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VASCULAR BIOLOGY

Shear stress increases expression of a K_ATP channel in rat and bovine pulmonary vascular endothelial cells

¹Institute for Environmental Medicine; and ²Institute for Medicine and Engineering, University of Pennsylvania, Medical Center, Philadelphia, Pennsylvania 19104; and ³Department of Cellular and Molecular Pharmacology, University of South Alabama, Mobile, Alabama 36688

Submitted 5 November 2002 ; accepted in final form 16 June 2003

We have shown previously that acute ischemia leads to depolarization of pulmonary microvascular endothelial cells that is prevented with cromakalim, suggesting the presence of ATP-sensitive K⁺ (K_ATP) channels in these cells. Thus K_ATP 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 inwardly rectifying K⁺ channel (K_IR) 6.2 subunit and fluorescent ligand binding for the sulfonylurea receptor (SUR). Low-level expression of a K_ATP 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 K_IR6.2 and SUR proteins) and inwardly rectified membrane current by patch clamp increased significantly when RPMVEC were adapted to flow at 10 dyn/cm² for 24 h in either a parallel plate flow chamber or an artificial capillary system. Induction of the K_ATP channel with flow adaptation was also 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 K_ATP channel opener and prevented by addition of cycloheximide to the medium during the flow adaptation period. These studies indicate the induction of K_ATP channels by flow adaptation in pulmonary endothelium and that the expression and activity of this channel are essential for the endothelial cell membrane depolarization response with acute decrease in shear stress.

flow adaptation; K_IR 6.2; sulfonylurea receptor; fluorescent glyburide; pulmonary microvascular endothelial cells

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