Motile Phenotypes & Functional Diversity of Microglia

描述（由申请人提供）：小胶质细胞是一类常驻大脑 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 脑组织损伤后小胶质细胞功能的细胞分子基础 损伤本申请将使用活体细胞的动态成像， 研究哺乳动物脑组织的分子基础，调节， 小胶质细胞运动功能。细胞运动性的变化 大多数脑损伤后小胶质细胞激活的模型，但很少 了解小胶质细胞在天然脑组织中的运动行为。我们 工作假设是实质小胶质细胞代表了一种异质性的 细胞群，以及运动行为和 活化的小胶质细胞的细胞间相互作用由以下决定： 细胞粘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。我们的主要目标是 阐明细胞对激活反应的多样性， 并表征不同的动机表型和细胞间的相互作用， 激活的小胶质细胞，并确定这些是否受 细胞因子，肿瘤坏死因子（TNF）-α，通过转录因子起作用， NF-κ B。将在活体大鼠和小鼠脑组织切片中观察小胶质细胞的运动。 通过活体荧光染色、3-D延时共聚焦成像和 计算机辅助定量图像分析。回顾性抗体染色 随后的时间推移观察将能够定量确定 与活化阶段和运动表型相关的免疫表型。 多通道延时成像将用于表征功能性 小胶质细胞之间的相互作用，以及小胶质细胞和死亡/垂死之间的相互作用 神经元最后，TNF受体敲除切片中的小胶质细胞行为 将评估小鼠以确定TNF是否调节基于运动性 小胶质细胞功能更全面地了解 小胶质细胞功能的研究应该有助于深入了解调节小胶质细胞功能的策略。 在各种CNS损伤条件下的小胶质细胞反应。包括创伤， 癫痫、中风、老年痴呆症和艾滋病。

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.