Abstract
Rat cortex synaptosomes have been successfully perforated with high concentrations (≥400 U/ml) of Staphylococcus aureus α-toxin. The free Ca 2+-concentration dependence of [ 3H]-noradrenaline release was similar to that observed for PC 12 and chromaffin cells. Release from the α-toxin perforated synaptosomes was not significantly inhibited by ω-conotoxin GVIA. Initially, Ca 2+-dependent release was independent of MgATP (for 0.5 min), but became increasingly dependent on MgATP with time. Lactate dehydrogenase efflux from α-toxin-perforated synaptosomes was not different than efflux from control synaptosomes, and an antibody to N-ethylmaleimide-sensitive fusion protein did not enter the synaptosomes. [ 3H]-noradrenaline release was temperature and α-toxin-concentration dependent. Ca 2+-dependent release was more resistant to rundown from α-toxin- than from streptolysin-O-perforated synaptosomes. This preparation of perforated synaptosomes should be useful for studies of regulated exocytosis from nerve endings.
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