A Novel Microscopic Assay Reveals Heterogeneous Regulation of Local Endothelial Barrier Function.
A Novel Microscopic Assay Reveals Heterogeneous Regulation of Local Endothelial Barrier Function.
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一种新的显微分析揭示了局部内皮屏障功能的异质调节
DOI：
10.1016/j.bpj.2019.02.008
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发表时间：
2019
影响因子：
3.4
通讯作者：
Jochen
中科院分区：
文献类型：
--
作者：
Klusmeier;Nadine;Schnittler;Hans-Joachim;Seebach;Jochen
血管内表面覆盖着内皮细胞，在血液和下面的组织之间形成选择性和半渗透性屏障。许多病理过程，如炎症或癌症转移，都伴随着血管通透性的增加。活细胞成像技术的进展最近揭示了内皮细胞接触的结构不断重组，并且内皮连接即使在一个细胞内也在亚细胞水平上显示出高度的异质性。虽然人们认为这种动态重塑与内皮屏障功能的局部变化有关，但由于缺乏适当的实验技术，缺少直接的证据。在这里，我们描述了一种动态测量局部内皮屏障功能的新检测方法，横向分辨率为± 15μm，时间分辨率为1分钟。在该设置中，将荧光标记的分子添加到内皮单层的顶室，并且通过利用所产生的倏逝场的全内反射荧光显微镜记录分子从顶室到基底室的渗透。通过这种技术，我们发现融合内皮细胞层内白蛋白的局部渗透性存在显著的异质性。在具有低渗透性的区域，用促炎剂组胺刺激导致细胞旁渗透性的瞬时增加。该效应显示沿着单个细胞的接触沿着的高变异性，表明内皮屏障功能的局部调节。在高基底通透性的区域，组胺没有明显的作用。相反，屏障增强药物毛喉素降低白蛋白和葡聚糖的渗透性均匀沿着细胞连接。由于这种新方法可以很容易地与其他活细胞成像技术相结合，它将有助于更好地了解伤口愈合，炎症和血管生成过程中亚细胞连接重组的机制。
Blood vessels are covered with endothelial cells on their inner surfaces, forming a selective and semipermeable barrier between the blood and the underlying tissue. Many pathological processes, such as inflammation or cancer metastasis, are accompanied by an increased vascular permeability. Progress in live cell imaging techniques has recently revealed that the structure of endothelial cell contacts is constantly reorganized and that endothelial junctions display high heterogeneities at a subcellular level even within one cell. Although it is assumed that this dynamic remodeling is associated with a local change in endothelial barrier function, a direct proof is missing mainly because of a lack of appropriate experimental techniques. Here, we describe a new assay to dynamically measure local endothelial barrier function with a lateral resolution of ∼15μm and a temporal resolution of 1 min. In this setup, fluorescence-labeled molecules are added to the apical compartment of an endothelial monolayer, and the penetration of molecules from the apical to the basal compartment is recorded by total internal reflection fluorescence microscopy utilizing the generated evanescent field. With this technique, we found a remarkable heterogeneity in the local permeability for albumin within confluent endothelial cell layers. In regions with low permeability, stimulation with the proinflammatory agent histamine results in a transient increase in paracellular permeability. The effect showed a high variability along the contact of one individual cell, indicating a local regulation of endothelial barrier function. In regions with high basal permeability, histamine had no obvious effect. In contrast, the barrier-enhancing drug forskolin reduces the permeability for albumin and dextran uniformly along the cell junctions. Because this new approach can be readily combined with other live cell imaging techniques, it will contribute to a better understanding of the mechanisms underlying subcellular junctional reorganization during wound healing, inflammation, and angiogenesis.
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影响因子：
6.7
作者：
J. Seebach;H. Mädler;B. Wojciak-Stothard;H. Schnittler
通讯作者：
J. Seebach;H. Mädler;B. Wojciak-Stothard;H. Schnittler
DOI：
10.1016/j.mrfmmm.2016.07.001
发表时间：
2016-08
期刊：
Mutation research
影响因子：
--
作者：
Petra Lakatos;C. Hegedűs;Nerea Salazar Ayestarán;Á. Juarranz;K. Kövér;É. Szabó;L. Virág
通讯作者：
Petra Lakatos;C. Hegedűs;Nerea Salazar Ayestarán;Á. Juarranz;K. Kövér;É. Szabó;L. Virág
使用电化学阻抗测量监测牛主动脉内皮细胞的 β-肾上腺素能刺激
DOI：
--
发表时间：
1999
期刊：
Pflügers Archiv
影响因子：
--
作者：
J. Wegener;S. Zink;P. Rösen;H. Franke
通讯作者：
H. Franke
用于上皮屏障功能横向解析多参数分析的大孔硅芯片。
影响因子：
6.1
作者：
S. Michaelis;Christina E. Rommel;J. Endell;P. Göring;R. Wehrspohn;C. Steinem;A. Janshoff;H. Franke;J. Wegener
通讯作者：
J. Wegener
CellBorderTracker，一种在亚细胞水平定量分析时空内皮连接动态的新工具
影响因子：
2.3
作者：
Seebach;Lindemann;Brinkmann;Bogdan;Schnittler
通讯作者：
Schnittler