Molecular mechanisms regulating local translation during axon growth and guidance

轴突生长和引导过程中调节局部翻译的分子机制

基本信息

批准号：
9171145
负责人：
Kristy Welshhans
金额：
$ 37.5万
依托单位：
KENT STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9171145
关键词：
Actins Adhesions Affect Behavior Biological Assay Brain Complex Cues Cytoskeletal Modeling Data Development Extracellular Matrix Genetic Translation Goals Growth Growth Cones Integrins Knowledge Link Location Mediating Messenger RNA Microfilaments Molecular Mutation Nature Nervous system structure Neurons Phosphorylation Process Protein Biosynthesis Proteins Regulation Reporter Research Role Scaffolding Protein Signal Pathway Signal Transduction Site Structure Synapses Testing Total Internal Reflection Fluorescent Transcript Translating Translations Tubulin axon growth axon guidance beta Actin graduate student insight live cell imaging member mutant nervous system development nervous system disorder neurodevelopment overexpression paxillin receptors for activated C kinase relating to nervous system small hairpin RNA trafficking undergraduate student

项目摘要

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.

项目概要这项研究的长期目标是了解该建立背后的分子机制发育过程中的神经元连接。具体来说，这项研究探讨了生长锥如何是发育中神经元的寻路结构，迁移到适当的目标并与之连接。神经元连接通过外部和内部线索进行指导，从而导致各种神经元的激活细胞信号通路以及最终的细胞骨架重组。本提案的重点是理解连接发育中的神经系统中的细胞信号传导和局部翻译的分子机制。它最近才发现轴突生长锥的局部翻译对于该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非常有限。本研究重点多功能支架蛋白 RACK1，它可能提供细胞信号传导之间的关键联系以及本地翻译的监管。我们有初步数据表明本地翻译发生在接触，这是生长锥内的粘附位点，对于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附位点，而且是关键的信号中心。在这里，我们假设生长锥内的点接触作为战略位置对于目标本地翻译，RACK1 对此过程至关重要。该提案将提供有价值的研究为本科生和研究生提供研究所发生的基本过程的机会在神经发育过程中。使用先进的活细胞成像、荧光翻译报告基因、shRNA 和轴突生长和引导测定，在这里我们将确定局部翻译是否发生在生长的点接触处锥体，并检查 RACK1 如何调节点接触动力学以及轴突生长和引导。采取总之，这项研究将使我们能够对 mRNA 贩运和本地化如何产生广泛的、机械的理解。翻译有助于神经元连接的建立，从而增加我们的知识大脑发育的复杂性。