Summary
Slices or particles from rat forebrain cortex were preloaded with [3H]choline, and the release of [3H]acetylcholine was evoked with potassium ions in a superfusion system. Release depended on the presence of calcium.
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1.
Incubation of the preloaded tissue preparation for 2 h with tetanus or botulinum A toxin did not change the [3H]acetylcholine content or the ratio [3H]acetylcholine/[3H]choline. Tetanus toxin diminished, dependent on dose and time, the release of [3H]acetylcholine evoked by 25 mM K+. It was about ten times more potent than botulinum A toxin. The effect of botulinum toxin was due to its neurotoxin content. Raising the potassium concentration partially overcame the inhibition by the toxins. Hemicholinium-3, applied to preloaded slices, left the subsequent [3H]acetylcholine release unchanged.
Pretreatment of particles with neuraminidase diminished the content of long-chain gangliosides to the detection limit. Such particles remained fully sensitive to tetanus toxin, and at least partially sensitive to botulinum A toxin.
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2.
The potassium or sea anemone toxin II stimulated uptake of 45Ca2+ into cortex synaptosomes or particles was not inhibited by either toxin. Both toxins appear to impede the Ca2+-dependent mobilization of an easily releasable acetylcholine pool, without inhibiting the transmembranal calcium fluxes.
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Ahnert G, Glossmann H, Habermann E (1979) Investigations on the mechanism of cyclic guanosine monophosphate increase due to depolarizing agents as studied with sea anemone toxin II in mouse cerebellar slices. Naunyn-Schmiedeberg's Arch Pharmacol 307:159–166
Bergmann C, Dubois JM, Rojas E, Rathmayer W (1976) Decreased rate of sodium conductance inactivation in the node of Ranivier induced by a polypeptide toxin from sea anemone. Biochim Biophys Acta 455:173–184
Bigalke H, Dimpfel W (1978) Kinetics of 3H-acetylcholine synthesis and release in primary cell cultures from mammalian CNS. J Neurochem 30:871–879
Bigalke H, Habermann E (1980a) Blockade by tetanus and botulinum A toxin of postganglionic cholinergic nerve endings in the myenteric plexus. Naunyn-Schmiedeberg's Arch Pharmacol 312:255–263
Bigalke H, Habermann E (1980b) Inhibition of 3H-choline uptake into and 3H-acetylcholine release from brain tissue by tetanus and botulinum toxin. In: Brzin M, Sket D, Kardelj E, Bachelard H (eds) Synaptic constituents in health and disease. Mladinska knjiga Ljubljana, Pergamon Press, London, p 137
Bigalke H, Dimpfel W, Habermann E (1978) Suppression of 3H-acetylcholine release from primary nerve cell cultures by tetanus and botulinum A toxin. Naunyn-Schmiedeberg's Arch Pharmacol 303:133–138
Blaustein MP (1975) Effects of potassium, veratridine and scorpion venom on calcium accumulation and transmitter release by nerve terminals in vitro. J Physiol 247:617–655
Burgen ASV, Dickens F, Zatman LJ (1949) The action of botulinum toxin on the neuromuscular junction. J Physiol (London) 109:10–24
Collingridge Gl, Collins GGS, Davies J, James TA, Neal MJ, Tongroach P (1980) Effect of tetanus toxin on transmitter release from the substantia nigra and striatum in vitro. J Neurochem 34:540–547
Cull-Candy SG, Lundh H, Thesleff S (1976) Effects of botulinum toxin on neuromuscular transmission in the rat. J Physiol (London) 260:177–203
Gundersen CB, Howard BD (1978) The effects of botulinum toxin on acetylcholine metabolism in mouse brain slices and synaptosomes. J Neurochem 31:1005–1013
Habermann E, Bigalke H, Heller I (1981) Inhibition of choline uptake by tetanus and botulinum A toxin. Partial dissociation of fixation and effect of tetanus toxin. Naunyn-Schmiedeberg's Arch Pharmacol 316:135–142
Habermann E, Dreyer F, Bigalke H (1980) Tetanus toxin blocks the neuromuscular transmission in vitro like botulinum A toxin. Naunyn-Schmiedeberg's Arch Pharmacol 311:33–40
Haga T, Noda H (1973) Choline uptake systems of rat brain synaptosomes. Biochim Biophys Acta 291:564–575
Hagenah R, Benecke R, Wiegand H (1977) Effects of type A botulinum toxin on the cholinergic transmission at spinal Renshaw cells and on the inhibitory action at Ia inhibitory interneurones. Naunyn-Schmiedeberg's Arch Pharmacol 299:267–272
Israel M, Dunant J, Manaranche R (1979) The present status of the vesicular hypothesis. Prog Neurobiol 13:237–275
Kessler PD, Marchbanks RM (1979) Choline transport is not coupled to acetylcholine synthesis. Nature 279:542–544
Leander S, Thesleff S (1980) On the mode of action of botulinum toxin. Acta Physiol Scand 108:195–196
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin-phenol reagent. J Biol Chem 193:265–275
Marchbanks RM, Wonnacott S (1979) Relationship of choline uptake to acetylcholine synthesis and release. Prog Brain Res 49:77–88
Moberg LJ, Sugiyama H (1978) Affinity chromatography purification of type A botulinum neurotoxin from crystalline toxic complex. Appl Environ Microbiol 35:878–880
Mulder AH, van den Berg WB, Stoff JC (1975) Calcium-dependent release of radiolabeled catecholamines and serotonin from rat brain synaptosomes in a superfusion system. Brain Res 99:419–424
Stevenson JW (1951) Studies of the effect of botulinum toxin upon the bacterial acetylation of choline. Can J Public Health 42:68
Tazieff-Depierre F, Métézeau P (1977) Action du venin de scorpion (Androctonus australis) sur la transmission neuromusculaire de grenouille inhibiée par la toxine botulique. CR Acad Sci (Paris) 285:1335–1338
Tazieff-Depierre F, Métézeau P, Wunderer G (1978) Action de la toxine II isolée des tentacules d'anhuemone de mer sur la transmission neuromusculaire de grenouille normale et inhibiée par la toxine botulique. CR Acad Sci (Paris) 286:655–658
Tonge DA, Gradidge TJ, Marchbanks RM (1975) Effects of botulinum and tetanus toxins on choline acetyltransferase activity in skeletal muscles in the mouse. J Neurochem 25:329–331
Wonnacott S (1980) Inhibition by botulinum toxin of acetylcholine release from synaptosomes: Latency of action and the role of gangliosides. J Neurochem 34:1567–1573
Wonnacott S, Marchbanks RM (1976) Inhibition by botulinum toxin of depolarization evoked release of 14C-acetylcholine from synaptosomes in vitro. Biochem J 156:701–712
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Bigalke, H., Ahnert-Hilger, G. & Habermann, E. Tetanus toxin and botulinum a toxin inhibit acetylcholine release from but not calcium uptake into brain tissue. Naunyn-Schmiedeberg's Arch. Pharmacol. 316, 143–148 (1981). https://doi.org/10.1007/BF00505308
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DOI: https://doi.org/10.1007/BF00505308