. Myelination insulates axons and facilitate saltatory conduction of action potentials. Within the central nervous system, oligodendrocytes are responsible for the formation of the myelin sheath. Differentiation of oligodendrocyte progenitor cells into mature oligodendrocytes is a prerequisite for myelination during development and remyelination after demyelination in many diseases. Although the crital role of oligodendrocyte transcription factor1 (oligo1) in the differentiation and maturation of oligodendrocyte precursor cells has been demonstrated, the underlying mechanism remains elusive. In a yeast two-hybrid screening, we identified eukaryotic translation initiation factor 3 subunit f (eIF3f) is a new binding partner of olig1, and our preliminary data strongly indicate that eIF3f could bind to olig1 to regulate the differentiation and maturation of oligodendrocyte precursor cells. In this proposal, we aim to further confirm the interaction between eIF3f and olig1, to characterize the interaction sites, and to explore their roles in regulation of the differentiation and maturation of oligodendrocyte precursor cells. The goal of this research is to establish the role of eIF3f in regulation of the differentiation and maturation of oligodendrocyte precursor cells, to open up new possibilities to achieve successful therapeutic explorations for myelin repair.

髓鞘是维持神经正常生理活动的重要结构基础。少突胶质前体细胞分化成熟是髓鞘形成的前提，也是髓鞘再生的关键。少突胶质细胞转录因子-1(olig1)在少突胶质前体细胞分化成熟以及髓鞘形成中发挥重要作用,然而其作用机制尚不清楚。本项目从源头入手，通过酵母双杂交筛选鉴定，发现真核细胞翻译起始因子3-亚基f(eIF3f)是olig1新的结合分子；我们的前期实验还显示，eIF3f极有可能通过与olig1结合来调控少突胶质前体细胞分化成熟。本项目拟在前期实验基础上，进一步确证eIF3f与olig1相互作用，明确其相互作用的结构基础，阐明其介导的生物学效应及其作用机制。从而深入揭示少突胶质前体细胞分化成熟的机制，并可能为促进髓鞘再生与修复提供新思路。

髓鞘是维持神经正常生理活动的重要结构基础。少突胶质前体细胞分化成熟是髓鞘形成的前提，也是髓鞘再生的关键。少突胶质细胞转录因子-1(olig1)在少突胶质前体细胞分化成熟以及髓鞘形成中发挥重要作用,然而其作用机制尚不清楚。本项目通过酵母双杂交筛选鉴定，发现真核细胞翻译起始因子3-亚基f(eIF3f)是olig1新的结合分子，进一步的免疫共沉淀等生化实验确证了olig1与eIF3f存在相互作用；通过运用蛋白质结构预测和小肽模拟实验，揭示了olig1与eIF3f存在相互作用的潜在位点; 运用免疫组织化学等方法，明确了eIF3f的表达与定位特征及其在脱髓鞘模型中的变化；通过在体外培养少突胶质细胞前体细胞中过表达和基因沉默实验，我们发现eIF3f能够促进少突胶质细胞前体细胞分化成熟，而且这种作用有可能通过eIF3f与Olig1结合竞争性抑制TIP30/Olig1复合体的形成，或者通过调节olig1蛋白的表达量来实现的；运用斑马鱼在体成像技术，我们发现在神经系统发育过程中，eIF3f能够促进少突胶质细胞前体细胞分化成熟和髓鞘形成。本项目研究不仅有助于深入揭示少突胶质前体细胞分化成熟的机制，而且还能够为促进髓鞘再生与修复提供新的理论依据。