Purpose: We tested the hypothesis that central and peripheral fatigue after constant-load cycling exercise would vary with exercise intensity and duration. Methods: Twelve well-trained male cyclists (V·O2max, 4.49 ± 0.35 L·min-1) completed three constant-load cycling trials to the limit of tolerance in a randomized crossover design. Exercise intensities were set according to the respiratory responses to a preliminary ramp test to elicit cardiorespiratory and metabolic responses consistent with exercise in the severe and heavy exercise domains: 1) at power at V·O2max (S+, 379 ± 31 W), 2) at 60% of the difference between gas exchange threshold and V·O2max (S-, 305 ± 23 W), and 3) at the respiratory compensation point (RCP, 254 ± 26 W). Pre- and postexercise twitch responses from the quadriceps to the electrical stimulation of the femoral nerve and magnetic stimulation of the motor cortex were recorded to assess neuromuscular and corticospinal function, respectively. Results: Exercise time was 3.14 ± 0.59, 11.11 ± 1.86, and 42.14 ± 9.09 min for S+, S-, and RCP, respectively. All trials resulted in similar reductions in maximum voluntary force (P = 0.61). However, the degree of peripheral fatigue varied in an intensity-dependent manner, with greater reductions in potentiated twitch force after S+ (-33% ± 9%) compared with both S- (-16% ± 9%, P <0.001) and RCP trials (-11% ± 9%, P <0.001) and greater after S- compared with RCP (P <0.05). For central fatigue, this trend was reversed, with smaller reductions in voluntary activation after S+ compared with RCP (-2.7% ± 2.2% vs –9.0% ± 4.7%, P <0.01). Conclusion: These data suggest the magnitude of peripheral and central fatigue after locomotor cycling exercise is exacerbated with exercise intensity and duration, respectively.