Intravital Imaging and Physiology Core

摘要 该计划项目的重点是研究潜在的分子和信号机制。 肺内皮龛在炎症性肺损伤中的免疫调节作用及消退 受伤。生命成像和生理学核心(核心D)对于精确定义动态 损伤和修复过程中肺内皮细胞的变化免疫细胞促进 或消炎性肺损伤。每个项目都清楚地表明，时空信号由 内皮细胞表面受体和线粒体在调节内皮细胞的功能中起着至关重要的作用。 免疫利基和定义肺损伤的结局和解决。因此，所有使用内皮细胞的项目- 特定的基因改变的活小鼠模型，将研究基础和激活的信号是如何发出的 内皮细胞改变中性粒细胞/单核/巨噬细胞命运以响应肺损伤并参与 决议。活体动物的呼吸运动是获得有意义的 对正常呼吸的肺进行活体成像研究的结果。我们开发了一种新的成像技术 在损伤和修复过程中监测肺微血管和免疫细胞运输的方法。因此，一个 核心D的主要功能将是提供先进的双光子活体内肺成像专业知识，以可靠地 定量血管内皮细胞的激活，测量肺血管渗漏，并确定激活的作用 肺血管生态位在调节活体小鼠肺内髓系细胞跨内皮细胞迁移中的作用。 每个项目将与核心D一起检查其感兴趣路径的功能影响 在炎症性肺损伤和修复阶段。成像分析将包括评估 吞噬细胞移位及免疫细胞吞噬和吞噬泡泡细胞的原位活性 内皮细胞内质网定位的S1PR1(鞘氨醇-1-磷酸受体-1)编程 项目1中的细胞；CHFR(一种E3连接酶，带有叉头和环指结构域的检查点)诱导的 项目2中VE-钙粘附素在内皮细胞中的降解；内皮细胞有丝分裂激活和 项目3中的线粒体生物发生。通过提供对内皮细胞活性及其 炎性损伤和修复过程中对髓系细胞功能和生理评估的影响 D将是该计划成功的关键。

ABSTRACT The focus of this Program Project is to investigate the molecular and signaling mechanisms underlying the immune regulatory role of the pulmonary endothelial niche during inflammatory lung injury and resolution of injury. The Intravital Imaging and Physiology Core (Core D) is essential to precisely define how dynamic changes in the pulmonary endothelium during injury and repair programs the immune cells to either promote or resolve inflammatory lung injury. Each Project makes clear that spatio-temporal signaling regulated by endothelial cell-surface receptors and mitochondria is fundamentally important in regulating the endothelial immune niche and defining the outcome of lung injury and resolution. Thus, all Projects, using endothelial- specific genetically altered living mouse models, will address how signals emanating from basal and activated endothelial cells alter neutrophil/monocyte/macrophage fate in response to lung injury and contribute to resolution. Respiratory motion in the live animal represents a major obstacle for obtaining meaningful results from in vivo imaging studies of the normally respiring lung. We have developed a novel imaging approach to monitor pulmonary microvessels and immune cell trafficking during injury and repair. Thus, a major Core D function will be to provide advanced two-photon intravital lung imaging expertise to reliably quantify the activation of endothelium, to measure lung vascular leak, and to identify the role of activated pulmonary vascular niche in regulating transendothelial migration of myeloid cells in the living mouse lung. Each Project in conjunction with Core D will examine the functional effects of their pathways of interest during inflammatory lung injury and repair phases. Imaging analyses will include the assessment of transmigration of phagocytic cells and in situ phagocytic and efferocytotic activities of immune cells as programmed by endoplasmic reticulum-localized S1PR1 (sphingosine-1-phosphate receptor-1) in endothelial cells in Project 1; CHFR (an E3 ligase, checkpoint with Forkhead and ring finger domains)-induced degradation of VE-cadherin in endothelial cells in Project 2; and activation of endothelial cell mitophagy and mitochondrial biogenesis in Project 3. By providing visual in vivo examination of endothelial activity and its impact on myeloid cell functions and physiological assessments during inflammatory injury and repair, Core D will be essential for the Program’s success.