Characterization of volume-activated chloride currents in endothelial cells from bovine pulmonary artery.
We have measured the kinetic and pharmacological properties of volume-activated Cl- currents (ICl, vol) in endothelial cells, and tried to correlate them with those of the already described volume-activated current ICln. Both conductances show a similar permeability sequence for monovalent anions, and they are blocked by extracellular ATP. In the present report, we demonstrate by Western blot and RT-PCR that cultured endothelial cells from bovine pulmonary artery (CPAE) contain pICln. The expression of this protein has been shown to be closely associated with the ICln current. ICl, vol showed however, in contrast with ICln, no striking inactivation at positive potentials. This property is also at variance with that of the volume-activated current related to MDR-1. Activation of ICl, vol at potentials more negative than -80 mV was not time dependent, which excludes a major contribution of a ClC-2 related current. The antiviral nucleoside analogue AZT (3'-azido-3'-deoxythymidine) inhibited ICl, vol by 21 +/- 2.7% (n = 10), at a concentration of 100 microM. Another antiviral drug, acyclovir (ACV, 9-[2-hydroxyethoxy) methyl]guanine) blocked ICl, vol by 27 +/- 6.2% at 100 microM (n = 11). Both blocking effects are much smaller than those reported for ICln. The phenol derivative gossypol, which blocks ICln-related currents, efficiently inhibited ICl, vol in CPAE cells (67 +/- 2.1% at 1 microM, n = 7, KI = 0.4 microns). The presence of pICln in CPAE cells and the similar qualitative pharmacological profile of ICl, vol and ICln support the hypothesis that pICln is a good molecular candidate for ICl, vol in endothelial cells. The discrepant kinetic properties may indicate that these time-dependent currents at high positive or negative potentials are not intrinsic properties of the channels, but are caused by time-dependent depletion/accumulation phenomena due to the large amplitudes of these currents.
Szücs
G
G
KU Leuven, Laboratorium voor Fysiologie, Belgium.
Buyse
G
G
Eggermont
J
J
Droogmans
G
G
Nilius
B
B
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D018118:Chloride Channels / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism*; D017931:DNA Primers; D004553:Electric Conductivity; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D006072:Gossypol / Q000494:pharmacology; D007473:Ion Channels; D007700:Kinetics; D008969:Molecular Sequence Data; D018408:Patch-Clamp Techniques; D011651:Pulmonary Artery
D018118:Chloride Channels; D002712:Chlorides; D017931:DNA Primers; D007473:Ion Channels; D006072:Gossypol