Waking up the nervous system: Molecular characterization of neuronal leader cells and their role in brain development

唤醒神经系统：神经元领导细胞的分子特征及其在大脑发育中的作用

• 批准号: 10315708
• 负责人: Nicole Ann Aponte-Santiago
• 金额: $ 7.51万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10315708
• 关键词: Affect; Atlases; Bioinformatics; Biology; Brain; CRISPR screen; Calcium; Cell Differentiation process; Cell Nucleus; Cell Proliferation; Cells; Communities; Complex; Data; Development; Developmental Process; Discipline; Distributional Activity; Drug or chemical Tissue Distribution; Embryo; Embryonic Development; Fluorescence Microscopy; Gap Junctions; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Genomics; Health; Human; In Vitro; Label; Learning; Light; Link; Maps; Mediating; Mentors; Methods; Molecular; Molecular Profiling; Motor; Motor Neurons; Nature; Nervous system structure; Neuraxis; Neuroglia; Neurologic; Neuronal Differentiation; Neurons; Outcome; Patients; Population Dynamics; Problem Solving; Process; Protocols documentation; Regulator Genes; Reporter; Resolution; Role; Shapes; Signal Pathway; Small Nuclear RNA; Techniques; Time; Tissue Engineering; Tissues; Transgenic Organisms; Work; Zebrafish; axon growth; cell type; computerized tools; embryo cell; experimental study; human model; in vivo; induced pluripotent stem cell; inhibitor/antagonist; insight; interest; nervous system disorder; neuropsychiatric disorder; neuropsychiatry; neurotransmission; new technology; optogenetics; programs; sensor; single-cell RNA sequencing; stem cell biology; stem cell therapy; success; synaptogenesis; transcriptome sequencing; transcriptomics

Project Summary/Abstract Learning how spontaneous neuronal activity shapes embryonic brain development is critical for understanding neurodevelopmental processes, with implications in neurological or neuropsychiatric disorders. Early sporadic activity is essential for the formation of mature correlated neuronal networks. To date, we have had success in identifying the motor circuit as the first circuit to function in a developing zebrafish embryo. In addition, a few neurons, the “leader cells”, are the first neurons to obtain spontaneous neuronal activity. This spontaneous activity is followed by the formation of small synchronized neuronal networks that elaborate into complex circuits as development progresses. However, due to their sparsity and transient nature, molecular characterization of leader cells has been a challenge. This proposal aims to combine zebrafish genetics with powerful new methods for single-cell transcriptomic profiling to deliver a high-resolution map of activity-dependent development and to use it to identify activity-dependent gene programs in a cell-type specific manner and profile leader cells. The proposal focuses on integrating new technologies into discovering how the embryonic vertebrate brain develops in an activity-dependent manner. The aims are to perform comprehensive molecular profiling of the leader cells as they acquire spontaneous activity in the developing vertebrate nervous system, identify potential genetic regulators (Aim 1), and assess how perturbations to leader cell activity shape circuit maturation and brain development (Aim 2). The proposed work leverages recent developments from several disciplines: single-cell RNA sequencing (scRNA-seq), zebrafish genetics, and optogenetics. Specifically assessing how the brain develops during health and when the motor circuit, the first circuit to function in the zebrafish nervous system, is disrupted. The ideal setting to carry out this proposal is with the co-mentoring of Dr. Wagner, who developed TRACERSEQ and STITCH to map the quantitative relationship between cell states and lineages in healthy and perturbed contexts using quantitative biology techniques and Dr. Kriegstein, an expert in development, genomics, and stem cell biology. Overall, these studies will lead to general insights into how transcriptional profiles of neuronal and non-neuronal cells coordinate to create a healthy brain with proper cell proliferation and differentiation in an activity-dependent manner.

项目总结/摘要 了解自发神经元活动如何塑造胚胎大脑发育对于理解 神经发育过程，涉及神经系统或神经精神障碍。早期散发 活性对于成熟相关神经元网络的形成是必不可少的。到目前为止，我们已经成功地 将所述运动回路识别为在发育中的斑马鱼胚胎中起作用的第一回路。此外，一些 神经元，即“领导细胞”，是获得自发神经元活动的第一批神经元。这种自发 活动之后是小型同步神经元网络的形成， 随着发展的进步。然而，由于它们的稀疏性和瞬时性， 领导细胞是一个挑战。这项提议旨在将联合收割机斑马鱼遗传学与强大的新方法相结合 用于单细胞转录组学分析，以提供活动依赖性发育的高分辨率图谱， 用它以细胞类型特异性的方式鉴定活性依赖性基因程序，并分析前导细胞。的 一项提案的重点是将新技术整合到发现胚胎脊椎动物大脑如何发育中 以依赖于活动的方式。其目的是对领导细胞进行全面的分子分析， 当它们在发育中的脊椎动物神经系统中获得自发活动时， 调节器（目标1），并评估如何扰动领导细胞活动形状电路成熟和大脑 发展（目标2）。拟议的工作利用了几个学科的最新发展：单细胞 RNA测序（scRNA-seq）、斑马鱼遗传学和光遗传学。特别是评估大脑是如何 当运动回路，第一个在斑马鱼神经系统中起作用的回路， 被打乱了执行此建议的理想环境是与瓦格纳博士共同指导，他开发了 TRACERSEQ和STITCH，以绘制健康和非健康人群中细胞状态和谱系之间的定量关系， 使用定量生物学技术和Kriegstein博士，一位发展专家， 基因组学和干细胞生物学。总的来说，这些研究将导致对转录如何 神经元和非神经元细胞的分布协调以产生具有适当细胞增殖的健康脑， 以活动依赖的方式分化。