Skip to main content
SpringerLink
Log in
Book cover

Bacterial Toxins: Methods and Protocols pp 259–286Cite as

In Vitro Physiological Studies on Clostridial Neurotoxins

Biological Models and Procedures for Extracellular and Intracellular Application of Toxins

  • Protocol

  • 1396 Accesses

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 145))

Abstract

Botulinum (BoNT, serotypes A-G, see also Chapter 2) and tetanus (TeNT) neurotoxins are known under the generic term of clostridial neurotoxins. These dichainal proteins comprise a light (Mr R~50) and a heavy (Mr R~100) chain that are disulfide linked. In mammals, these proteins are the causative agents of two severe neuroparalytic diseases, botulism and tetanus. Botulism manifests as a flaccid muscle paralysis caused by a near irreversible and selective inhibition of acetylcholine release at the skeletal neuromuscular junction. Tetanus is characterized by a spastic neuromuscular paralysis that results from motoneuron disinhibition following the specific blockage of inhibitory glycinergic or Γ-aminobutyric acid-ergic (GABAergic) synapses by TeNT in the central nervous system (CNS). The cellular action of BoNT and TeNT can be depicted according to several steps. After binding to specific acceptors located at the nerve ending membrane, TeNT and BoNT are endocytosed. Subsequently, their active moiety (the light chain) is translocated from the endocytic compartment into the cytosol. Here, it cleaves one among three synaptic proteins (viz. the vesicle-associated membrane protein [VAMP]/synaptobrevin, syntaxin, and synaptosomal-associated protein of Mr 25 kDa [SNAP-25] which are also known under the collective term of SNAREs) involved in docking and fusion of synaptic vesicles at the active zone (i.e., release site) (for reviews see [1,2]).

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Niemann H., Blasi J., and Jahn R. (1994) Clostridial neurotoxins: new tools for dissecting exocytosis. Trends Cell Biol. 4, 179–185.

    Article  PubMed  CAS  Google Scholar 

  2. Montecucco C. and Schiavo G. (1995) Structure and function of tetanus and botulinum neurotoxins. Q. Rev. Biophys. 28, 423–472.

    Article  PubMed  CAS  Google Scholar 

  3. Schmitt A., Dreyer F., and John C. (1981) At least three sequential steps are involved in the tetanus toxin-induced block of neuromuscular transmission. Naunyn Schmiedebergs Arch. Pharmacol. 317, 326–330.

    Article  PubMed  CAS  Google Scholar 

  4. Rabasseda X., Blasi J., Marsal J., Dunant Y., Casanova A., and Bizzini B. (1988) Tetanus and botulinum toxins block the release of acetylcholine from slices of rat striatum and from the isolated electric organ of Torpedo at different concentrations. Toxicon 26, 329–336.

    Article  PubMed  CAS  Google Scholar 

  5. Schiavo G., Benfenati F., Poulain B., Rossetto O., Polverino de Laureto P., DasGupta B. R., and Montecucco C. (1992) Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin. Nature 359, 832–835.

    Article  PubMed  CAS  Google Scholar 

  6. Patarnello T., Bargelloni L., Rossetto O., Schiavo G., and Montecucco C. (1993) Neurotransmission and secretion. Nature 364, 581–582.

    Article  PubMed  CAS  Google Scholar 

  7. Bruns D., Engers S., Yang C., Ossig R., Jeromin A., and Jahn R. (1997) Inhibition of transmitter release correlates with the proteolytic activity of tetanus toxin and botulinus toxin A in individual cultured synapses of Hirudo medicinalis. J. Neurosci. 17, 1898–1910.

    PubMed  CAS  Google Scholar 

  8. Sadoul K., Berger A., Niemann H., Weller U., Roche P. A., Klip A., Trimble W. S., Regazzi R., Catsicas S., and Halban P. A. (1997) SNAP-23 is not cleaved by botulinum neurotoxin E and can replace SNAP-25 in the process of insulin secretion. J. Biol. Chem. 272, 33,023-33,027.

    Article  Google Scholar 

  9. Galli T., Zahraoui A., Vaidyanathan V. V., Raposo G., Tiańy J. M., Karińy M., Niemann H., and Louvard D. (1998) A novel tetanus neurotoxin insensitive vesicle associated membrane protein (TI-VAMP) in SNARE complexes of the apical plasma membrane of epithelial cells. Mol. Biol. Cell 9, 1437–1448.

    PubMed  CAS  Google Scholar 

  10. Doussau F., Clabecq A., Henry J. P., Darchen F., and Poulain B. (1998) Calcium-dependent regulation of rab3 in short-term plasticity. J. Neurosci. 18, 3147–3157.

    PubMed  CAS  Google Scholar 

  11. Van der Kloot W. and Molgó J. (1994) Quantal acetylcholine release at the vertebrate neuromuscular junction. Physiol. Rev. 74, 899–991.

    PubMed  Google Scholar 

  12. Angleson J. K. and Betz W. J. (1997) Monitoring secretion in real time: capacitance, amperometry and fluorescence compared. Trends Neurosci. 20, 281–287.

    Article  PubMed  CAS  Google Scholar 

  13. Raciborska D. A., Trimble W. S., and Charlton M. P. (1998) Presynaptic protein interactions in vivo: evidence from botulinum A, C, D and E action at frog neuromuscular junction. Eur. J. Neurosci. 10, 2617–2628.

    Article  PubMed  CAS  Google Scholar 

  14. Lomneth R., Suszkiw J. B., and DasGupta B. R. (1990) Response of the chick ciliary ganglion-iris neuromuscular preparation to botulinum neurotoxin. Neurosci. Lett. 113, 211–216.

    Article  PubMed  CAS  Google Scholar 

  15. Ginsborg B. L. and Warriner J. (1960) The isolated chick biventer cervitis nerve-muscle preparation. Br. J. Pharmacol. 15, 410–411.

    CAS  Google Scholar 

  16. Simpson L. L. (1982) The interaction between aminoquinolines and presynaptically acting neurotoxins.J. Pharmacol. Exp. Ther. 222, 43–48.

    PubMed  CAS  Google Scholar 

  17. Diamond J. and Mellanby J. (1971) The effect of tetanus toxin in the goldfish. J. Physiol. (Lond.) 215, 727–741.

    CAS  Google Scholar 

  18. MacKenzie I., Burnstock G., and Dolly J. O. (1982) The effects of purified botulinum neurotoxin type A on cholinergic, adrenergic and non-adrenergic, atro-pine-resistant autonomic neuromuscular transmission. Neuroscience 7, 997–1006.

    Article  PubMed  CAS  Google Scholar 

  19. Dunant Y., Esquerda J. E., Loctin F., Marsal J., and Muller D. (1987) Botulinum toxin inhibits quantal acetylcholine release and energy metabolism in the Torpedo electric organ. J. Physiol. (Lond.) 385, 677–692.

    CAS  Google Scholar 

  20. Herreros J., Blasi J., Arribas M., and Marsal J. (1995) Tetanus toxin mechanism of action in Torpedo electromotor system: a study on different steps in the intoxication process. Neuroscience 65, 305–311.

    Article  PubMed  CAS  Google Scholar 

  21. Sweeney S. T., Broadie K., Keane J., Niemann H., and O’Kane C. J. (1995) Targeted expression of tetanus toxin light chain in Drosophila specifically eliminates synaptic transmission and causes behavioral defects. Neuron 14, 341–351.

    Article  PubMed  CAS  Google Scholar 

  22. Martinez-Padron M. and Ferrus A. (1997) Presynaptic recordings from Drosophila: correlation of macroscopic and single-channel K+ currents. J. Neurosci. 17, 3412–3424.

    PubMed  CAS  Google Scholar 

  23. Poulain B., Tauc L., Maisey E. A., Wadsworth J. D., Mohan P. M., and Dolly J. O. (1988) Neurotransmitter release is blocked intracellularly by botulinum neurotoxin, and this requires uptake of both toxin polypeptides by a process mediated by the larger chain. Proc. Natl. Acad. Sci. USA 85, 4090–4094.

    Article  PubMed  CAS  Google Scholar 

  24. Poulain B., Wadsworth J. D., Shone C. C., Mochida S., Lande S., Melling J., Dolly J. O., and Tauc L. (1989) Multiple domains of botulinum neurotoxin contribute to its inhibition of transmitter release in Aplysia neurons. J. Biol. Chem. 264, 21,928-21, 933.

    Google Scholar 

  25. Poulain B., De Paiva A., Deloye F., Doussau F., Tauc L., Weller U., and Dolly J. O. (1996) Differences in the multiple step process of inhibition of neurotransmitter release induced by tetanus toxin and botulinum neurotoxins type A and B at Aplysia synapses. Neuroscience 70, 567–576.

    Article  PubMed  CAS  Google Scholar 

  26. Mochida S., Poulain B., Weller U., Habermann E., and Tauc L. (1989) Light chain of tetanus toxin intracellularly inhibits acetylcholine release at neuro-neu-ronal synapses, and its internalization is mediated by heavy chain. FEBS Lett. 253, 47–51.

    Article  PubMed  CAS  Google Scholar 

  27. Kurazono H., Mochida S., Binz T., Eisel U., Quanz M., Grebenstein O., Poulain B., Tauc L., and Niemann H. (1992) Minimal essential domains specifying toxicity of the light chains of tetanus toxin and botulinum neurotoxin type A. J. Biol. Chem. 267, 14, 721-14,729.

    Google Scholar 

  28. Hunt J. M., Bommert K., Charlton M. P., Kistner A., Habermann E., Augustine G. J., and Betz H. (1994) A post-docking role for synaptobrevin in synaptic vesicle fusion. Neuron 12, 1269–1279.

    Article  PubMed  CAS  Google Scholar 

  29. Llinas R., Sugimori M., Chu D., Morita M., Blasi J., Herreros J., Jahn R., and Marsal J. (1994) Transmission at the squid giant synapse was blocked by tetanus toxin by affecting synaptobrevin, a vesicle-bound protein. J. Physiol. (Lond.) 477, 129–133.

    CAS  Google Scholar 

  30. Marsal J., Ruiz-Montasell B., Blasi J., Moreira J. E., Contreras D., Sugimori M., and Llinas R. (1997) Block of transmitter release by botulinum C1 action on syntaxin at the squid giant synapse. Proc. Natl. Acad. Sci. USA 94, 14, 871-14, 876.

    Article  Google Scholar 

  31. Habermann E. and Dreyer F. (1986) Clostridial neurotoxins: handling and action at the cellular and molecular level. Curr. Top. Microbiol. Immunol. 129, 93–179.

    PubMed  CAS  Google Scholar 

  32. Wellhöner H. H. (1992) Tetanus and botulinum neurotoxins, in Selective Neurotoxicity, Handbook of Experimental Pharmacology, Vol. 102 (Herken H. and Hucho F., eds.), Springer-Verlag, Berlin, pp. 357–417.

    Google Scholar 

  33. Beise J., Hahnen J., Ansersen-Beckh B., and Dreyer F. (1994) Pore formation by tetanus toxin, its chain and fragments in neuronal membranes and evaluation of the underlying motifs in the structure of the toxin molecule. Naunyn Schmiedebergs Arch. Pharmacol. 349, 66–73.

    Article  PubMed  CAS  Google Scholar 

  34. Owe-Larsson B., Kristensson K., Hill R. H., and Brodin L. (1997) Distinct effects of clostridial toxins on activity-dependent modulation of autaptic responses in cultured hippocampal neurons. Eur. J. Neurosci. 9, 1773–1777.

    Article  PubMed  CAS  Google Scholar 

  35. Gähwiler B. H., Capogna M., Debanne D., McKinney R. A., and Thompson S. M. (1997) Organotypic slice cultures: a technique has come of age. Trends Neurosci. 20, 471–477.

    Article  PubMed  Google Scholar 

  36. Stanley E. F. and Mirotznik R. R. (1997) Cleavage of syntaxin prevents G-pro-tein regulation of presynaptic calcium channels. Nature 385, 340–343.

    Article  PubMed  CAS  Google Scholar 

  37. Borst J.G. and Sakmann B. (1996) Calcium influx and transmitter release in a fast CNS synapse. Nature 383, 431–434.

    Article  PubMed  CAS  Google Scholar 

  38. Lawrence G. W., Foran P., Mohammed N., DasGupta B. R., and Dolly J. O. (1997) Importance of two adjacent C-terminal sequences of SNAP-25 in exocytosis from intact and permeabilized chromaffin cells revealed by inhibition with botulinum neurotoxin A and E. Biochemistry 36, 3061–3067.

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  40. Bittner M. A., DasGupta B. R., and Holz R. W. (1989) Isolated light chains of botulinum neurotoxins inhibit exocytosis. Studies in digitonin-permeabilized chromaffin cells. J. Biol. Chem. 264, 10,354-10, 360.

    Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  42. Bittner M. A., and Holz R. W. (1992) Kinetic analysis of secretion from permeabilized adrenal chromaffin cells reveals distinct components. J. Biol. Chem. 267, 16,219-16, 225.

    Google Scholar 

  43. Vitale N., Gensse M., Chasserot-Golaz S., Aunis D., and Bader M. F. (1996) Trimeric G proteins control regulated exocytosis in bovine chromaffin cells: sequential involvement of Go associated with secretory granules and Gi3 bound to the plasma membrane. Eur. J. Neurosci. 8, 1275–1285.

    Article  PubMed  CAS  Google Scholar 

  44. Lawrence G. W., Weller U., and Dolly J. O. (1994) Botulinum A and the light chain of tetanus toxin inhibit distinct stages of MgATP-dependent catecholamine exocytosis from permeabilized chromaffin cells. Eur. J. Biochem. 222, 325–333.

    Article  PubMed  CAS  Google Scholar 

  45. Nemoz-Gaillard E., Bosshard A., Regazzi R., Bernard C., Cuber J. C., Takahashi M., Catsicas S., Chayvialle J. A., and Abello J. (1998) Expression of SNARE proteins in enteroendocrine cell lines and functional role of tetanus toxin-sensitive proteins in cholecystokinin release. FEBS Lett. 425, 66–70.

    Article  PubMed  CAS  Google Scholar 

  46. Stecher B., Ahnert-Hilger G., Weller U., Kemmer T. P., and Gratzl M. (1992) Amylase release from streptolysin O-permeabilized pancreatic acinar cells. Effects of calcium, guanosine 5′-[Γ-thio]triphosphate, cyclic AMP, tetanus toxin and botulinum A toxin. Biochem. J. 283, 899–904.

    PubMed  CAS  Google Scholar 

  47. Chasserot-Golaz S., Vitale N., Sagot I., Delouche B., Dirrig S., Pradel L. A., Henry J. P., Aunis D., and Bader M. F. (1996) Annexin II in exocytosis: catecholamine secretion requires the translocation of p36 to the subplasmalemmal region in chromaffin cells. J. Cell Biol. 133, 1217–1236.

    Article  PubMed  CAS  Google Scholar 

  48. Maksymowych A. B. and Simpson L. L. (1998) Binding and transcytosis of botulinum neurotoxin by polarized human colon carcinoma cells. J. Biol. Chem. 273, 21, 950-21, 957.

    Article  Google Scholar 

  49. Simpson L. L. (1980) Kinetic studies on the interaction between botulinum toxin type A and the cholinergic neuromuscular junction. J. Pharmacol. Exp. Ther. 212, 16–21.

    PubMed  CAS  Google Scholar 

  50. Cull-Candy S. G., Lundh H., and Thesleff S. (1976) Effects of botulinum toxin on neuromuscular transmission in the rat. J. Physiol. (Lond.) 260, 177–203.

    CAS  Google Scholar 

  51. Molgó J., Siegel L. S., Tabti N., and Thesleff S. (1989) A study of synchronization of quantal transmitter release from mammalian motor endings by the use of botulinal toxins type A and D. J. Physiol. (Lond.) 411, 195–205.

    Google Scholar 

  52. Duchen L. W. and Tonge D. A. (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. (Lond.) 228, 157–172.

    CAS  Google Scholar 

  53. Bevan S. and Wendon L. M. (1984) A study of the action of tetanus toxin at rat soleus neuromuscular junctions. J. Physiol. (Lond.) 348, 1–17.

    CAS  Google Scholar 

  54. Sarafian T., Aunis D., and Bader M. F. (1987) Loss of proteins from digitonin-permeabilized adrenal chromaffin cells essential for exocytosis. J. Biol. Chem. 262, 16, 671-16, 676.

    Google Scholar 

  55. Sontag J. M., Aunis D., and Bader M. F. (1988) Peripheral actin filaments control calcium-mediated catecholamine release from streptolysin-O-permeabilized chromaffin cells. Eur. J. Cell Biol. 46, 316–326.

    PubMed  CAS  Google Scholar 

  56. Sarafian T., Pradel L. A., Henry J. P., Aunis D., and Bader, M. F. (1991) The participation of annexin II (calpactin I) in calcium-evoked exocytosis requires protein kinase C. J. Cell Biol. 114, 1135–1147.

    Article  PubMed  CAS  Google Scholar 

  57. Livett B. G. (1984) Adrenal medullary chromaffin cells in vitro. Physiol. Rev. 64, 1103–1161.

    PubMed  CAS  Google Scholar 

  58. Fohr K. J., Warchol W., and Gratzl M. (1993) Calculation and control of free divalent cations in solutions used for membrane fusion studies. Methods Enzymol. 221, 149–157.

    Article  PubMed  CAS  Google Scholar 

  59. Vitale N., Mukai H., Rouot B., Thierse D., Aunis D., and Bader M. F. (1993) Exocytosis in chromaffin cells. Possible involvement of the heterotrimeric GTP-binding protein G(o). J. Biol. Chem. 268, 14, 715-14, 723.

    Google Scholar 

  60. Mólgo J., Comella J. X., Angaut-Petit D., Pécot-Dechavassine M., Tabti N., Faille L., Mallart A., and Thesleff S. (1990) Presynaptic actions of botuli-nal neurotoxins at vertebrate neuromuscular junctions. J. Physiol. (Paris) 84, 152–166.

    Google Scholar 

  61. Simpson L. L. and DasGupta B. R. (1983) Botulinum neurotoxin type E: studies on mechanism of action and on structure-activity relationships. J. Pharmacol. Exp. Ther. 224, 135–140.

    PubMed  CAS  Google Scholar 

  62. De Paiva A. and Dolly J. O. (1990) Light chain of botulinum neurotoxin is active in mammalian motor nerve terminals when delivered via liposomes. FEBS Lett. 277, 171–174.

    Article  PubMed  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  64. Cornille F., Goudreau N., Ficheux D., Niemann H., and Roques B. P. (1994) Solid-phase synthesis, conformational analysis and in vitro cleavage of synthetic human synaptobrevin II 1-93 by tetanus toxin L chain. Eur. J. Biochem. 222, 173–181.

    Article  PubMed  CAS  Google Scholar 

  65. Lamana C. and Carr C. J. (1967) The botulinal, tetanal, and enterostaphylococcal toxins: a review. Clin. Pharmacol. Ther. 8, 286–332.

    Google Scholar 

  66. Bulbring E. (1997) Observations on the isolated phrenic nerve diaphragm preparation of the rat, 1946. Br. J. Pharmacol. 120(4 Suppl.), 3–26.

    PubMed  CAS  Google Scholar 

  67. Simpson L. L. and Tapp J. T. (1967) Actions of calcium and magnesium on the rate of onset of botulinum toxin paralysis of the rat diaphragm. Int. J. Neuropharmacol. 6, 485–492.

    Article  PubMed  CAS  Google Scholar 

  68. Clark A. W., Bandyopadhyay S., and DasGupta B. R. (1987) The plantar nerves-lumbrical muscles: a useful nerve-muscle preparation for assaying the effects of botulinum neurotoxin. J. Neurosci. Methods 19, 285–295.

    Article  PubMed  CAS  Google Scholar 

  69. McArdle J. J., Angaut-Petit D., Mallart A., Bournaud R., Faille L., and Brigant J. L. (1981) Advantages of the triangularis sterni muscle of the mouse for investigations of synaptic phenomena. J. Neurosci. Methods 4, 109–115.

    Article  PubMed  CAS  Google Scholar 

  70. Dreyer F., Mallart A., and Brigant J. L. (1983) Botulinum A toxin and tetanus toxin do not affect presynaptic membrane currents in mammalian motor nerve endings. Brain Res. 270, 373–375.

    Article  PubMed  CAS  Google Scholar 

  71. Angaut-Petit D., Molgó J., Connold A. L., and Faille L. (1987) The levator auris longus muscle of the mouse: a convenient preparation for studies of short-and long-term presynaptic effects of drugs or toxins. Neurosci. Lett. 82, 83–88.

    Article  PubMed  CAS  Google Scholar 

  72. Angaut-Petit D., Molgó J., Comella J. X., Faille L., and Tabti N. (1990) Terminal sprouting in mouse neuromuscular junctions poisoned with botulinum type A toxin: morphological and electrophysiological features. Neuroscience 37, 799–808.

    Article  PubMed  CAS  Google Scholar 

  73. Lambert H., Pankov R., Gauthier J., and Hancock R. (1990) Electroporation-mediated uptake of proteins into mammalian cells. Biochem. Cell Biol. 68, 729–734.

    Article  PubMed  CAS  Google Scholar 

  74. Bartels F. and Bigalke H. (1992) Restoration of exocytosis occurs after inactivation of intracellular tetanus toxin. Infect. Immun. 60, 302–307.

    PubMed  CAS  Google Scholar 

  75. Erdal E., Bartels F., Binscheck T., Erdmann G., Frevert J., Kistner A., Weller U., Wever J., and Bigalke H. (1995) Processing of tetanus and botulinum A neu-rotoxins in isolated chromaffin cells. Naunyn Schmiedebergs Arch. Pharmacol. 351, 67–78.

    Article  PubMed  CAS  Google Scholar 

  76. Martin T. F. and Walent J. H. (1989) A new method for cell permeabilization reveals a cytosolic protein requirement for Ca2+-activated secretion in GH3 pituitary cells. J. Biol. Chem. 264, 10, 299-10, 308.

    Google Scholar 

  77. Lomneth R., Martin T. F., and DasGupta B. R. (1991) Botulinum neurotoxin light chain inhibits noradrenaline secretion in PC 12 cells at an intracellular membranous or cytoskeletal site. J. Neurochem. 57, 1413–1421.

    Article  PubMed  CAS  Google Scholar 

  78. Aguado F., Gombau L., Majo G., Marsal J., Blanco J., and Blasi J. (1997) Regulated secretion is impaired in AtT-20 endocrine cells stably transfected with botulinum neurotoxin type A light chain. J. Biol. Chem. 272, 26, 005-26, 008.

    Article  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  80. Kalz H. J. and Wellhöner H. H. (1996) Acidification of the cytosol inhibits the uptake of tetanus toxin in NG108-15 and NBr-10A neurohybridoma cells. Naunyn Schmiedebergs Arch. Pharmacol. 353, 606–609.

    Article  PubMed  CAS  Google Scholar 

  81. Barth H., Hofmann F., Olenik C., Just I., and Aktories K. (1998) The N-termi-nal part of the enzyme component (C2I) of the binary Clostridium botulinum C2 toxin interacts with the binding component C2II and functions as a carrier system for aRho ADP-ribosylating C3-like fusion toxin. Infect. Immun. 66, 1364–1369.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Neurobiologie Cellulaire, Strasbourg, France

    Bernard Poulain

  2. INSERM U338, Strasbourg, France

    Marie-France Bader

  3. Laboratoire de Neurobiologie Cellulaire et Moléculaire, Gif-sur-Yvette, France

    Jordi Molgó

Authors
  1. Bernard Poulain
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marie-France Bader
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jordi Molgó
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Research Center Borstel, Germany

    Otto Holst

Rights and permissions

Reprints and permissions

Copyright information

© 2000 Humana Press Inc.

About this protocol

Cite this protocol

Poulain, B., Bader, MF., Molgó, J. (2000). In Vitro Physiological Studies on Clostridial Neurotoxins. In: Holst, O. (eds) Bacterial Toxins: Methods and Protocols. Methods in Molecular Biology™, vol 145. Humana Press. https://doi.org/10.1385/1-59259-052-7:259

Download citation

  • DOI: https://doi.org/10.1385/1-59259-052-7:259

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-604-8

  • Online ISBN: 978-1-59259-052-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation