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

Watson, CJ; Rowland, M; Warhurst, G

Authors

CJ Watson

M Rowland

G Warhurst



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.

Citation

Watson, C., Rowland, M., & Warhurst, G. (2001). Functional modeling of tight junctions in intestinal cell monolayers using polyethylene glycol oligomers. American Journal of Physiology - Cell Physiology, 281(2), C388-C397

Journal Article Type Article
Publication Date Aug 1, 2001
Deposit Date Aug 20, 2007
Journal American Journal of Physiology - Cell Physiology
Print ISSN 0363-6143
Publisher American Physiological Society
Peer Reviewed Peer Reviewed
Volume 281
Issue 2
Pages C388-C397