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

项目摘要/摘要 了解自发的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，与神经或神经精神障碍有关。早期零星分布 活动对于形成成熟的相关神经网络是必不可少的。迄今为止，我们在以下方面取得了成功 确定马达电路是斑马鱼胚胎发育中第一个起作用的电路。此外，还有几个 神经元是“先导细胞”，是最早获得神经元自发活动的神经元。这是自发的 活动之后是形成小型同步神经网络，形成复杂的电路 随着发展的进步。然而，由于它们的稀疏性和瞬变性质，分子表征 领头羊细胞一直是一个挑战。这项提议旨在将斑马鱼的遗传学与强大的新方法相结合。 用于单细胞转录图谱，以提供活动依赖的发育的高分辨率地图，并 使用它以细胞类型特定的方式识别活性依赖的基因程序，并描述领导细胞。这个 提案的重点是整合新技术来发现胚胎脊椎动物的大脑是如何发育的 以一种依赖活动的方式。其目的是对先导细胞进行全面的分子图谱分析 当它们在发育中的脊椎动物神经系统中获得自发活动时，识别潜在的基因 调节器(目标1)，并评估对领导细胞活动的扰动如何塑造电路成熟和大脑 发展(目标2)。这项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。专门评估大脑是如何 在健康期间发展，当运动回路，斑马鱼神经系统中第一个起作用的回路，是 打乱了。执行这项提议的理想环境是在瓦格纳博士的共同指导下，他开发了 TRACERSEQ和SINTCH绘制健康人和正常人细胞状态和谱系之间的定量关系图 使用定量生物学技术和发展专家克里格斯坦博士的扰动背景， 基因组学和干细胞生物学。总体而言，这些研究将导致对转录如何 神经元和非神经元细胞的轮廓协调一致，以创造一个健康的大脑，具有适当的细胞增殖和 以依赖活动的方式进行分化。

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.