The Role of ERM Proteins in Axonal Development

ERM 蛋白在轴突发育中的作用

• 批准号: 6927891
• 负责人: Cyrus David Mintz
• 金额: $ 3.07万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-04 至 2006-08-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6927891
• 关键词: actin binding protein; axon; cell adhesion molecules; central nervous system; cytoskeletal proteins; developmental neurobiology; electroporation; embryo /fetus tissue /cell culture; genetic manipulation; green fluorescent proteins; immunocytochemistry; laboratory rat; membrane proteins; nervous system regeneration; polymerase chain reaction; protein binding; protein structure function; site directed mutagenesis; synaptogenesis; western blottings

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.

描述（由申请人提供）：Ezrin，radixin和Moesin（统称为ERM蛋白）作为膜相关分子和基础细胞骨架之间的接头。最近，已经发现了ERM蛋白，Ezrin和发育重要的粘附分子L1之间的结合相互作用。此外，已证明ERM蛋白功能的破坏会影响细胞培养模型中的轴突生长。由于无法在体外概括粘附分子的底物，因此目前无法通过与轴突发育中的粘附分子和细胞骨架的同时相互作用，目前无法通过与粘附分子和细胞骨架的同时相互作用来充分探索ERM蛋白的假定作用。在本提案的目标1中，将通过免疫组织化学和蛋白质印迹测试与发育中的大脑皮层中轴突发芽有关的粘附分子与ERMS之间相互作用的合理性。在AIM 2中，ERM主动性主动性ERM和对照将在神经元中表达，其轴突通过子宫基因转移在轴突中投射到大脑皮层的中间区域。表达功能丧失，功能增益和控制构建体的动物中轴突发育标记和形态的比较显微镜分析将阐明轴突生长中的ERM功能。对ERM功能的完全了解不仅会促进发育神经生物学，而且还会有助于知识体体，如果轴突再生通过概括发育而出于疾病，例如中风，阿尔茨海默氏病和脊髓损伤的治疗原因而进行。