The purpose of the present study was to determine whether some of the age-related changes that occur in binding to the NMDA receptor complex can be accounted for by changes in subunit expression during the aging process. In situ hybridization for the NMDA subunits zeta1, epsilon1, and epsilon2, and receptor autoradiography, using the agonist glutamate and the competitive antagonist [(+/-)-2-carboxypiperazin-4-yl] propyl-1-phosphonic acid (CPP), were performed on sections from C57Bl/6 mice representing three different age groups (3, 10, and 30 months of age). There was a significant overall decrease between 3 and 30 month olds in the density of mRNA for the zeta1 subunit in the cortex and hippocampus, but only a few individual brain regions exhibited significant declines. The mRNA for the epsilon2 subunit was significantly decreased in a majority of cortical regions and in the dentate granule cells. Emulsion analysis indicated that the change in the density of epsilon2 subunit mRNA in the inner frontal cortex was primarily attributable to a decrease in the amount of messages per cell. Age-related changes in mRNA density of the epsilon2 subunit correlated with changes in NMDA-displaceable [(3)H]glutamate binding, and mRNA density changes in the zeta1 subunit showed a significant relationship with changes in [(3)H]CPP binding in the 30-month-old mice. These results suggest that changes during aging in the expression of different subunits of the NMDA receptor may account for the differential effects of aging on agonist versus antagonist binding to the NMDA binding site.