The sensorimotor striatum is critical for the acquisition and consolidation of skilled learning-related motor sequences. Excitatory corticostriatal synapses undergo neuroplastic changes that impact signal transmission efficacy. Modification of N-methyl d-aspartate (NMDA) and α-amino-3-hydroxyl-5-methyl-4-isoxazolepropionic acid (AMPA) glutamate receptor subunit composition and phosphorylation is critical for bidirectional experience-driven plasticity observed at these synapses. Metaplastic regulation of the ratio of NR2A to NR2B subunits of the NMDA receptor controls the threshold for the induction of subsequent plasticity. However, little is known about how repeated practice effects the differential regulation of glutamate receptors during the acquisition of a unilateral motor skill. Using immunoblot analysis, we assessed changes in NMDA and AMPA receptors during the associative stage of skill acquisition in synaptoneurosome preparations from the rat sensorimotor striatum. We found that the NR2A/B subunit ratio in the striatum contralateral to the trained limb decreased during skill acquisition optimizing the threshold for inducing subsequent synaptic plasticity during learning of the lateralized motor skill. In contrast, there was a significant increase in the NR2A/B subunit ratio in the ipsilateral striatum making the induction of subsequent plasticity more difficult. In addition, there was a selective decrease in AMPAR phosphorylation levels at serine site 831 but not 845 on the GluR1 subunit ipsilaterally with a trend toward a decrease contralaterally. These findings suggest that the successful acquisition of a lateralized motor skill necessitates the integration of motor programs in both striata, each of which reflects unique changes in the NR2A/B ratio that modulate the different task demands on the associated limb.