Molecular Mechanisms of Synapse Coordination

项目摘要/摘要 大脑是一个极其复杂的器官。这种复杂性并不是从它估计的820亿个神经元中产生的， 但是相反，在这些神经元之间建立的数万亿个特定的连接中 发育称为突触。然而，许多不同类型的突触是如何形成的，从根本上说是未知的 在发育过程中，也不知道突触群体是如何协调形成工作的 神经回路中的内部平衡。这是对早期大脑发育的理解上的一个关键差距 许多神经发育障碍，如自闭症和癫痫，被认为是由不当引起的 突触的形成会导致回路失衡。 突触被广泛地定义为两类：首先形成并持续到成年期的电性突触； 以及在电突触之后发展起来的化学突触，通过化学突触进行交流 神经递质。虽然电突触和化学突触的生化组成是不同的，但这表明 不同的过程调节它们的形成，自闭症和癫痫相关基因Neurobeachin是 电突触和化学突触的形成都需要。本提案的目标是1)确定 Neurobeachin用于调节电和化学突触形成的分子机制，以及2) 确定这些机制是相似还是不同。为了实现这一目标，斑马鱼 Mauner电路将被使用，因为它是在早期发育过程中解剖Neurobeachin功能的理想工具 活着。莫特纳神经元可以很容易地产生可识别的、刻板的电子和化学突触 在发育中的胚胎中观察到的，并可用于细胞生物学、遗传学和生化研究的。 总而言之，这项提议将解锁引导电和化学突触形成的共同机制 并为一种对大脑发育至关重要的分子提供了新的线索，该分子与多种 神经发育障碍最终导致合理的治疗干预措施的设计。 该提案旨在提供有关CRISPR的尖端编辑、动态实时成像和 斑马鱼早期大脑模型系统中复杂的细胞生物学和生物化学技术 俄勒冈大学的发展。俄勒冈大学是斑马鱼的发源地 模范生物和卓越的机构学习创新的斑马鱼技术 研究人员的协作和支持环境。这一经历将培养博士后研究人员 在高级研究方法和定量分析、统计、手稿写作和指导方面 成功成长为独立学术研究岗位所需的技术。

PROJECT SUMMARY / ABSTRACT The brain is an exquisitely complex organ. This complexity is not born out of its estimated 82 billion neurons, but instead in the trillions of specific connections these neurons make between one another during development called synapses. Yet, it is fundamentally unknown how the many different types of synapses form during development, nor is it known how synapse populations coordinate formation to create a working balance within in a neural circuit. This is a critical gap in the understanding of early brain development as numerous neurodevelopmental disorders, such as autism and epilepsy, are thought to arise from improper synapse formation resulting in circuit imbalances. Synapses are broadly defined into two groups: electrical synapses which form first and persist into adulthood; and chemical synapses which develop after electrical synapses and communicate via chemical neurotransmitters. While the biochemical makeup of electrical and chemical synapses are distinct, suggesting different processes regulate their formation, the autism- and epilepsy-associated gene Neurobeachin is required for both electrical and chemical synapse formation. The goal of this proposal is to 1) identify the molecular mechanisms used by Neurobeachin to regulate electrical and chemical synapse formation, and 2) determine if these mechanisms are similar to or distinct from one another. To realize this goal, the zebrafish Mauthner circuit will be used as it is ideal for dissection of Neurobeachin function during early development in vivo. Mauthner neurons make identifiable, stereotyped electrical and chemical synapses that are easily observed in developing embryos and accessible to cell biological, genetic, and biochemical investigation. Together, this proposal will unlock the shared mechanisms guiding electrical and chemical synapse formation and shed new light onto a molecule critical for brain development and implicated in various neurodevelopmental disorders ultimately leading to the design of rational therapeutic interventions. This proposal is designed to provide training in cutting edge CRISPR editing, dynamic live imaging, and sophisticated cell biology and biochemistry techniques within the zebrafish model system across early brain development at the University of Oregon. The University of Oregon is the birthplace of the zebrafish as a model organism and the preeminent institution to learn innovative zebrafish techniques among a highly collaborative and supportive environment of researchers. This experience will train the postdoctoral researcher in advanced research methodology and quantitative analysis, statistics, manuscript writing, and mentoring techniques necessary for successful growth into an independent academic research position.