Diversity Supplement (Monica Quinones-Frias): Roles of Recycling Endosomes in Neuronal Extracellular Vesicle Cargo Traffic

多样性补充剂（Monica Quinones-Frias）：回收内体在神经元细胞外囊泡货物运输中的作用

• 批准号: 10782371
• 负责人: Avital Adah Rodal
• 金额: $ 6.63万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2025-05-01
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10782371
• 关键词: Alzheimer' s Disease; Binding; Biochemistry; Brain; Cell membrane; Cellular Membrane; Chemicals; Clathrin; Communication; Complex; Disease; Drosophila genus; Endocytosis; Endosomes; Foundations; Genetic; Goals; Human; Knowledge; Link; Mediating; Membrane; Membrane Protein Traffic; Microscopy; Morphogenesis; Morphology; Nature; Nervous System; Nervous System Physiology; Neuroglia; Neurons; Parkinson Disease; Play; Presynaptic Terminals; Process; Proteins; Quinones; Recycling; Regulation; Research; Resolution; Role; Route; Signal Transduction; Sorting; Synapses; Synaptic Membranes; Synaptic Transmission; Testing; Therapeutic; Vesicle; cell type; experimental study; extracellular vesicles; in vivo; insight; live cell imaging; nervous system disorder; novel; retrograde transport; tool; trafficking; vesicular release

PROJECT SUMMARY The goal of this proposal is to understand how cellular membrane trafficking machinery controls the packaging and release of extracellular vesicle (EV) cargoes from synapses in vivo. EVs are small membrane-bound vesicles released by numerous cell types including neurons, carrying cargoes critical for signaling and disease. However, we understand very little about how EV cargo traffic is spatially and temporally regulated within the polarized and complex morphology of neurons. We have developed tools to track and manipulate EV traffic at Drosophila presynaptic terminals in vivo, and discovered that flux of cargoes through a plasma membrane-recycling endosome route determines whether they are locally sorted for packaging and release in EVs, rather than depleted from synapses by retrograde transport. Recycling endosomes have primarily been studied in non-neuronal cells, and very little is known about their lifetime, functions, or dynamics at presynaptic terminals. We do know that recycling endosomes play critical roles in signaling, neuronal morphogenesis, EV traffic, and synaptic transmission. Understanding and therapeutically intervening in these important processes will require a deeper knowledge of the mechanisms of neuronal recycling endosome function. In this proposal, we will elucidate the mechanisms of synaptic EV cargo and recycling endosome traffic in vivo. To achieve these goals, we will use Drosophila genetics, biochemistry, high-resolution microscopy, and live cell imaging. 1) We will determine the functions, dynamics, and regulation of different types of synaptic recycling endosomes. To this end, we will develop new tools and approaches to define and control functionally distinct recycling compartments at synapses. Using these tools, we will test novel mechanistic hypotheses for how membrane traffic machinery sorts cargoes at synaptic recycling compartments. 2) We will determine how EV cargo traffic depends on distinct modes of synaptic endocytosis: clathrin-mediated endocytosis, which operates under low neuronal activity and activity-dependent bulk endocytosis, which operates during intense neuronal activity. These experiments will ascertain if EV fate is determined by different modes of internalization, how recycling endosomes contribute to these functions, and provide new mechanisms to link activity, endosomal traffic, and EV release. Given the conserved nature of synaptic membrane trafficking machinery, our findings and tools will lay the foundation for new insights into EV traffic in many aspects of nervous system function, including in human neurological disease.

项目概要 该提案的目标是了解细胞膜运输机制如何控制 体内突触中细胞外囊泡（EV）货物的包装和释放。电动汽车很小 由包括神经元在内的多种细胞类型释放的膜结合囊泡，携带对生命至关重要的货物 信号传导和疾病。然而，我们对电动汽车货运交通的空间和分布知之甚少。 在神经元的极化和复杂形态中进行时间调节。我们开发了工具来 跟踪和操纵果蝇体内突触前终端的 EV 流量，并发现 通过质膜回收内体途径的货物决定它们是否被局部分类 用于在电动汽车中包装和释放，而不是通过逆行运输从突触中耗尽。回收 核内体主要在非神经元细胞中进行研究，对其寿命知之甚少， 功能，或突触前末梢的动态。我们确实知道回收内体在 信号传导、神经元形态发生、EV 交通和突触传递。理解和 对这些重要过程进行治疗干预需要更深入地了解其机制 神经元回收内体功能。在本提案中，我们将阐明突触 EV 的机制 体内的货物和回收内体运输。为了实现这些目标，我们将使用果蝇遗传学， 生物化学、高分辨率显微镜和活细胞成像。 1）我们将确定功能， 不同类型突触回收内体的动力学和调节。为此，我们将开发 新的工具和方法来定义和控制突触功能上不同的回收隔间。 使用这些工具，我们将测试关于膜交通机械如何分类的新机制假设 突触回收隔间的货物。 2) 我们将确定电动汽车货运量如何取决于不同的 突触内吞作用的模式：网格蛋白介导的内吞作用，在低神经元活性下运行 以及活动依赖性大量内吞作用，其在强烈的神经元活动期间起作用。这些 实验将确定电动汽车的命运是否由不同的内化模式决定，如何回收 内体有助于这些功能，并提供新的机制来连接活动、内体运输、 和电动汽车发布。鉴于突触膜运输机制的保守性，我们的发现和 工具将为在神经系统功能的许多方面对电动汽车交通的新见解奠定基础， 包括人类神经系统疾病。