Acetylcholine induces Ca-dependent K currents in rabbit endothelial cells.

Effects of acetylcholine (ACh) on the membrane potential and current recorded from endothelial cells dispersed from the rabbit aorta were investigated using the patch-clamp technique. ACh hyperpolarized the endothelial cell membrane. Using the whole-cell voltage-clamp procedure, ACh (10(-6) M) induced an outward current, and this current was blocked by atropine (10(-6) M). Application of either pirenzepine (3 x 10(-7) M) or AF-DX 116 (3 x 10(-6) M) slightly inhibited the ACh-induced outward current, and simultaneous application of these two blockers markedly inhibited the outward current. Application of caffeine (2 x 10(-2) M), ryanodine (10(-5) M) or heparin (10(-5) g/ml) reduced the amplitude of the ACh-induced outward current. A single-channel current recording using the patch-clamp technique revealed that ACh opens a Ca-dependent K-channel with a single-channel current conductance of 9 pS. These results indicate that both M1 and M2 receptor subtypes are present in endothelial cells of the rabbit aorta and that ACh activates the Ca-dependent K channel via release of Ca from intracellular store sites. In addition, methylene blue inhibited the ACh-induced outward current from the outside membrane.