CYTOSKELETON AND NEURONAL REGENERATION

细胞骨架和神经元再生

• 批准号: 2714604
• 负责人: LINDA M PARYSEK
• 金额: $ 16.29万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2714604
• 关键词: cytoskeletal proteins; gene induction /repression; genetically modified animals; laboratory mouse; nerve injury; nervous system regeneration; nucleic acid sequence; peripheral nervous system

THIS IS A SHANNON AWARD PROVIDING PARTIAL SUPPORT FOR THE RESEARCH PROJECTS THAT FALL SHORT OF THE ASSIGNED INSTITUTE'S FUNDING RANGE BUT ARE IN THE MARGIN OF EXCELLENCE. THE SHANNON AWARD IS INTENDED TO PROVIDE SUPPORT TO TEST THE FEASIBILITY OF THE APPROACH; DEVELOP FURTHER TESTS AND REFINE RESEARCH TECHNIQUES; PERFORM SECONDARY ANALYSIS OR AVAILABLE DATA SETS; OR CONDUCT DISCRETE PROJECTS THAT CAN DEMONSTRATE THE PI'S RESEARCH CAPABILITIES OR LEND ADDITIONAL WEIGHT TO AN ALREADY MERITORIOUS APPLICATION. THE ABSTRACT BELOW IS TAKEN FROM THE ORIGINAL DOCUMENT SUBMITTED BY THE PRINCIPAL INVESTIGATOR. DESCRIPTION: (adapted from Applicant's Abstract) Peripheral nerve regeneration is essential to the restoration of muscle movement, touch sensation, and pain sensation in denervated or damaged tissues. Nerve regeneration, however, is not always successful. To knowledgeably optimize the regeneration process and enhance the success rate, it is essential to understand how genes encoding neuronal components that are key to successful regeneration are activated by nerve injury. The overall goals of the proposed studies are to determine how genes respond to peripheral nerve injury and to determine the importance of one of those genes to the process of nerve regeneration. These studies focus on a gene encoding a subunit of the intermediate filament cytoskeleton, the peripherin gene, the expression of which increases robustly after nerve injury. The proposed experiments will define specific subsequences in the peripherin gene that are required for this response and will reveal mechanisms by which peripherin mRNA increases in response to nerve injury. These nerve injury responses of peripherin will be defined initially by analyzing, in transgenic mice, the nerve injury response of peripherin transgenes that contain deletions in regions suspected to be essential. These regions then will be tested for their ability to confer nerve injury-responsiveness on a heterologous reporter gene. Subsequences in these regions will be further delineated by DNA mobility shift assays that compare the mobility of the subsequences in the presence of nuclear extracts from non-injured and injured neurons. To determine whether the nerve injury response sequences are found in other genes that are similarly activated after nerve injury, several genes within the Genbank database that contain these sequences will be selected and their expression will be tested in nerve-crush studies. Finally, the proposed studies seek to clearly define the requirement for the intermediate filament cytoskeleton during regeneration by determining whether unmyelinated axons that are depleted of the peripherin intermediate filament network in transgenic mice can grow new daughter axons after nerve injury. Identification of factors that bind to the conserved sequences and identification of components that are key to regeneration ultimately will permit enhancement of the regenerative process.

这是香农奖，为研究提供部分支持 与指定研究所的资金范围相差的项目，但 处于卓越的范围。香农奖旨在提供 支持测试方法的可行性；开发进一步的测试 并完善研究技术；进行次要分析或可用 数据集；或执行可以证明PI的离散项目 研究能力或增加本来有功的重量 应用。下面的摘要取自原始文档 由首席调查员提交。 描述：（改编自申请人的摘要）外围神经 再生对于恢复肌肉运动至关重要，触摸 在被取消或受损的组织中的感觉和疼痛感。 神经 但是，再生并不总是成功的。 知识 优化再生过程并提高成功率，这是 了解如何编码神经元成分的基因 成功再生的关键通过神经损伤激活。 拟议研究的总体目标是确定基因 应对周围神经损伤并确定的重要性 这些基因之一是神经再生过程的。 这些研究 专注于编码中间细丝亚基的基因 细胞骨架，外周基因，其表达增加 神经损伤后坚固。 提出的实验将定义 外围蛋白基因中需要的特定子序 反应并将揭示外周mRNA增加的机制 响应神经损伤。 这些神经损伤反应的外围蛋白 最初将通过在转基因小鼠中分析神经来定义 外围蛋白转基因中包含缺失的损伤反应 怀疑是必不可少的地区。 然后将测试这些区域 因为他们能够在异源上赋予神经损伤响应性 记者基因。 这些地区的子序列将进一步划定 通过DNA移动性转移测定法，比较了 在存在未造成的核提取物的情况下的子序列 受伤的神经元。 确定神经损伤是否反应 序列是在其他类似激活之后被激活的基因中发现的 神经损伤，GenBank数据库中的几个基因 这些序列将被选中，并将其表达式测试 神经灌溉研究。 最后，拟议的研究试图清楚 定义对中间细丝细胞骨架的要求 通过确定是否耗尽的不髓鞘轴突来再生 转基因小鼠中的外围蛋白中间细丝网络可以 神经损伤后成长新女儿轴突。识别因素 与保守序列结合并识别组件 这是再生的关键，最终将允许增强 再生过程。