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Distinct targets for tetanus and botulinum A neurotoxins within the signal transducing pathway in chromaffin cells

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Summary

Tetanus and botulinum A neurotoxins inhibited exocytosis evolved by various secretagogues in intact and permeabilized chromaffin cells. The block of exocytosis in intact chromaffm cells due to botulinum A neurotoxin could partially be overcome by enhancing nicotine- and veratridine-induced stimulation, whereas the block due to tetanus toxin persisted under the same conditions. The receptor-mediated restoration of 3H-noradrenaline release was specific for nicotinic stimulation, because exocytosis did not occur during muscarinic stimulation. Depolarization of intact chromaffin cells with increasing concentration of K+ failed to restore exocytosis that had been blocked by either toxin.

When chromaffin cells, treated with tetanus or botulinum A neurotoxins, were exposed to the Ca2+-ionophore A 23187 or permeabilized by staphylococcal α-toxin, Ca2+-stimulated exocytosis was also inhibited. The inhibition was unaffected by increasing concentrations of free Ca2+. Activation of proteinkinase C and of G-proteins by phorbolester and GMPPNHP, respectively, increased Ca2+-induced exocytosis in control cells as well as in cells treated with tetanus and botulinum A neurotoxins. The block, however, could not be relieved by these manipulations, and it could not be relieved by activating the cGMP or cAMP pathways with analoga of cyclic nucleotides, phosphodiesterases inhibitors, and forskolin either.

It is concluded that nicotine and veratridine trigger a mechanism within the sequence of events leading to exocytosis that is located beyond the increase in intracellular Ca2+-concentration. This pathway may not be affected by botulinum A neurotoxin. The target of tetanus toxin is probably located even closer to the fusion process, i. e. beyond the step upon which botulinum A neurotoxin acts.

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References

  • Ahnert-Hilger G, Glossmann H, Habermann E (1979) Effects of ion channel toxins on the cyclic nucleotide content of cerebellar slices, primary brain cultures and neuronal cell lines. Naunyn-Schmiedeberg's Arch Pharmacol 307:151 -157

    Google Scholar 

  • Ahnert-Hilger G, Mach W, Föhr KJ, Gratzl M (1989a) Poration by alpha-toxin and streptolysin O: An approch to analyze intracellular processes. Methods Cell Biol 31:63–90

    Google Scholar 

  • Ahnert-Hilger G, Weller U, Dauzenroth ME, Habermann E, Gratzl M (1989b) The tetanus toxin light chain inhibits exocytosis. FEBS Lett 242:245–248

    Google Scholar 

  • Ashton AC, Dolly JO (1991) Microtubule-dissociating drugs and A 23187 reveal differences in the inhibition of synaptosomal transmitter release by botulinum neurotoxins types A and B. J Neurochem 56:827–835

    Google Scholar 

  • Aunis D, Bader M (1988) The cytoskeleton as a barrier to exocytosis. J Exp Biol 139:253–266

    Google Scholar 

  • Bader M-F, Sontag J-M, Thierse D, Aunis D (1989) A reassessment of guanine nucleotide effects on catecholamine secretion from permeabilized chromaffin cells. J Biol Chem 264:16426–16434

    Google Scholar 

  • Bartels F, Marxen P, Bigalke H (1990) Translocation of tetanus toxin into intact and permeabilized chromaffin cells: a comparison. Biol Chem Hoppe-Seyler S371:1035–1036

    Google Scholar 

  • Bergey GK, Bigalke H, Nelson P (1987) Differential effects of tetanus toxin on inhibitory and excitatory synpatic transmission in mammalian spinal cord neurons in culture: A presynaptic locus of action. J Neurophysiol 57:121–131

    Google Scholar 

  • Bigalke H, Ahnert-Hilger G, Habermann E (1981 a) 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

    Google Scholar 

  • Bigalke H, Heller I, Bizzini B, Habermann E (1981 b) Tetanus and botulinum A toxin inhibit release and uptake of various transmitters, as studied with particulate preparations from rat brain and spinal cord. Naunyn-Schmiedeberg's Arch Pharmacol 316:244–251

    Google Scholar 

  • Bigalke H, Dreyer F, Bergey G (1985) Botulinum A neurotoxin inhibits non-cholinergic transmission in mouse spinal cord neurons in culture. Brain Res 360:318–324

    Google Scholar 

  • Bigalke H, Müller H, Dreyer F (1986) Botulinum A neurotoxin unlike tetanus toxin acts via a neuraminidase sensitive structure. Toxicon 24:1065–1074

    Google Scholar 

  • Bigalke H, Marxen P, Ahnert-Hilger G (1989) Restoration of noradrenaline release by increasing concentrations of carbachol in botulinum A neurotoxin blocked chromaffin cells in culture. Biol Chem Hoppe-Seyler S370:993

    Google Scholar 

  • Binz T, Eisel U, Kurazono H, Binscheck T, Bigalke H, Niemann H (1990) Tetanus and botulinum A toxins: Comparison of sequences and microinjection of tetanus toxin A subunit-specific mRNA into bovine chromaffin cells. In: Rappuoli R, Alouf JE, Falmagne P, Fehrenbach FJ, Gross J, Jeljaszewicz J, Montecucco C, Tomasi M, Wadstr6m T, Witholt B (eds) Bacterial protein toxins. ZbI Bakt [Suppl] 19:105–108

  • Bittner MA, Holz RW (1988) Effects of tetanus toxin on catecholamine release from intact and digitonin permeabilized chromaffin cells. J Neurochem 51:451–456

    Google Scholar 

  • Bittner MA, Holz RW, Neubig RR (1986) Guanine nucleotide effects on catecholamine secretion from digitonin-permeabilized chromaffin cells. J Biol Chem 261:10182–10188

    Google Scholar 

  • Bittner MA, DasGupta BR, Holz RW (1989) Isolated light chains of botulinum neurotoxins inhibit exocytosis. J Biol Chem 264:10354–10360

    Google Scholar 

  • Burgoyne RD, Morgan A, O'Sullivan AJ (1988) A major role for protein kinase C in calcium-activated exocytosis in permeabilized adrenal chromaffin cells. FEBS Lett 238:151–155

    Google Scholar 

  • Dreyer F (1989) Peripheral actions of tetanus toxin. In: Simpson LL (ed) Botulinum neurotoxin and tetanus toxin. Academic Press, London, pp179–202

    Google Scholar 

  • 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

    Google Scholar 

  • Dreyer F, Rosenberg F, Becker C, Bigalke H, Penner R (1987) Differential effects of various secretagogues on quantal transmitter release from mouse nerve terminals treated with botulinum A and tetanus toxin. Naunyn-Schmiedeberg's Arch Pharmacol 335:1–7

    Google Scholar 

  • Föhr KJ, Warchol W, Gratzl M (1991) Calculation and control of the divalent cations in solutions used for membrane fusion studies. In: Duzdiingers N (ed) Methods in enzymology: techniques in membrane fusion. Academic Press, London, in press

    Google Scholar 

  • Gansel M, Penner R, Dreyer F (1987) Distinct sites of action of clostridial neurotoxins revealed by double-poisoning of mouse motor nerve terminals. Pflüger's Arch 409:533–539

    Google Scholar 

  • Habermann E (1989) Clostridial neurotoxins and the central nervous system: Functional studies on isolated preparations. In: Simpson LL (ed) Botulinum neurotoxin and tetanus toxin. Academic Press, London, pp 255–279

    Google Scholar 

  • Halpern JL, Habig WH, Trenchard H, Russel JT (1990) Effect of tetanus toxin on oxytocin and vasopressin release from nerve endings of the neurohypophysis. J Neurochem 55:2072–2078

    Google Scholar 

  • Knight DE (1986) Botulinum toxin type A, B and D inhibit catecholamine secretion from bovine adrenal medullary cells. FEBS Lett 207:222–229

    Google Scholar 

  • Knight DE, Baker PF (1985) Guanine nucleotides and Ca2+-dependent exocytosis. Studies on two adrenal cell preparations. FEBS Lett 189:345–349

    Google Scholar 

  • Knight DE, Sudgen D, Baker PF (1988) Evidence implicating protein kinase C in exocytosis from electropermeabilized bovine chromaffin cells. J Membr Biol 104:21–34

    Google Scholar 

  • Lind I, Ahnert-Hilger G, Fuchs G, Gratzl M (1987) Purification of α-toxin from Staphylococcus aureus and application to cell permeabilization. Anal Biochem 164:84–89

    Google Scholar 

  • Marley PD (1988) Desensitization of the nicotinic secretory response of adrenal chromaffin cells. TIPS 9:102–107

    Google Scholar 

  • Marxen P, Bigalke H (1989) Tetanus toxin: Inhibitory action in chromaffin cells is initiated by specified types of gangliosides and promoted in low ionic strength solution. Neurosci Lett 107:261–266

    Google Scholar 

  • Marxen P, Bigalke H (1991) Tetanus and botulinum A toxins inhibit stimulated F-actin rearrangement in chromaffin cells. Neuroreport 2:33–37

    Google Scholar 

  • Marxen P, Fuhrmann U, Bigalke H (1989) Gangliosides mediate inhibitory effects of tetanus and botulinum A neurotoxin on exocytosis in chromaffin cells. Toxicon 27:849–859

    Google Scholar 

  • Marxen P, Ahnert-Hilger G, Wellhöner HH, Bigalke H (1990) Tetanus antitoxin binds to intracellular tetanus toxin in permeabilized chromaffin cells without restoring Ca2+-induced exocytosis. Toxicon 28:1077–1082

    Google Scholar 

  • Marxen P, Erdmann G, Bigalke H (1991) The translocation of botulinum A neurotoxin by chromaffin cells is promoted in low ionic strength solution and is insensitive to trypsin. Toxicon 29:181–189

    Google Scholar 

  • Nakov R, Habermann E, Hertting G, Wurster S, Allgaier C (1989) Effects of botulinum A toxin on presynaptic modulation of evoked transmitter release. Eur J Pharmacol 164:45–53

    Google Scholar 

  • Pocotte SL, Frye RA, Senter DR, TerBush DR, Lee SA, Holz RW (1985) Effects of phorbol ester on catecholamine secretion and protein phosphorylation in adrenal chromaffin cells. Proc Natl Acad Sci USA 82:930–934

    Google Scholar 

  • Penner R, Neher E, Dreyer F (1986) Intracellularly injected tetanus toxin inhibits exocytosis in bovine adrenal chromaffin cells. Nature 324:76–78

    Google Scholar 

  • Sandberg K, Berry CJ, Eugster E, Rogers TB (1989) A role of cGMP during tetanus toxin blockade of acetylcholine release in the rat pheochromocytoma (PC 12) cell line. J Neurosci 9:3946–3954

    Google Scholar 

  • Sasakawa N, Nakaki T, Yamamoto S, Kato R (1987) Inositol triphosphate accumulation by high potassium stimulation in cultured adrenal chromaffin cells. FEBS Lett 223:413–416

    Google Scholar 

  • Sontag JM, Aunis D, Bader M-F (1988) Peripheral actinfilaments control calcium-mediated catecholamine release from streptolysin-O-permeabilized chromaffin cells. J Cell Biol 46:316–326

    Google Scholar 

  • Stecher B, Gratzl M, Ahnert-Hilger G (1989a) Reductive chain separation of botulinum A toxin a prerequisite to its inhibitory action on exocytosis in chromaffin cells. FEBS Lett 248:23–27

    Google Scholar 

  • Stecher B, Weller U, Habermann E, Gratzl M, Ahnert-Hilger G (1989b) The light chain but not the heavy chain of botulinum A toxin inhibits exocytosis from permeabilized adrenal chromaffin cells. FEBS Lett 255:391–394

    Google Scholar 

  • Stoer SJ, Smolen JE, Holz RW, Agranoff BW (1986) Inositol triphosphate mobilizes intracellular calcium in permeabilized adrenal chromaffin cells. J Neurochem 46:637–640

    Google Scholar 

  • Stryer C, Bourne H (1986) G-proteins: a family of signal transducers. Ann Rev Cell Biol 2:391–419

    Google Scholar 

  • TerBush D, Holz RW (1986) Effects of phorbolesthers, diglyceride, and cholinergic agonists on the subcellular distribution of protein kinase C in intact or digitonin-permeabilized adrenal chromaffin cells. J Biol Chem 261:17099–17106

    Google Scholar 

  • Vesterberg O, Wadstrom T, Vesterberg K, Svennson H, Malmgren B (1967) Studies on extracellular proteins from Staphylococcus aureus. I. Separation and characterization of enzymes and toxins by isoelectric focusing. Biochim Biophys Acta 133:435–445

    Google Scholar 

  • Wellhöner HH, Neville D (1987) Tetanus toxin binds with high affinity to neuroblastoma × glioma NG 108–15 and impairs their stimulated acetylcholine release. J Biol Chem 292:17374–17378

    Google Scholar 

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Authors and Affiliations

  1. Institut für Peptidforschung, W-3000, 61, Hannover, Federal Republic of Germany

    P. Marxen

  2. Medizinische Hochschule Hannover, Zentrum Pharmakologie und Toxikologie, W-3000, 61, Hannover, Federal Republic of Germany

    F. Bartels & H. Bigalke

  3. Institut für Neuropsychopharmakologie der Freien Universität Berlin, Ulmenallee 30, W-1000, 19, Berlin, Federal Republic of Germany

    G. Ahnert-Hilger

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  1. P. Marxen
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  2. F. Bartels
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  3. G. Ahnert-Hilger
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  4. H. Bigalke
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Marxen, P., Bartels, F., Ahnert-Hilger, G. et al. Distinct targets for tetanus and botulinum A neurotoxins within the signal transducing pathway in chromaffin cells. Naunyn-Schmiedeberg's Arch Pharmacol 344, 387–395 (1991). https://doi.org/10.1007/BF00172577

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  • DOI: https://doi.org/10.1007/BF00172577

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