In vivo studies have shown that sympathetic nerve stimulation improves the transmural distribution of myocardial perfusion by increasing the endocardial/epicardial flow ratio; however, the mechanism of this effect is unknown. During nerve stimulation both norepinephrine (NE) and neuropeptide Y (NPY) are released, either or both of which may exert vasoconstrictor effects. The present studies were performed to examine the effects of these two cotransmitters on the transmural distribution of myocardial perfusion in a canine model. In anesthetized open-chest dogs, during maximal coronary vasodilation with intracoronary adenosine, both neuropeptide Y (29.7 micrograms/min) and norepinephrine (0.5-2.0 micrograms/min) reduced myocardial perfusion to a greater extent in the epicardium than in the subendocardium. The endo/epi ratio with adenosine alone was 1.11 +/- 0.02. Norepinephrine increased this by 80%, neuropeptide Y by 20%, and the combination of the two by 76% (P < 0.05 for all three vs. adenosine). Neuropeptide Y alone constricted the coronary vasculature but did not alter transmural flow. Thus neuropeptide Y preferentially reduces myocardial perfusion in the epicardium. We speculate that neuronally released neuropeptide Y contributes importantly to the transmural distribution of myocardial perfusion during sympathetic nerve stimulation.