Time-lapse microscopy of macrophages during embryonic vascular development
Time-lapse microscopy of macrophages during embryonic vascular development
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DOI：
10.1002/dvdy.23835
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发表时间：
2012-09-01
影响因子：
2.5
通讯作者：
Jones, Elizabeth A. V.
中科院分区：
文献类型：
--
作者：
Al-Roubaie, Sarah;Hughes, Jasmine H.;Jones, Elizabeth A. V.
背景：巨噬细胞在血流开始前就存在了，但对其在血管发育中的功能知之甚少。我们开发了一种同时标记内皮细胞和巨噬细胞的技术，用于延时显微镜，使用荧光偶联乙酰化低密度脂蛋白（AcLDL）和吞噬染料PKH26-PCL。结果：我们对双标记细胞进行了表征，确认了巨噬细胞的特异性标记。双标记细胞循环，沿内皮滚动，并从血管外渗。大多数观察到的巨噬细胞整合到血管壁上，呈现内皮样形态。我们使用表达由内皮特异性启动子Tie1与吞噬染料结合驱动的荧光蛋白的转基因鹌鹑来分析这些细胞。循环pkh26 - pcl标记的细胞大部分是Tie1-，但那些已经整合到血管壁的细胞大部分是Tie1+。内皮样吞噬细胞在正常血管发育过程中一般是静止的。因此，我们诱导了血管重塑，并发现这些细胞可以被招募到重塑的部位。结论：内皮细胞和巨噬细胞的积极相互作用支持了这些细胞参与血管重塑的假设。吞噬性内皮样细胞的存在表明某些内皮细胞起源于髓系，或者循环内皮细胞/造血干细胞具有吞噬能力。科学通报，2012(1):1 - 4。(c) 2012 Wiley期刊有限公司
Background: Macrophages are present before the onset of blood flow, but very little is known about their function in vascular development. We have developed a technique to concurrently label both endothelial cells and macrophages for time-lapse microscopy using co-injection of fluorescently conjugated acetylated low-density lipoprotein (AcLDL) and phagocytic dye PKH26-PCL. Results: We characterize double-labeled cells to confirm specific labeling of macrophages. Double-labeled cells circulate, roll along the endothelium, and extravasate from vessels. Most observed macrophages are integrated into the vessel wall, showing an endothelial-like morphology. We used transgenic quail that express a fluorescent protein driven by the endothelial-specific promoter Tie1 in conjugation with the phagocytic dye to analyze these cells. Circulating PKH26-PCL-labeled cells are mostly Tie1-, but those which have integrated into the vessel wall are largely Tie1+. The endothelial-like phagocytic cells were generally stationary during normal vascular development. We, therefore, induced vascular remodeling and found that these cells could be recruited to sites of remodeling. Conclusions: The active interaction of endothelial cells and macrophages support the hypothesis that these cells are involved in vascular remodeling. The presence of phagocytic endothelial-like cells suggests either a myeloid-origin to certain endothelial cells or that circulating endothelial cells/hematopoietic stem cells have phagocytic capacity. Developmental Dynamics 241:1423-1431, 2012. (c) 2012 Wiley Periodicals, Inc.