Abstract
Claudin-5 is a transmembrane tight junction protein highly expressed in brain endothelial cells, the site of the blood–brain barrier. The properties of the brain endothelial tight junction complex are considered to be dependent on claudin-5 cell–cell interaction, putting this protein in a position to play a major role in the maintenance of brain endothelial barrier integrity. Thus, alterations in claudin-5 function can lead to “opening” of the paracellular route and increased brain endothelial barrier permeability. Recent work from the authors’s laboratory has established that caveolae-dependent internalization/recycling of claudin-5 is a mechanism underlying transient increases in brain endothelial paracellular permeability in the presence of pro-inflammatory mediators. The biochemical and microscopic techniques presented here were used to investigate trafficking of claudin-5 during those changes in paracellular permeability.
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Acknowledgments
We expresses our appreciation to Dr Michael Wang for his enthusiasm for sharing with us the details of his protocols for GFP-tagging membrane proteins. In addition, we are grateful to the Molecular Imaging Laboratory in the Department of Cell and Developmental Biology at the University of Michigan for their support for the Live Cell Imaging technique. This work is supported by Grant (A.V.A) NS 044907 from the National Institutes of Health and Office of the Vice President for Research Faculty Grants and Awards Program, University of Michigan.
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Stamatovic, S.M., Keep, R.F., Andjelkovic, A.V. (2011). Tracing the Endocytosis of Claudin-5 in Brain Endothelial Cells. 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_22
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DOI: https://doi.org/10.1007/978-1-61779-185-7_22
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