title: Shear stress increases expression of a KATP channel in rat and bovine pulmonary vascular endothelial cells.
abstract: 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+ (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 inwardly rectifying K+ channel (KIR) 6.2 subunit and fluorescent ligand binding for the sulfonylurea receptor (SUR). 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 KIR6.2 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 KATP 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 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 are essential for the endothelial cell membrane depolarization response with acute decrease in shear stress.
authors.affiliation: Institute for Environmental Medicine, University of Pennsylvania Medical Center, Philadelphia, 19104, USA.
authors.forename: Shampa
authors.initials: S
authors.lastname: Chatterjee
authors.forename: Abu-Bakr
authors.initials: AB
authors.lastname: Al-Mehdi
authors.forename: Irena
authors.initials: I
authors.lastname: Levitan
authors.forename: Troy
authors.initials: T
authors.lastname: Stevens
authors.forename: Aron B
authors.initials: AB
authors.lastname: Fisher
mesh_terms: D018528:ATP-Binding Cassette Transporters; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D019806:Cromakalim / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D004594:Electrophysiology; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D005905:Glyburide / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008833:Microcirculation / Q000502:physiology; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000187:drug effects / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D024661:Potassium Channels, Inwardly Rectifying / Q000187:drug effects / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011652:Pulmonary Circulation / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011955:Receptors, Drug / Q000187:drug effects / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D013314:Stress, Mechanical; D064233:Sulfonylurea Receptors
chemical_list: D018528:ATP-Binding Cassette Transporters; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D012333:RNA, Messenger; D011955:Receptors, Drug; D064233:Sulfonylurea Receptors; D019806:Cromakalim; D000255:Adenosine Triphosphate; D003513:Cycloheximide; D005905:Glyburide