In this study we investigated effects of acute and chronic ethanol exposure on carbachol-induced activator protein-1 (AP-1) DNA binding in rat cerebellar granule cells. Acute ethanol application did not alter, whereas chronic ethanol exposure potentiated the carbachol-induced AP-1 DNA binding. The protein composition of the AP-1 transcription factor complex activated by carbachol stimulation of muscarinic receptors was analysed in control and chronic ethanol-exposed cells using a supershift assay with specific antibodies against c-Fos, Fos B, c-Jun, Jun B and Jun D proteins. Supershift analysis revealed that the carbachol-induced AP-1 complex was composed predominantly of Jun D and c-Fos. The composition of the AP-1 complex activated by carbachol in chronic ethanol-exposed cells did not differ from control. These findings indicate that chronic ethanol treatment can modulate carbachol-induced AP-1 DNA binding activity in cerebellar granule cells.