Potassium conductance of smooth muscle cells from rabbit aorta in primary culture
- PMID: 1945762
- DOI: 10.1007/BF00373748
Potassium conductance of smooth muscle cells from rabbit aorta in primary culture
Abstract
Vascular smooth muscle cells were obtained from rabbit aorta and were studied in primary culture on days 1-7 after seeding with electrophysiological techniques. In impalement experiments a mean membrane potential difference (PD) of -50 +/- 0. 3 mV (n = 387) was obtained with Ringer-type solution in the bath. PD was depolarized by 6 +/- 0. 3 mV (n = 45) and 16 +/- 2 mV (n = 5) when the bath K+ concentration was increased from the control value of 3. 6 mmol/l to 13. 6 and 23. 6 mmol/l, respectively. Ba2+ (0. 1-1 mmol/l) depolarized PD. Tetraethylammonium (TEA, 10 mmol/l) depolarized PD only slightly but significantly. Verapamil (0. 1 mmol/l) and charybdotoxin (10 nmol/l) had no effect on PD. The conductance properties of these cells were further examined with the patch-clamp technique. K+ channels were spontaneously present in cell-attached patches. When the pipette was filled with 145 mmol/l KCl, a mean conductance (gK) of 209. 6 +/- 4. 6 mV (n = 17) was read from the current/voltage curves at a clamp voltage (Vc) of 0 mV. After excision K+ channels were found in 129 patches with inside-out and in 50 with outside-out configuration. With KCl on one and NaCl on the other side the mean gK at a Vc of 0 mV was 134. 6 +/- 3. 9 pS (n = 179). The mean permeability was 0. 89 +/- 0. 03 x 10(-12) cm3/s. With symmetrical KCl solution the mean gK was 227 +/- 6 pS (n = 17). The conductance sequence was gK much greater than gRb = gCs = gNa = 0. TEA blocked dose-dependently only from the outside (1-10 mmol/l). Lidocaine (5 mmol/l) quinidine (0. 01-1 mmol/l) and quinine (0 VSports手机版. 01-1 mmol/l) blocked from both sides. Charybdotoxin (0. 5-5 nmol/l) blocked only from the extracellular side. Ba2+ blocked from the cytosolic side and the inhibition was increased by depolarization and reduced by hyperpolarization. At a Vc of 0 mV a half-maximal inhibition (IC50) of 2 mumol/l was obtained. Verapamil and diltiazem blocked from both sides, verapamil with an IC50 of 2 mumol/l and diltiazem with an IC50 of 10 mumol/l. The open probability of this channel was increased by CA2+ on the cytosolic side at activities greater than 0. 1 mumol/l. Half-maximal activation occurred at Ca2+ activities exceeding 1 mumol/l. The present data indicate that the vascular smooth muscle cells of rabbit aorta in primary culture possess a K+ conductance. In excised patches only a maxi K+ channel was detected. This channel has properties different from the macroscopic K+ conductance. (ABSTRACT TRUNCATED AT 400 WORDS) .
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