CFTR limits F ‐actin formation and promotes morphological alignment with flow in human lung microvascular endothelial cells

Cystic fibrosis transmembrane conductance regulator (CFTR) in human lung microvascular endothelial cells (HLMVEC) appears to limit cytosolic actin polymerization while maintaining a cortical rim filamentous actin (F-actin) distribution, as demonstrated by staining F-actin with rhodamine phalloidin (red) under static conditions in the absence and presence of the CFTR inhibitor GlyH-101. The CFTR inhibitor induced loss of the cortical rim F-actin leading to loss of cell-cell contacts and reduced alignment with flow, without effects on endothelial nitrite or endothelin-1 production. AbstractMicro- and macrovascular endothelial dysfunction in response to shear stress has been observed in cystic fibrosis (CF), and has been associated with inflammation and oxidative stress. We tested the hypothesis that the cystic fibrosis transmembrane conductance regulator (CFTR) regulates endothelial actin cytoskeleton dynamics and cellular alignment in response to flow. Human lung microvascular endothelial cells (HLMVEC) were cultured with either the CFTR inhibitor GlyH-101 (20  µM) or CFTRinh-172 (20 µM), tumor necrosis factor (TNF)-α (10 ng/ml) or a vehicle control (0.1% dimethyl sulfoxide) during 24 and 48 h of exposure to shear stress (11.1 dynes/cm2) or under static control conditions. Cellular morphology and filamentous actin (F-actin) were assessed using immunocytochemistry. [Nitrite] and endothelin-1 ([ET-1]) were determined in cell culture supernatant by ozone-based chemilumine...
Source: Physiological Reports - Category: Physiology Authors: Tags: ORIGINAL ARTICLE Source Type: research