Primary nerve cell cultures from the brainstem of embryonic mice take up [3H]noradrenaline. Release can be evoked by high K+ or sea anemone toxin II and depends on Ca2+. The cultures allow neurochemical studies on the long-term actions of clostridial neurotoxins. Tetanus and botulinum A and C neurotoxins partially inhibit the absolute and fractional release evoked by high K+, as well as the fractional basal release. The detection limit for the toxins is below 5 pM. Total radioactivity is higher in the poisoned cultures, although the initial velocity of uptake is not measurably influenced by tetanus or botulinum A toxin. Pretreatment with neuraminidase prevents the effects of botulinum A toxin and diminishes those of botulinum C and tetanus toxins. Within 6 days, the cultures partially recover from tetanus toxin poisoning. Antitoxin prevents the actions of the toxin, but only slightly promotes recovery. The data indicate close pharmacological analogies between the clostridial neurotoxins.
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