Abstract
When blood‐brain barrier (BBB) endothelial cells are exposed to hypoxic stress for prolonged periods of time, they undergo a morphological change and round up, disrupting cell‐cell junctions which leads to increased paracellular permeability. In this study, we investigated the hypothesis that myosin light chain kinase (MLCK) is integral to this process, and investigated alterations in tight junction structure after hypoxic stress. iTRAQ proteomic analysis identified a number of proteins in BBB endothelial cells that are involved in modulation of the actin cytoskeleton that were significantly altered after 6 hrs of hypoxia. Western blotting confirmed the changes in actin, moesin and vasodilatory stimulated phosphoprotein (VASP). We treated cells with ML‐7, a selective MLCK inhibitor, and assessed barrier function; ML‐7 completely blocked the hypoxia‐induced barrier disruption seen in untreated monolayers. This protection of BBB integrity was correlated with a prevention of stress fiber formation and decreased ZO‐1 expression at tight junction sites, implicating contraction of the actin‐myosin cytoskeleton in BBB disruption following hypoxic stress.
Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
