Occludin Content Modulates Hydrogen Peroxide-Induced Increase in Renal Epithelial Paracellular Permeability

Danielle Janosevic, Josephine Axis, Robert Bacallao, Kurt Amsler

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The ability of hydrogen peroxide (H2O2) to increase paracellular permeability of renal epithelial cell monolayers was examined and the role of occludin in this regulation was investigated. H2O2 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-PK1, in a concentration-dependent manner. At the same concentrations, H2O2 did not alter transepithelial resistance (TER) nor increase cell death. The magnitude of the H2O2-induced increase in leak pathway permeability was inversely related to cellular occludin protein content. H2O2 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 H2O2 treatment. The most dramatic effect of H2O2 was on the dynamic mobility of GFP-occludin into the tight junction region. H2O2 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 H2O2 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 H2O2. These data indicate that the H2O2-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.

Original languageEnglish (US)
Pages (from-to)769-779
Number of pages11
JournalJournal of Cellular Biochemistry
Volume117
Issue number3
DOIs
StatePublished - Mar 1 2016

Fingerprint

Occludin
Hydrogen Peroxide
Permeability
Kidney
Tight Junctions
Epithelial Cells
Proteins
Monolayers
Octoxynol
Cell death
Cell Death

Keywords

  • FRAP
  • HYDROGEN PEROXIDE
  • OCCLUDIN
  • RENAL EPITHELIAL CELL
  • TIGHT JUNCTION

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Occludin Content Modulates Hydrogen Peroxide-Induced Increase in Renal Epithelial Paracellular Permeability. / Janosevic, Danielle; Axis, Josephine; Bacallao, Robert; Amsler, Kurt.

In: Journal of Cellular Biochemistry, Vol. 117, No. 3, 01.03.2016, p. 769-779.

Research output: Contribution to journalArticle

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AB - The ability of hydrogen peroxide (H2O2) to increase paracellular permeability of renal epithelial cell monolayers was examined and the role of occludin in this regulation was investigated. H2O2 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-PK1, in a concentration-dependent manner. At the same concentrations, H2O2 did not alter transepithelial resistance (TER) nor increase cell death. The magnitude of the H2O2-induced increase in leak pathway permeability was inversely related to cellular occludin protein content. H2O2 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 H2O2 treatment. The most dramatic effect of H2O2 was on the dynamic mobility of GFP-occludin into the tight junction region. H2O2 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 H2O2 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 H2O2. These data indicate that the H2O2-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.

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