Motile Phenotypes & Functional Diversity of Microglia

描述(申请人提供)：小胶质细胞是常驻脑的一类 在组织对损伤或感染的反应中起关键作用的细胞 哺乳动物的大脑。这项研究计划的长期目标是了解 脑组织小胶质细胞功能的细胞和分子基础 受伤。本申请将使用活细胞的动态成像 以研究哺乳动物脑组织的分子基础、调控和 小胶质细胞的运动功能。细胞运动性的变化被纳入其中 大多数脑损伤后小胶质细胞激活的模型，但很少有 已知小胶质细胞在自然脑组织中的运动行为。我们的 工作假说是实质小胶质细胞代表异质性 以及运动行为和运动行为的差异 激活的小胶质细胞的细胞间相互作用由以下因素决定 细胞黏附分子表达的动态模式，为 小胶质细胞群体中的功能多样性。我们在这里的主要目标 是为了阐明细胞对激活反应的多样性，以确定 并表征了不同的Moti表型和细胞间的相互作用 激活的小胶质细胞，并确定这些细胞是否受 细胞因子，肿瘤坏死因子-a，通过转录因子发挥作用， 核因子-KB。活体大鼠和小鼠脑组织切片中的小胶质细胞运动 通过生命荧光染色、3-D延时共聚焦成像和 计算机辅助定量图像分析。回溯性抗体染色 随后的时间推移观测将使定量确定 免疫表型与激活阶段和运动表型的关系。 将使用多通道时间推移成像来表征功能 小胶质细胞之间以及小胶质细胞与死亡/死亡之间的相互作用 神经元。最后，肿瘤坏死因子受体基因敲除切片中的小胶质细胞行为 将对小鼠进行评估以确定肿瘤坏死因子是否调节基于运动的 小胶质细胞功能。对分子机制有了更全面的了解 对小胶质细胞功能的研究应该有助于洞察调节 小胶质细胞在各种中枢神经系统损伤条件下的反应。包括创伤， 癫痫、中风、阿尔茨海默氏症和艾滋病。

DESCRIPTION (provided by applicant): Microglia are a class of resident brain cells that play key roles in the tissue response to injury or infection of the mammalian brain. The long-term goal of this research program is to understand the cellular and molecular bases of microglial function following brain tissue injury. The present application will use dynamic imaging of cells in live mammalian brain tissues to investigate the molecular basis, regulation, and function of microglial motility. Changes in cellular motility are incorporated into most models of microglial activation following brain injury, but little is known about the motile behaviors of microglia within native brain tissue. Our working hypothesis is that parenchymal microglia represent a heterogeneous population of cells, and that differences in the motile behaviors and intercellular interactions of activated microglial cells are determined by dynamic patterns of cell adhesion molecule expression, providing a basis for functional diversity within the microglial population. Our primary goals here are to elucidate the diversity in cellular response to activation, to define and characterize distinct moti' phenotypes and intercellular interactions of activated microglia, and to determine whether these are regulated by the cytokine, tumor necrosis factor (TNF)-a, acting via the transcription factor, NF-KB. Microglial movements in live rat and mouse brain tissue slices will be analyzed by vital fluorescent staining, 3-D time-lapse confocal imaging, and computer-assisted quantitative image analysis. Retrospective antibody staining following time-lapse observation will enable a quantitative determination of immunophenotypes in relation to the stages of activation and motile phenotypes. Multi-channel time-lapse imaging will be used to characterize functional interactions among microglial cells, and between microglia and dead/dying neurons. Finally, microglial behaviors in slices from TNF receptor knockout mice will be assessed to determine whether TNF regulates motility-based microglial functions. A more complete understanding of the molecular mechanisms of microglial function should yield insight into strategies for regulating the microglial response under a variety of CNS injury conditions. including trauma, epilepsy, stroke, Alzheimer's Disease, and AIDS.