Visualizing axonal delivery and removal of tubulin

可视化轴突输送和微管蛋白去除

• 批准号: 10647032
• 负责人: Shaul Yogev
• 金额: $ 20.94万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10647032
• 关键词: Aging; Axon; Brain; Caenorhabditis elegans; Cells; Consensus; Cytoskeletal Proteins; Cytoskeleton; Dendrites; Development; Diffusion; Disease; Disease model; Elements; Excision; Fluorescence; Frequencies; Functional disorder; Genetic; Genome engineering; Goals; Homeostasis; Hour; Image; Immobilization; Knowledge; Length; Maintenance; Mass Spectrum Analysis; Measures; Methodology; Microtubule Polymerization; Microtubules; Modeling; Movement; Nature; Neurons; Outcome; Photobleaching; Physiologic pulse; Physiological; Play; Polymers; Production; Proteins; Radiolabeled; Reagent; Research; Resolution; Role; Spectrin; Speed; Study models; System; Time; Tubulin; Visualization; axon growth; dimer; experimental study; imaging study; in vivo; insight; nervous system disorder; neurodevelopment; novel; pharmacologic; prevent; slow axonal transport; stem; superresolution microscopy; tool

Project Summary Tubulin plays fundamental roles in axon formation, maintenance, and dysfunction. Despite this, how tubulin is delivered to axons remains highly controversial, and how tubulin is removed from axons is entirely unknown. This is due to technical challenges in visualizing tubulin movement, that stem from the fact that tubulin is highly abundant in immobile polymers and that when movement occurs, it is extremely slow. We recently overcame similar obstacles to the visualization of spectrin delivery to axons. Here we propose to modify, adapt and validate our methodology in order to generate a new probe for visualizing the axonal delivery and removal of endogenous tubulin, in vivo, with single-axon resolution. Successful completion of this project will reveal how tubulin is delivered and removed from axons, and how delivery and removal are balanced to generate a steady state cytoskeletal mass under different physiological conditions. This knowledge and tools will move the field forward by opening the way for more mechanistic and disease-focused studies. Furthermore, the approach could be adapted to visualize the axonal delivery and removal of other slow axonal transport cargo.

项目概要 微管蛋白在轴突形成、维持和功能障碍中发挥重要作用。尽管如此，微管蛋白如何 递送到轴突仍然存在很大争议，并且微管蛋白如何从轴突中去除也是完全未知的。 这是由于微管蛋白运动可视化的技术挑战，这是由于微管蛋白高度 富含固定聚合物，当发生运动时，速度极其缓慢。我们最近克服了 血影蛋白传递至轴突的可视化存在类似的障碍。在此我们建议修改、调整和 验证我们的方法，以生成一种新的探针，用于可视化轴突传递和去除 体内内源性微管蛋白，具有单轴突分辨率。该项目的成功完成将揭示如何 微管蛋白从轴突中递送和去除，以及如何平衡递送和去除以产生稳定的微管蛋白 状态不同生理条件下的细胞骨架质量。这些知识和工具将推动该领域的发展 通过为更多的机制和以疾病为重点的研究开辟道路来向前发展。此外，该方法 可以适应轴突输送和其他缓慢轴突运输货物的移除的可视化。