To examine the electrophysiologic characteristics of the subvalvular mitral region, we retrospectively searched for the presence of subvalvular diastolic potentials (DP) in 91 patients (mean age, 46.9 ± 16.6 years) who underwent catheter ablation of left-sided accessory pathways (AP). We detected low-amplitude (0.19 ± 0.09 mV) DP in 14 patients (15.4%), including 8 with overt preexcitation and 6 patients with concealed AP. The mean interval between ventricular electrogram and DP was 383 ± 46 ms (range, 306-475). DP were detected in 4 of 20 patients with antero-lateral, 3 of 38 with lateral, 4 of 12 with postero-lateral, 2 of 14 with posterior, and 3 of 10 patients with postero-septal AP. In 6 of 14 patients, DP were detected before ablation. In 4 of 8 patients with overt preexcitation, DP were consistently recorded after elimination of the delta wave, suggesting that they were not associated with AP conduction. In 6 of 11 patients, DP were observed during both sinus rhythm and ventricular pacing, suggesting that they were not artifacts. The electrophysiologic characteristics of clinically relevant DP around the mitral annulus suggest that, in normal human hearts, an anatomical substrate may be present around the mitral annulus.