The functional plasticity of the nervous system may result in part from the direct modulation of the effectiveness of the release machinery of synaptic terminals. To date, direct modulation of secretion in neurons has proven difficult to study because of the lack of a suitable tool to probe the release machinery independently of calcium influx. We report that the polyvalent cation ruthenium red (RR) directly evokes rapid and reversible calcium-independent quantal secretion in hippocampal neurons by binding to external sites on the presynaptic terminal membrane. This binding can be displaced by heparin and is not associated with ultrastructural damage to the synaptic terminals. The use of RR-evoked release as a tool has allowed us to detect a direct modulation of the secretory apparatus after activation of A1 adenosine receptors on hippocampal neurons.