1. The effects of the polyamines, spermine and spermidine on neuronal N-type voltage-sensitive calcium channels were investigated using the binding and function of the ligand omega-conotoxin GVIA (omega-CT). 2. Spermine and spermidine enhanced (EC50 approximately 0.16 and 0.45 microM) and, at higher concentrations, inhibited (IC50 of 9 and 240 microM) the binding of [125I]omega-CT to rat hippocampal synaptosomes. 3. Spermine and, less potently, spermidine inhibited the neurotransmitter-mediated, omega-CT-sensitive, electrical-field-stimulated contractile responses of the rat vas deferens. 4. The polyamines also inhibited the phenylephrine-evoked contractile responses of the vas deferens with the same rank order, consistent with a postsynaptic mechanism of inhibition. 5. However, pre-exposure to spermine prevented the irreversible inhibition of vas deferens twitch responses by omega-CT (previously found to be presynaptic). The prevention of inhibition by omega-CT demonstrates that the neuronal binding of spermine and omega-CT is mutually exclusive. Thus spermine (and presumably spermidine at higher concentrations) appears to modulate the actions of omega-CT at N-type voltage-sensitive calcium channels.