Claudin-5 controls intercellular barriers of human dermal microvascular but not human umbilical vein endothelial cells.
Claudin-5 controls intercellular barriers of human dermal microvascular but not human umbilical vein endothelial cells.
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DOI：
10.1161/atvbaha.112.300893
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发表时间：
2013-03
期刊：
影响因子：
--
通讯作者：
Pober JS
中科院分区：
文献类型：
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作者：
Kluger MS;Clark PR;Tellides G;Gerke V;Pober JS
为了评估claudin-5（一种内皮细胞（EC）紧密连接（TJ）蛋白）在建立人体基础渗透性水平中的作用，通过比较claudin-5的原位表达水平并分析两种广泛使用的培养EC模型（即人真皮微血管（HDM）EC和人脐静脉（HUV）EC）中的连接组织和功能。通过免疫荧光显微镜，EC在人真皮毛细血管与毛细血管后小静脉中以及在脐动脉和冠状动脉与静脉中更高地表达紧密连接蛋白-5（但等效地表达VE-钙粘蛋白），这与TJ频率和渗透性屏障中的已知节段性差异相关。融合后培养的HDMEC表达更多的claudin-5（但等效的VE-钙粘蛋白），并显示出更高的跨内皮电阻（TEER）和更低的大分子通量比类似培养的HUVEC。通过透射电子显微镜观察，HDMEC连接更复杂，并且显示出比HUVEC连接更连续的claudin-5免疫荧光。钙螯合或显性负性VE-钙粘蛋白过表达降低TEER并破坏HUVEC中的连接，但不在HDMEC中。HUVEC中的紧密连接蛋白-5过表达不能增加TEER或紧密连接蛋白-5连续性，而HDMEC中而不是HUVEC中的紧密连接蛋白-5敲低降低了TEER并增加了抗体对连接蛋白的可及性。紧密连接蛋白-5表达和连接组织控制HDMEC和小动脉-毛细血管旁细胞屏障，而HUVEC和小静脉连接利用VE-钙粘蛋白。
To assess the role claudin-5, an endothelial cell (EC) tight junction (TJ) protein, plays in establishing basal permeability levels in humans by comparing claudin-5 expression levels in situ and analyzing junctional organization and function in two widely used models of cultured ECs, namely human dermal microvascular (HDM)ECs and human umbilical vein (HUV)ECs. By immunofluorescence microscopy, ECs more highly express claudin-5 (but equivalently express VE-cadherin) in human dermal capillaries versus post-capillary venules and in umbilical and coronary arteries versus veins, correlating with known segmental differences in TJ frequencies and permeability barriers. Post-confluent cultured HDMECs express more claudin-5 (but equivalent VE-cadherin) and show higher transendothelial electrical resistance (TEER) and lower macromolecular flux than similarly cultured HUVECs. HDMEC junctions are more complex by transmission electron microscopy and show more continuous claudin-5 immunofluorescence than HUVEC junctions. Calcium chelation or dominant negative VE-cadherin overexpression decreases TEER and disrupts junctions in HUVECs, but not in HDMECs. Claudin-5 overexpression in HUVECs fails to increase TEER or claudin-5 continuity while claudin-5 knockdown in HDMECs, but not HUVECs, reduces TEER and increases antibody accessibility to junctional proteins. Claudin-5 expression and junctional organization control HDMEC and arteriolar-capillary paracellular barriers whereas HUVEC and venular junctions utilize VE-cadherin.