Molecular and genetic decoding of neuron-glial interactions

项目摘要 正常的神经系统功能依赖于神经元和非神经元细胞之间的适当沟通。 在人类大脑中，大多数非神经细胞被称为胶质细胞，占脑细胞总数的近一半。 尽管几十年来的研究主要集中在神经元和神经胶质细胞上，但它们之间的交流方式仍然存在 人们对此知之甚少。这在一定程度上是由于缺乏全面分析分子的方法 在动态细胞-细胞相互作用过程中，在神经元-神经胶质界面上丰富。这项提案旨在开发一种 基因引导的蛋白质组学工具箱用于时空分析富含在神经元-神经胶质细胞中的关键分子 活体内的界面，允许对神经元-神经胶质相互作用的分子和遗传进行剖析。我们将首先设计 并产生时空精确度的神经胶质细胞表面邻近标记探针。我们会申请 少突胶质细胞是中枢神经系统(CNS)中唯一产生髓鞘的细胞，这个平台可以 确定少突胶质细胞-轴突包膜启动的分子机制。我们会 利用这一新方法，以及单细胞RNA测序、全基因组CRISPR筛查和新颖的 转基因小鼠品系，询问一个神秘的细胞阶段(髓鞘前少突胶质细胞)在 发育和适应性髓鞘形成。为了扩展神经胶质表面邻近标记工具包，我们将开发一个 神经元-神经胶质互补邻近标记系统可用于瞬时神经元-神经胶质细胞的可视化 在活体中，分子的相互作用和捕获只在神经元-神经胶质界面上丰富。使用这个系统，我们将 解决控制少突胶质细胞亚群之间髓鞘选择性的分子密码和 在功能上不同的神经元亚群。这项拟议的工作将填补髓鞘生物学方面的知识空白， 这将对理解神经元-胶质细胞和胶质细胞-胶质细胞相互作用机制具有更广泛的意义。 本工作建立的方法和试剂也将极大地丰富细胞多样性研究的方法。 细胞通讯，包括神经免疫和癌症免疫相互作用。

Project Summary Normal nervous system function depends on proper communications between neurons and non-neuronal cells. In the human brain, the majority of non-neuronal cells are called glia that consist of nearly half of total brain cells. Despite decades of work focusing on neurons and glia alone, how they communicate with each other remains poorly understood. This is partly due to a dearth of methods to comprehensively profile molecules that are enriched at the neuron-glial interface during dynamic cell-cell interactions. This proposal aims to develop a genetically-guided proteomic toolbox to spatiotemporally profile critical molecules enriched at neuron-glial interface in vivo, allowing for molecular and genetic dissection of neuron-glial interactions. We will first design and generate glial-specific cell surface proximity-labeling probes with the spatiotemporal precision. We will apply this platform in the oligodendrocytes, the sole myelin-producing cells in the central nervous system (CNS), to determine the molecular mechanisms governing the initiation of oligodendrocyte-axon ensheathment. We will leverage this new method, along with single-cell RNA sequencing, genome-wide CRISPR screens, and novel transgenic mouse strains, to interrogate a mysterious cell stage (the pre-myelinating oligodendrocytes) during developmental and adaptive myelination. To extend the glial surface proximity labeling toolkit, we will develop a neuron-glial complementary proximity labeling system allowing for visualization of transient neuron-glial interactions and capture of molecules only enriched at neuron-glial interface in vivo. Using this system we will address the molecular codes governing myelination selectivity between subsets of oligodendrocytes and functionally distinct neuronal subpopulations. This proposed work will fulfill the knowledge gap in myelin biology, and will have broader implications in understanding neuron-glial and glia-glial interaction mechanisms in general. The methods and reagents established by the work will also greatly enrich the methods in studying diverse cell- cell communications, including neuro-immune and cancer-immune interactions.