Molecular mechanisms regulating local translation during axon growth and guidance

项目总结 这项研究的长期目标是了解其形成的分子机制。 在发育过程中神经元的连通性。具体地说，这项研究考察了增长锥体是如何 是发育中神经元的寻路结构，迁移到适当的靶点并与之相连。 神经元的连接是通过外部和内部线索来引导的，这导致了各种 细胞信号通路和最终的细胞骨架重组。这项建议侧重于理解 神经系统发育过程中细胞信号和局部翻译的分子机制。它 直到最近才发现轴突生长锥体中的局部平移对于这些过程是必要的 轴突的生长和引导。因此，我们对本地翻译的理解是非常有限的。这项研究的重点是 在多功能支架蛋白RACK1上，RACK1可能在细胞信号之间提供关键联系 以及对当地翻译的规范。我们有初步数据表明，这种本地翻译发生在 接触，这是生长锥体内的粘连部位，对适当的轴突生长和 指导。因此，我们的初步数据表明，点接触不仅是粘连部位，而且是至关重要的 信号中心。这里，我们假设生长锥体内的点接触是一个战略位置。 对于目标本地翻译和RACK1，RACK1对此过程至关重要。这项建议将提供有价值的研究 本科生和研究生都有机会学习发生的基本过程 在神经发育过程中。使用先进的活细胞成像、荧光翻译记者、shRNA和 轴突生长和引导分析，这里我们将确定局部平移是否发生在生长中的点接触处 并研究RACK1如何调节点接触动力学以及轴突生长和引导。已被占用 总之，这项研究将使人们能够广泛地、机械性地理解信使核糖核酸的贩运和当地 翻译有助于建立神经元的连接，从而增加我们对 大脑发育的复杂性。

PROJECT SUMMARY The long-term goal of this research is to understand the molecular mechanisms underlying the establishment of neuronal connectivity during development. Specifically, this research examines how the growth cone, which is the pathfinding structure of the developing neuron, migrates to and connects with its appropriate targets. Neuronal connectivity is directed via both external and internal cues, which results in the activation of various cellular signaling pathways and ultimately, cytoskeletal reorganization. This proposal focuses on understanding the molecular mechanisms linking cellular signaling and local translation in the developing nervous system. It has only recently been discovered that local translation in axonal growth cones is necessary for the processes of axon growth and guidance. As such, our understanding of local translation is very limited. This study focuses on a multifunctional scaffolding protein, RACK1, which likely provides a critical link between cellular signaling and the regulation of local translation. We have preliminary data that this local translation takes place at point contacts, which are adhesion sites within growth cones that are essential for appropriate axon growth and guidance. Thus, our preliminary data suggests that point contacts are not just adhesion sites, but also critical signaling centers. Here, we hypothesize that point contacts within growth cones serve as a strategic location for targeted local translation and RACK1 is critical to this process. This proposal will provide valuable research opportunities for both undergraduate and graduate students to study fundamental processes that take place during neural development. Using advanced live cell imaging, fluorescent translation reporters, shRNA and axon growth and guidance assays, here we will determine if local translation occurs at point contacts in growth cones, and examine how RACK1 regulates point contact dynamics and axon growth and guidance. Taken together, this research will enable a broad, mechanistic understanding as to how mRNA trafficking and local translation contributes to the establishment of neuronal connectivity, thereby increasing our knowledge about the complex nature of brain development.