Characterization of the cellular and molecular mechanisms that mediate glial tiling

开发具有功能和效率的神经系统需要经过精心设计的规范、迁移和 胶质细胞分化。在中枢和外周神经系统(CNS和PNS， 分别)，指定了大量的胶质细胞种群，它们必须迁移并分化为不仅功能 神经胶质细胞，但也协调它们的发育，使它们占据离散的、不重叠的区域 邻近的神经胶质细胞。这种神经胶质细胞间隔或平铺的现象可能发生在中枢神经系统、三叉神经节或 在神经胶质细胞之间，其中一个细胞位于中枢神经系统，另一个细胞位于三叉神经节。尽管我们知道 这些平铺事件发生时，我们不知道这些相互作用的分子性质，也不知道它们是否被 全是瓦片状的神经胶质。此外，局部胶质细胞-胶质细胞相互作用如何在整体胶质细胞瓦片中发挥作用尚不清楚。在这 建议，以斑马鱼为模式生物，我们将研究细胞和分子机制 在运动出口点(MEP)、移行区(TZ)和发育中的脊髓中介导的髓鞘前胶质细胞瓦片 电源线。在这个项目的目标1中，我们将使用表情分析、多色、在体、延时相结合的方法 新发现的介体的成像、电子显微镜、药理和遗传操作(CRISPR) 目的：阐明MEP TZS髓鞘胶质瓦片形成的细胞和分子机制。在目标2中，我们 将确定我们在目标1中描述的相同机制是否也调节少突胶质细胞之间的平铺 发育中的脊髓中的祖细胞(OPC)。为了更好地了解、诊断和治疗许多 中枢神经系统和三叉神经节的退行性疾病，我们需要了解其细胞和分子机制 它介导了神经胶质细胞之间的相互作用和拼贴。斑马鱼提供了一个独特的机会来直接观察和 操纵细胞群以深入了解神经胶质细胞在正常生理条件下如何相互作用，以及 如果这些最终导致神经胶质瓦片的相互作用在疾病中受到干扰。

Development of a functional and efficient nervous system requires the orchestrated specification, migration and differentiation of glia. During gliogenesis in both the central and peripheral nervous systems (CNS and PNS, respectively), large populations of glia are specified that must migrate and differentiate into not only functional glial cells, but also coordinate their development so that they occupy discrete, non-overlapping territories with neighboring glia. This phenomenon of glial spacing, or tiling, can occur between glia found in the CNS, PNS, or between glial cells where one cell resides in the CNS and the other resides in the PNS. Although we know that these tiling events occur, we don’t know the molecular nature of these interactions or whether they are used by all tiling glia. Additionally, how local glial-glial interactions play a role in global glial tiling is unknown. In this proposal, using zebrafish as a model organism, we will investigate the cellular and molecular mechanisms that mediate pre-myelinating glial tiling at motor exit point (MEP) transition zones (TZ) and in the developing spinal cord. In Aim 1 of this project, we will use a combination of expression analysis, multi-color, in vivo, time-lapse imaging, electron microscopy, pharmacological and genetic manipulation (CRISPR) of newly identified mediators to elucidate the cellular and molecular mechanisms that govern myelinating glial tiling at MEP TZs. In Aim 2, we will determine if the same mechanisms we characterize in Aim 1 also mediate tiling between oligodendrocyte progenitor cells (OPC) in the developing spinal cord. To better understand, diagnose and treat the many degenerative disorders of the CNS and PNS, we need to comprehend the cellular and molecular mechanisms that mediate glial-glial interactions and tiling. Zebrafish provide a unique opportunity to directly observe and manipulate cell populations to gain insight into how glial cells interact under normal physiological conditions, and if those interactions that ultimately result in glial tiling are perturbed in disease.