title: Intercellular localization of occludins and ZO-1 as a solute transport barrier of the mesothelial monolayer.
abstract: A hyperpermeable state has been observed in patients on long-term peritoneal dialysis. To understand the causes of the structural or functional changes and the progression of the fibrotic process, it is important to determine which region of the peritoneum exhibits these changes. The objectives of this study were to determine the solute permeability associated with cell-cell adhesion of human peritoneal mesothelial cells (HPMCs), to study the relationship between solute permeability and localizations of tight junction-associated proteins (TJPs: occludins and ZO-1), and to assess the effect of exogenous H2O2 supplementation. HPMCs were cultured on a Transwell until the transmesothelial electrical resistance (TER) reached a plateau. Solute permeation tests were conducted using fluorescein isothiocyanate - labeled dextran (molecular weight: 4, 10, 70, and 150 kDa) to calculate the solute permeability coefficient (SPC). Localization of TJPs was observed by a confocal laser scanning microscope after immunofluorescent staining. TER levels increased steadily, beginning at 97.5 +/- 0.7 ohms.cm2 and leveling off at 128 +/- 3.6 ohms.cm2 (n = 4). This was accompanied by the confluence of cells and the appearance of localized TJPs. SPC levels of the HPMC monolayer on the Transwell were reduced compared to those of the Transwell itself, indicating that the HPMC monolayer provided resistance against solute permeation. Exogenous H2O2 supplementation revealed an increased permeability accompanied with delocalization of TJPs, particularly occludins. The delocalization of occludins and ZO-1 at the intercellular space led to a decrease in intercellular binding capacity and thus triggered an increase in the solute permeability.
authors.affiliation: Department of Chemistry for Materials, Graduate School of Engineering, Mie University, Tsu, Mie 514-8507, Japan.
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Kaneda
authors.forename: Keiichi
authors.initials: K
authors.lastname: Miyamoto
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Nomura
authors.forename: Takashi
authors.initials: T
authors.lastname: Horiuchi
mesh_terms: D002478:Cells, Cultured; D017097:Electric Impedance; D004847:Epithelial Cells / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D008567:Membranes, Artificial; D018613:Microscopy, Confocal; D062793:Occludin; D018384:Oxidative Stress; D010530:Peritoneal Dialysis; D010539:Permeability; D010750:Phosphoproteins / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D062826:Zonula Occludens-1 Protein
chemical_list: D008565:Membrane Proteins; D008567:Membranes, Artificial; C568985:OCLN protein, human; D062793:Occludin; D010750:Phosphoproteins; D017382:Reactive Oxygen Species; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein