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1 Department of Cell Physiology and Metabolism, University of Geneva Medical School, Geneva, Switzerland
2 Department of Cell Biology, Faculty of Sciences, University of Geneva, Geneva, Switzerland
3 Laboratoire Central de Chimie Clinique, Hopital Cantonal Universitaire, Geneva, Switzerland
4 Departamento de Bioquimica, Instituto de Investigaciones Biomedicas, Madrid, Spain
5 Service de Nephrologie, Fondation pour Recherches Medicales, Geneva, Switzerland
* To whom correspondence should be addressed. E-mail: Roberto.Montesano{at}medecine.unige.ch.
Constitutive expression of the transcription factor Snail was previously shown to trigger complete epithelial-mesenchymal transition. The aim of this study was to determine whether inducible expression of Snail could modify epithelial properties without eliciting full mesenchymal conversion. For this purpose, we expressed mouse Snail cDNA (mSnail) in MDCK cells under the control of a doxycycline-repressible transactivator. Inducible expression of Snail did not result in overt EMT, but induced a number of phenotypic alterations of MDCK cells, the most significant of which was the absence of fluid-filled blister-like structures called "domes". To understand the mechanisms responsible for dome suppression, we assessed the effect of mSnail expression on epithelial barrier function. Although mSnail did not alter tight junction (TJ) organization and permeability to uncharged solutes, it markedly decreased the transepithelial electrical resistance. In light of these findings, we evaluated the ability of MDCK cell monolayers to maintain ionic gradients, and found that expression of mSnail selectively increases Na+ and Cl- permeability. Analysis of the expression of claudins, transmembrane proteins that regulate TJ ionic permeability, showed that mSnail induces a moderate decrease in claudin-2 and a substantial decrease in claudin-4 and claudin-7 expression. Taken together, these results suggest that induction of mSnail selectively increases the ionic permeability of TJs by differentially modulating the expression of specific claudins.
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