Visualizing axonal delivery and removal of tubulin

项目摘要 微管蛋白在轴突的形成、维持和功能障碍中起着重要作用。尽管如此，微管蛋白是如何 向轴突输送微管蛋白仍然存在很大争议，而且微管蛋白是如何从轴突中移除的完全未知。 这是由于在可视化微管蛋白运动方面的技术挑战，这源于微管蛋白高度 富含固定的聚合物，当运动发生时，它是极其缓慢的。我们最近克服了 向轴突传递幽灵蛋白的可视化遇到了类似的障碍。在这里，我们建议修改、适应和 验证我们的方法，以便产生一个新的探针，用于可视化轴突的传递和移除 内源性微管蛋白，体内，具有单一轴突分辨率。这个项目的成功完成将揭示 微管蛋白是从轴突传递和移除的，以及传递和移除如何平衡以产生稳定的 不同生理条件下细胞骨架质量的状态。这些知识和工具将推动这一领域的发展 通过为更机械化和以疾病为重点的研究开辟道路。此外，这种方法 可用于可视化轴突输送和移除其他缓慢的轴突运输货物的过程。

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.