Dopamine (DA) and noradrenaline (NA) modulate responses to nociceptive stimuli, within the dorsal horn of the spinal cord. Both neurotransmitters may play a role in supraspinal regulation in response to proprioceptive afferences to the dorsal horn. However, direct evidence of changes in neurotransmitter release within the dorsal horn due to non-noxious stimuli is lacking. The present study was designed to determine, whether non-nociceptive exercise produces changes in release of DA and NA within the dorsal horn, and whether these changes are associated with long-lasting inhibition after the exercise stops. Microdialysis probes, implanted in layers 2-5 of Rexed, in combination with high-performance liquid chromatography coupled to electrochemical detection (HPLC-EC) were used to measure concentrations of DA and NA metabolite (MHPG) in lumbar spinal cords of rats. Microdialysate was sampled before, during, and after a treadmill exercise of one hour. Results indicate that DA and NA releases are inhibited during non-nociceptive motor activity. At rest, DA concentration was 204 ± 10.5 pg/10 μl and was significantly decreased during exercise to -11.4% (P ≤ 0.05). Greater decrease occurred after 30 min of exercise and was of -31.4% (P ≤ 0.05). Similarly, MHPG was significantly decreased of -18% during exercise (P ≤ 0.05). When exercise stopped, both systems showed long-lasting inhibition. Exercise post-release lasted 30 min for DA and 90 min for MHPG. MHPG greatest decrease of -47.8% occurred 30 min after stopping the exercise (P ≤ 0.001). Thus, DA and NA systems seem to respond to exercise-induced proprioceptive afferent stimuli to the dorsal horn.