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Functional modeling of tight junctions in intestinal cell monolayers using polyethylene glycol oligomers

Functional modeling of tight junctions in intestinal cell monolayers using polyethylene glycol... Abstract Despite significant advances in the characterization of tight junction (TJ) proteins, little is known about how molecular changes relate to function due primarily to the limitations of conventional paracellular probes. To address this, the paracellular pathway in Caco-2 and T84 cell lines was profiled by measuring the permeabilities of 24 polyethylene glycols (PEG) of increasing molecular radius (3.5–7.4 Å) analyzed by mass spectrometry. When combined with a paracellular sieving model, these data provided quantitative descriptors of the pathway under control conditions and after exposure to TJ modulators. PEG profiles in both cell lines conformed to a biphasic process involving a restrictive pore (radius 4.3–4.5 Å) and a nonrestrictive component responsible for permeability of larger molecules. PEG profiling revealed significant differences between the effects of EGTA and sodium caprate (C10). The restrictive component of EGTA-treated cells lost all size discrimination due to an increase in pore radius. Sodium caprate had no effect on pore radius but increased permeability via a different mechanism possibly involving increased numbers of functional pores. PEG profiling provides a useful tool for probing the functional regulation of the paracellular route. paracellular permeability Caco-2 T84 transepithelial resistance tight junction modulation Footnotes This work was supported by AstraZeneca. Address for reprint requests and other correspondence: G. Warhurst, Section of Gastrointestinal Science, Clinical Sciences Bldg., Hope Hospital, Salford, UK, M6 8HD (E-mail: gwarhurs@fs1.ho.man.ac.uk ). The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “ advertisement ” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Copyright © 2001 the American Physiological Society http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png AJP - Cell Physiology The American Physiological Society

Functional modeling of tight junctions in intestinal cell monolayers using polyethylene glycol oligomers

AJP - Cell Physiology , Volume 281 (2): C388 – Aug 1, 2001

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Publisher
The American Physiological Society
Copyright
Copyright © 2010 the American Physiological Society
ISSN
0363-6143
eISSN
1522-1563
Publisher site
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Abstract

Abstract Despite significant advances in the characterization of tight junction (TJ) proteins, little is known about how molecular changes relate to function due primarily to the limitations of conventional paracellular probes. To address this, the paracellular pathway in Caco-2 and T84 cell lines was profiled by measuring the permeabilities of 24 polyethylene glycols (PEG) of increasing molecular radius (3.5–7.4 Å) analyzed by mass spectrometry. When combined with a paracellular sieving model, these data provided quantitative descriptors of the pathway under control conditions and after exposure to TJ modulators. PEG profiles in both cell lines conformed to a biphasic process involving a restrictive pore (radius 4.3–4.5 Å) and a nonrestrictive component responsible for permeability of larger molecules. PEG profiling revealed significant differences between the effects of EGTA and sodium caprate (C10). The restrictive component of EGTA-treated cells lost all size discrimination due to an increase in pore radius. Sodium caprate had no effect on pore radius but increased permeability via a different mechanism possibly involving increased numbers of functional pores. PEG profiling provides a useful tool for probing the functional regulation of the paracellular route. paracellular permeability Caco-2 T84 transepithelial resistance tight junction modulation Footnotes This work was supported by AstraZeneca. Address for reprint requests and other correspondence: G. Warhurst, Section of Gastrointestinal Science, Clinical Sciences Bldg., Hope Hospital, Salford, UK, M6 8HD (E-mail: gwarhurs@fs1.ho.man.ac.uk ). The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “ advertisement ” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Copyright © 2001 the American Physiological Society

Journal

AJP - Cell PhysiologyThe American Physiological Society

Published: Aug 1, 2001

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