Purpose. This study investigated maturation-related differences in neuromuscular fatigue after a short-term maximal run. Methods. Eight male children, eight adolescents, and eight adults performed a maximal ca. 50-s run (300/350/400 m, respectively). Mechanisms of neuromuscular fatigue were assessed through isometric plantar flexor tests, electrical stimulation of the posterior tibial nerve, soleus electromyography, and blood tests. Results. All the groups showed a decrease in the running speed (children: -12.2 ± 6.5%; adolescents: -9.8 ± 5.1%; adults: -12.2 ± 3.1%), but only adults revealed a decline in the maximal isometric plantar flexor torque (-16.1 ± 13.0%). On the other hand, the relative pre- to post-fatigue change in the maximal isometric plantar flexor torque differed only between adults and adolescents. The peak torque in the passive twitch test decreased in adolescents (-19.2 ± 12.2%) and adults (-23.7 ± 13.7%). Moreover, post-fatigue minimum blood pH (children: 7.18 ± 0.03; adolescents: 7.14 ± 0.07; adults: 6.97 ± 0.06) differed between the groups. No changes were observed in the neural drive or mechanisms at the spinal level. Conclusions. Despite the loss of running speed, children showed no post-exercise fatigue, whereas adolescents and adults demonstrated fatigue at peripheral sites. Central fatigue could not be established for the studied groups.