1. The effect of chronic antidepressant administration on CRE-, SP1- and GRE-binding activity was studied in rat hippocampus and frontal cortex. 2. Fluoxetine and desipramine (3 and 10 mg/kg/day respectively) were given to rats for 21 consecutive days. The animals were killed 3 hr after the last injection and nuclear extracts were prepared to perform the DNA-protein reaction with consensus CRE, SP1 and GRE oligonucleotides. 3. Gel-shift assays showed that CRE-binding activity was increased in both frontal cortex and hippocampus by chronic fluoxetine treatment. Desipramine, however, only enhanced this activity in the frontal cortex. 4. Chronic fluoxetine decreased SP1-binding activity in the two selected brain regions. Again, desipramine only produced a significant reduction in the frontal cortex. 5. GRE-binding in the hippocampus was only enhanced by desipramine. Since chronic desipramine, and not fluoxetine, is able to increase hippocampal glucocorticoid receptor (GR) expression, interactions of GR with CREB and SP1 may determine the lack of effect of desipramine on binding activity of the two latter transcription factors in this brain region. 6. Overall, the results show a differential and region-specific effect of chronic, and not acute, antidepressant treatment on the DNA-binding activities studied and are consonant with the possible role of changes in gene expression in the mechanism of antidepressant action.