Cellular Imaging Core

摘要 该计划项目的总体目标是测试血管内皮细胞(EC)服务的新假设 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 用于准确识别和跟踪特定分子的时空组织和表达 在欧共体中调节免疫利基。核心的功能是提供先进的试剂和成像 体外和体内免疫功能分析所需的方法学，以及能够测试 整个计划中的假设。我们将提供工具来解决如何主动调节内皮细胞 细胞表面受体信号转导和吞噬细胞对吞噬细胞的识别促进天然 肺部的免疫信号。为了获得每个项目成功所需的机械信号洞察， 核心将提供常规和超分辨率水平的方法，以识别和审问 信号通路。利用多通道显微镜和超分辨率活动成像，我们将研究 酪氨酸磷酸化的鞘氨醇-1-磷酸内化诱导的时空信号 EC中的受体1(S1PR1)及其表达的泛素连接酶CFHR在调节VE-钙粘附素中的作用 表情和功能。这些分析将允许对时间序列和空间进行细粒度的评估 肺内皮细胞免疫信号的组织及其对肺功能的影响。通过提供无缝的可视 检测上述EC受体下游单个EC的广泛信号活性 在炎症损伤和修复过程中，核心对于支持和测试分子和 由项目提出的功能假设，对该计划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.