An animal model of peripheral neuropathy resulting in a unilateral hyperalgesia has recently been developed. The N-methyl-D-aspartate (NMDA) antagonist MK-801 reduces the thermal hyperalgesia observed in this model. The goal of the present study was to determine whether the immunohistochemical changes in dorsal horn peptides shown by neuropathic animals could also be modified by MK-801. Changes in immunostaining densities of substance P (SP) and calcitonin gene-related peptide (CGRP) within the spinal cord of untreated (reference population) neuropathic rats and that of neuropathic rats treated for 7 days with MK-801 were quantified and compared. The reference neuropathic animals demonstrated thermal hyperalgesia and an ipsilateral decrease in SP staining density without an accompanying change in CGRP staining density. MK-801-treated animals showed a dose-dependent attenuation of the thermal hyperalgesia. The expected ipsilateral decrease in SP was prevented in neuropathic animals treated with a low dose (0.5 mg/kg) of MK-801, while a higher dose of MK-801 (1 mg/kg) resulted in an increase in SP staining ipsilateral to the injury. MK-801 treatment in naive rats caused a global increase in both SP and CGRP staining in the dorsal horn. However, this global increase failed to mask the changes in staining density in neuropathic animals following MK-801 treatment. The results suggest a functional interaction between excitatory amino acids (EAAs) and SP, with activation of NMDA receptors mediating depletion of SP in neuropathic animals. It is suggested that SP-containing interneurons are a target of the EAAs in the dorsal horn.