caa a22 4500 475774930 CHVBK 20180406123628.0 cr unu---uuuuu 170329e20000501xx s 000 0 eng 10.1007/s004180000143 doi (NATIONALLICENCE)springer-10.1007/s004180000143 Disruption of epithelial tight junctions is prevented by cyclic nucleotide-dependent protein kinase inhibitors [Elektronische Daten] [Cécile Klingler, Uwe Kniesel, Simon D. Bamforth, Hartwig Wolburg, Britta Engelhardt, Werner Risau] Abstract.: Tight junctions (TJs), the most apical of the intercellular junctions, prevent the passage of ions and molecules through the paracellular pathway. Intracellular signalling molecules are likely to be involved in the regulation of TJ integrity. In order to specifically investigate the role of protein kinaseA (PKA) in the maintenance of epithelial TJ integrity, calcium-switch experiments were performed, in which calcium was removed from EpH4 and MDCK culture medium, in the absence or presence of the PKA inhibitors H-89 or HA-1004. Removal of calcium from the culture media of the epithelial cells resulted in disruption of the TJs, characterised by a loss of membrane association of the TJ-associated proteins occludin, ZO-1 and ZO-2, by a loss of TJ strands, by a marked decrease in the transepithelial electrical resistance and by a dramatic increase in the transepithelial permeability to tracers. The association of occludin, ZO-1 and ZO-2 with the actin cytoskeleton is not affected. In contrast, when the removal of calcium was performed in the presence of either the PKA inhibitor H-89 or HA-1004, all barrier characteristics were preserved. Our data indicate that following the removal of calcium from the culture medium of epithelial cells in vitro, PKA is activated and subsequently is involved in the disruption of TJs. Springer-Verlag, 2000 Tight junction Calcium switch Epithelial cells Protein kinaseA nationallicence Klingler Cécile Max Planck Institute for Physiological and Clinical Research, Department of Molecular Cell Biology, Parkstrasse 1, 61231 Bad Nauheim, Germany aut Kniesel Uwe Department of Pathology, University of Tübingen, 72076 Tübingen, Germany aut Bamforth Simon D. Max Planck Institute for Physiological and Clinical Research, Department of Molecular Cell Biology, Parkstrasse 1, 61231 Bad Nauheim, Germany aut Wolburg Hartwig Department of Pathology, University of Tübingen, 72076 Tübingen, Germany aut Engelhardt Britta Max Planck Institute for Physiological and Clinical Research, Department of Molecular Cell Biology, Parkstrasse 1, 61231 Bad Nauheim, Germany aut Risau Werner Max Planck Institute for Physiological and Clinical Research, Department of Molecular Cell Biology, Parkstrasse 1, 61231 Bad Nauheim, Germany aut https://doi.org/10.1007/s004180000143 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s004180000143 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Klingler Cécile Max Planck Institute for Physiological and Clinical Research, Department of Molecular Cell Biology, Parkstrasse 1, 61231 Bad Nauheim, Germany aut NATIONALLICENCE 700 1- Kniesel Uwe Department of Pathology, University of Tübingen, 72076 Tübingen, Germany aut NATIONALLICENCE 700 1- Bamforth Simon D. Max Planck Institute for Physiological and Clinical Research, Department of Molecular Cell Biology, Parkstrasse 1, 61231 Bad Nauheim, Germany aut NATIONALLICENCE 700 1- Wolburg Hartwig Department of Pathology, University of Tübingen, 72076 Tübingen, Germany aut NATIONALLICENCE 700 1- Engelhardt Britta Max Planck Institute for Physiological and Clinical Research, Department of Molecular Cell Biology, Parkstrasse 1, 61231 Bad Nauheim, Germany aut NATIONALLICENCE 700 1- Risau Werner Max Planck Institute for Physiological and Clinical Research, Department of Molecular Cell Biology, Parkstrasse 1, 61231 Bad Nauheim, Germany aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer