Evidence for calcium-dependent vesicular transmitter release insensitive to tetanus toxin and botulinum toxin type F

Whether exocytosis evoked by a given releasing stimulus from different neuronal families or by different stimuli from one neuronal population occurs through identical mechanisms is unknown. We studied the release of 3H]noradrenaline, 3H]acetylcholine and 3H]dopamine induced by different stimuli from superfused rat brain synaptosomes pretreated with tetanus toxin or botulinum toxin F, known to block exocytosis by cleaving VAMP/synaptobrevin. The external Ca2(+)-dependent 3H]transmitter overflows evoked by KCl were similarly inhibited by tetanus toxin or botulinum toxin F; the toxins cleaved similar amounts of synaptosomal synaptobrevin, as determined by western blot analysis, suggesting prevalent involvement of synaptobrevin-II. GABA uptake-mediated release of the three 3H]transmitters was that differentially sensitive to the toxins: only the release of 3H]noradrenaline, which is dependent on external Ca2+, but not of 3H]acetylcholine and 3H]dopamine was blocked. Neither toxin affected the 3H]transmitter overflows evoked by the Ca2(+) ionophore ionomycin. Cadmium blocked the K(+)-evoked release of all 3H]transmitters and the GABA-evoked release of 3H]noradrenaline; the GABA-evoked releases of 3H]acetylcholine and 3H]dopamine and those elicited by ionomycin were insensitive to cadmium. The results suggest that tetanus toxin and botulinum toxin F selectively affect exocytosis linked to activation of voltage-sensitive Ca2(+) channels; the Ca2(+)-dependent, exocytotic-like release induced by stimuli not leading to activation of voltage-sensitive Ca2+ channels seems insensitive to these clostridial toxins.