Identification of candidate juvenile protective factors in neuron, glia, and vascular cells of human and mouse brain

项目总结/摘要 神经胶质细胞（星形胶质细胞、小胶质细胞和少突胶质细胞）和血管细胞对发育至关重要。 和中枢神经系统的功能。神经胶质和血管缺陷与衰老有关 和神经退化青少年的大脑表现出非凡的可塑性和弹性， 大脑成熟。胶质细胞和血管细胞对幼年脑可塑性的潜在贡献 对成年人和老年人大脑再生潜能的减少仍然知之甚少。 研究幼年和成熟神经胶质细胞和血管细胞之间的分子差异， 识别大脑中的保护因素。我们最近开发了免疫淘选方法， 从人和小鼠中纯化星形胶质细胞、小胶质细胞、少突胶质细胞、神经元和血管细胞 大脑通过免疫淘选分离的细胞群具有高纯度，并产生丰富的RNA， 通过RNA测序（RNA-seq）进行转录组谱分析。纯化的细胞群的测序提供了 与替代方法如单一的基因表达检测相比， 细胞RNA测序使用免疫淘洗，我们从一系列发育阶段纯化星形胶质细胞， 人类和老鼠的大脑。在这项拟议的研究中，我们将首先对我们的 幼年和成熟星形胶质细胞数据集，并确定在每个阶段差异表达的基因（目标1）。 人类和老鼠的进化大约在1亿年前分开。翻译发现在 将小鼠模型应用于临床一直是一个挑战。我们的人类和小鼠星形胶质细胞数据集将使我们能够 鉴定经过一亿年保存下来的发育调节分子通路， 进化，因此可能是必不可少的。这些分析将产生候选的星形胶质细胞少年 这些保护因素可以在未来的研究中进行测试。此外，我们将扩大我们的RNA-seq比较， 幼稚和成熟细胞的神经元，小胶质细胞，少突胶质细胞和内皮细胞， 免疫淘洗纯化的细胞群体（目的2）。这些系统分析有可能揭示 候选的青少年保护因子在每种细胞类型，提高我们的理解所发挥的作用， 每种细胞类型在大脑成熟和衰老中的作用，并揭示候选分子通路，以靶向大脑中的 衰老和神经变性的治疗。拟议的研究将为更大规模的研究奠定基础。 一项测试候选保幼因子在神经元、神经胶质和血管细胞中功能的研究。

Project Summary/Abstract Glia (astrocytes, microglia, and oligodendrocytes) and vascular cells are critical for the development and function of the central nervous system. Glial and vascular defects are associated with aging and neurodegeneration. Juvenile brains exhibit remarkable plasticity and resilience that diminish when brains mature. The potential contribution of glia and vascular cells to the plasticity of juvenile brains and the diminution of regenerative potentials in adult and aging brains remain poorly understood. Investigating molecular differences between juvenile and mature glia and vascular cells holds promise for the identification of protective factors in the brain. We recently developed immunopanning methods to purify astrocytes, microglia, oligodendrocytes, neurons, and vascular cells from both human and mouse brains. Cell populations isolated by immunopanning have high purity and produce abundant RNA for transcriptome profiling by RNA-sequencing (RNA-seq). Sequencing of purified populations of cells provides higher sensitivity for the detection of differential gene expression than alternative methods such as single cell RNA-seq. Using immunopanning, we purified astrocytes from a series of developmental stages from both human and mouse brains. In this proposed study, we will first perform bioinformatics analysis of our juvenile and mature astrocyte datasets and identify differentially expressed genes at each stage (Aim 1). Human and mouse evolution separated about 100 million years ago. Translating discoveries made in mouse models into clinics has been challenging. Our human and mouse astrocyte datasets will allow us to identify developmentally regulated molecular pathways preserved through a hundred million years of evolution and therefore likely to be essential. These analyses will generate candidate astrocytic juvenile protective factors that can be tested in future studies. Furthermore, we will expand our RNA-seq comparison of juvenile and mature cells to neurons, microglia, oligodendrocytes, and endothelial cells using immunopanning purified cell populations (Aim 2). These systematic analyses have the potential to reveal candidate juvenile protective factors in each cell type, improve our understanding of the roles played by each cell type in brain maturation and aging, and uncover candidate molecular pathways to target in the treatment of aging and neurodegeneration. The proposed study will build the foundation for a larger scale study that tests the function of candidate juvenile protective factors in neurons, glia, and vascular cells.