Detection of tight junction barrier function in vivo by biotin.
Detection of tight junction barrier function in vivo by biotin.
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DOI：
10.1007/978-1-61779-185-7_7
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Chen, Yan-Hua
中科院分区：
文献类型：
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作者：
Ding, Lei;Zhang, Yuguo;Tatum, Rodney;Chen, Yan-Hua
紧密连接(TJ)是哺乳动物上皮细胞连接复合体中最顶端的成分，形成选择性的细胞旁屏障，限制溶质和离子通过上皮层。Claudins是一个TJ整合膜蛋白家族，在调节细胞旁屏障通透性方面起着关键作用。在体外细胞培养系统中，跨上皮阻抗(TER)的测量和不同大小的放射性同位素或荧光标记分子的通量已被广泛用于确定TJ屏障功能。在体内系统中，示踪分子Sulfo-NHS-Biotin最初用于非洲爪哇胚胎系统，随后在实验条件下成功地应用于许多动物组织和不同生物中，由几个实验室检测TJs的功能完整性。在本章中，我们将描述将生物素作为细胞旁示踪分子应用于不同体内系统以检测TJ屏障功能的详细程序。
Tight junctions (TJs) are the most apical component of the junctional complexes in mammalian epithelial cells and form selective paracellular barriers restricting the passage of solutes and ions across the epithelial sheets. Claudins, a TJ integral membrane protein family, play a critical role in regulating paracellular barrier permeability. In the in vitro cell culture system, transepithelial electrical resistance (TER) measurement and the flux of radioisotope or fluorescent labeled molecules with different sizes have been widely used to determine the TJ barrier function. In the in vivo system, the tracer molecule Sulfo-NHS-Biotin was initially used in Xenopus embryos system and subsequently was successfully applied to a number of animal tissues in situ and in different organisms under the experimental conditions to examine the functional integrity of TJs by several laboratories. In this chapter, we will describe the detailed procedures of applying biotin as a paracellular tracer molecule to different in vivo systems to assay TJ barrier function.