High Throughput Clonal Analyses of Gliogenesis in Neocortical and Paleocortical areas of the Mouse Brain

小鼠大脑新皮质和古皮质区域胶质生成的高通量克隆分析

• 批准号: 10536298
• 负责人: Hooman Troy Ghashghaei
• 金额: $ 41.16万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10536298
• 关键词: 3-Dimensional; Address; Adult; Alexander Disease; Alleles; Alzheimer' s Disease; Area; Artificial Intelligence; Atlases; Behavior; Behavioral; Biological; Birth; Brain; Brain Injuries; Brain Neoplasms; CNS degeneration; Cell Lineage; Cells; Cerebral cortex; Classification; Color; Computer Analysis; Custom; Data; Defect; Detection; Development; Disease; Embryo; Engineering; Ensure; Epidermal Growth Factor Receptor; Equilibrium; Event; Experimental Models; Future; Genes; Genetic; Genotype; Glioma; Goals; Hippocampal Formation; Homeostasis; Image; Imaging Device; Individual; Injury; Knowledge; Label; Lead; Life; Light; Manuals; Maps; Methodology; Methods; Microscope; Microscopy; Mitotic Recombination; Mosaicism; Motor; Multiple Sclerosis; Mus; Neocortex; Neurobiology; Neurodevelopmental Disorder; Neuroglia; Neurons; Pathologic; Pattern; Physiological; Plant Roots; Population; Population Sizes; Positioning Attribute; Process; Production; Property; Prosencephalon; Publishing; Regulation; Research; Research Proposals; Resolution; Rodent; Siblings; Source; Spinal cord injury; Stroke; Structure; Structure-Activity Relationship; Techniques; Technology; Testing; Three-Dimensional Imaging; Time; Tissues; Trees; Wild Type Mouse; Work; analytical method; analytical tool; base; brain tissue; cell behavior; cell growth; cell type; central nervous system injury; dosage; falls; glial cell development; gliogenesis; in vivo; innovation; mathematical model; neocortical; nerve stem cell; nervous system disorder; neurogenesis; new technology; postnatal development; progenitor; public health relevance; regenerative approach; regional difference; repaired; response; response to brain injury; somatosensory; stem cell expansion; tool

Project Summary The brain 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 clonal principles underlying 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 in neocortical and paleocortical areas. This effort requires the development of optimized imaging and analytical tools to ensure reliable and repeatable interpretation of quantitative data. To this end we are developing light sheet microscopy and AI-based automated quantification methods to facilitate unbiased and precise imaging and quantification of clonal data in the brain. 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 repair and regenerative approaches in the cortex in the context of various neurological disorders and brain injury. 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.

项目摘要 大脑在胚胎和早期产后发育过程中依赖神经元和神经胶质的平衡产生。 最近开发的克隆谱系分析揭示了神经干细胞（NSC）的行为，从而引起神经元的神经元 具有前所未有的单细胞分辨率的大脑皮层。但是，形成的基础的克隆原则 NSC的Glia尚不清楚，尚未使用这些新技术进行系统的研究。神经胶质发生是 对于适当的神经元功能至关重要，并且在破坏时可能导致各种神经系统疾病。重建 在开发过程中，如何从单个NSC产生神经胶质并在皮层中组织起来是必不可少的 结构功能关系以及如何通过克隆特异性因素调节它们。我们已经建立了 利用MADM（带有双标记的马赛克分析）小鼠的基于遗传的单细胞谱系示踪技术 在开发皮层中标记NSC，并开始解决此知识差距。使用这种方法，我们发现了两个 占颗粒的不同种群，占据了皮质的不同领土及其相关结构海马 形成。拟议的研究的目的是重建，量化和数学建模 在新皮质和古生物区域中单独标记为NSC体内的NSC。这项工作需要发展 优化的成像和分析工具，以确保对定量数据的可靠且可重复的解释。为此，我们 正在开发光片显微镜和基于AI的自动定量方法，以促进无偏和精确的 大脑克隆数据的成像和定量。成功完成我们的研究将导致全面 单个NSC及其神经胶质后代的图。我们的方法还将建立一个平台 神经胶质发生，神经胶质多样性及其修复潜力的详细定量和计算分析 在各种神经系统疾病和脑损伤的背景下，皮质中的再生方法。 潜力更广泛 我们了解大脑，神经胶质细胞的重要成分的方法具有广泛的影响。 神经胶质发育的破坏是大脑中一系列病理条件的根源。因此，了解 控制神经胶质发生的基本原理和细胞机制，不仅要欣赏健康的健康 开发可以通过系统地产生神经胶质细胞来控制，而且神经胶质发生异常 导致毁灭性神经发育障碍和脑肿瘤。