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: 10350606
• 负责人: Meike Lobb-Rabe
• 金额: $ 4.6万
• 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/10350606
• 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.

项目概要/摘要 该项目的总体目标是确定作用于神经元和/或其靶标的分子决定因素 促进适当功能所需的特定环境突触树的精细化和维护。 神经元找到它们的突触伙伴，突触末端必须扩展到适当的大小以维持 虽然将轴突引导至目标区域的引导线索已得到很好的表征，但还不清楚。 了解产生每个突触独特的刻板生长模式的分子机制 末端乔木。电路维护/突触传递功能障碍与 神经系统疾病，例如自闭症和精神分裂症，强调了描述这些疾病的紧迫性 为研究这一重要的发育过程而提出的策略是检查。 Ig 结构域细胞表面蛋白 (CSP) 的 Dpr 和 DIP 亚家族如何指导突触末端 我将利用果蝇神经肌肉回路和功能的相对简单性。 强大的遗传工具可以通过单突触分辨率来检查这些问题。 假设是这些 CSP 最初决定目标特定的突触树状结构，同时， 为了检验这个假设，我将重点关注 DIP-α，因为我的初步数据。 提出了 DIP-α 在这两个过程中的新作用。在目标 1 中，我将利用细胞特异性生产者来检查所有过程。 表达 DIP-α 的运动神经元确定 DIP-α 的作用是指导特定突触的形成 我将采用基于候选的遗传筛选来揭示潜在的机制。 目标 2，我将描述 DIP-α 在通过本地化形成新的和/或塑造现有活动区域方面的作用 此外，我将同时进行电生理学检查。 我预计从这些研究中可以通过记录和光遗传学来分析运动神经元的特异性反应。 揭示 DIP 在突触特异性加工和突触发生中的新作用 该提案具有创新性。 它结合了跨学科方法，包括电生理学、生物化学、单细胞遗传学 操作和显微镜，以阐明控制突触乔木扩张和 这项研究意义重大，因为它解决了发育中长期存在的问题。 神经生物学：同一神经元的离散突触乔木如何获得独特的形态。 项目提供了一个理想的培训机会来支持我成为一名 独立研究者专注于阐明神经回路背后的新分子机制 发展，重要的是，有助于我们理解这些分子途径如何 破坏以揭示疾病病因。