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Permeability of Endothelial Barrier: Cell Culture and In Vivo Models

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Permeability Barrier

Part of the book series: Methods in Molecular Biology ((MIMB,volume 763))

Abstract

The methods for assessment of endothelial barrier permeability are vital tools of experimental biology. They allow us to measure permeability of endothelial monolayer in cell culture and in lung vessels or to determine formation of tissue edema resulting from increased permeability of vasculature. This chapter provides an overview of the most common protocols.

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References

  1. Komarova, Y., and A.B. Malik. (2010). Regulation of endothelial permeability via paracellular and transcellular transport pathways. Annu Rev Physiol 72:463–493.

    Article  PubMed  CAS  Google Scholar 

  2. Mehta, D., and A.B. Malik. (2006). Signaling mechanisms regulating endothelial permeability. Physiol Rev 86:279–367.

    Article  PubMed  CAS  Google Scholar 

  3. Parker, J.C., and M.I. Townsley. (2004). Evaluation of lung injury in rats and mice. Am J Physiol Lung Cell Mol Physiol 286:L231–246.

    Article  PubMed  CAS  Google Scholar 

  4. Tiruppathi, C., A.B. Malik, P.J. Del Vecchio, C.R. Keese, and I. Giaever. (1992). Electrical method for detection of endothelial cell shape change in real time: assessment of endothelial barrier function. Proc Natl Acad Sci U S A 89:7919–7923.

    Article  PubMed  CAS  Google Scholar 

  5. Wegener, J., C.R. Keese, and I. Giaever. (2000). Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces. Exp Cell Res 259:158–166.

    Article  PubMed  CAS  Google Scholar 

  6. Ellis, C.A., C. Tiruppathi, R. Sandoval, W.D. Niles, and A.B. Malik. (1999). Time course of recovery of endothelial cell surface thrombin receptor (PAR-1) expression. Am J Physiol 276:C38–45.

    PubMed  CAS  Google Scholar 

  7. Yalow, R.S., and S.A. Berson (1960) Immunoassay of endogenous plasma insulin in man J Clin Invest39:1157–75.

    Google Scholar 

  8. Du, T., and M.J. Alfa. (2004). Translocation of Clostridium difficile toxin B across polarized Caco-2 cell monolayers is enhanced by toxin A. Can J Infect Dis 15:83–88.

    PubMed  Google Scholar 

  9. John, T.A., S.M. Vogel, C. Tiruppathi, A.B. Malik, and R.D. Minshall. (2003). Quantitative analysis of albumin uptake and transport in the rat microvessel endothelial monolayer. Am J Physiol Lung Cell Mol Physiol 284:L187–196.

    PubMed  CAS  Google Scholar 

  10. Siflinger-Birnboim, A., P.J. Del Vecchio, J.A. Cooper, F.A. Blumenstock, J.M. Shepard, and A.B. Malik. (1987). Molecular sieving characteristics of the cultured endothelial monolayer. J Cell Physiol 132:111–117.

    Article  PubMed  CAS  Google Scholar 

  11. Bhattacharya, J. (1988). Hydraulic conductivity of lung venules determined by split-drop technique. J Appl Physiol 64:2562–2567.

    PubMed  CAS  Google Scholar 

  12. Parker, J.C., T. Stevens, J. Randall, D.S. Weber, and J.A. King. (2006). Hydraulic conductance of pulmonary microvascular and macrovascular endothelial cell monolayers. Am J Physiol Lung Cell Mol Physiol 291:L30–37.

    Article  PubMed  CAS  Google Scholar 

  13. Solodushko, V., J.C. Parker, and B. Fouty. (2008). Pulmonary microvascular endothelial cells form a tighter monolayer when grown in chronic hypoxia. Am J Respir Cell Mol Biol 38:491–497.

    Article  PubMed  CAS  Google Scholar 

  14. Hemingway, A. (1950). A method of chemical analysis of guinea pig lung for the factors involved in pulmonary edema. J Lab Clin Med 35:817–822.

    PubMed  CAS  Google Scholar 

  15. Pearce, M.L., J. Yamashita, and J. Beazell. (1965). Measurement of Pulmonary Edema. Circ Res 16:482–488.

    PubMed  CAS  Google Scholar 

  16. Fukuda, N., H.G. Folkesson, and M.A. Matthay. (2000). Relationship of interstitial fluid volume to alveolar fluid clearance in mice: ventilated vs. in situ studies. J Appl Physiol 89:672–679.

    Google Scholar 

  17. Kobayashi, H., R. Hataishi, H. Mitsufuji, M. Tanaka, M. Jacobson, T. Tomita, W.M. Zapol, and R.C. Jones. (2001). Antiinflammatory properties of inducible nitric oxide synthase in acute hyperoxic lung injury. Am J Respir Cell Mol Biol 24:390–397.

    PubMed  CAS  Google Scholar 

  18. Klinzing, S., T. Lesser, H. Schubert, M. Bartel, and U. Klein. (2000). Wet-to-dry ratio of lung tissue and surfactant outwash after one-lung flooding. Res Exp Med (Berl) 200:27–33.

    CAS  Google Scholar 

  19. Boutoille, D., X. Marechal, M. Pichenot, C. Chemani, B. Guery, and K. Faure. (2009). FITC-albumin as a marker for assessment of endothelial permeability in mice: comparison with 125I-albumin. Exp Lung Res 35:263–271.

    Article  PubMed  CAS  Google Scholar 

  20. Patterson, C.E., R.A. Rhoades, and J.G. Garcia. (1992). Evans blue dye as a marker of albumin clearance in cultured endothelial monolayer and isolated lung. J Appl Physiol 72:865–873.

    Article  PubMed  CAS  Google Scholar 

  21. Moitra, J., S. Sammani, and J.G. Garcia. (2007). Re-evaluation of Evans Blue dye as a marker of albumin clearance in murine models of acute lung injury. Transl Res 150:253–265.

    Article  PubMed  CAS  Google Scholar 

  22. Dodd-o, J.M., M.L. Hristopoulos, N. Faraday, and D.B. Pearse. (2003). Effect of ischemia and reperfusion without airway occlusion on vascular barrier function in the in vivo mouse lung. J Appl Physiol 95:1971–1978.

    PubMed  Google Scholar 

  23. Standiford, T.J., S.L. Kunkel, N.W. Lukacs, M.J. Greenberger, J.M. Danforth, R.G. Kunkel, and R.M. Strieter. (1995). Macrophage inflammatory protein-1 alpha mediates lung leukocyte recruitment, lung capillary leak, and early mortality in murine endotoxemia. J Immunol 155:1515–1524.

    PubMed  CAS  Google Scholar 

  24. Vogel, S.M., X. Gao, D. Mehta, R.D. Ye, T.A. John, P. Andrade-Gordon, C. Tiruppathi, and A.B. Malik. (2000). Abrogation of thrombin-induced increase in pulmonary microvascular permeability in PAR-1 knockout mice. Physiol Genomics 4:137–145.

    PubMed  CAS  Google Scholar 

  25. Parker, J.C., M.N. Gillespie, A.E. Taylor, and S.L. Martin. (1999). Capillary filtration coefficient, vascular resistance, and compliance in isolated mouse lungs. J Appl Physiol 87:1421–1427.

    PubMed  CAS  Google Scholar 

  26. Kern, D.F., D. Levitt, and D. Wangensteen. (1983). Endothelial albumin permeability measured with a new technique in perfused rabbit lung. Am J Physiol 245:H229–236.

    PubMed  CAS  Google Scholar 

  27. Kern, D.F., and A.B. Malik. (1985). Microvascular albumin permeability in isolated perfused lung: effects of EDTA. J Appl Physiol 58:372–375.

    PubMed  CAS  Google Scholar 

  28. Rippe, B., and A. Taylor. (2001). NEM and filipin increase albumin transport in lung microvessels. Am J Physiol Heart Circ Physiol 280:H34–41.

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Pharmacology, University of Illinois, Chicago, IL, USA

    Alexander N. Garcia, Stephen M. Vogel, Yulia A. Komarova & Asrar B. Malik

Authors
  1. Alexander N. Garcia
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  2. Stephen M. Vogel
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  3. Yulia A. Komarova
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  4. Asrar B. Malik
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Corresponding author

Correspondence to Asrar B. Malik .

Editor information

Editors and Affiliations

  1. Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Smyth Road 501, Ottawa, K1Y 8L6, Canada

    Kursad Turksen

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Garcia, A.N., Vogel, S.M., Komarova, Y.A., Malik, A.B. (2011). Permeability of Endothelial Barrier: Cell Culture and In Vivo Models. In: Turksen, K. (eds) Permeability Barrier. Methods in Molecular Biology, vol 763. Humana Press. https://doi.org/10.1007/978-1-61779-191-8_23

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  • DOI: https://doi.org/10.1007/978-1-61779-191-8_23

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-190-1

  • Online ISBN: 978-1-61779-191-8

  • eBook Packages: Springer Protocols

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