CYTOSKELETON AND NEURONAL REGENERATION

细胞骨架和神经元再生

• 批准号: 2892085
• 负责人: LINDA M PARYSEK
• 金额: $ 21.35万
• 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/2892085
• 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.

这是为研究提供部分支持的香农奖 未达到指定机构资助范围但 处于卓越边缘。香农奖旨在提供 支持测试该方法的可行性；开展进一步的测试 并完善研究技术；进行二次分析或可用 数据集；或者进行可以展示 PI 的离散项目 研究能力或对已有的功绩给予额外的重视 应用。以下摘要摘自原始文件 由首席研究员提交。 描述：（改编自申请人的摘要）周围神经 再生对于恢复肌肉运动、触觉至关重要 失去神经或受损组织的感觉和疼痛感。 神经 然而，再生并不总是成功。 知识渊博 优化再生流程，提高成功率， 对于理解基因如何编码神经元成分至关重要 成功再生的关键是由神经损伤激活。 拟议研究的总体目标是确定基因如何 应对周围神经损伤并确定其重要性 这些基因之一与神经再生过程有关。 这些研究 关注编码中间丝亚基的基因 细胞骨架，外周蛋白基因，其表达增加 神经损伤后恢复强劲。 拟议的实验将定义 外周蛋白基因中为此所需的特定子序列 反应并将揭示外周蛋白 mRNA 增加的机制 以应对神经损伤。 外周蛋白的这些神经损伤反应 最初将通过分析转基因小鼠的神经来定义 含有缺失的外周蛋白转基因的损伤反应 被怀疑是必要的区域。 然后将测试这些区域 因为它们能够赋予异源神经损伤反应性 报告基因。 这些区域的后续序列将被进一步描绘 通过 DNA 迁移率变动分析比较 存在来自未受伤和的核提取物的子序列 受伤的神经元。 判断是否有神经损伤反应 在其他基因中也发现了类似的序列 神经损伤，Genbank 数据库中的几个基因包含 将选择这些序列并测试它们的表达 神经挤压研究。 最后，拟议的研究旨在明确 定义过程中对中间丝细胞骨架的要求 通过确定无髓鞘轴突是否耗尽来再生 转基因小鼠的外周蛋白中间丝网络的 神经损伤后长出新的子轴突。因素识别 与保守序列结合并识别成分 这对再生至关重要，最终将增强 再生过程。