DIP-ÃÂñ is required for synaptic elaboration and function

DIP-àªàà± 是突触精细化和功能所必需的

• 批准号: 10477294
• 负责人: Meike Lobb-Rabe
• 金额: $ 3.58万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2023-04-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10477294
• 关键词: Abdominal Muscles; Address; Area; Axon; Biochemistry; Biological Assay; Cell Surface Proteins; Cells; Complex; Confocal Microscopy; Cues; Data; Defect; Development; Developmental Process; Disease; Drosophila genus; Electrophysiology (science); Environment; Etiology; Functional disorder; Genetic; Genetic Screening; Glutamates; Goals; Growth; Health; Human; Image; Immunoglobulins; Link; Maintenance; Malignant Neoplasms; Mediating; Methods; Microscopy; Modeling; Molecular; Morphology; Motor Neurons; Muscle; Neuroglia; Neuromuscular Junction; Neurons; Orthologous Gene; Output; Pathway Analysis; Pathway interactions; Pattern; Presynaptic Terminals; Process; Proteins; Reporter; Research; Research Personnel; Resolution; Role; Scaffolding Protein; Schizophrenia; Signal Pathway; Site; Stereotyping; Synapses; Synaptic Transmission; Testing; Training; Work; autism spectrum disorder; base; density; developmental neurobiology; experimental study; genetic approach; genetic manipulation; human disease; innovation; insight; interdisciplinary approach; nervous system development; nervous system disorder; neural circuit; neuromuscular; neurotransmission; novel; optogenetics; programs; receptor; response; skills; synaptic function; synaptogenesis; tool; training opportunity; translational impact

Project Summary/ Abstract The overall goal of this project is to identify the molecular determinants that act in the neuron and/or its target to promote the context-specific synaptic arbor elaboration and maintenance required for proper function. Once neurons find their synaptic partners, synaptic terminals must expand to an appropriate size to maintain synaptic fidelity. While the guidance cues that direct axons to their target area are well characterized, less is known about the molecular mechanisms that produce the stereotyped growth patterns unique to each synaptic terminal arbor. The clear link between dysfunction in circuit maintenance/synaptic transmission and neurological disease, such as autism and schizophrenia, underscores the urgency in delineating these molecular pathways. The strategy proposed to investigate this important developmental process is to examine how the Dpr and DIP subfamilies of Ig-domain cell-surface proteins (CSPs), instruct synaptic terminal elaboration and function. I will leverage the relative simplicity of the Drosophila neuromuscular circuit and the powerful genetic tools available to examine these questions with single-synapse resolution. My central hypothesis is that these CSPs initially determine target-specific synaptic arbor elaboration, and concurrently, organize synaptic active zones. To test this hypothesis, I will focus on DIP-α since my preliminary data suggests novel roles for DIP-α in both processes. In Aim 1, I will utilize cell-specific reporters to examine all motor neurons that express DIP-α to determine DIP-α’s role is in instructing elaboration of specific synaptic terminal arbors. I will employ a candidate based genetic screen in order to uncover potential mechanisms. In Aim 2, I will characterize DIP-α’s role in forming new and/or sculpting existing active zones through localization of a core active zone scaffolding protein. Additionally, I will perform simultaneous electrophysiological recordings and optogenetics to analyze motor neuron specific responses. From these studies, I anticipate unraveling new roles for DIPs in synapse specific elaboration and synaptogenesis. This proposal is innovative in that it combines interdisciplinary approaches, including electrophysiology, biochemistry, single-cell genetic manipulations, and microscopy, to elucidate fundamental mechanisms governing synaptic arbor expansion and neurotransmission. This research is significant because it addresses a long-held question in developmental neurobiology: how do discrete synaptic arbors of the same neuron acquire unique morphologies. Lastly, this project provides an ideal training opportunity to support my long-term professional goal of becoming an independent investigator focused on elucidating novel molecular mechanisms that underlie neural circuit development, and importantly, contributes to our understanding of how these molecular pathways can be disrupted to reveal disease etiologies.

项目摘要/摘要 该项目的总体目标是确定在神经元和/或其目标中起作用的分子确定词 促进适当功能所需的特定上下文特定的突触植物阐述和维护。一次 神经元发现其突触伙伴，突触终端必须扩展到适当的大小才能维持 突触忠诚。虽然将轴突引向其目标区域的指导提示的特征是 知道产生刻板印象的生长模式的分子机制已知 终端植物。电路维护/突触传输中的功能障碍与 神经疾病，例如自闭症和精神分裂症，强调了描述这些疾病的紧迫性 分子途径。提出的旨在调查这一重要发展过程的策略是检查 Ig-域细胞表面蛋白（CSP）的DPR和浸入下属如何指示突触终端 阐述和功能。我将利用果蝇神经肌肉电路的相对简单性和 有力的遗传工具可用于以单节解析来检查这些问题。我的中央 假设是这些CSP最初确定靶标特异性合成植物轴的阐述，并同时确定 组织合成活性区。为了检验该假设，我将重点放在DIP-α上，因为我的初步数据 在这两个过程中都提出了DIP-α的新作用。在AIM 1中，我将利用特定细胞的记者检查所有 表达DIP-α以确定DIP-α的作用的运动神经元在指导特定突触 末端乔木。我将采用基于候选的遗传筛查，以发现潜在的机制。在 AIM 2，我将表征DIP-α在通过本地化形成新的和/或雕刻现有活动区域中的作用 核心活性区支架蛋白。此外，我将同时进行电生理学 记录和光遗传学分析运动神经元特定反应。从这些研究中，我期望 在突触特异性阐述和突触发生中阐明新作用的新作用。该建议是创新的 因为它结合了跨学科方法，包括电生理学，生物化学，单细胞遗传 操纵和显微镜，以阐明有关突触阵列扩张和的基本机制 神经传递。这项研究很重要，因为它解决了发展中长期存在的问题 神经生物学：同一神经元的离散突触街道如何获得独特的形态。最后，这个 项目提供了理想的培训机会，以支持我成为一个长期专业目标 独立研究者着重于阐明基于神经回路的新型分子机制 发展，重要的是，我们对这些分子途径的理解有助于我们的理解 被破坏以揭示病因。