OBJECTIVE Left ventricular (LV) diastolic function is an important determinant of aerobic fitness. The purpose of this paper was to investigate the relationship between aerobic fitness and the rate and extent of isovolumic LV relaxation. METHODS Two series of experiments were performed utilizing both human and animal models. In the first series of experiments, the relationship between LV diastolic time intervals and exercise capacity was assessed in two groups of collegiate men (N = 18) with variable peak run times (Bruce protocol). In the second series of experiments, the extent of LV relaxation was examined in sedentary and exercise-trained rats (treadmill running), using an isolated, isovolumic heart preparation. Subsequent morphological assessment was also performed in rats. RESULTS At rest, men with greater peak treadmill time had a shorter resting LV isovolumic relaxation time (R-R interval adjusted 1000 ms) (long duration runners, 84+/-5 ms vs short duration runners, 105+/-7 ms, P < 0.05) despite a similar LV diastolic interval. Peak treadmill time was inversely correlated to LV isovolumic relaxation time (R-R interval adjusted 1000 ms) (r = -0.55; P < 0.02). In animal studies (N = 26), the LV pressure-volume relationship was shifted rightward in exercise-trained rats (P = 0.003). Exercise-trained rats had an increased LV inner diameter (sedentary, 5.1+/-0.35 mm vs exercise-trained, 6.1+/-0.28 mm, P < 0.05) and a thicker interventricular septum (sedentary, 1.52+/-0.06 mm vs exercise-trained, 1.72+/-0.09 mm, P < 0.05). CONCLUSIONS This study suggests that both the rate and extent of LV isovolumic relaxation is enhanced with exercise training. Further study is required to understand the interrelationship between exercise and diastolic function.