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Contribution of claudin-5 to barrier properties in tight junctions of epithelial cells

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Abstract

Claudin-5 is a transmembrane protein reported to be primarily present in tight junctions of endothelia. Unexpectedly, we found expression of claudin-5 in HT-29/B6 cells, an epithelial cell line derived from human colon. Confocal microscopy showed colocalization of claudin-5 with occludin, indicating its presence in the tight junctions. By contrast, claudin-5 was absent in the human colonic cell line Caco-2 and in Madin–Darby canine kidney cells (MDCK sub-clones C7 and C11), an epithelial cell line derived from the collecting duct. To determine the contribution of claudin-5 to tight junctional permeability in cells of human origin, stable transfection of Caco-2 with FLAG-claudin-5 cDNA was performed. In addition, clone MDCK-C7 was transfected. Synthesis of the exogenous FLAG-claudin-5 was verified by Western blot analysis and confocal fluorescent imaging by employing FLAG-specific antibody. FLAG-claudin-5 was detected in transfected cells in colocalization with occludin, whereas cells transfected with the vector alone did not exhibit specific signals. Resistance measurements and mannitol fluxes after stable transfection with claudin-5 cDNA revealed a marked increase of barrier function in cells of low genuine transepithelial resistance (Caco-2). By contrast, no changes of barrier properties were detected in cells with a high transepithelial resistance (MDCK-C7) after stable transfection with claudin-5 cDNA. We conclude that claudin-5 is present in epithelial cells of colonic origin and that it contributes to some extent to the paracellular seal. Claudin-5 may thus be classified as a tight-junctional protein capable of contributing to the “sealing” of the tight junction.

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Authors and Affiliations

  1. Department of Clinical Physiology, Charité-Universitary Medicine Berlin, Campus Benjamin Franklin, 12200, Berlin, Germany

    Salah Amasheh, Thomas Schmidt, Michaela Mahn, Peter Florian, Alfred H. Gitter & Michael Fromm

  2. Department of Gastroenterology, Infectious Diseases, and Rheumatology, Charité-Universitary Medicine Berlin, Campus Benjamin Franklin, 12200, Berlin, Germany

    Joachim Mankertz, Shida Tavalali & Jörg-Dieter Schulzke

  3. Department of Medical Engineering, Jena University of Applied Sciences, Carl-Zeiss-Promenade 2, 07745, Jena, Germany

    Alfred H. Gitter

Authors
  1. Salah Amasheh
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  2. Thomas Schmidt
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  3. Michaela Mahn
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  4. Peter Florian
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  5. Joachim Mankertz
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  9. Michael Fromm
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Correspondence to Michael Fromm.

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We thank Anja Fromm and Sieglinde Lüderitz for excellent technical assistance. The work was supported by the Deutsche Forschungsgemeinschaft (DFG Fr 652/4) and by funds of the Campus Benjamin Franklin, Charité-Universitary Medicine Berlin.

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Amasheh, S., Schmidt, T., Mahn, M. et al. Contribution of claudin-5 to barrier properties in tight junctions of epithelial cells. Cell Tissue Res 321, 89–96 (2005). https://doi.org/10.1007/s00441-005-1101-0

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