In this study, we investigated the effects of different skills of sport on electromyography (EMG) of limb muscles in sportsmen. Two different skill groups of sport consisted of 10 soccer players who use extensively lower limb, and 10 handball, basketball and volleyball players with selectively high usage of the upper limb. Surface EMG (sEMG) were recorded from the abductor pollicis brevis and gastrocnemius muscles of subjects. In addition, Hoffmann reflex (H-reflex) were recorded from the lower limb of participants. EMG findings of two groups were compared with each other and age-sex matched sedentary controls. Amplitude and area of sEMG recorded from gastrocnemius muscle of handball -basketball-volleyball players were significantly higher than those of soccer players (p < 0.01 and p < 0.05), whereas not significantly different based on the abductor pollicis brevis (p > 0.05 in both). F-response of both muscles and H-reflex of handball-basketball-volleyball players were also significantly higher from those of soccer players and control group (p < 0.05 for all), whereas only F-response of gastrocnemius muscle of soccer player was significantly lower than that of control group (p < 0.05). We conclude that EMG amplitude of gastrocnemius muscle, F-response and H-reflex times of lower limbs increase in handball-basketball-volleyball players due to the nature of training skills. Volleyball, handball or basketball training contributes to neuromuscular differences in both upper and lower extremities more than football training because both extremities are extensively used in these sport categories.