THE ROLE OF AGRIN IN THE DEVELOPMENT OF CENTRAL NEURONS

AGRIN 在中枢神经元发育中的作用

• 批准号: 6874473
• 负责人: ADRIANA B. FERREIRA
• 金额: $ 24.69万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-15 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6874473
• 关键词: agrin; axon; biological signal transduction; dendrites; electron microscopy; fluorimetry; gene induction /repression; gene mutation; growth factor receptors; immunocytochemistry; immunoprecipitation; laboratory mouse; laboratory rat; morphometry; nerve /myelin protein; neurogenesis; phenotype; polymerase chain reaction; posttranslational modifications; protein localization; protein structure function; protein tyrosine kinase; pyramidal cells; synaptogenesis; western blottings

DESCRIPTION (provided by applicant): In the past two decades, a great deal of attention has been directed to the study of synaptogenesis in the mammalian central nervous system. Many of these studies are driven by the belief that the diagnosis and the prevention of synapse loss associated with several neurodegenerative diseases can be achieved through a better understanding of the basic mechanisms of synapse formation in central neurons. Although these mechanisms are not completely understood, it is tempting to speculate that proteins, such as agrin, that play a key role in the formation of the neuromuscular junction might also play an important role in the formation of synaptic contacts between neurons. Recently, we have shown that agrin differentially regulates the rates of axonal and dendritic elongation as well as synapse formation in hippocampal neurons. However, the agrin receptor(s) has yet to be identified in these neurons. The experiments proposed here are intended to test the following hypothesis: ror proteins, two tyrosine kinase receptors considered "orphan receptors", could mediate the effects of agrin on early stages of development in central neurons. A combination of techniques including: Western blot analysis, immunocytochemistry, RT-PCR, the generation of null mutations, binding assays, and immunoprecipitation will be used to analyze: 1) the pattern of expression and localization of ror 1 and ror 2 in central neurons; 2) the role of these tyrosine kinase receptors in neurite elongation and synapse formation; and 3) the participation of ror 1 and ror 2 in the agrin-signaling pathway.

描述（由申请人提供）：在过去的二十年中，大量的注意力已经指向哺乳动物中枢神经系统中突触发生的研究。这些研究中的许多是由以下信念驱动的：通过更好地理解中枢神经元中突触形成的基本机制，可以实现与几种神经退行性疾病相关的突触丧失的诊断和预防。虽然这些机制尚未完全理解，但很容易推测在神经肌肉接头形成中起关键作用的蛋白质（如聚集蛋白）也可能在神经元之间突触接触的形成中起重要作用。最近，我们已经表明，聚集蛋白差异调节率的轴突和树突的伸长，以及突触的形成在海马神经元。然而，聚集蛋白受体在这些神经元中尚未被鉴定。本文提出的实验旨在验证以下假设：ror蛋白，两种被认为是“孤儿受体”的酪氨酸激酶受体，可以介导聚集蛋白对中枢神经元发育早期的影响。将使用蛋白质印迹分析、免疫细胞化学、RT-PCR、无效突变的产生、结合试验和免疫沉淀等技术组合来分析：1）ror 1和ror 2在中枢神经元中的表达和定位模式; 2）这些酪氨酸激酶受体在神经突延伸和突触形成中的作用;和3）ROR 1和ROR 2参与聚集蛋白信号通路。