1. In rabbit and human hearts there are significant differences in the action potential configuration in atrium and ventricle, and the action potential waveform exhibits marked frequency dependence in both tissues. To study the ionic mechanism(s) of these phenomena, the size and time course of the potassium (K+) currents responsible for repolarization have been recorded from single cells using a whole-cell microelectrode voltage clamp method. 2. At physiological heart rates, the action potential in atrial cells has a short plateau phase; however, the rapid early repolarization is strongly frequency dependent. Ventricular myocytes have a long plateau (400-700 ms at 23 degrees C), and the late repolarizing phase of the action potential is much faster in ventricle than in atrium. 3. In both cell types, four different outward currents can be recorded: (i) a large transient outward current, It; (ii) IK(Ca), a smaller Ca2+-dependent K+ current; (iii) IK, a small, maintained time- and voltage-dependent delayed rectifier K+ current; (iv) IK1, an inwardly rectifying K+ current. 4. It, which is responsible for early repolarization, is much larger in atrium than in ventricle. It has very rapid activation and inactivation kinetics but a very slow time course of recovery from inactivation (tau = 5.4 s at 23 degrees C). Our results show that the reactivation kinetics of It are responsible for the pronounced dependence of the shape of the atrial action potential on stimulus frequency. 5. IK(Ca) is variable from cell to cell and is larger in atrium than in ventricle. In both cell types, IK(Ca) is much smaller than It. 6. The delayed rectifier current, IK, is very small and turns on relatively slowly in both cell types. It is therefore not activated strongly during the relatively short plateau of the atrial action potential. Even in ventricle, it contributes only a small repolarizing current. 7. IK1, the inward rectifier K+ current, is much larger in ventricle than in atrium. The current-voltage relationship for IK1 in ventricle exhibits a negative slope conductance between -50 and 0 mV. IK1 is the outward current which generates the resting membrane potential and it modulates the final repolarization phase of the action potential in both cell types. 8. These data strongly suggest that the action potential configuration and its frequency dependence in rabbit atrial and ventricular cells are mainly due to the differences in sizes and kinetics of It and IK1.