The Role of ERM Proteins in Axonal Development

描述(由申请人提供)：Ezrin、Radioxin和Moesin统称为ERM蛋白，作为膜相关分子和底层细胞骨架之间的连接物。最近，ERM蛋白Ezrin和发育重要的黏附分子L1之间的结合作用已被发现。此外，在细胞培养模型中，ERM蛋白功能的中断已被证明影响轴突生长。由于黏附分子的底物不能在体外重述，目前还不可能通过ERM蛋白与黏附分子和细胞骨架的同时相互作用来充分探索ERM蛋白在轴突发育中的可能作用。在这项提议的目标1中，将用免疫组织化学和Western blotting来测试黏附分子和ERM之间相互作用的可能性，这与发育中的大脑皮层中的轴突萌发有关。在目标2中，通过宫内基因转移，在轴突投射到大脑皮层中间带的神经元中表达ERM显性阴性、成分活跃的ERM和对照。对表达功能丧失、功能获得和控制结构的动物的轴突发育标志物和形态进行比较显微分析，将阐明ERM在轴突生长中的作用。对ERM功能的全面了解不仅有助于发育神经生物学，而且有助于掌握可能需要的知识，如果为了治疗中风、阿尔茨海默病和脊髓损伤等疾病而通过概括发育来进行轴突再生的话。

DESCRIPTION (provided by applicant): Ezrin, radixin, and moesin, collectively known as the ERM proteins, serve as linkers between the membrane associated molecules and the underlying cytoskeleton. Recently, a binding interaction between the ERM protein, ezrin, and the developmentally important adhesion molecule, L1, has been discovered. Furthermore, disruption of ERM protein function has been shown to affect axonal outgrowth in cell culture models. Because the substrate for adhesion molecules cannot be recapitulated in vitro, it is not currently possible to fully explore the putative role of ERM proteins, via their simultaneous interactions with adhesion molecules and the cytoskeleton, in axonal development. In Aim 1 of this proposal the plausibility of an interaction between adhesion molecules and ERMs, which is relevant to axonal sprouting in the developing cerebral cortex, will be tested with immunohistochemistry and Western blotting. In Aim 2, an ERM dominant negative, a constitutively active ERM, and controls will be expressed in neurons whose axons project into the intermediate zone of the cerebral cortex via in utero gene transfer. Comparative microscopic analysis of axonal development markers and morphology in animals expressing loss of function, gain of function, and control constructs will elucidate ERM function in axon outgrowth. A complete understanding of ERM function will contribute not only to developmental neurobiology, but also to the body of knowledge that will likely be needed if axonal regeneration by recapitulation of development is to be undertaken for therapeutic reasons in diseases such as stroke, Alzheimer's disease and spinal cord injury.