Understanding the Developmental Mechanisms that Ensure Robustness in Neuronal Patterning

了解确保神经元模式稳健性的发育机制

• 批准号: 10251027
• 负责人: Pavak Kirit Shah
• 金额: $ 24.4万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2023-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10251027
• 关键词: 3-Dimensional; Ablation; Adult; Advisory Committees; Alleles; Award; Behavior; Brain; Caenorhabditis elegans; Calcium; Cells; Cellular biology; Cilia; Communities; Complement; Development; Developmental Process; Dorsal; Embryo; Embryonic Development; Embryonic Nervous System; Ensure; Environment; Equilibrium; Feedback; Financial compensation; Fluorescence Microscopy; Future; Generations; Genetic; Glutamates; Goals; Growth; Head; Head Movements; Human Development; Image; Individual; Institutes; Interleukin-1; Knowledge; Label; Laboratories; Lasers; Learning; Measurement; Measures; Mechanics; Memory; Mental disorders; Mentors; Mentorship; Methods; Modeling; Motor; Motor Neurons; Motor output; Movement; Muscle; Nature; Nematoda; Nervous System Physiology; Nervous system structure; Neurodevelopmental Disorder; Neuromuscular Junction; Neurons; Optics; Output; Paper; Pattern; Phase; Physical Stimulation; Play; Postdoctoral Fellow; Process; Property; Publishing; Quantitative Microscopy; Reagent; Regulation; Reporter; Resolution; Sensory; Source; Specificity; Speed; Structure; Synapses; Synaptic Transmission; Synaptic plasticity; System; Testing; Time; Tissues; Training; Training Support; Universities; Vertebrates; Work; analysis pipeline; automated image analysis; base; career; career development; cholinergic; design; experience; flexibility; human disease; improved; in vivo; insight; interest; medical schools; motor behavior; mutant; neuronal circuitry; neuronal patterning; optogenetics; preservation; relating to nervous system; response; sensory feedback; sensory input; transgene expression

Abstract: Establishing a balance between the activities of competing neuronal circuits is essential to the coordination of behaviors. We propose to investigate the developmental strategies that ensure the establishment of this balance in the nervous system. Studies of the vertebrate embryonic nervous system have shown that correlated activity patterns the structure and output nascent circuits through mechanisms akin to learning. The scale and complexity of vertebrate brains has made it challenging to identify the rules which govern this learning or the mechanisms which drive optimization of the circuit toward a balanced output. The small size and invariance of the Caenorhabditis elegans nervous system make it possible to study these processes with specificity not possible in vertebrates. Using the development of one of the first functional circuits in the C. elegans embryo as a model, I will study how functional balance is established in the developing nervous system and identify the developmental mechanisms that ensure the robustness of this balance. To accomplish this goal, I propose the following specific aims: 1) To identify the models of IL1 neuron function in the patterning of embryonic head movement. 2) To automate and improve an approach for controlling transgene expression with single cell resolution by infrared laser-induced heatshock. And 3) To determine the mechanisms by which the IL1 circuit establishes and maintains a balanced output to produce coordinated movements. To enable this, I have developed a real-time cell tracking system which automates the identification of targeted cells during embryonic development and can control the conduct of single cell optical perturbations such as laser ablation as described in a recently published paper in Developmental Cell or, as will be developed in Aim 2, single cell heatshock. I have also developed an automated image analysis pipeline which allows me to track and measure movement patterns in the early embryo with which I have identified the neuronal origins of patterned head movement and, using genetic mutants of neuronal function, shown that this patterning is defined by neuronal input. I believe that this work will provide important insights into the patterning of the embryonic nervous system with implications for our understanding of the circuit-level origins of some neurodevelopmental disorders. In order to expand my training as a biologist and complement my broad technical training and expertise, I will carry out the mentored phase of this award as a postdoctoral Research Associate in the laboratory of Dr. Zhirong Bao, an expert on C. elegans embryonic development, and under the mentorship of a well-accomplished advisory committee comprised of experienced biologists and neuroscientists. I have designed a structured plan to further my scientific training and support my career development to prepare for an independent career. The Sloan Kettering Institute and the tri- institutional community at Weill Cornell Medical College and The Rockefeller University provide an unparalleled intellectual environment to support my scientific growth and progress towards independence.

摘要：在竞争性神经元回路的活动之间建立平衡对于 行为协调。我们建议研究确保发展战略 在神经系统中建立这种平衡。对脊椎动物胚胎神经系统的研究 研究表明，相关活动通过类似于以下的机制对结构和输出新生电路进行模式化： 学习。脊椎动物大脑的规模和复杂性使得识别大脑的规则变得具有挑战性。 控制这种学习或驱动电路优化以实现平衡输出的机制。这 秀丽隐杆线虫神经系统的小尺寸和不变性使得研究这些成为可能 脊椎动物中不可能实现的特异性过程。使用第一个功能的开发 以秀丽隐杆线虫胚胎中的回路为模型，我将研究如何在 发育神经系统并确定确保其稳健性的发育机制 平衡。为了实现这个目标，我提出以下具体目标：1）确定IL1神经元的模型 在胚胎头部运动模式中发挥作用。 2）自动化和改进方法 通过红外激光诱导的热激以单细胞分辨率控制转基因表达。以及 3) 至 确定 IL1 电路建立和维持平衡输出以产生的机制 协调的动作。为了实现这一点，我开发了一个实时细胞跟踪系统，可以自动执行 胚胎发育过程中目标细胞的识别并可控制单细胞的行为 最近发表在《发展》杂志上的一篇论文中描述了光学扰动，例如激光烧蚀 细胞或，如目标 2 中将开发的，单细胞热休克。我还开发了一个自动图像 分析管道使我能够跟踪和测量早期胚胎的运动模式 已经确定了模式化头部运动的神经元起源，并利用神经元的基因突变体 函数，表明这种模式是由神经元输入定义的。我相信这项工作将提供重要 深入了解胚胎神经系统的模式对我们理解胚胎神经系统的意义 一些神经发育障碍的回路水平起源。为了扩大我作为生物学家的培训和 为了补充我广泛的技术培训和专业知识，我将作为一名 秀丽隐杆线虫胚胎专家鲍志荣博士实验室博士后研究员 发展，并在由经验丰富的顾问委员会组成的出色顾问委员会的指导下进行 生物学家和神经科学家。我设计了一个结构化计划来进一步加强我的科学培训和支持 我的职业发展是为独立的职业生涯做准备。斯隆凯特琳研究所和三 威尔康奈尔医学院和洛克菲勒大学的机构社区提供了无与伦比的 支持我的科学成长和走向独立的智力环境。