Chronic administration of antidepressants produces region-specific adaptive changes in the radioligand binding properties ofN-methyl-d-aspartate (NMDA) receptors. We hypothesized that this effect of chronic antidepressant administration was owing to an alteration in NMDA receptor subunit composition. This hypothesis was examined usingin situ hybridization with [³⁵S]-labeled riboprobes to quantify the impact of chronic (16 d) injection with either imipramine (15 mg/kg) or citalopram (20 mg/kg) on the levels of transcripts encoding NMDA receptor subunits in mouse brain. These antidepressants altered the levels of mRNA encoding the ζ-subunit in a parallel fashion, with both drugs either reducing transcript levels (e.g., in the cortex, cerebellum, thalamus, and striatum) or producing no substantial effects (e.g., hippocampus). In contrast, these antidepressants often produced distinct, region-specific effects on mRNA levels encoding the ɛ family of subunits. For example, citalopram treatment produced widespread reductions in ɛ1-subunit mRNA levels (e.g., in frontal cortex, CA2 of hippocampus, and amygdala), whereas imipramine reduced levels of this transcript only in the amygdala. Conversely, imipramine treatment produced widespread reductions in ɛ2-subunit mRNA levels (e.g., in cortex, CA1-4 of hippocampus, and amygdala), whereas the effects of citalopram on levels of this transcript were largely restricted to amygdala. These findings indicate that long-term antidepressant treatment produces region-specific changes in expression of transcripts for NMDA receptor subunits, presumably altering NMDA receptor composition. Because subunit composition determines the physiological and pharmacological properties of NMDA receptors, these changes may play a critical role in the therapeutic actions of structurally diverse antidepressants.