NADPH oxidase regulates growth cone guidance

NADPH氧化酶调节生长锥引导

• 批准号: 10437838
• 负责人: DANIEL Marcel SUTER
• 金额: $ 35.88万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10437838
• 关键词: Actins; Address; Adhesions; Adult; Affect; Antioxidants; Aplysia; Axon; Behavior; Behavioral Assay; Biological Assay; Biological Models; Biosensor; Cell Proliferation; Cell physiology; Cells; Cues; Data; Development; Dyes; Embryo; Environment; Evolution; Eye Development; Fishes; Fluorescent Dyes; Functional disorder; Goals; Growth; Growth Cones; Hydrogen Peroxide; Image; Imaging Techniques; Immunity; In Vitro; Knowledge; Measures; Mediating; Molecular; Movement; NADP; NADPH Oxidase; Nerve Regeneration; Nervous System Trauma; Nervous system structure; Neurodegenerative Disorders; Neurons; Oxidases; Physiological; Physiology; Play; Positioning Attribute; Process; Proteins; Publishing; Rattus; Reactive Oxygen Species; Research; Retinal Ganglion Cells; Role; Second Messenger Systems; Signal Transduction; Signaling Molecule; Source; Structure; Testing; Transgenic Organisms; Visual; Work; Zebrafish; axon growth; axon guidance; axon regeneration; axonal pathfinding; cell motility; central nervous system injury; experience; extracellular; improved; in vivo; inhibitor; innovation; insight; live cell imaging; migration; mutant; nerve supply; nervous system development; neurite growth; neuron development; neuron regeneration; neuronal growth; novel; regeneration following injury; response; retinotectal; small molecule; src-Family Kinases; therapy design

Reactive oxygen species (ROS) can act as signaling molecules mediating physiological functions in immunity, cell proliferation, differentiation, and migration. Whether ROS have a major signaling function as second messengers in axonal growth and guidance is currently unclear. The neuronal growth cone is a highly motile structure at the tip of neuronal processes, guiding them to appropriate target cells during development and regeneration of the nervous system. The growth cone integrates molecular information from the environment and transduces it via multiple signaling cascades to affect underlying cytoskeletal dynamics. Whereas most major second messenger systems have been implicated in regulating directional growth cone movement, such a role has not been established for ROS. The present study has two major objectives focusing on ROS produced by nicotinamide adenine dinucleotide phosphate-(NADPH) oxidase (Nox): (1) to determine the cellular and molecular mechanism by which ROS control neurite growth; and (2) to determine whether ROS act as second messengers downstream of specific guidance cues to control axonal growth and guidance. The four central hypotheses state that (1) a physiological level of ROS is optimal and required for adhesion-mediated neurite growth; (2) Src tyrosine kinase is a key target of ROS signaling in neuronal growth cones; (3) neuronal Nox2-derived ROS regulate axonal pathfinding; and (4) specific axon guidance cues such as slit2 control axonal pathfinding via Nox2-derived ROS both in vitro and in vivo. This project will take advantage of two excellent model systems to test these hypotheses: large Aplysia growth cones for quantitative live cell imaging of growth cone motility and intracellular ROS in vitro and developing zebrafish embryos for imaging and manipulating axonal development in vivo. In vitro growth cone guidance assays, novel fluorescent dyes and biosensors specific for hydrogen peroxide and Src activity, respectively, advanced imaging techniques, chimeric analysis of Nox2-deficient zebrafish lines as well as retinal ganglion cell-specific Nox2-mutant fish lines will be used to address the following two Specific Aims: (1) The first aim is to determine the cellular and molecular mechanism by which ROS in control neurite growth. (2) The second aim is to determine the role of neuronal Nox2 in axonal pathfinding of retinal ganglion cells. The proposed work is highly innovative because it investigates ROS as a novel group of signaling molecules in axonal growth and guidance and develops several new zebrafish lines suitable for studying Nox function in the nervous system. In summary, these studies have the potential of leading to breakthrough findings in the field of neuronal development and regeneration. Furthermore, since basic mechanisms of axonal growth and guidance are highly conserved across species, these studies will impact the development of antioxidant treatments for neurodegenerative diseases and central nervous system injuries.

活性氧（ROS）可以作为介导免疫生理功能的信号分子， 细胞增殖、分化和迁移。 ROS是否具有次要的信号传导功能 轴突生长和引导中的信使目前尚不清楚。神经元生长锥是一个高度活动的 神经元过程尖端的结构，在发育和发育过程中引导它们到达适当的靶细胞 神经系统的再生。生长锥整合了环境中的分子信息 并通过多个信号级联转导它来影响潜在的细胞骨架动力学。而大多数 主要的第二信使系统与调节定向生长锥运动有关，例如 尚未为 ROS 建立角色。本研究有两个主要目标，重点关注产生的 ROS 通过烟酰胺腺嘌呤二核苷酸磷酸-（NADPH）氧化酶（Nox）：（1）测定细胞和 ROS控制神经突生长的分子机制； (2) 确定 ROS 是否充当第二个 特定引导线索下游的信使，控制轴突的生长和引导。四大中心 假设指出 (1) ROS 的生理水平是最佳的，并且是粘附介导的神经突所必需的 生长; (2) Src酪氨酸激酶是神经元生长锥中ROS信号传导的关键靶点； (3)神经元Nox2衍生的ROS调节轴突寻路； (4)特定的轴突引导线索，例如slit2控制轴突 通过 Nox2 衍生的 ROS 在体外和体内进行寻路。该项目将利用两个优秀的 测试这些假设的模型系统：用于定量活细胞生长成像的大海兔生长锥 体外视锥细胞运动和细胞内 ROS 以及用于成像和操作的斑马鱼胚胎发育 体内轴突发育。体外生长锥引导测定、新型荧光染料和生物传感器 分别针对过氧化氢和 Src 活性，先进的成像技术，嵌合分析 Nox2 缺陷斑马鱼品系以及视网膜神经节细胞特异性 Nox2 突变鱼品系将用于 解决以下两个具体目标：（1）第一个目标是确定细胞和分子机制 ROS通过它控制神经突的生长。 (2)第二个目的是确定神经元Nox2在轴突中的作用 视网膜神经节细胞的寻路。拟议的工作具有高度创新性，因为它将 ROS 作为一种 轴突生长和引导中的一组新型信号分子，并开发了几种新的斑马鱼品系 适合研究神经系统中的Nox功能。总而言之，这些研究有可能引领 神经元发育和再生领域的突破性发现。此外，由于基本 轴突生长和引导的机制在物种之间高度保守，这些研究将影响 开发针对神经退行性疾病和中枢神经系统损伤的抗氧化疗法。