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1 Institute for Environmental Medicine, University of Pennsylvania, Philadelphia, PA, USA
2 Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, PA, USA
* To whom correspondence should be addressed. E-mail: abf{at}mail.med.upenn.edu.
We have shown previously that acute ischemia leads to depolarization of pulmonary microvascular endothelial cells that is prevented with cromakalim suggesting the presence of KATP channels in these cells, Thus, KATP channel expression and activity were evaluated in rat pulmonary microvascular endothelial cells (RPMVEC) by whole cell current measurements, dot blot (mRNA) and immunoblot (protein) for the KIR 6.2 subunit, and fluorescent ligand binding for the sulfonylurea receptor. Low level expression of a KATP channel was detected in endothelial cells in routine (static) culture and led us to examine whether its expression is inducible when endothelial cells are adapted to flow. Channel expression (mRNA and both KIR 6.2 and SUR proteins) and inwardly rectified membrane current by patch clamp increased significantly when RPMVEC were adapted to flow at 10 dynes/cm2 for 24 in either a parallel plate flow chamber or an artificial capillary system. Induction of the KATP channel with flow-adaptation also was observed in bovine pulmonary artery endothelial cells. Flow-adapted but not static RPMVEC showed cellular plasma membrane depolarization upon stop of flow that was inhibited by a KATP channel opener and prevented by addition of cycloheximide to the medium during the flow-adaptation period. These studies indicate the induction of KATP channels by flow-adaptation in pulmonary endothelium and that the expression and activity of this channel is essential for the endothelial cell membrane depolarization response with acute decrease in shear stress.
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