Abstract
(1) We investigated the effects of single- and double-poisoning with tetanus toxin (TeTx), botulinum neurotoxin type A (BoTx A) and botulinum neurotoxin type B (BoTx B) on spontaneous and nerve-evoked quantal transmitter release at motor endplates of the triangularis sterni preparation of the mouse. (2) Inhibitory effects of TeTx and BoTx B on spontaneous and nerve-evoked transmitter release were very similar, except that the action of BoTx B required 500-fold lower concentrations and was less dependent on temperature. BoTx A caused stronger inhibition of quantal release than TeTx or BoTx B, but was comparatively much easier counteracted by 4-aminopyridine (4-AP). (3) In contrast to BoTx A, with TeTx or BoTx B the increase of transmitter release following onset of 50 Hz nerve stimulation was delayed for a few seconds and synaptic latencies of quanta showed large variations. This release pattern was also evident in all double-poisoning experiments, regardless of intoxication sequence. (4) Inhibition of evoked release was found to be slightly stronger with TeTx than with BoTx B, so the amount of nerve-evoked quanta released after double-poisoning with any sequence of these toxins always approached that of TeTx. In no case supraadditive actions were observed. (5) A strong reduction of evoked quanta was observed when BoTx A was applied in addition to either of the two other toxins. With reversed poisoning sequences (BoTx A-TeTx or BoTx A-BoTx B) the resulting values remained at the extremely low level of BoTx A. (6) In the presence of 4-AP double-poisoning with any combination between BoTx A and TeTx or BoTx B (regardless of intoxication sequence) revealed supra-additive effects, since the number of quanta released was considerably lower than that obtained with any of the toxins alone (in the presence of 4-AP). (7) Our results indicate that tetanus toxin and botulinum toxin type B have a common site of action which is different and independent from that of botulinum toxin type A.
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References
Borochov-Neori H, Yavin E, Montal M (1984) Tetanus toxin forms channels in planar lipid bilayers containing gangliosides. Biophys J 45:83–85
Cull-Candy SG, Lundh H, Thesleff S (1976) Effects of botulinum toxin on neuromuscular transmission in the rat. J Physiol 260:177–203
Dreyer F, Schmitt A (1981) Different effects of botulinum A toxin and tetanus toxin on the transmitter releasing process at the mammalian neuromuscular junction. Neurosci Lett 26:307–311
Dreyer F, Mallart A, Brigant JL (1983) Botulinum A toxin and tetanus toxin do not effect presynaptic membrane currents in mammalian motor nerve endings. Brain Res 270:373–375
Dreyer F, Becker C, Bigalke H, Funk J, Penner R, Rosenberg F, Ziegler M (1984) Action of botulinum A toxin and tetanus toxin on synaptic transmission. J Physiol (Paris) 79:252–258
Dreyer F, Rosenberg F, Becker C, Bigalke H, Penner R (1986) Differential effects of various secretagogues on quantal transmitter release from mouse motor nerve terminals treated with botulinum A and tetanus toxin. Naunyn-Schmiedeberg's Arch Pharmacol 335:1–7
Duchen LW, Tonge DA (1973) The effects of tetanus toxin on neuromuscular transmission and on the morphology of motor end-plates in slow and fast skeletal muscle of the mouse. J Physiol 228:157–172
Grasso A, Alemá S, Rufini S, Senni MI (1980) Black widow spider toxin-induced calcium fluxes and transmitter release in a neurosecretory cell line. Nature 283:774–776
Gundersen CB, Katz B, Miledi R (1982) The antagonism between botulinum toxin and calcium in motor nerve terminals. Proc Roy Soc B 216:369–376
Habermann E, Dreyer F (1986) Clostridial neurotoxins: Handling and action at the cellular and molecular level. Curr Top Microbiol Immunol 129:93–179
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
Harris AJ, Miledi R (1971) The effect of type D botulinum toxin on frog neuromuscular junctions. J Physiol 217:497–515
Hoch DH, Romero-Mira M, Ehrlich BE, Finkelstein A, DasGupta BR, Simpson LL (1985) Channels formed by botulinum, tetanus and diphtheria toxins in planar lipid bilayers: Relevance to translocation of proteins across membranes. Proc Natl Acad Sci USA 82:1692–1696
Kao I, Drachman DB, Price DL (1976) Botulinum toxin: Mechanism of presynaptic blockade. Science 193:1256–1258
Kryzhanovsky GN (1973) The mechanism of action of tetanus toxin: Effect on synaptic processes and some particular features of toxin binding by the nervous tissue. Naunyn-Schmiedeberg's Arch Pharmacol 276:247–270
Lundh H, Leander S, Thesleff S (1977) Antagonism of the paralysis produced by botulinum toxin in the rat. Neurol Sci 32:29–43
McArdle JJ, Angaut-Petit D, Mallart A, Bournaud R, Faille L, Brigant JL (1981) Advantages of the triangularis sterni muscle of the mouse for investigations of synaptic phenomena. Neurosci Lett 4:109–115
Mellanby J (1984) Comparative activities of tetanus and botulinum toxins. Neurosci 11:29–34
Moberg LJ, Sugiyama H (1978) Affinity chromatography purification of type A botulinum neurotoxin from crystalline toxic complex. Appl Environ Microbiol 35:878–880
Molgo J, Thesleff S (1984) Studies on the mode of action of botulinum toxin type A at the frog neuromuscular junction. Brain Res 297:309–316
Nicholls DG, Rugolo M, Scott IG, Meldolesi J (1982) Latrotoxin of black widow spider venom depolarizes the plasma membrane, induces massive calcium influx, and stimulates transmitter release in guinea pig brain synaptosomes. Proc Natl Acad Sci USA 79:7924–7928
Penner R, Neher E, Dreyer F (1986) Intracellularly injected tetanus toxin inhibits exocytosis in bovine adrenal chromaffin cells. Nature 324:76–78
Schmitt A, Dreyer F, John Chr (1981) At least three sequential steps are involved in the tetanus toxin-induced block of neuromuscular transmission. Naunyn-Schmiedeberg's Arch Pharmacol 317:326–330
Sellin LC, Kauffman JA, Way JF, Siegel LS (1983a) Comparison of the action of types A and F botulinum toxin at the rat neuromuscular junction. Soc Neurosci Abstr 9
Sellin LC, Thesleff S, DasGupta BR (1983b) Different effects of types A and B botulinum toxin on transmitter release at the rat neuromuscular junction. Acta Physiol Scand 119:127–133
Simpson LL (1986) Molecular pharmacology of botulinum toxin and tetanus toxin. Annu Rev Pharmacol Toxicol 26:427–453
Spitzer N (1972) Miniature end-plate potentials at mammalian neuromuscular junctions poisoned by botulinum toxin. Nature 237:26–27
Sugiyama H (1980) Clostridium botulinum neurotoxin. Microbiol Rev 44:419–448
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This is part of the thesis of M. G. to be presented to the Fachbereich Humanmedizin, Justus-Liebig-Universität Gießen
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Gansel, M., Penner, R. & Dreyer, F. Distinct sites of action of clostridial neurotoxins revealed by double-poisoning of mouse motor nerve terminals. Pflugers Arch. 409, 533–539 (1987). https://doi.org/10.1007/BF00583812
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DOI: https://doi.org/10.1007/BF00583812