Journal
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY
Volume 303, Issue 12, Pages C1278-C1291Publisher
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/ajpcell.00434.2011
Keywords
claudin-4; claudin-2; paracellular transport; opossum kidney cells
Categories
Funding
- Kidney Foundation of Canada
- Canadian Institutes of Health Research
- KRESCENT New Investigator Award
- Alberta Innovates Health Solutions Clinical Investigator Award
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Borovac J, Barker RS, Rievaj J, Rasmussen A, Pan W, Wevrick R, Alexander RT. Claudin-4 forms a paracellular barrier, revealing the interdependence of claudin expression in the loose epithelial cell culture model opossum kidney cells. Am J Physiol Cell Physiol 303: C1278-C1291, 2012. First published October 17, 2012; doi:10.1152/ajpcell.00434.2011.-The effect of claudins on paracellular fluxes has been predominantly studied in either Madin-Darby canine kidney (MDCK) or LLCPK cells. Neither model system has a very low transepithelial resistance (TER) as observed in leaky epithelia. Moreover, results from one model system are not always consistent with another. Opossum kidney (OK) cells form tight junctions yet have a very low TER. We therefore set out to characterize the paracellular transport properties of this cell culture model. Ussing chamber dilution potential measurements revealed that OK cells exhibit a very low TER (11.7 +/- 1.4 Omega.cm(2)), slight cation selectivity (P-Na/P-Cl > 1.10 +/- 0.01), and the Eisenman permeability sequence IV; the permeability of monovalent cations ranking K+ > Cs+ > Rb+ > Na+ > Li+. Quantitative real-time PCR studies found that OK cells endogenously express claudin-4 > -1 > -6 > -20 > -9 > -12 > -11 > -15. Overexpression of claudin-4 significantly increased TER, decreased Na+ and Cl- permeability, and increased levels of claudin-1, -6, and -9 mRNA. Knockdown of claudin-4 in the overexpressing cells significantly decreased TER without altering claudin expression; thus claudin-4 forms a barrier in OK cells. Knockdown of endogenous claudin-4 decreased claudin-1, -9, and -12 expression without altering TER. Claudin-2 overexpression decreased TER, significantly increased Na+ and Cl- permeability, and decreased claudin-12 and -6 expression. Together these results demonstrate that claudin expression is tightly coupled in OK cells.
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