Organization and function of neuronal endosomes

神经元内体的组织和功能

• 批准号: 9324369
• 负责人: Bettina R Winckler
• 金额: $ 34.56万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9324369
• 关键词: AGFG1 gene; Address; Affect; Alzheimer' s Disease; Autistic Disorder; Axon; Brain Diseases; Cell Adhesion Molecules; Data; Defect; Dendrites; Development; Disease; Down-Regulation; Early Endosome; Electron Microscopy; Endocytosis; Endosomes; Felis catus; Fibroblasts; Fluorescence Microscopy; Functional disorder; Future; Gene Family; Goals; Golgi Apparatus; Grant; Growth; Health; Image; Integral Membrane Protein; Intervention; Knowledge; Lead; Length; Link; Location; Lysosomes; Mass Spectrum Analysis; Mediating; Membrane; Membrane Protein Traffic; Membrane Proteins; Modeling; Molecular; Mus; Mutation; Names; Nervous system structure; Neural Cell Adhesion Molecule L1; Neurons; Outcome; Pathology; Physiological; Play; Process; Proteins; Regulation; Research; Resolution; Rodent; Role; Signal Transduction; Site; Sorting - Cell Movement; Surface; System; Testing; Vesicle; Work; Yeasts; arm; axon growth; axon guidance; base; functional outcomes; innovation; insight; member; neurodevelopment; neurological pathology; postsynaptic; prevent; public health relevance; receptor; relating to nervous system; synaptic function; trafficking

DESCRIPTION (provided by applicant): Endosomal trafficking of receptors plays fundamental roles in neuronal function and in neurodevelopment, as well as in disease states of the nervous system. The current understanding of endosome organization and function is based primarily on work in yeast and fibroblasts. However, our recent work showed that neurons employ neuronal-specific endocytic and endosomal machinery. There is thus presently a fundamental gap in the understanding of how endosomal function is adapted to cater to the specific physiological needs of neurons. Since the particular organization of the endosome determines postendocytic trafficking of receptors (i.e. signaling, processing, and/or degradation) and thereby specific functional outcomes, the field's ignorance of the neuronal adaptations of the endosomal system constitutes a significant barrier to progress in both basic and disease-oriented fields. Our long-term goals are to uncover the functional contributions of endosomes to neurodevelopment, and to the healthy and diseased brain. The rationale motivating this proposal is that vertebrate neurons express neuronal-specific proteins in their endosomes (in particular NEEP21/Nsg-1 and P19/Nsg-2) that control the endosomal trafficking of crucial neuronal membrane proteins, such as receptors important in development (the axonal adhesion molecule L1/NgCAM), synaptic function (GluA2), and disease (bAPP). Nsg-proteins are members of a gene family of transmembrane proteins found specifically in the neuronal Golgi and in poorly characterized somatodendritic endosomes ("Nsg-endosomes"). The central concept of this application is that crucial neuronal functions depend on Nsg-endosomes. Our preliminary data suggest the specific hypothesis that Nsg-proteins maintain a specific subset of endocytosed receptors in a distinct non-degradative compartment from where the cargo can recycle to multiple locations to regulate axon growth and synaptic function. We will use innovative approaches including quantitative single vesicle live imaging, super-resolution fluorescence microscopy, and electron microscopy in combination with functional interference approaches in primary neurons and in rodent cortex to address three specific aims: Aim 1) How are Nsg-endosomes formed and how do they relate to other somatodendritic endosomes? With which other proteins does NEEP21/Nsg-1 interact and which rabs regulate Nsg-endosomal organization? Aim 2) How does loss of NEEP21/Nsg-1 and P19/Nsg-2 affect endosomal organization and cargo trafficking? Aim 3) Does loss of NEEP21/Nsg-1 and P19/Nsg-2 affect axon and dendrite development in the cortex? The proposed research is significant because it will discover the contribution of endosomes to neuronal function in health and disease. The new insights gained will not only lead to fundamental advances in understanding the regulation of neuronal membrane traffic, but also raise the possibility of new targets for tailoring translational strateges in the future.

描述（由申请方提供）：受体的内体运输在神经元功能和神经发育以及神经系统疾病状态中起着重要作用。目前对内体组织和功能的理解主要基于酵母和成纤维细胞中的工作。然而，我们最近的工作表明，神经元采用神经元特异性内吞和内体机制。因此，目前在理解内体功能如何适应神经元的特定生理需求方面存在根本性的差距。由于内体的特定组织决定了受体的胞内后运输（即信号传导、加工和/或降解），从而决定了特定的功能结果，因此该领域对内体系统的神经元适应的忽视构成了基础领域和疾病导向领域进展的重大障碍。我们的长期目标是揭示内体对神经发育以及健康和患病大脑的功能贡献。 该提议的基本原理是脊椎动物神经元在其内体中表达神经元特异性蛋白（特别是NEEP 21/Nsg-1和P19/Nsg-2），其控制关键神经元膜蛋白的内体运输，例如在发育（轴突粘附分子L1/NgCAM）、突触功能（GluA 2）和疾病（bAPP）中重要的受体。Nsg-蛋白是在神经元高尔基体和特征不明显的体树突内体（“Nsg-内体”）中特异性发现的跨膜蛋白基因家族的成员。本申请的中心概念是关键的神经元功能依赖于Nsg-内体。我们的初步数据表明，特定的假设，NSG蛋白保持一个特定的内吞受体的子集在一个独特的非降解区室，货物可以回收到多个位置，以调节轴突生长和突触功能。我们将使用创新的方法，包括定量单囊泡活成像，超分辨率荧光显微镜，电子显微镜结合功能干扰的方法在初级神经元和啮齿类动物皮层，以解决三个具体的目标：1）Nsg-内体是如何形成的，他们如何与其他体树突内体？NEEP 21/Nsg-1与哪些其他蛋白相互作用，哪些rab调节Nsg-内体组织？目的2）NEEP 21/Nsg-1和P19/Nsg-2的缺失如何影响内体组织和货物运输？目的3）NEEP 21/Nsg-1和P19/Nsg-2的缺失是否影响皮层轴突和树突的发育？这项研究意义重大，因为它将发现内体对健康和疾病中神经元功能的贡献。获得的新见解不仅将导致理解神经元膜交通调控的根本性进展，而且还提高了未来定制翻译策略的新靶点的可能性。