Discovery for the Target Molecules to Control. Glial Scar Formation after Spinal Cord Injury

发现要控制的目标分子。

• 批准号: 16390433
• 负责人: YAMAMOTO Shinichi
• 金额: $ 8.7万
• 依托单位: Research Institute, National Rehabilitation Center
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390433/
• 关键词: spinal cord injury; oligodendrocyte; precursor cells; primary culture; differentiation; gene transfer

Spinal cord injury is one of the most severe trauma in central nervous systems, resulting several thousand new patients every year. From the view point of innovating new therapeutic approach for this injury, the proper control of "Glial Scar" is very important because the scar is believed to interrupt regenerating axons. In peripheral nervous systems, which can regenerate after injury, Schwann cells, myelinating cell in peripheral nerve, proliferate after injury and they occupy the damaged area. The regenerating axons go though "Schwann cell packed area". Therefore, we put focus on controlling oligodendrocyte precursor cells, myelinating cells in central nervous systems, so that they occupy the damaged area in spinal cord and help regenerating axons.In this research project, we first worked on the establishment of primary culture of mouse oligodendrocyte, which enable us to reveal molecular function by in vitro experiments. Oligodendrocytes are known to proliferate in immature form if … More the culture condition contain enough growth factors. It is not known whether growth factors convey any specific effect to keep the cells in immature status. From the protein expression analysis, we chose Cyclin D1 molecule, well known cell cycle promoting molecule, because its protein level shows strong correlation with immature status of oligodendrocyte. Molecular function of Cyclin D1 on oligodendrocyte maturation is investigated by cloning Cyclin D1, generating mutant forms of the molecule, gene transfer to the cells and analyzing differentiation markers. We found that Cyclin D1 has distinct mechanism to control oligodendrocyte maturation apart from its cell-cycle regulating function. For the future studies, we are planning to introduce these finding to animal model of spinal cord injury to see whether we can control proliferating oligodendrocyte in after spinal cord injury and prevent glial scar formation.We also perform DNA microarray analysis to survey the molecules which is relevant to immature status of oligodendrocyte. Our primary culture of mouse oligodendrocyte is helpful and we could identified 20 genes which is possibly related to oligodendrocyte maturation. We are planning to explore molecular function of each molecules in the future. Less

脊髓损伤是中枢神经系统最严重的创伤之一，每年都会导致数千名新患者。从创新治疗这种损伤的新方法的角度来看，正确控制“胶质疤痕”非常重要，因为疤痕被认为会中断轴突的再生。在周围神经系统中，受伤后可以再生，周围神经中的髓鞘细胞雪旺细胞在受伤后增殖并占据受损区域。再生轴突穿过“施万细胞聚集区”。因此，我们重点关注控制中枢神经系统中的少突胶质前体细胞、髓鞘细胞，使其占据脊髓受损区域，帮助轴突再生。在本研究项目中，我们首先致力于小鼠少突胶质细胞原代培养物的建立，使我们能够通过体外实验揭示分子功能。众所周知，如果培养条件含有足够的生长因子，少突胶质细胞就会以不成熟的形式增殖。目前尚不清楚生长因子是否具有任何特定作用以保持细胞处于未成熟状态。从蛋白表达分析来看，我们选择了众所周知的细胞周期促进分子Cyclin D1分子，因为它的蛋白水平与少突胶质细胞的未成熟状态有很强的相关性。通过克隆 Cyclin D1、产生分子突变形式、将基因转移至细胞并分析分化标记，研究了 Cyclin D1 对少突胶质细胞成熟的分子功能。我们发现Cyclin D1除了具有细胞周期调节功能外，还具有独特的控制少突胶质细胞成熟的机制。在未来的研究中，我们计划将这些发现引入到脊髓损伤的动物模型中，看看是否可以控制脊髓损伤后少突胶质细胞的增殖，预防胶质疤痕的形成。我们还进行DNA微阵列分析，以调查与少突胶质细胞未成熟状态相关的分子。我们对小鼠少突胶质细胞的原代培养很有帮助，我们可以鉴定出 20 个可能与少突胶质细胞成熟相关的基因。我们计划将来探索每个分子的分子功能。较少的