AbstractBackgroundNeuromuscular electrical stimulation (NMES) can generate muscle contractions and elicit excitability of neural circuits. However, the optimal stimulation frequency for effective neuromodulation remains unclear.MethodsEleven able‐bodied individuals participated in our study to examine the effects of: (1) low‐frequency NMES at 25 Hz, (2) high‐frequency NMES at 100 Hz; and (3) mixed‐frequency NMES at 25 and 100 Hz switched every second. NMES was delivered to the right tibialis anterior (TA) muscle for 1 min in each condition. The order of interventions was pseudorandomized between participants with a washout of at least 15 min between conditions. Spinal reflexes were elicited using single‐pulse transcutaneous spinal cord stimulation applied over the lumbar enlargement to evoke responses in multiple lower‐limb muscles bilaterally and maximum motor responses (Mmax) were elicited in the TA muscle by stimulating the common peroneal nerve to assess fatigue at the baseline and immediately, 5, 10, and 15 min after each intervention.ResultsOur results showed that spinal reflexes were significantly inhibited immediately after the mixed‐frequency NMES, and for at least 15 min in follow‐up. Low‐frequency NMES inhibited spinal reflexes 5 min after the intervention, and also persisted for at least 10 min. These effects were present only in the stimulated TA muscle, while other contralateral and ipsilateral muscles were unaffected. Mmax responses were not affected by any intervention.ConclusionsOur results indicate that even a short‐duration (1 min) NMES intervention using low‐ and mixed‐frequency NMES could inhibit spinal reflex excitability of the TA muscle without inducing fatigue.