1. Intracellular recording techniques were used to study neurotransmitter release mechanisms in postganglionic sympathetic nerve terminals of the rat isolated anococcygeus muscle. 2. Low concentrations of the N-type calcium channel blocker omega-conotoxin GVIA (omega-CgTX GVIA) irreversibly abolished excitatory junction potentials (EJPs) evoked by trains of < or = five stimuli at 10 Hz. When the frequency of stimulation was increased (10-50 Hz) trains of stimuli evoked EJPs even in the presence of 1 microM omega-CgTX GVIA. We have termed this omega-CgTX GVIA-resistant release 'residual release'. EJP amplitude in the presence of omega-CgTX GVIA depended on both the frequency and number of stimuli in a train. 3. Residual release was inhibited by the P-type calcium channel blocker omega-agatoxin IVA (100 nM). However, even in the presence of both toxins, longer trains of stimuli could still evoke neurotransmitter release. 4. Residual release was abolished by omega-conotoxin MVIIC and by the non-specific calcium channel antagonist omega-grammotoxin SIA. Therefore, it would appear that a heterogeneous population of calcium channels is involved in mediating neurotransmitter release from these sympathetic nerve terminals.