CELL SURFACE MOLECULES INVOLVED IN NEURITE EXTENSION

参与神经突延伸的细胞表面分子

基本信息

批准号：
3412409
负责人：
SUSANNAH S CHANG
金额：
$ 13万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
1988
资助国家：
美国
起止时间：
1988-09-01 至 1991-08-31
项目状态：
已结题

项目摘要

During development of the nervous system, neuronal growth cones extend through a complex environment to accurately find their proper target area. Among the features of the environment which growth cones encounter are the axons of other, earlier differentiating neurons. Since growth cones are frequently observed in vivo to associate with other axons, we have chosen to focus our research on growth cone-axon interactions. The long term objective of this research is to isolate cell surface molecules which are localised on neuronal processes and which promote neurite extension. A bioassay which measures neurite extension on axons has been developed. This assay will be used to functionally screen monoclonal antibodies which have first been selected because they are localised on the cell surface and are different from known molecules. Monoclonal antibodies will be generated against crude membrane preparations, or against membrane proteins which have been fractionated by biochemical methods. In conjunction with monoclonal antibody generation, the bioassay will be expanded to include other neural tissues, and new bioassays will be developed. To directly identify membrane proteins which support neurite outgrowth, brain membrane proteins will be fractionated by gel electrophoresis and blotted onto nitrocellulose. Neural cells will be cultured on the blots and assessed for neurite outgrowth.

在神经系统的发展过程中，神经元生长锥遍历复杂的环境，以准确找到他们的适当的目标区域。在环境的特征中生长锥相遇是其他早期的轴突区分神经元。由于生长锥经常在体内观察到与其他轴突相关，我们选择了将我们的研究集中在生长锥形相互作用上。这项研究的长期目标是分离细胞表面位于神经元过程的分子，这些分子促进神经突延伸。测量神经突的生物测定已经开发了轴突的延伸。该测定将习惯在功能上筛选单克隆抗体，首先是之所以选择，是因为它们位于细胞表面，并且是与已知分子不同。单克隆抗体将是针对粗膜制剂或反对膜蛋白已被生化分馏方法。与单克隆抗体的产生结合生物测定将扩展到包括其他神经组织，新的生物测定将开发。直接识别膜支持神经突生长的蛋白质，脑膜蛋白质将被凝胶电泳分馏并印迹进入硝酸纤维素。神经细胞将在印迹上培养，评估神经突生长。