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Occludin Content Modulates Hydrogen Peroxide‐Induced Increase in Renal Epithelial Paracellular Permeability
Author(s) -
Janosevic Danielle,
Axis Josephine,
Bacallao Robert L.,
Amsler Kurt
Publication year - 2016
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.25362
Subject(s) - occludin , paracellular transport , tight junction , microbiology and biotechnology , claudin , chemistry , cell junction , biophysics , biochemistry , biology , permeability (electromagnetism) , cell , membrane
The ability of hydrogen peroxide (H 2 O 2 ) to increase paracellular permeability of renal epithelial cell monolayers was examined and the role of occludin in this regulation was investigated. H 2 O 2 treatment increased the paracellular movement of calcein, a marker for the leak pathway permeability, across monolayers of two renal epithelial cell lines, MDCK and LLC‐PK 1 , in a concentration‐dependent manner. At the same concentrations, H 2 O 2 did not alter transepithelial resistance (TER) nor increase cell death. The magnitude of the H 2 O 2 ‐induced increase in leak pathway permeability was inversely related to cellular occludin protein content. H 2 O 2 treatment did not produce any major change in total cellular content or Triton X‐100‐soluble or –insoluble fraction content of occludin protein. Occludin protein staining at the tight junction region was diminished following H 2 O 2 treatment. The most dramatic effect of H 2 O 2 was on the dynamic mobility of GFP‐occludin into the tight junction region. H 2 O 2 treatment slowed lateral movement of GFP‐occludin into the tight junction region but not on the apical membrane. Further, removal of the cytoplasmic C‐terminal region of occludin protein eliminated the effect of H 2 O 2 on GFP‐occludin lateral movement into the tight junction region. An increase in the mobile fraction of GFP‐occludin was associated with a loss of response to H 2 O 2 . These data indicate that the H 2 O 2 ‐induced increase in renal epithelial cell paracellular permeability is mediated, at least in part, through occludin protein, possibly through a slowing of the rate of occludin movement into the tight junction region. J. Cell. Biochem. 117: 769–779, 2016. © 2015 Wiley Periodicals, Inc.