Purpose: This study aimed to investigate effect of ankle instability on peripheral muscle activation among female ballet dancers to provide information on the development of prevention programs for ankle injury rehabilitation. Methods: 32 female ballet dancers were randomly divided into two groups: experience ankle sprain group (n=16, age, $20.7{\pm}0.8years$, BMI $18.6{\pm}1.2kg/m2$) and non-experience ankle sprain group (n=16, $age=21.0{\pm}0.8years$, BMI $19.6{\pm}2.0kg/m2$). Activation of the peroneus longus, tibialis anterior muscle, and gastrocnemius during vertical landing, half pointe, and gait between the two groups were measured. Body composition analyzer was used to examine skeletal muscle mass and body fat mass. Results: A total of 32 patients were included. In the experience ankle sprain group (n=16: left sprain 14, right sprain 2), average ankle sprain injury occurred 7.5 months before the study. The average age of the dancers in the experience ankle sprain group and non-experience ankle sprain group was $20.7{\pm}0.8$ and $21.0{\pm}0.8years$, major period was $64.5{\pm}23.8$ and $71.6{\pm}25.8months$, BMI was $18.6{\pm}1.2$ and $19.5{\pm}2.0kg/m2$, respectively. No significant differences were found on body composition between the two groups (p>0.05). The experience ankle sprain group showed significantly lower tibialis anterior and peroneus longus muscle activation (p<0.5), while gastrocnemius muscle activation appeared to be significantly higher (p<0.05) during landing, half pointe, and normal gait. Conclusion: Ankle sprain can cause a decline in peripheral muscle activation and coordination, which increased the risk for repetitive ankle sprain in the future. Moreover, ankle peripheral muscle selective strength training, coordination program development, and application need to be considered to prevent ankle sprain.