Epicardial electrodes were applied to 12 thoractomized dogs to determine the effects of trains of subthreshold conditioning stimuli (TSc) on ventricular refractoriness when delivered preceding a premature suprathreshold stimulus (S2). Several factors were analyzed: (1) the influence of TSc pulse frequency (100-900 Hz); (2) the delay between TSc and S2 (1 or 10 ms); (3) the distance between the electrodes for the emission of TSc and S2 (same electrodes or different electrodes at 3 mm); and (4) S2 current intensity (two- or fourfold diastolic threshold). The TSc (mean current intensity 0.33 mA, range 0.1-0.7) were found to significantly prolong the effective ventricular refractory period (EVRP) at all train pulse frequencies. The EVRP increment was progressively greater as pulse frequency was increased, the maximum EVRP increment being at 900 Hz (mean 50.8 +/- 32.3 ms; maximum increment 130 ms). On increasing S2 current intensity, the EVRP increment was less (maximum value 35 ms) and less consistent (in four of six dogs); in two cases the EVRP was shortened. The increase in delay between TSc and S2 attenuated the EVRP prolongation, which was present in only three of six dogs tested, and the EVRP was shortened in two dogs. There was no EVRP prolongation at any TSc pulse frequency when TSc and S2 were delivered at different electrodes. Thus TSc decreases myocardial ventricular excitability, prolonging EVRP in direct proportion to TSc pulse frequency. However, this property is limited by S2 current intensity as well as the time and distance between TSc and S2.