Regional uptake of iodine-125-metaiodobenzylguanidine ([125I]MIBG) was evaluated in normal (n = 12) and reserpinized (n = 12) rat hearts. At 15 min and 1, 3 and 6 h after injection of [125I]MIBG, tissue activities were calculated for the right ventricular myocardium (RV), the whole left ventricular myocardium (whole LV), the epicardial layer of the left ventricular myocardium (Ep LV), the endocardial layer of the left ventricular myocardium (En LV), the basal segment of the left ventricular myocardium and the apical segment of the left ventricular myocardium. The uptake of [125I]MIBG at 6 h after injection in the normal rat heart was higher in RV than in whole LV (0.45 +/- 0.09% vs 0.30 +/- 0.06% kg dose/g), and in Ep LV than in En LV (0.32 +/- 0.07% vs 0.25 +/- 0.05%). In the reserpinized rat heart, the difference in the uptake between Ep LV and En LV was smaller. This suggests that the difference in the regional [125I]MIBG uptake might reflect different intravesicular uptake in the layers of the heart. To our knowledge, the low uptake in the endocardial layer was a new finding which seems to indicate a difference in innervation between the epicardial and endocardial layers of the left ventricle in the rat heart. Autoradiographic study also showed the low uptake of [125I]MIBG in the endocardial layer, while homogeneous perfusion was observed with thallium-201, supporting the tissue uptake study. Thus, the endocardial and epicardial layers of the left ventricle in the rat heart were considered to be differently innervated.