An estimated 23,000 ankle injuries occur every day in the U.S. Ankle sprains account for 85% of all ankle injuries and inversion ankle sprains account for 85% of all ankle sprains. There is growing evidence that suggests gender and fatigue may increase the risk for inversion ankle sprains. Investigating the effects of fatigue and gender on peroneal reflex response after ankle inversion may help explain the differences in sprain rates with fatigue and gender. Therefore, the purpose of this study was to investigate the effects of fatigue and gender on peroneus brevis and peroneus longus reflexes after ankle inversion. A "trap-door" platform was used to elicit peroneal reflexes from sixteen males and fifteen females by suddenly inverting the ankle to 20°. Five unfatigued peroneal reflex measurements were performed before and after a fatigue protocol that attempted to fatigue the ankle evertors over 12 minutes to 75% of the unfatigued MVC torque. Results showed that reflex delay was not affected by fatigue, gender, or their interaction. PL reflex amplitude was not affected by fatigue or gender but was affected by their interaction. Results showed that PL reflex amplitude decreased by 11.3% in males and increased 22.1% in females with fatigue. A secondary analysis attempted to rule out extraneous factors that could have contributed to the differences in reflex response, but no experimental explanations were found. The differences in PL reflex amplitude were attributed to biomechanical, physiological, and anatomical differences between males and females.