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Am J Physiol Gastrointest Liver Physiol 277: G1138-G1148, 1999;
0193-1857/99 $5.00
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Vol. 277, Issue 6, G1138-G1148, December 1999

Copper treatment alters the permeability of tight junctions in cultured human intestinal Caco-2 cells

Simonetta Ferruzza1, Maria-Laura Scarino1, Giuseppe Rotilio1,2, Maria Rosa Ciriolo3, Paolo Santaroni1, Andrea Onetti Muda4, and Yula Sambuy1

1 Istituto Nazionale della Nutrizione, 00178 Rome; 2 Dipartimento di Biologia, Università di Roma Tor Vergata, 00133 Rome; 3 Dipartimento di Scienze Biomediche, Università di Chieti, 66013 Chieti; and 4 Dipartimento di Medicina Sperimentale e Patologia, Università di Roma La Sapienza, 00161 Rome, Italy

The effects of copper on tight-junction permeability were investigated in human intestinal Caco-2 cells, monitoring transepithelial electrical resistance and transepithelial passage of mannitol. Apical treatment of Caco-2 cells with 10-100 µM CuCl2 (up to 3 h) produced a time- and concentration-dependent increase in tight-junction permeability, reversible after 24 h in complete medium in the absence of added copper. These effects were not observed in cells treated with copper complexed to L-histidine [Cu(His)2]. The copper-induced increase in tight-junction permeability was affected by the pH of the apical medium, as was the apical uptake of 64CuCl2, both exhibiting a maximum at pH 6.0. Treatment with CuCl2 produced a concentration-dependent reduction in the staining of F actin but not of the junctional proteins zonula occludens-1, occludin, and E-cadherin and produced ultrastructural alterations to microvilli and tight junctions that were not observed after treatment with up to 200 µM Cu(His)2 for 3 h. Overall, these data point to an intracellular effect of copper on tight junctions, mediated by perturbations of the F actin cytoskeleton.

CuCl2; copper-histidine; uptake; pH; F actin organization; microvilli; junctional proteins; recovery


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