Transmural heterogeneity of myocardial metabolism and function are present in the left ventricle under normal and ischemic conditions. To determine if endocardial versus epicardial differences of [Ca2+]i are also present, perfused rat heart studies using indo-1 fluorescence as an index of [Ca2+]i were performed in the left ventricular epicardium and endocardium. Hearts were studied during control conditions and low-flow ischemia. Results demonstrated the following: 1) At a pacing rate of 1.5 Hz, endocardial levels of diastolic and systolic [Ca2+]i (470 +/- 40 and 1,240 +/- 170 nM) were higher than epicardial levels (290 +/- 30 and 920 +/- 150 nM). 2) At a more physiological pacing rate of 5 Hz, endocardial levels of diastolic and systolic [Ca2+]i (680 +/- 50 and 1,230 +/- 70 nM) were also higher than epicardial levels (390 +/- 20 and 950 +/- 60 nM. 3) During low-flow ischemia, endocardial levels of diastolic [Ca2+]i rose to a greater degree (from 680 +/- 50 to 1,050 +/- 70 nM at 10% of control coronary flow) compared with epicardial levels (from 390 +/- 20 to 580 +/- 40 nM at 10% of control flow), suggesting that the endocardium is more susceptible to low-flow ischemia. 4) The amplitude of the [Ca2+]i transient was the same at the endocardium (540 +/- 50 nM) and epicardium (560 +/- 50 nM) and did not change during low-flow ischemia, despite marked contractile dysfunction.(ABSTRACT TRUNCATED AT 250 WORDS)