Neuropharmacology
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The recent development of peptide antagonists that selectively block subtypes of neuronal calcium channel has provided tools to study the role of presynaptic calcium channels in triggering exocytosis. A variety of methods have consistently demonstrated that multiple channel types participate in exocytosis. We have studied the subsecond kinetics of [3H]glutamate release from rat cortical synaptosomes as an assay for presynaptic calcium channel activity. ⋯ These results suggest that the N type channel in nerve terminals is distinct from that found in hippocampal somata, since it appears to be resistant to by omega-conotoxin MVIIC. The combination of omega-conotoxin GVIA (100 nM) and either omega-agatoxin IVA or omega-conotoxin MVIIC (1 microM each) blocked approx 90% of release when the Ca2+ concentration was reduced (0.46 mM or less), but 30-40% of release remained when the concentration of Ca2+ in the stimulus buffer was 1 mM or greater, indicating that a resistant channel type(s) also participates in exocytosis. Specific inhibitors of this resistant phenotype will be useful for further refinement of our understanding of the role of presynaptic calcium channels in mediating neurosecretion.