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和膜突蛋白，统称为ERM蛋白，作为膜相关分子和底层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，已显示ERM蛋白功能的破坏影响细胞培养模型中的轴突生长。由于粘附分子的底物不能在体外重现，因此目前还不能完全探索ERM蛋白在轴突发育中通过与粘附分子和细胞骨架同时相互作用的假定作用。在本提案的目标1中，将用免疫组织化学和蛋白质印迹法测试粘附分子和ERMs之间的相互作用的可行性，这与发育中的大脑皮层中的轴突发芽有关。在目标2中，ERM显性阴性、组成型活性ERM和对照将在神经元中表达，所述神经元的轴突通过子宫内基因转移投射到大脑皮层的中间区中。在表达功能丧失、功能获得和对照构建体的动物中轴突发育标志物和形态学的比较显微镜分析将阐明ERM在轴突生长中的功能。对ERM功能的完整理解不仅有助于发育神经生物学，而且有助于知识体系，如果通过发育的重演进行轴突再生以用于诸如中风、阿尔茨海默病和脊髓损伤等疾病的治疗原因，则可能需要该知识体系。