Accessory atrioventricular (AV) pathways responsible for the Wolff-Parkinson-White syndrome have been treated successfully by surgical and radiofrequency catheter ablation techniques. In this study, we explored the feasibility of permanent chemical ablation of accessary pathways by direct injection of 100% ethanol into the canine AV groove. Right coronary artery blood flow and AV compartment pressure were measured in 5 adult mongrel dogs (part I) during injection of saline solution (10 to 15 mL) into the right AV groove. Atrioventricular groove compartment pressure increased from 1 ± 1 to 31 ± 12 mm Hg ( p <0.0001) and right coronary artery blood flow increased slightly from 117 ± 17 to 138 ± 44 mL/min ( p = not significant). In 8 additional dogs (part II), 100% ethanol (10 mL) was injected into the left AV groove. After 6 weeks (n = 6) or 12 weeks (n = 2), histologic examination of the left AV groove demonstrated localized fibrous replacement of injured epicardial muscle, with only scant inflammatory infiltrate. Mild intimal fibrosis and intimal proliferation were present in the circumflex artery and its larger branches, but no significant luminal narrowing was evident. Angiograms at 12 weeks (n = 2) demonstrated no significant narrowing of the circumflex artery. In summary, injection of 100% ethanol into the AV groove is capable of locally ablating myocardial tissue, presumably including conductive tissues, while sparing the coronary arteries.