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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概要大脑在胚胎和产后早期发育过程中严重依赖于神经元和神经胶质细胞的平衡产生。最近开发的克隆谱系分析揭示了神经干细胞（NSC）产生神经元的行为大脑皮层具有前所未有的单细胞分辨率。然而，形成的克隆原理神经干细胞产生的神经胶质细胞仍不清楚，并且尚未使用这些新技术进行系统研究。胶质细胞生成是对于正常的神经元功能至关重要，当它受到干扰时，可能会导致各种神经系统疾病。重建神经胶质细胞是如何从单个神经干细胞产生并在发育过程中在皮层中组织的，对于了解这一点至关重要结构-功能关系以及它们如何通过克隆特异性因素进行调节。我们已经建立了一个基于基因的单细胞谱系追踪技术，利用 MADM（双标记马赛克分析）小鼠标记发育中皮层中的 NSC，并开始解决这一知识差距。使用这个方法我们发现了两个占据皮质不同区域的不同神经胶质细胞群及其相关结构海马体形成。拟议研究的目标是重建、量化和数学建模在体内新皮质和古皮质区域单独标记的 NSC。这项努力需要发展优化的成像和分析工具，确保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人工智能的自动定量方法，以促进公正和精确大脑中克隆数据的成像和量化。成功完成我们的研究将带来全面的各个皮质区域的单个 NSC 及其神经胶质后代的图谱。我们的方法还将建立一个平台胶质生成、胶质多样性及其修复和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情况下，皮层的再生方法。产生更广泛影响的潜力我们了解大脑的重要组成部分——神经胶质细胞——如何发育的方法具有广泛的意义。神经胶质发育的破坏是大脑中一系列病理状况的根源。因此，了解控制胶质生成的基本原理和细胞机制不仅对于了解健康状况至关重要发育可能是通过神经胶质细胞的系统产生来控制的，但神经胶质生成的异常也可能是如何控制的导致毁灭性的神经发育障碍和脑肿瘤。