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

项目摘要/摘要 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。 以及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显著的可塑性和韧性，当 大脑是成熟的。神经胶质细胞和血管细胞对幼年脑可塑性的潜在贡献 成人和老年大脑的再生潜力减弱的原因仍然知之甚少。 研究幼年和成熟胶质细胞和血管细胞之间的分子差异有望为 识别大脑中的保护性因素。我们最近开发了免疫泛素方法来 从人和小鼠中提纯星形胶质细胞、小胶质细胞、少突胶质细胞、神经元和血管细胞 大脑。通过免疫沉淀分离的细胞群体具有高纯度并产生丰富的RNA用于 转录组测序(rna-seq)。对纯化的细胞群体进行测序提供了 与单一方法相比，检测差异基因表达的灵敏度更高 细胞RNA序列使用免疫泛素，我们从一系列发育阶段的星形胶质细胞中提纯 人类和老鼠的大脑都有。在这项拟议的研究中，我们将首先对我们的 收集幼年和成熟星形胶质细胞数据，并确定每个阶段差异表达的基因(目标1)。 人类和老鼠的进化在大约1亿年前就分开了。翻译在以下方面的发现 将老鼠模型应用于临床一直是一项挑战。我们的人类和小鼠星形胶质细胞数据集将使我们能够 识别在一亿年前保存下来的发育调节分子通路 进化，因此很可能是必不可少的。这些分析将产生候选的星形细胞青少年 可以在未来的研究中测试的保护性因素。此外，我们将扩大我们的rna-seq比较。 将幼年和成熟细胞分化为神经元、小胶质细胞、少突胶质细胞和内皮细胞 免疫泛素纯化的细胞群体(目标2)。这些系统的分析有可能揭示 候选青少年保护因子中的每种细胞类型，提高我们对所起作用的认识 每种细胞类型在大脑成熟和衰老中的作用，并发现靶向于 治疗衰老和神经退行性变。拟议的研究将为更大规模的研究奠定基础 测试候选幼体保护因子在神经元、神经胶质细胞和血管细胞中的功能的研究。

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.