Cellular Imaging Core

细胞成像核心

• 批准号: 10494614
• 负责人: Gary CH Mo
• 金额: $ 33.14万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10494614
• 关键词: Address; Binding; Biochemical; Biochemical Process; Biological Response Modifiers; Biosensing Techniques; Calcium; Calcium Channel; Cell Surface Receptors; Cell membrane; Cells; Complement; Complex; Data Analyses; Development; Diffusion; Endoplasmic Reticulum; Endothelial Cells; Endothelium; Event; Fluorescence Resonance Energy Transfer; Goals; Grain; Image; Immune; Immune signaling; In Vitro; Individual; Inflammatory; Injury; Innate Immune System; Instruction; Link; Lung; Lysine; Mediating; Membrane; Membrane Proteins; Methodology; Methods; Microscopy; Molecular; Molecular Chaperones; Monitor; Phagocytes; Physiology; Play; Protein translocation; Proteins; Reagent; Receptor Cell; Receptor Signaling; Regulation; Resolution; Role; Second Messenger Systems; Signal Pathway; Signal Transduction; Signaling Protein; Techniques; Testing; Time; Training; Tyrosine; Ubiquitin; Ubiquitination; Vascular Endothelial Cell; Visual; Visualization; cadherin 5; cellular imaging; combinatorial; depolymerization; experimental study; immune function; immunoreaction; in vivo; in vivo imaging; injury and repair; insight; intravital imaging; novel; novel strategies; programs; protein degradation; protein protein interaction; pulmonary function; response; single molecule; small molecule; spatiotemporal; sphingosine 1-phosphate; success; tool; ubiquitin ligase; ubiquitin-protein ligase; vascular injury

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.

抽象的 该计划项目的总体目标是检验血管内皮细胞（EC）服务的新假设 在受伤和修复过程中，作为肺部的免疫科活性小众。细胞成像芯（核心C）是必不可少的 为了准确鉴定和跟踪时空组织和特定分子的表达 在EC调节中，免疫利基。核心的功能是提供高级试剂和成像 体外和体内免疫功能分析所需的方法学都需要测试 整个程序中的假设。我们将提供工具来解决内皮的主动调节 细胞表面受体信号传导和线粒体介导的对吞噬细胞的识别EC促进先天 肺中的免疫信号传导。为了获得每个项目成功所需的机械信号洞察， 核心将在常规和超分辨率级别上提供方法，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 酪氨酸磷酸化的鞘氨醇1-磷酸诱导的时空信号传导诱导的时空信号传导 EC中的受体1（S1PR1）和EC表达的泛素连接酶CFHR在调节VE-cadherin中的作用 表达和功能。这些分析将允许对时间序和空间进行细粒度评估 肺EC中免疫信号的组织及其如何影响肺功能。通过提供无缝的视觉 检查上述EC受体下游的单个EC中的各种信号传导活动 在炎症性损伤和修复期间，核心对于支持和测试分子和 项目提出的功能假设对该计划的整体成功至关重要。