The purpose of this investigation was to examine the effects of voluntary muscular fatigue in one lower limb and determine whether a 'cross-over' of fatigue is evident in the contralateral limb. Twenty-eight subjects (13 males and 15 females) performed a series of voluntary and evoked isometric contractions of both the dominant (exercised) and non-dominant (non-exercised) leg extensor muscles, prior to and after a fatigue protocol consisting of a 100-s sustained maximal isometric contraction (MVC) performed by the dominant limb only. Force values and surface electromyography (EMG) from the vastus lateralis muscle were obtained allowing for the determination of twitch and compound action potential (M-wave) values. Maximal twitch tension and peak-to-peak amplitude were significantly decreased after the fatigue test in the dominant limb, as was maximal voluntary force (~65 N reduction), EMG activity (~0.1 mV decrease) and voluntary activation (~17% decline). However, no significant changes were observed in the non-dominant limb with respect to twitch and M-wave properties nor in MVC force. The voluntary activation of the non-dominant limb decreased significantly by 8.7% after the fatigue test, which was performed only on the dominant limb. The results of the present study suggest that the decrease in force production in the exercised limb was primarily related to peripheral fatigue mechanisms, with central fatigue making a lesser contribution. Centrally mediated mechanisms appear to be the sole contributor to fatigue in the non-exercised limb suggesting an anticipatory fatigue response and a 'cross-over' of central fatigue between the exercised and non-exercised contralateral limb.