Inhibition of exocytosis in bovine adrenal medullary cells by botulinum toxin type D

Abstract

Botulinum toxins are known to block transmitter release at peripheral cholinergic synapses¹, producing muscular weakness and paralysis. The toxins may also block adrenergic transmission^2–5, although this effect is less well understood¹. The mechanisms by which toxins act are unclear. They are proteins of relative molecular mass ∼150,000 and are structurally similar to tetanus toxin. It is generally accepted that a rise in intracellular calcium concentration is sufficient to trigger secretion by exocytosis^6–13, but it is not known whether the toxins block secretion by preventing this Ca transient^14,15 or whether they act downstream from Ca entry by interfering with the process of exocytosis itself. We have attempted to resolve these questions in the case of the adrenergic system by studying the effects of botulinum toxins (types A, B, D and E) on the secretory response of isolated bovine adrenal medullary cells maintained in culture¹⁶. The cells were either challenged with various secretagogues or rendered leaky and challenged directly with Ca buffers. We report here that botulinum toxin type D inhibits secretion in a time- and dose-dependent manner, the results being entirely consistent with the idea that the toxin acts at or near the site of exocytosis rather than at the sites controlling the rise in free Ca.