Cellular Imaging Core

摘要 本计划项目的总体目标是测试新的假设，即血管内皮细胞（EC） 在损伤和修复过程中作为肺中的免疫活性小生境。细胞成像核心（核心C）是必不可少的 用于精确识别和跟踪特定分子的时空组织和表达 在调节免疫生态位的EC。核心的功能是提供先进的试剂和成像 免疫功能的体外和体内分析所需的方法，并使测试免疫功能成为可能。 整个计划的假设。我们将提供工具，以解决如何积极调节内皮细胞 细胞表面受体信号传导和EC介导的吞噬细胞的线粒体吞噬识别促进先天性 肺中的免疫信号。为了获得每个项目成功所需的机械信号洞察力， “核心”计划将提供常规和超分辨率水平的方法，以识别和询问 信号通路使用多通道显微镜和超分辨率活动成像，我们将研究 通过酪氨酸磷酸化鞘氨醇-1-磷酸内化诱导的时空信号传导 EC中S1 PR 1受体的表达以及EC表达的泛素连接酶CFHR在调节VE-钙粘蛋白中的作用 表达和功能。这些分析将允许对时间序列和空间分布进行细粒度评估。 肺EC中免疫信号传导的组织及其如何影响肺功能。通过提供无缝的视觉效果， 检查上述EC受体下游的单个EC中的广泛信号传导活性 在炎性损伤和修复过程中，核心蛋白对于支持和检测分子和 项目提出的功能假设，对项目的整体成功至关重要。

ABSTRACT The overall goal of this Program Project is to test the novel hypothesis that vascular endothelial cells (EC) serve as an immune active niche in the lungs during injury and repair. The Cellular Imaging Core (Core C) is essential for accurate identification and tracking of the spatiotemporal organization and expression of specific molecules in the EC regulating the immune niche. The function of the Core is to provide advanced reagents and imaging methodologies required for both in vitro and in vivo analysis of immune function and to enable testing of the hypotheses across the Program. We will provide the tools to address how the active regulation of endothelial cell-surface receptor signaling and mitophagy-mediated recognition of phagocytes by EC promote innate immune signaling in the lung. To gain the mechanistic signaling insights needed for the success of each Project, the Core will provide methods at both the conventional and super-resolution levels to identify and interrogate the signaling pathways. Using multi-channel microscopy and super-resolution activity imaging, we will study spatiotemporal signaling induced through internalization of tyrosine phosphorylated sphingosine-1-phosphate receptor1 (S1PR1) in EC and the role of the EC-expressed ubiquitin ligase CFHR in regulating VE-cadherin expression and function. These analyses will permit a fine-grained assessment of the time-sequence and spatial organization of the immune signaling in lung EC and how it impacts lung function. By providing a seamless visual examination of a wide range of signaling activities in individual EC downstream of above listed EC-receptors during inflammatory injury and repair, the Core is essential for supporting and testing the molecular and functional hypotheses advanced by the Projects, and crucial to the Program’s overall success.