Membrane currents and the resting membrane potential in cultured bovine pulmonary artery endothelial cells.
1. We have used the whole-cell patch-clamp technique to characterize the ionic conductances that determine the resting membrane potential in cultured endothelial cells from calf pulmonary artery (CPAE cells). 2. Resting membrane potentials were scattered between -88 and +5 mV with a mean +/- S.E.M. of -26 +/- 3 mV (n = 104). 3. The most prominent membrane current in resting cells was an inwardly rectifying K+ current. This current showed Na(+)-dependent inactivation and was efficiently blocked by external Ba2+ (EC50 = 2.2 microM), but was relatively insensitive to quinine, quinidine and TEA. 4. Hypertonic cell shrinkage inhibited an outwardly rectifying Cl- current, which was also efficiently blocked by 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB; 100 microM), quinine (500 microM) and quinidine (500 microM). 5. A linear, time-independent background current remained after elimination of these two currents. This current was dependent on extracellular monovalent cations with a permeability sequence of Cs+ > Na+ > Li+ >> N-methyl-D-glucamine. It was partially blocked by millimolar concentrations of the divalent cations Ca2+, Ni2+ and Ba2+. Gd3+ (200 microM) had no significant effect on this background current. 6. Continuous measurements of the membrane potential confirm that the three described conductances are the major determinants of the membrane potential. Due to the low slope conductance in the region between -70 and 0 mV, small changes in one of the current components can evoke large depolarizations or hyperpolarizations, which explains the large scattering of the resting membrane potentials.
Voets
T
T
Laboratory of Physiology, Catholic University of Leuven, Belgium.
Droogmans
G
G
Nilius
B
B
D000818:Animals; D001464:Barium / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D002586:Cesium / Q000494:pharmacology; D002614:Chelating Agents / Q000494:pharmacology; D002712:Chlorides / Q000378:metabolism; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D004594:Electrophysiology; D004730:Endothelium, Vascular / Q000378:metabolism*; D005942:Gluconates / Q000494:pharmacology; D007473:Ion Channels / Q000378:metabolism; D008094:Lithium / Q000494:pharmacology; D008536:Meglumine / Q000031:analogs & derivatives / Q000494:pharmacology; D008564:Membrane Potentials; D009532:Nickel / Q000494:pharmacology; D009579:Nitrobenzoates / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism; D011651:Pulmonary Artery / Q000378:metabolism / Q000502:physiology; D012964:Sodium / Q000378:metabolism / Q000494:pharmacology
D002614:Chelating Agents; D002712:Chlorides; D005942:Gluconates; D007473:Ion Channels; D009579:Nitrobenzoates; C051643:N-methylglucamine gluconate; D002586:Cesium; D001464:Barium; C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; D004533:Egtazic Acid; D008536:Meglumine; D009532:Nickel; D008094:Lithium; D012964:Sodium; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D011188:Potassium