Regenerative and degenerative responses to axonal injury

对轴突损伤的再生和退行性反应

• 批准号: 10739408
• 负责人: KENNETH M CADIGAN
• 金额: $ 3.66万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10739408
• 关键词: Axon; Biochemical; Biomedical Research; Brain; Collaborations; Drosophila genus; Exposure to; Glutamates; Goals; Grant; In Vitro; Mediating; Mediator; Mentorship; Methods; Michigan; Mission; Molecular Probes; Mus; Neurons; Outcome; Pathway interactions; Pharmacology; Phenotype; Phosphotransferases; Postbaccalaureate; Regulation; Research; Research Support; Signal Transduction; Stress; Students; Synapses; Universities; Work; axon injury; axon regeneration; biological adaptation to stress; career development; cellular imaging; disadvantaged background; experimental study; genetic approach; genetic manipulation; in vivo; medical schools; neuron loss; parent grant; pharmacologic; regenerative; response

Project Summary/Abstract: Axons form connections between neurons over great distances in the brain and body, hence are vulnerable to damage and stress. This project studies an evolutionarily conserved stress response pathway that becomes activated in multiple scenarios of axonal damage and stress. The pathway, governed by the dileucine zipper kinase DLK, known as Wallenda (Wnd) in Drosophila, engages structural plasticity mechanisms in neurons that allow circuits to adapt to axon damage. These responses include axonal regeneration, neuronal death, and, newly discovered in this project, synapse loss. The long-term goals of this project are (1) to understand the mechanisms that lead to DLK signaling activation, and (2) to understand the cellular pathways that are regulated by DLK. The parent grant uses genetic manipulations of DLK/Wnd in mice and Drosophila, with a focus on in vivo phenotypes. However, to understand the mechanism(s) of DLK’s regulation (an overarching goal of Aim 1) and how it mediates diverse outcomes (a central goal of Aim 2), we sought to develop an in vitro paradigm for studying DLK that is amenable to biochemical, pharmacological and genetic approaches. As discussed in the April 2022 RRPR, we have now established robust methods to activate DLK signaling within primary cortical neuron cultures. The new methods provide new inroads to advancing the advancing both aims of the original grant. Pharmacology experiments are used to probe the molecular mediators of DLK signaling activation (Aim 1), and cellular imaging and biochemical experiments are used to probe how DLK activation can mediate distinct outcomes in the cultures (Aim 2). This endeavor is carried out in collaboration with Sami Barmada at the University of Michigan (UM) School of Medicine. The supplement application seeks to hire Yelitzza Aguilar, whose postbaccalaureate work on this project (in the Barmada lab at UM) will study the mechanism by which DLK mediates neuronal death in cortical neuron cultures exposed to excess glutamate. The proposed supplemental work will advance the mission of increasing diversity in biomedical research by supporting the mentorship and career development of a promising student from a disadvantaged background in biomedical research.

项目摘要/摘要： 轴突在大脑和身体的距离上形成神经元之间的连接，因此很容易受到影响 损害和压力。该项目研究了一种进化组成的压力响应途径，该途径成为 在轴突损伤和压力的多种情况下激活。该路径由Dileucine拉链管理 激酶DLK，被称为果蝇中的wallenda（WND），参与神经元中的结构可塑性机制 这使电路可以适应轴突损伤。这些反应包括轴突再生，神经元死亡， 而且，在这个项目中新发现的突触损失。该项目的长期目标是（1）了解 导致DLK信号激活的机制，以及（2）了解细胞途径 由DLK调节。父母赠款在小鼠和果蝇中使用DLK/WND的遗传操作， 专注于体内表型。但是，要了解DLK法规的机制（总体 目标的目标1）及其如何介导潜水员的结果（目标2的主要目标），我们试图开发体外 用于研究DLK的范式，该DLK适合生化，药物和遗传方法。作为 在2022年4月的RRPR中讨论，我们现在已经建立了强大的方法来激活DLK信号传导 原发性皮质神经元培养物。新方法为促进前进的目标提供了新的侵害 原始赠款。药理学实验用于探测DLK信号传导的分子介质 激活（AIM 1），细胞成像和生化实验用于探测DLK如何激活 可以调节文化中的不同结果（AIM 2）。这项工作是与萨米合作进行的 密歇根大学（UM）医学院的Barmada。补充申请旨在雇用 Yelitzza Aguilar，他的后库后工作（在UM的Barmada Lab）将研究 DLK在暴露于谷氨酸过多的皮质神经元培养物中介导神经元死亡的机制。 拟议的补充工作将推动通过增强生物医学研究多样性的使命 支持来自灾难背景的承诺学生的精神训练和职业发展 在生物医学研究中。