Dopaminergic neurons and receptors are widely distributed throughout the Drosophila CNS. However, the role of dopamine in modulating Drosophila central synaptic transmission has not been investigated. By using electrophysiological techniques, I explored dopamine modulation of cholinergic and GABAergic transmission in primary neuronal cultures. Cholinergic synaptic currents were suppressed by dopamine. This effect was mimicked by the mammalian D1-like receptor agonists SKF38393 and (±)-6-chloro-APB. GABAergic transmission was enhanced by dopamine and the D2-like receptor agonist (-)-quinpirole. These results demonstrated that there are two distinct dopamine signaling pathways modulating cholinergic and GABAergic transmission through D1- and D2-like receptors, respectively. Forskolin modulated cholinergic and GABAergic transmission exactly opposite to dopamine modulation, suggesting D1- and D2-like receptor actions are predominantly independent of the increase in intracellular cAMP levels. This study reports dopamine modulation of Drosophila central synaptic transmission for the first time and will help to elucidate mechanisms of dopamine modulation underlying behaviors.