The question whether activation of the ATP-regulated K channel is responsible for macroscopic anoxia-induced outward currents was examined in ventricular cells isolated enzymatically from guinea-pig heart. Gigaseal patch-clamp electrodes were used for a whole-cell voltage clamp. Membrane currents were compared in the same cell while the cell interior was dialysed by perfusing the electrode with different solutions. When the cell was dialysed with various ATP-deficient (less than or equal to 2 mM) internal solutions, the Ca current decreased in a dose-dependent manner to less than 10% of control at 0.5 mM-ATP. A slight (ca. 25%) decrease of the slope conductance for hyperpolarizing current was observed. When a delayed rectification on depolarization followed by a marked outward current tail on repolarization was present under control conditions, this time-dependent outward current was also depressed. An increase in a time-independent outward current was observed accompanied by marked current fluctuations. The outward current showed a reversal potential near the K equilibrium potential, inward rectification, and no relaxation on voltage jumps. The power density spectrum of the current fluctuations showed a pattern similar to the spectrum calculated from the single-channel currents of ATP-regulated K channels. The amplitude of the single-channel current, estimated from the fluctuations, was almost equal to that of the single-channel current. The total number of channels within one cell was estimated as 2000-3000. It is concluded that the ATP-regulated K channels are responsible for the increase in the outward current and the shortening of the action potential duration under various anoxic conditions.