Cellular and Genetic Origins of Astrocytes

星形胶质细胞的细胞和遗传起源

基本信息

批准号：
8214621
负责人：
DAVID H ROWITCH
金额：
$ 72.57万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-02-01 至 2014-01-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The human brain is made up of neurons and glia. Glia comprise about 90% of brain cells and are involved in devastating disorders such as multiple sclerosis, seizures and Alzheimer's disease. We know little about development of the most prevalent glial cells called astrocytes. This proposal will identify fundamental mechanisms by which neural stem cells give rise to astrocytes. It is also intended to generate useful markers to investigate roles for astrocytes during development and in human disease. The goal of the proposed research is to identify a set of genetic markers for development of astrocytes and to map their cellular origins in the central nervous system. The underlying hypothesis to be tested is that astrocytes develop from heterogeneous locations in the developing CMS. We have three specific aims: Specific Aim 1 is to identify and characterize activity of transcription factors that uniquely mark and may regulate astrocyte development. In preliminary work, we have interrogated the mammalian transcriptome to identify transcription factors that co-localize with astrocytes in the developing spinal cord. Expression analysis and gain- and Ioss-of-function screens will be conducted to prioritize generation of antibodies to transcription factors that mark and may regulate astrocyte lineage heterogeneity in the CNS. Specific Aim 2 is to develop cell type- and stage-specific markers for fibrous and protoplasmic astrocytes. We have developed fluorescent activated cell sorting (FACS) protocols for acute harvest of gray matter astrocytes and analysis by expression profiling. By sorting cells from Gensat transgenic mice that express green fluorescent protein (GFP) in (1) pan-astroglial and (2) white matter astrocyte compartments, we will identify markers specific for fibrous astrocytes, protoplasmic astrocytes and astrocyte precursors in various CNS regions. Specific Aim 3 is to determine whether astrocyte diversity correlates with developmental site of origin in the embryonic spinal cord and brain. We will use a cohort of cre/lox transgenic mice to fate map heterogeneous progenitor domains for astrocytes and their ultimate cellular origins in the embryonic CNS. Based on information obtained in Aims1/2, we will generate a floxed conditional reporter transgenic mouse Iine that will express the FACS-selectable marker GFP exclusively in astroglia. This will enable acute purification of astrocyte subsets for further discrimination of astrocyte heterogeneity. PUBLIC HEALTH RELEVANCE These studies will elucidate the mechanisms by which neural stem cells give rise to astrocytes and provide useful genetic tools for understanding the diverse roles of astrocytes in human development, with practical implications for diagnosis and treatment of human disorders such as epilepsy, brain cancer and neurodegeneration.

描述（由申请人提供）：人脑由神经元和神经胶质组成。神经胶质细胞约为90％的脑细胞，并参与毁灭性疾病，例如多发性硬化症，癫痫发作和阿尔茨海默氏病。我们对最普遍的神经胶质细胞的发展知之甚少。该建议将确定神经干细胞引起星形胶质细胞的基本机制。它还旨在产生有用的标记，以研究在发育和人类疾病中星形胶质细胞的作用。拟议的研究的目的是确定一组遗传标记，以开发星形胶质细胞，并绘制其在中枢神经系统中的细胞起源。要测试的基本假设是，星形胶质细胞从发育中的CMS中的异质位置发展。我们有三个特定的目的：特定目的1是识别和表征唯一标记并可能调节星形胶质细胞发育的转录因子的活性。在初步工作中，我们询问了哺乳动物转录组，以识别与发育中脊髓中星形胶质细胞共定位的转录因子。表达分析以及功能屏幕将进行，以优先考虑标记并可能调节CNS中星形胶质细胞谱系异质性的转录因子的抗体。具体目的2是开发纤维和原生质星形胶质细胞的细胞类型和特异性标记。我们已经开发了荧光活化的细胞分选（FACS）方案，用于通过表达分析进行灰质星形胶质细胞的急性收获和分析。通过对（1）泛腹和（2）白质星形胶质细胞室中表达绿色荧光蛋白（GFP）的Gensat转基因小鼠的细胞进行分类，我们将确定针对各种CNS地区的纤维胶质胶质细胞，原生质星形胶质细胞和星形胶质细胞的特异性标记物。具体目的3是确定星形胶质细胞多样性是否与胚胎脊髓和大脑中原产的发育部位相关。我们将使用一组CRE/LOX转基因小鼠来命运地图，用于星形胶质细胞的异质祖细胞及其在胚胎中枢神经系统中的最终细胞起源。基于在AIMS1/2中获得的信息，我们将生成一个有条件的记者转基因小鼠Iine，该鼠标将在Astroglia中独家表达FACS可选择的标记GFP。这将使星形胶质细胞子集的急性纯化以进一步歧视星形胶质细胞异质性。公共卫生相关性这些研究将阐明神经干细胞引起星形胶质细胞的机制，并为理解星形胶质细胞在人类发育中的多种作用提供了有用的遗传工具，对诊断和治疗人类疾病（如癫痫，脑癌，脑癌和神经变性）的诊断和治疗具有实际意义。