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，充当膜相关分子和底层细胞骨架之间的连接体。最近，人们发现了 ERM 蛋白 ezrin 和发育上重要的粘附分子 L1 之间的结合相互作用。此外，ERM 蛋白功能的破坏已被证明会影响细胞培养模型中的轴突生长。由于粘附分子的底物无法在体外重现，因此目前不可能通过 ERM 蛋白与粘附分子和细胞骨架的同时相互作用来充分探索 ERM 蛋白在轴突发育中的假定作用。在该提案的目标 1 中，将通过免疫组织化学和蛋白质印迹来测试粘附分子和 ERM 之间相互作用的合理性，这与发育中的大脑皮层中的轴突萌芽相关。在目标 2 中，ERM 显性失活、组成型活性 ERM 和对照将在轴突通过子宫内基因转移投射到大脑皮层中间区的神经元中表达。对表达功能丧失、功能获得和对照结构的动物的轴突发育标志物和形态进行比较显微镜分析将阐明轴突生长中的 ERM 功能。对 ERM 功能的完整了解不仅有助于发育神经生物学，而且有助于建立知识体系，如果要通过发育重演进行轴突再生来治疗中风、阿尔茨海默病和脊髓损伤等疾病，则可能需要这些知识体系。