Study Design Controlled laboratory study. Background Joint mobilization and manipulation decrease pain and improve patient function. Yet, the processes underlying these changes are not well understood. Measures of corticospinal excitability provide insight into potential mechanisms mediated by the central nervous system. Objectives To investigate the differential effects of joint mobilization and manipulation at the talocrural joint on corticospinal excitability in individuals with resolved symptoms following ankle sprain. Methods Twenty-seven participants with a history of ankle sprain were randomly assigned to the control, joint mobilization, or thrust manipulation group. The motor-evoked potential (MEP) and cortical silent period (CSP) of the tibialis anterior and gastrocnemius were obtained with transcranial magnetic stimulation at rest and during active contraction of the tibialis anterior. The slopes of MEP/CSP input/output curves and the maximal MEP/CSP values were calculated to indicate corticospinal excitability. Behavioral measures, including ankle dorsiflexion and dynamic balance, were evaluated. Results A repeated-measures analysis of variance of the MEP slope showed a significant group-by-time interaction for the tibialis anterior at rest (P = .002) and during active contraction (P = .042). After intervention, the thrust manipulation group had an increase in corticospinal excitability, while the corticospinal excitability decreased in the mobilization group. The thrust manipulation group, but not other groups, also demonstrated a significant increase in the maximal MEP amplitude of the tibialis anterior after intervention. Conclusion The findings suggest that joint manipulation and mobilization have different effects on corticospinal excitability. The increased corticospinal excitability following thrust manipulation may provide a window for physical therapists to optimize muscle recruitment and subsequently movement. The trial was registered at ClinicalTrials.gov (NCT00847769). J Orthop Sports Phys Ther 2016;46(7):562-570. Epub 6 Jun 2016. doi:10.2519/jospt.2016.6602.