Regulation of protein targeting in axon guidance and neuronal morphogenesis

轴突引导和神经元形态发生中蛋白质靶向的调节

• 批准号: 9069619
• 负责人: MARY C HALLORAN
• 金额: $ 32.73万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9069619
• 关键词: Afferent Neurons; Alzheimer' s Disease; Axon; Axonal Transport; Behavior; Binding; Biosensor; Carrier Proteins; Cell membrane; Cells; Charcot-Marie-Tooth Disease; Complex; Cues; Data; Defect; Dendrites; Development; Disease; Dynein ATPase; Embryo; Endosomes; Environment; Event; Goals; Health; Human; Image; Individual; Kinesin; Label; Learning; Life; Ligands; Link; Location; Maintenance; Mediating; Mediator of activation protein; Membrane; Membrane Protein Traffic; Memory; Microtubules; Modeling; Molecular; Molecular Target; Morphogenesis; Morphology; Motor; Natural regeneration; Neurodegenerative Disorders; Neurons; Neurotrophic Tyrosine Kinase Receptor Type 3; Neurotrophin 3; Niemann-Pick Diseases; PHluorin; Pain Disorder; Peripheral; Process; Protein Family; Proteins; Regulation; Role; Route; Semaphorin-3A; Semaphorins; Sensory; Signal Transduction; Slide; Structure; System; Testing; Time; Vesicle; Vesicle Transport Pathway; Zebrafish; axon growth; axon guidance; axon injury; extracellular; in vivo; in vivo Model; knock-down; mutant; neural circuit; neuron development; neuronal transport; neurotrophic factor; novel; painful neuropathy; polymerization; protein function; protein transport; receptor; research study; response; synaptogenesis; targeted imaging; temporal measurement; trafficking

 DESCRIPTION (provided by applicant): Proper development of neuronal morphology and neural circuits, as well as neuronal maintenance, requires tightly controlled subcellular localization of proteins such as axon guidance cue receptors. Proteins are targeted to specific cell locations by elaborate membrane trafficking and axonal transport systems. Neurons are particularly dependent on high fidelity protein trafficking because of their highly polarized and complex structure. Defects in trafficking and transport underlie multiple human developmental and neurodegenerative diseases, including Alzheimer's disease, Charcot-Marie-Tooth, and Niemann Pick disease. Despite their importance, the mechanisms regulating trafficking processes in neurons are poorly understood, in part due to a paucity of in vivo models in which the machinery and mechanisms of axon transport can be studied. A major challenge to the field and the long term goal of this project is to understand the mechanisms controlling axonal transport and protein localization as neurons develop in their natural environment, where they must integrate multiple extracellular cues. We established a model in which we can image dynamics of neuronal cargo transport, protein localization, and microtubule behavior in the intact zebrafish embryo. Vertebrate sensory neurons extend distinct central and peripheral axons to form the sensory circuit. We found that these axons show distinct responses to axon guidance cues. Moreover, we discovered roles for endosomal trafficking and the kinesin adaptor Calsyntenin-1 (Clstn-1) in differential guidance of sensory axons. In Aim 1 we propose to determine how calsyntenins regulate endosome transport routes to different axon compartments. In Aim 2 we will investigate mechanisms regulating specific localization of receptors for Neurotrophin-3 and Semaphorin3d. We will test the hypothesis that Calsyntenins and another class of kinesin adaptors, the Collapsin response mediator proteins (CRMPs) function to target receptors to specific axon compartments. In Aim 3 we will determine how Clstn-1 and CRMPs organize microtubule polarity and dynamics, processes essential for accurate trafficking and axon growth. Our unique model allows us to connect the molecular events of axonal transport, guidance receptor localization and microtubule organization to specific axon guidance decisions at the time and place they naturally occur. Elucidation of the molecular signals regulating sensory axon growth, guidance, and protein trafficking is critical for understanding neurodegenerative disorders, neuropathic pain disorders and the conditions under which regeneration after axon injury can occur. Our experiments will uncover such mechanisms and thus may help to identify molecular targets for disease treatment.

 描述（由应用提供）：正确发展神经元形态和神经元电路以及神经元维护，需要严格控制蛋白质的亚细胞定位，例如轴突引导提示受体。蛋白质通过精致的膜运输和轴突运输系统针对特定的细胞位置。神经元由于其高度极化和复杂的结构而特别取决于高保真蛋白的运输。贩运和运输的缺陷是多种人类发育和神经退行性疾病的基础，包括阿尔茨海默氏病，charcot-marie-tooth和Niemann Pick病。尽管它们的重要性，但这些机制调节神经元中的运输过程的理解很少，部分原因是由于体内模型很少，可以研究轴突运输的机理和机制。该项目的主要挑战和该项目的长期目标是了解控制轴突运输和蛋白质定位的机制，因为在其自然环境中发展的神经元，它们必须整合多个细胞外提示。我们建立了一个模型，在该模型中，我们可以在完整的斑马鱼胚胎中图像神经元货物传输，蛋白质定位和微管行为的动力学。脊椎动物感觉神经元延伸不同的中央和周围轴突以形成感觉电路。我们发现这些轴突对轴突引导提示显示不同的响应。此外，我们在感觉轴突的差异指导中发现了内体贩运和驱动蛋白适配器Calsyntenin-1（CLSTN-1）的作用。在AIM 1中，我们建议确定Calsyntenins如何调节向不同轴突隔室的内体运输路线。在AIM 2中，我们将研究调节神经营养蛋白3和Semaphorin3d受体特定定位的机制。我们将检验以下假设：Calsyncinns和另一类的动力蛋白适配器，Collapsin响应介质蛋白（CRMP）功能对特定轴突室的靶向受体。在AIM 3中，我们将确定CLSTN-1和CRMP如何组织微管极性和动力学，这对于准确的运输和轴突生长至关重要。我们独特的模型使我们能够将轴突传输，引导受体定位和微管组织的分子事件与它们自然发生的时间和地点联系起来。阐明减轻感觉轴突生长，引导和蛋白质运输的分子信号对 了解神经退行性疾病，神经性疼痛障碍以及轴突损伤后再生的疾病。我们的实验将发现这种机制，因此可能有助于确定疾病治疗的分子靶标。