FUNCTIONS OF BETA 1 INTEGRINS IN GROWTH CONE MOTILITY

这项建议的长期目标是增加我们对 神经系统发育。这样的理解对于 神经损伤和神经性疾病的治疗设计。这个 实验将(1)检验这一假设：β1整合素是主要的 类细胞外基质受体(ECM)，具有明显的 细胞外基质分子层粘连蛋白(LN)和细胞外基质(ECM)在生长锥运动中的作用 (Fn)和(2)β1整合素之间功能关系的检验 以及Beta1结合蛋白talin。这些实验将使用 发色团辅助激光灭活(CALL)，这是一种独特的方法，将 允许在生长锥体中的Beta1整合素失活 扰乱了细胞上其他地方的Beta1功能。实验策略： (1)我们将证明(CALI)可以用来具体地 灭活β1整合素。(2)我们将使用CALI在以下位置停用Beta1 LN和FN上DRG神经元生长锥的丝状足孔。中的更改 丝状径向运动将由时间推移显微镜和 对重要性进行统计分析。(3)我们将使用CALI来 同时灭活β1整合素和talin，比较 所产生的表型分别为Beta1和talin的失活。这 实验将直接检验两者之间的功能关系 Beta1整合素和talin，一种长期被怀疑的关系，基于 Beta1-talin结合和共定位。

The long term goal of this proposal is to increase our understanding of nervous system development. Such an understanding will be important for the design of treatments for nerve injury and neurological disorders. The experiments will (1) test the hypothesis that beta1 integrins, a major class of receptors for extracellular matrix (ECM), have distinct functions in growth cone motility on the ECM molecules laminin (LN) and (FN) and (2) test for a functional relationship between beta1 integrins and the beta1-binding protein, talin. The experiments will use chromophore assisted laser inactivating (CALl), a unique method that will allow inactivation of beta1 integrins in the growth cone without disrupting beta1 function elsewhere on the cell. Experimental strategy: (1) We will demonstrate that (CALI) can be used to specifically inactivate beta1 integrins. (2) We will use CALI to inactivate beta1 in the filopodia of growth cones of DRG neurons on LN and FN. Changes in filopodial motility will be recorded by time lapse microscopy and analyzed statistically for significance. (3) We will use CALI to inactivate beta1 integrins and talin simultaneously and compare the resulting phenotype to inactivation of beta1 and talin individually. This experiment will be a direct test of the functional relationship between beta1 integrin and talin, a relationship long suspected on the basis of beta1-talin binding and colocalization.