Action potential conduction through the atrium and the ventricle of the heart depends on the membrane properties of the atrial and ventricular cells, particularly with respect to the determinants of the initiation of action potentials in each cell type. We have utilized both current- and voltage-clamp techniques on isolated cells to examine biophysical properties of the two cell types at physiological temperature. The resting membrane potential, action potential amplitude, current threshold, voltage threshold, and maximum rate of rise measured from atrial cells (-80 +/- 1 mV, 109 +/- 3 mV, 0.69 +/- 0.05 nA, -59 +/- 1 mV, and 206 +/- 17 V/s, respectively; means +/- SE) differed significantly (P < 0.05) from those values measured from ventricular cells (-82.7 +/- 0.4 mV, 127 +/- 1 mV, 2.45 +/- 0.13 nA, -46 +/- 2 mV, and 395 +/- 21 V/s, respectively). Input impedance, capacitance, time constant, and critical depolarization for activation also were significantly different between atrial (341 +/- 41 M omega, 70 +/- 4 pF, 23.8 +/- 2. 3 ms, and 19 +/- 1 mV, respectively) and ventricular (16.5 +/- 5.4 M omega, 99 +/- 4.3 pF, 1.56 +/- 0.32 ms, and 36 +/- 1 mV, respectively) cells. The major mechanism of these differences is the much greater magnitude of the inward rectifying potassium current in ventricular cells compared with that in atrial cells, with an additional difference of an apparently lower availability of inward Na current in atrial cells. These differences in the two cell types may be important in allowing the atrial cells to be driven successfully by normal regions of automaticity (e.g., the sinoatrial node), whereas ventricular cells would suppress action potential initiation from a region of automaticity (e.g., an ectopic focus).