Build them up and break them down: Tight junctions of cell lines expressing typical hepatocyte polarity with a varied repertoire of claudins.
Build them up and break them down: Tight junctions of cell lines expressing typical hepatocyte polarity with a varied repertoire of claudins.
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DOI：
10.4161/tisb.25210
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发表时间：
2013-10-01
期刊：
影响因子：
3.1
通讯作者：
Cassio D
中科院分区：
文献类型：
--
作者：
Grosse B;Degrouard J;Jaillard D;Cassio D
表达简单上皮极性的细胞的紧密连接（TJs）已被广泛研究，但对表达复杂极性的细胞的TJs知之甚少。在本文中，我们分析了形成胆小管（BC）并表达典型肝细胞极性的四个不同品系的TJ; WIF-B 9、11-3、Can 3-1、Can 10。在紧密连接蛋白表达中观察到显著差异。这些细胞系均不产生密蛋白-1。WIF-B 9和11-3仅表达密蛋白-2，而Can 3-1和Can 10表达密蛋白-2、密蛋白-3、密蛋白-4、密蛋白-5。这两类系的TJ在其超微结构、细胞旁渗透性和稳健性方面不同。表达大的紧密连接蛋白库的系，特别是Can 10，具有复杂且有效的TJ，当细胞耗尽钙时，所述TJ得以维持。WIF-B 9和11-3的TJs是不稳定的，泄漏的，允许的，并且由于钙耗尽而被拆除。有趣的是，我们发现，在极化过程中，TJ蛋白表达的所有线依次解决在一个特定的顺序：首先occludin，ZO-1和cingulin，然后JAM-A和ZO-2，最后claudin-2。仅在Can系中表达的claudin也依次被固定：claudin-3是第一个被固定的。抑制claudin-3表达可延迟Can 10中BC的形成，并诱导单纯上皮极性的表达。这些结果突出了紧密连接蛋白在表达典型肝细胞极性的细胞系中的沉降和TJ的效率中的作用。Can 10似乎是这些细胞系中最有前途的，因为其紧密连接蛋白库接近肝细胞的紧密连接蛋白库，并且其能够形成由有效TJ密封的延伸的管状BC。
Tight junctions (TJs) of cells expressing simple epithelial polarity have been extensively studied, but less is known about TJs of cells expressing complex polarity. In this paper we analyzed, TJs of four different lines, that form bile canaliculi (BC) and express typical hepatocyte polarity; WIF-B9, 11–3, Can 3–1, Can 10. Striking differences were observed in claudin expression. None of the cell lines produced claudin-1. WIF-B9 and 11–3 expressed only claudin-2 while Can 3–1 and Can 10 expressed claudin-2,-3,-4,-5. TJs of these two classes of lines differed in their ultra-stucture, paracellular permeability, and robustness. Lines expressing a large claudin repertoire, especially Can 10, had complex and efficient TJs, that were maintained when cells were depleted in calcium. Inversely, TJs of WIF-B9 and 11–3 were leaky, permissive and dismantled by calcium depletion. Interestingly, we found that during the polarization process, TJ proteins expressed by all lines were sequentially settled in a specific order: first occludin, ZO-1 and cingulin, then JAM-A and ZO-2, finally claudin-2. Claudins expressed only in Can lines were also sequentially settled: claudin-3 was the first settled. Inhibition of claudin-3 expression delayed BC formation in Can10 and induced the expression of simple epithelial polarity. These results highlight the role of claudins in the settlement and the efficiency of TJs in lines expressing typical hepatocyte polarity. Can 10 seems to be the most promising of these lines because of its claudin repertoire near that of hepatocytes and its capacity to form extended tubular BC sealed by efficient TJs.