PURPOSE Despite growing interest in task-dependent alterations of central and peripheral fatigue after endurance exercise, little is known about the effect of rowing on quadriceps muscle fatigue. This study aimed to investigate central and peripheral mechanisms of fatigue after a 2000-m rowing time trial. METHODS Eight competitive rowers (four males and four females, 20 ± 4 yr) performed a 2000-m time trial on an indoor rower and a control condition (sitting). The neuromuscular function of the knee extensors was analyzed before and 3 min after each experimental condition. Maximal voluntary torque, voluntary activation, and normalized root-mean-square of the EMG signal were measured during isometric and concentric contractions. Furthermore, knee extensor twitch torque and maximal M-wave amplitudes in response to electrical nerve stimulation were assessed. RESULTS After the 2000-m rowing, there were significant reductions in isometric and concentric maximal voluntary torque of the knee extensors (-20% ± 9% and -18% ± 7%, respectively, P < 0.01). Both the voluntary activation of the knee extensors during isometric and concentric contractions decreased by 18% ± 15% (P < 0.05, respectively). The normalized muscle activity of rectus femoris was significantly reduced after rowing (P = 0.007), whereas vastus medialis and vastus lateralis muscle activities did not significantly differ from baseline values. No significant change was observed for knee extensor twitch torque in response to paired electrical stimuli after rowing. CONCLUSION The 2000-m rowing time trial resulted in significant knee extensor strength loss. Quadriceps muscle fatigue after high-intensity rowing exercise was explained primarily by central factors that lead to large reductions in voluntary drive.