Gait disturbances similar to those of human Parkinson's disease (PD) can be observed in animals after administration of neurotoxin 6-hydroxydopamine (6-OHDA) to induce unilateral nigrostriatal dopamine depletion. However, the relationship between gait disturbances and dopamine depletion following 6-OHDA infusion has not been determined. The present study investigated the longitudinal changes of spatiotemporal gait patterns using a walkway system to acquire footprints and lateral limb images over a 6-week period following unilateral 6-OHDA injection into the medial forebrain bundle of rats. Our results indicated that hemiparkinsonian rats exhibited changes in gait patterns, as compared to normal controls, and pre-lesion levels, including a significantly decreased walking speed and step/stride length as well as an increased base of support and foot angle. The relative percentage of the gait cycle was also altered, showing an increase in the stance to swing ratio, which was more evident in the affected hindlimb. Time-course observations showed that these gait disturbances occurred as early as 4 days post-lesion and gradually increased up to 42 days post-injury. The extents of gait disturbances were compared with conventional apomorphine-induced turning behavior and akinesia bar tests, which were also apparent at 4 days post-lesion but remained relatively unchanged after 28 days. Our time-course gait analysis of a unilateral 6-OHDA rodent model provides insight into the compensatory changes of motor functions during the 6-week development of a nigrostriatal lesion, which might be useful for future objective assessment of novel treatments for human PD subjects. Crown Copyright (C) 2011 Published by Elsevier B.V. All rights reserved.