Abstract
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 embryo 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.
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Acknowledgments
We would like to thank Joani T. Zary and Beverly G. Jeansonne for their technical assistance. Part of this work was supported by the North Carolina Biotechnology Center grant and the National Institutes of Health grant HL085752 to Y.H.C.
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Ding, L., Zhang, Y., Tatum, R., Chen, YH. (2011). Detection of Tight Junction Barrier Function In Vivo by Biotin. In: Turksen, K. (eds) Claudins. Methods in Molecular Biology, vol 762. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-185-7_7
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DOI: https://doi.org/10.1007/978-1-61779-185-7_7
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