Purpose: Critical torque (CT) and work done above it (Wʹ) are key predictors of exercise performance associated with neuromuscular fatigue. The aim of the present study was to understand the role of the metabolic cost of exercise in determining exercise tolerance, CT and Wʹ and the mechanisms of neuromuscular fatigue. Methods: Twelve subjects performed four knee extension time-trials (6, 8, 10, and 12-minutes) using eccentric, isometric, or concentric contractions (3s-on/2s-off at 90° or 30°/s) to modulate the metabolic cost of exercise. Exercise performance was quantified by total impulse and mean torque. CT and Wʹ were determined using the linear relationship between total impulse and contraction time. Cardiometabolic, neuromuscular, and ventilatory responses were quantified. Neuromuscular function was evaluated by maximal voluntary contraction, resting potentiated single/doublet electrical stimulations, and superimposed single electrical stimulation to quantify neuromuscular, peripheral, and central fatigue, respectively. Results: Compared to isometric exercise, total impulse (+36±21%; P<0.001), CT (+27±30%; P<0.001), and Wʹ (+67±99%; P<0.001) were increased during eccentric exercise whereas total impulse (-25±7%; P<0.001), critical torque (-26±15%; P<0.001) and Wʹ (-18±19%; P<0.001) were reduced in concentric exercise. Conversely, the metabolic response and the degree of peripheral fatigue were reduced during eccentric exercise whereas they were increased during concentric exercise. CT was negatively associated with oxygen consumption gain ( R2=0.636; P<0.001) and Wʹ was negatively associated with rates of neuromuscular and peripheral fatigue indices ( R2=[0.252-0.880]; P<0.001). Conclusion: The contraction mode influenced both CT and Wʹ, and consequently exercise tolerance, indicating that the metabolic cost of contraction played a key role. This work was supported by the University of Strasbourg Initiative of Excellence (2021-022-10 and ANR-10-IDEX-0002-02) This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.