Fine-wire electromyography (EMG) is a traditional laboratory technique to estimate muscle activity of the small foot muscles, however, recordings have not been reported from extensor hallucis brevis (EHB). As an extensor of the great toe, EHB is an important muscle when studying physiological changes associated with foot pathologies such as hallux valgus. The purpose of this study was to develop an ultrasound-guided fine-wire EMG protocol to record EHB muscle activity and report normative EMG profiles of healthy young adults during locomotion. Sixteen asymptomatic young adults completed 20 walking trials at a self-selected velocity. Ensemble averages were calculated from the time normalized linear envelopes and represented from 0% to 100% of the single stance phase of gait. EHB muscle bursts were observed between 0% and 20% of the stance phase of gait in all participants. A second burst of EMG was observed between 80% and 100% of stance in 50% of the participants. This study introduces a novel ultrasound-guided EMG protocol and normative data from EHB recordings suggest a synergistic role to anterior compartment musculature at contact. These results provide preliminary insights into understanding the functional role of EHB and may help elucidate the biomechanical factors exacerbating the progression of hallux pathologies.