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Molecular mechanism of axonal regeneration-Role of receptor of repulsive factor for neurite elongation-

轴突再生的分子机制-神经突伸长排斥因子受体的作用-

基本信息

批准号：
06454699
负责人：
MURAKAMI Fujio
金额：
$ 3.01万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

Axons in the central nervous system of mature mammals generally fail toregenerate following injury. Although the reason for this regenerative failure remains unknown, several lines of evidence suggest that it is dueto non-permissiveness of oligodendrocytes for axonal elongation. In recentyears, repulsive factors derived from oligodendrocytes have been thought to be important for studying this issue. It was shown that application of monoclonal antibodies to these molecules made growth cones to retract or avoid on contact with oligodendrocytes. This finding suggests the existence of receptor for repulsive factors.The aim of this project was to elucidate the mechanisms of axonal regeneration by identifying this presumable receptor. As a strategy, we used monoclonal antibody method and functional screening method.We cultured dorsal root ganglia of the chick embryo. Growth cones were isolated, homogenized and growth cone-specific fractions were isolated by gel electrophoresis or column chromatography. Mise were immunized with this antigen. Spleen cells were hybridized with myeloma and hybridomas secreting specific antibodies were screened. We applied monoclonal antibodies produced by the hybridomas and clones whose produced antibodies made growth cones to retract were selected. We have developed time-lapse video analysis system and used this system effectively in the present study.

成熟哺乳动物的中枢神经系统中的轴突在受伤后通常不能再生。尽管再生失败的原因尚不清楚，但一些证据表明这是由于少突胶质细胞不允许轴突延长所致。近年来，来自少突胶质细胞的排斥因子被认为是研究这一问题的重要因素。结果表明，应用抗这些分子的单抗可使生长锥体收缩或避免与少突胶质细胞接触。这一发现表明排斥因子受体的存在。本项目的目的是通过鉴定这种可能的受体来阐明轴突再生的机制。作为策略，我们采用了单抗方法和功能筛选方法，培养了鸡胚胎背根节。分离生长锥，匀浆，用凝胶电泳法或柱层析法分离生长锥特异组分。用该抗原免疫MISE。将脾细胞与骨髓瘤细胞杂交，筛选出分泌特异性抗体的杂交瘤细胞。我们应用杂交瘤产生的单抗，筛选出产生抗体使生长锥缩回的克隆。我们开发了延时视频分析系统，并在本研究中有效地使用了该系统。