The aim of the present study was to determine the magnitude of the maximal level of peripheral fatigue attainable (fatigue threshold) during an all-out intermittent isometric knee-extensor protocol in both younger (24 ± 1 yr, n = 12) and older (60 ± 2 yr, n = 12) participants to provide new insights into the effects of aging on neuromuscular function. Participants performed two experimental sessions, in which they performed 60 maximal voluntary contractions (MVCs; 3 s of contraction, 2 s of relaxation). One trial was performed in the unfatigued state (CTRL) and one other following fatiguing neuromuscular electrical stimulation of the quadriceps (FNMES). Peripheral fatigue was quantified via pre/postexercise decrease in quadriceps twitch force (∆Ptw). Critical force (CF) was determined as the mean force output of the last 12 contractions, whereas W′ was calculated as the area above CF. Although FNMES led to a significant decrease in Ptw before performing the 60-MVCs protocol ( P = 0.024), ∆Ptw was not different between CTRL and FNMES for both the young group ( P = 0.491) and the old group ( P = 0.523). However, this peripheral fatigue threshold was significantly greater in young versus old participants (∆Ptw = −48 ± 10% vs. −29 ± 13%, respectively, P = 0.028). In CTRL, W′ was 55 ± 13% lower in the old group than in the young group ( P < 0.001), but CF was similar (326 ± 10 N vs. 322 ± 12 N, respectively, P = 0.941). ∆Ptw was correlated with W′, independently of age ( r2 = 0.84, P < 0.001). Exercise performance decreases with aging consequent to a lower tolerance to peripheral fatigue. However, the peripheral fatigue threshold mechanism persists with healthy aging and continues to play a protective role in preserving locomotor muscle function during exercise.