The purpose of the present study was to examine the neuromuscular modifications of cyclists to changes in grade and posture. Eight subjects were tested on a computerized ergometer under three conditions with the same work rate (250 W): pedaling on the level while seated, 8% uphill while seated, and 8% uphill while standing (ST). High-speed video was taken in conjunction with surface electromyography (EMG) of six lower extremity muscles. Results showed that rectus femoris, gluteus maximus (GM), and tibialis anterior had greater EMG magnitude in the ST condition. GM, rectus femoris, and the vastus lateralis demonstrated activity over a greater portion of the crank cycle in the ST condition. The muscle activities of gastrocnemius and biceps femoris did not exhibit profound differences among conditions. Overall, the change of cycling grade alone from 0 to 8% did not induce a significant change in neuromuscular coordination. However, the postural change from seated to ST pedaling at 8% uphill grade was accompanied by increased and/or prolonged muscle activity of hip and knee extensors. The observed EMG activity patterns were discussed with respect to lower extremity joint moments. Monoarticular extensor muscles (GM, vastus lateralis) demonstrated greater modifications in activity patterns with the change in posture compared with their biarticular counterparts. Furthermore, muscle coordination among antagonist pairs of mono- and biarticular muscles was altered in the ST condition; this finding provides support for the notion that muscles within these antagonist pairs have different functions.