Endothelin and vasopressin activate low conductance chloride channels in aortic smooth muscle cells.
The non-contractile aortic smooth muscle cell line A7r5 was used to study the membrane events involved in the effect of vasoconstrictor peptides. Whole-cell voltage-clamp and membrane potential recording techniques were used to demonstrate the contribution of an increased Cl- conductance to the late depolarization induced by endothelin-1 and vasopressin. During cell-attached patch recording with N-methyl-D-glucamine in the pipette, bath application of endothelin or vasopressin induced single-channel inward currents in the following minutes. The current/potential (I/V) curve of the most frequently observed channel type--a small conductance Cl- (SCl) channel--reversed near the cell membrane potential and showed a single-channel conductance of 1.8 pS for inward currents. After patch excision in an extracellular solution containing CaCl2 (2 mM), the frequency of SCl channel openings increased. Patch excision in the absence of peptide stimulation also produced this channel activity. Replacement of CaCl2 by a Ca2+ chelator on the intracellular face of a patch reversibly inhibited the channel activity, indicating that these SCl channels are Ca(2+)-activated Cl- channels. The single-channel I/V characteristic showed outward rectification above +50 mV. An analysis of the gating kinetics of the SCl channel is given. Another channel type was recorded less frequently after peptide stimulation. It had a lower conductance (1.0-1.3 pS) and slower kinetics and was designated a very small conductance Cl- channel. It is concluded that activation of two types of Cl- channels (at least one of which is Ca2+ dependent) is involved in the late depolarization produced by vasoconstrictor peptides in vascular smooth muscle cells of the aortic cell line A7r5.
Van Renterghem
C
C
Institut de Pharmacologie Moléculaire et Cellulaire, Valbonne, France.
Lazdunski
M
M
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001011:Aorta; D001127:Arginine Vasopressin / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D018118:Chloride Channels / Q000187:drug effects / Q000502:physiology*; D004553:Electric Conductivity; D004622:Embryo, Mammalian; D016232:Endothelins / Q000494:pharmacology*; D006160:Guanosine Triphosphate / Q000494:pharmacology; D007700:Kinetics; D008564:Membrane Potentials; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000502:physiology*; D051381:Rats
D018118:Chloride Channels; D016232:Endothelins; D001127:Arginine Vasopressin; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D002118:Calcium