CYTOSKELETAL REGULATION DURING GROWTH CONE GUIDANCE

描述(申请人提供)：在胚胎发育过程中，大量神经元必须与其适当的目标形成联系。轴突由生长锥引导到它们的目标，生长锥整合引导信号并将其转化为运动所需的机械力。运动产生于细胞骨架聚合物和相关蛋白质的动态调节。Ena/Vasp蛋白家族在多种轴突导向通路中发挥作用，并调节肌动蛋白细丝的组装。Ena/Vasp蛋白集中在生长锥丝状孢子的顶端，在那里发生对引导信号的初始反应。轴突引导中涉及的几个信号通路调节Ena/Vasp功能。因此，Ena/Vasp蛋白处于有利地位，可以作为引导通路信号和肌动蛋白细胞骨架之间的关键聚合点。我们将检验Ena/Vasp蛋白调节肌动蛋白重塑响应引导信号的假设。我们建议使用高分辨率光学显微镜和电子显微镜来分析细胞骨架的动力学和几何构型，以研究Ena/Vasp在生长锥引导和移位中的作用。我们还将使用基因操作来扰乱Ena/Vasp的功能，并使用生化方法来研究Ena/Vasp在信号通路中的作用。这项工作应该对神经系统是如何发展的以及如何设计损伤后修复的方法提供有价值的见解。

DESCRIPTION (provided by applicant): During embryonic development, vast numbers of neurons must form connections with their proper targets. Axons are guided to their targets by growth cones, which integrate and convert guidance signals into mechanical forces required for locomotion. Movement arises from dynamic regulation of cytoskeletal polymers and associated proteins. The Ena/VASP protein family functions in various axon guidance pathways and is known to regulate the assembly of actin filaments. Ena/VASP proteins are concentrated in the tips of growth cone filopodia, where the initial response to guidance signals occurs. Several signaling pathways implicated in axon guidance regulate Ena/VASP function. Therefore, Ena/VASP proteins are well positioned to act as key convergence points between signals from guidance pathways and the actin cytoskeleton. We will test the hypothesis that Ena/VASP proteins regulate actin remodeling in response to guidance signals. We propose to examine the role of Ena/VASP in growth cone guidance and translocation using high-resolution light and electron microscopy to analyze cytoskeletal dynamics and geometry. We will also employ genetic manipulation to perturb Ena/VASP function and biochemical approaches to study Ena/VASP's role in signaling pathways. This work should yield valuable insight into how the nervous system develops and how to design methods to repair it after injury.