Clonal analysis of gliogenesis in the cerebral cortex

大脑皮层在胚胎期和早期严重依赖于神经元和神经胶质的平衡产生。 产后发育最近发展的克隆谱系分析揭示了神经干的行为 神经干细胞（NSC）以前所未有的单细胞分辨率在大脑皮层中产生神经元。然而，在这方面， 神经干细胞形成神经胶质细胞的作用尚不清楚，还有待于用这些新的方法进行系统的研究。 技术.胶质细胞生成对于适当的神经元功能是至关重要的，并且当被破坏时，它可以导致各种神经元损伤。 神经系统疾病重建神经胶质细胞是如何从单个神经干细胞中产生并在神经干细胞中组织的 发育过程中的皮质对于理解结构-功能关系以及它们如何被 由克隆特异性因子调节。我们已经建立了一个基于遗传学的单细胞谱系追踪 利用MADM（双标记镶嵌分析）小鼠标记发育中皮层中的NSC的技术 并开始填补这一知识空白。使用这种方法，我们发现了两种不同的神经胶质细胞， 占据皮层及其相关结构海马结构的不同区域。的目标 建议的研究是重建，量化和数学模型的行为，个别标记 体内NSC。我们将利用这种标记方法的力量来筛选神经胶质克隆的基因表达 在单细胞分辨率，这一切都将帮助我们破译一般原则，组织胶质细胞克隆， 皮层，并定义克隆兄弟姐妹如何相互作用。我们将测试一些已确定的 基因在皮层神经胶质细胞克隆的产生，这将进一步帮助确定生物系统的基础 神经胶质生成的原理成功完成我们的研究将导致单一神经干细胞的全面地图 以及它们的神经胶质后代。我们的方法还将建立一个平台， 定量和计算分析胶质细胞生成、胶质细胞多样性及其再生潜力 在大脑皮层中接近。 更广泛影响的潜力 我们的方法来了解如何重要的组成部分的大脑，神经胶质细胞，发展有广泛的 影响神经胶质发育的中断是脑中一系列病理状况的根源。 因此，了解控制胶质细胞生成的基本原理和细胞机制对于 我不仅理解神经胶质细胞的系统性产生是如何控制健康发育的，而且 胶质细胞生成异常如何导致毁灭性的神经发育障碍和脑肿瘤。

The cerebral cortex critically relies on balanced production of neurons and glia during embryonic and early postnatal development. Recently developed clonal lineage analysis has revealed the behavior of neural stem cells (NSCs) giving rise to neurons in the cerebral cortex with unprecedented single-cell resolution. However, the formation of glia by NSCs remains unclear and has yet to be systematically investigated using these new technologies. Gliogenesis is critical for proper neuronal functions and when disrupted, it can result in various neurological diseases. Reconstructing how glia are generated from individual NSCs and organized in the cortex during development is essential to understand the structure-function relationships and how they can be modulated by clone-specific factors. We have established a genetically-based single-cell lineage tracing technique utilizing MADM (Mosaic Analysis with Double Markers) mice to label NSCs in the developing cortex and begin to address this knowledge gap. Using this method we have found two distinct populations of glia that occupy different territories of the cortex and its related structure the hippocampal formation. The goal of the proposed research is to reconstruct, quantify, and mathematically model the behavior of individually labeled NSCs in vivo. We will use the power of this labeling method to also screen for gene expression of glial clones at single cell resolution, which all together will help us decipher the general principles organizing glial clones in the cortex, and define how clonal siblings interact with each other. We will test the role of some of the identified genes in generation of glial clones in the cortex, which will further help define the biological system underlying principles of gliogenesis. Successful completion of our study will result in a comprehensive map of single NSCs and their glial progeny in various cortical regions. Our approach will also establish a platform for detailed quantitative and computational analysis of gliogenesis, glial diversity, and their potential for regenerative approaches in the cortex. Potential for Broader Impact Our approaches to understand how important constituents of the brain, the glial cells, develop have wide implications. Disruption of glial development is the root of a range of pathological conditions in the brain. Therefore, understanding the basic principles and cellular mechanisms that control gliogenesis is critical to appreciate not only how healthy development may be controlled by systematic production of glial cells, but also how abnormalities in gliogenesis may lead to devastating neurodevelopmental disorders and brain tumors.