Comparison of the intracellular effects of clostridial neurotoxins on exocytosis from streptolysin O-permeabilized rat pheochromocytoma (PC 12) and bovine adrenal chromaffin cells

Abstract

The inhibitory effects of tetanus toxin, botulinum toxin A, their constituent light chains, and botulinum toxin B were compared using streptolysin O-permeabilized rat pheochromocytoma (PC 12) and bovine adrenal chromaffin cells in primary culture. In both types of chromaffin cells exocytosis can be triggered by micromolar amounts of free Ca2+, bovine adrenal chromaffin cells in addition require ATP. In PC 12 cells the isolated tetanus toxin light chain alone blocks exocytosis without any additive. The time-course of the inhibitory action of tetanus toxin light chain in permeabilized PC 12 cells in the absence of ATP is similar to the one obtained with permeabilized bovine adrenal chromaffin cells, in the presence of ATP. Thus, ATP does not seem to be crucial for tetanus toxin (two-chain form) poisoning. Botulinum toxin B (two-chain form), if preactivated by dithiothreitol, also inhibits exocytosis from permeabilized PC 12 cells up to 90% in the absence of ATP. By contrast, botulinum toxin A (two-chain form) or its isolated light chain, which are highly potent in permeabilized bovine adrenal chromaffin cells, causes only a weak inhibition in PC 12 cells. In streptolysin O-permeabilized bovine adrenal chromaffin cells omission of ATP during the incubation with the toxin increases the potency of botulinum toxin A light chain. Under the same conditions the effect of tetanus toxin light chain remains unchanged. Tetanus toxin and botulinum toxin B (two-chain forms) probably block a step which occurs during exocytosis from both PC 12 cells and adrenal chromaffin cells and which could be closely related to the final fusion event.(ABSTRACT TRUNCATED AT 250 WORDS)