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Am J Physiol Cell Physiol 289: C1002-C1014, 2005. First published June 1, 2005; doi:10.1152/ajpcell.00175.2005
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MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

Inducible expression of Snail selectively increases paracellular ion permeability and differentially modulates tight junction proteins

Fabio Carrozzino,1 Priscilla Soulié,1 Denise Huber,2 Noury Mensi,3 Lelio Orci,1 Amparo Cano,4 Eric Féraille,5 and Roberto Montesano1

1Department of Cell Physiology and Metabolism, University of Geneva Medical Center, 2Department of Cell Biology, Faculty of Sciences, University of Geneva, and 3Laboratoire Central de Chimie Clinique, Hôpital Cantonal Universitaire, Geneva, Switzerland; 4Departamento de Bioquimica, Universidad Autónoma de Madrid (UAM), Instituto de Investigaciones Biomedicas "Alberto Sols" Consejo Superior de Investigaciones Científicas-UAM, Madrid, Spain; and 5Service de Néphrologie, Fondation pour Recherches Médicales, Geneva, Switzerland

Submitted 12 April 2005 ; accepted in final form 30 May 2005

Constitutive expression of the transcription factor Snail was previously shown to trigger complete epithelial-mesenchymal transition (EMT). 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 (mSnail) cDNA in Madin-Darby canine kidney (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 blisterlike 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 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 -7 expression. Together, these results suggest that induction of mSnail selectively increases the ionic permeability of TJs by differentially modulating the expression of specific claudins.

epithelium; Madin-Darby canine kidney cells; claudin; dome


Address for reprint requests and other correspondence: R. Montesano, Dept. of Cell Physiology and Metabolism, Univ. of Geneva Medical Center, Rue Michel-Servet 1, CH-1211 Geneva 4, Switzerland (e-mail: Roberto.Montesano{at}medecine.unige.ch)




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