BackgroundWalking requires constant adjustments to the changing environment. An asymmetrical perturbation can affect the gait symmetry, cause gait adaptations, and potentially induce retention of the adapted gait after removal of the perturbation. A unilateral ankle load has the potential to create asymmetry and facilitate the emergence of new gait patterns. However, few studies have examined the effect of unilateral loading on muscular adjustments during walking. The purpose of this study was to investigate gait adaptations and muscular adjustments after unilaterally loading or unloading the ankle. Research questionWhat are the effects of unilateral loading and unloading on gait spatiotemporal parameters and muscle activation in young adults? MethodsTwenty young adults (10 M/10 F) walked on a treadmill at their preferred walking speeds in 3 conditions: 1) a 2-minute baseline trial; 2) three 5-minute trials with a load (3 % of bodyweight) on the dominant ankle (Loading); and 3) one 5-minute trial with the load removed (Unloading). Inertial measurement units (IMUs) and electromyography sensors (EMGs) were used for data collection. Early and late adaptation and post-adaptation were assessed using the first 5 strides and the last 30 strides of loading and unloading conditions. Outcome measures included symmetry index (SI) of spatiotemporal parameters, range-of-motion (ROM) of the lower body joints, and EMG integrals of leg muscles. Repeated measures ANOVA was conducted for statistical analysis (α = 0.05). ResultsSI of swing phase percentage demonstrated rapid adaptation after unilateral loading or unloading. Stride length demonstrated an aftereffect following unloading. Young adults reduced ankle ROMs bilaterally in early adaptation and increased loaded-side knee and hip ROMs in late adaptation. Additionally, they increased the tibialis anterior activity bilaterally immediately after unilateral loading. SignificanceYoung adults showed an aftereffect in some variables after unilateral unloading, signifying that unilateral ankle loading can induce short term learning of a new gait pattern.