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Increased claudin‐5 increases lung epithelial permeability and is associated with disruption of tight junction assembly
Author(s) -
Overgaard Christian,
Mitchell Leslie A.,
Ward Christina,
Guidot David M.,
Koval Michael
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.1063.12
Subject(s) - tight junction , claudin , barrier function , occludin , microbiology and biotechnology , chemistry , biology
Chronic alcohol abuse increases the risk of developing the Acute Respiratory Distress Syndrome (ARDS) 3–4‐fold. Based on experimental models, alcohol disrupts alveolar epithelial permeability and is associated with an increase in claudin‐5 (cldn‐5). Here we used viral expression systems to alter cldn‐5 expression by cultured primary rat alveolar epithelial cells (AECs). AECs transduced to express increasing levels of cldn‐5 showed decreased transepithelial resistance (TER), which plateaued at 80% of control levels. Conversely, decreasing cldn‐5 using shRNA increased TER of normal and “alcoholic” AECs by 25–35%, thus improving barrier function. By immunofluorescence, cells expressing increased cldn‐5 showed disrupted localization of cldn‐18 and ZO‐1, where cell junctions had an increase in strand breaks and inward projections (“spikes”) suggesting decreased stability. Analysis of differential Triton X‐100 solubility was used to biochemically confirm these observations. This suggests a model for tight junction assembly, where bidirectional interplay between claudin and scaffold protein composition determine tight junction morphology and function.