Tetanus Toxin Biotinylation and Localization of Binding Sites in Catecholaminergic Cultures and Granules

  • Philip Lazarovici
  • Alexander Fedinec
  • Bernard Bizzini


Tetanus toxin (TT), the 150-kD protein secreted by Clostridium tetani bacteria, is a very powerful neurotoxin which produces spastic paralysis and death in humans in the range of ng/ kg body weight.1 The pharmacokinetics of TT entry to the central nervous system target2include the following main steps: (1) selective binding of TT on the surface of the peripheral motor neuron terminals;3 (2) TT internalization into a vesicular compartment which delivers TT by retrograde axonal transport to the spinal cord perikarya and dendrites;4 (3) TT release from the motor neuron in the spinal cord;5 (4) trans-synaptic transfer of TT and uptake into the presynaptic terminals of the inhibitory intemeurons.5 (5) Thereafter, the constitutive release of gamma amino butyric acid and/or glycine is blocked, resulting with a syndrome of motor neuron dysinhibition;6 (6) the results are muscular spastic contractions, spasms, and convulsive seizures leading to musculature rigidity, respiratory failure, and death.7 It is believed that the binding of TT to neuronal G 1 b polysialogangliosides is involved in these pharmacokinetic steps and is the initial step triggering axonal transport and inhibition of neurotransmitter release.8–9 For many years this TT-specific recognition has been used as a diagnostic and basic research tool for marking central and peripheral neuronal,10 as well as neurosecretory cells such as thyroid C, anterior pituitary, pancreatic islet cells, and derived tumors (insulinoma, pheochromocytoma, neuroblastoma).11–13 All these cells express on their surface G1b polysialogangliosides of different sugar compositions, therefore binding TT.


PC12 Cell Nerve Growth Factor Gold Particle Neurite Outgrowth Chromaffin Cell 
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  1. 1.
    Gill, DM. Bacterial toxins: a table of lethal amounts. Microbiol Rev 1982; 46: 86.PubMedGoogle Scholar
  2. 2.
    Lazarovici P, Yavin E, Bizzini B, Fedinec AA. Retrograde transport in sciatic nerves of gangliosideaffinity-purified tetanus toxins. In: Dolly JO, ed. Neurotoxins in Neurochemistry. Chichester: Ellis Horwood Ltd., 1988: 100.Google Scholar
  3. 3.
    Schwab ME, Thoenen H. Selective binding, uptake, and retrograde transport of tetanus toxin by nerve terminals in the rat iris. An electron microscope study using colloidal gold as a tracer. J Cell Biol 1978; 77: 1.PubMedCrossRefGoogle Scholar
  4. 4.
    Stoeckel K, Schwab M, Thoenen H. Comparison between the retrograde axonal transport of nerve growth factor and tetanus toxin in motor, sensory and adrenergic neurons. Brain Res 1975; 99: 1.CrossRefGoogle Scholar
  5. 5.
    Schwab, ME, Suda K, Thoenen H. Selective retrograde transsynaptic transfer of a protein, tetanus toxin, subsequent to its retrograde axonal transport. J Cell Biol 1979; 82: 798.PubMedCrossRefGoogle Scholar
  6. 6.
    Curtis DR, De Groat WC. Tetanus toxin and spinal inhibition. Brain Res 1968; 10: 208.PubMedCrossRefGoogle Scholar
  7. 7.
    Kryzhanovsky GN. Pathophysiology. In: Veronesi R, ed. Tetanus Important New Concepts. Excerpta Medica. Amsterdam: Casparie, 1981: 109.Google Scholar
  8. 8.
    Mellanby J, Greene J. How does tetanus toxin act? Neuroscience 1981; 6: 281.PubMedCrossRefGoogle Scholar
  9. 9.
    Wellhoner HH. Tetanus neurotoxin. Rev Physiol Biochem Phannacol 1982; 93: 1.CrossRefGoogle Scholar
  10. 10.
    Minsky R. The use of antibodies to define and study major cell types in the central and peripheral nervous system. In: Brookes J, ed. Neuroimmunology. New York: Plenum Press, 1982: 141.Google Scholar
  11. 11.
    Pearse AGE. The cytochemistry and ultrastructure of polypeptide hormone-producing cells of the APUD series and the embryologic, physiologic and pathologic implications of the concept. J Histochem Cytochem 1969; 17: 303.PubMedCrossRefGoogle Scholar
  12. 12.
    Heymann J, Neumann K, Haremann K. Tetanus toxin as a marker for small-cell lung cancer cell lines. J Cancer Res Clin Oncol. 1989; 115: 537.CrossRefGoogle Scholar
  13. 13.
    Berliner P, Unsicker K. Tetanus toxin labeling as a novel, rapid and highly specific tool in human neuroblastoma differential diagnosis. Cancer 1985; 56: 419.PubMedCrossRefGoogle Scholar
  14. 14.
    Rosenheck K, Lelkes P. Stimulus-secretion coupling in chromaffin cells. VoL I, II. Boca Raton, FL: CRC Press, 1987.Google Scholar
  15. 15.
    Lazarovici P, Fujita K, Contreras ML, Diorio JP, Lelkes PI. Affinity purified tetanus toxin binds to isolated chromaffin granules and inhibits catecholamine release in digitonin-permeabilized chromaffin cells. FEBS Lett 1989; 253: 121.PubMedCrossRefGoogle Scholar
  16. 16.
    Bittner M, Holz R. Effects of tetanus toxin on catecholamine release from intact and digitoninpermeabilized chromaffin cells. J Neurochem 1988; 51: 451.PubMedCrossRefGoogle Scholar
  17. 17.
    Penner R, Neher E, Dreyer F. lntracellularly injected tetanus toxin inhibits exocytosis in bovine adrenal chromaffin cells. Nature 1986; 324: 76.PubMedCrossRefGoogle Scholar
  18. 18.
    Lazarovici P. Characterization and visualization of tetanus toxin acceptors on adrenal chromaffin granules. J Physiol (Paris) 1990; 84: 197.Google Scholar
  19. 19.
    Sandberg K, Berry CJ, Rogers T. Studies on the intoxication pathway of tetanus toxin in the rat pheochromocytoma (PC12) cell line. Binding, internalization, and inhibition of acetylcholine release. J Biol Chem 1989; 264: 5679.PubMedGoogle Scholar
  20. 20.
    Fujita K, Guroff G, Yavin E, Goping G, Orenberg R, Lazarovici P. Preparation of affinity-purified, biotinylated tetanus toxin, and characterization and localization of cell surface binding sites on nerve growth factor-treated PC12 cells. Neurochem Res. 1990; 15: 373.PubMedCrossRefGoogle Scholar
  21. 21.
    Bayer EA, Wilchek M. The use of avidin-biotin complex as a tool in molecular biology. In: Glick D, ed. Methods of Biochemical Analysis. New York: John Wiley & Sons, 1980; 26: 1.CrossRefGoogle Scholar
  22. 22.
    Kenimer JG, Habig WH, Hardegree MC. Monoclonal antibodies as probes of tetanus toxin structure and function. Infect Immun 1983; 42: 942.PubMedGoogle Scholar
  23. 23.
    Bizzini B Tetanus toxin structure as a basis for elucidating its immunological and neuropharmacological activities. In: Cuatrecasas P, ed. Receptors and Recognition. London: Chapman and Hall, 1977; 1: 175.Google Scholar
  24. 24.
    Lazarovici P, Yavin E. Affinity-purified tetanus neurotoxin interaction with synaptic membranes: properties of a protease-sensitive receptor component. Biochemistry 1986; 25: 7047.PubMedCrossRefGoogle Scholar
  25. 25.
    Yavin E, Lazarovici P, Nathan A. Molecular interactions of ganglioside receptors with tetanus toxin on solid supports, aqueous solutions and natural membranes. In: Wirtz KWA, ed. Membrane Receptors, Dynamics, and Energetics. New York: Plenum Publishing Co., 1987: 135.CrossRefGoogle Scholar
  26. 26.
    Pollard HB, Zinder O, Hoffman PG, Nikodijevic O. Regulation of the transmembrane potential of isolated chromaffin granules by ATP, ATP analogs, and external pH. J Biol Chem 1976; 251: 4544.PubMedGoogle Scholar
  27. 27.
    Wagner JA. Structure of catecholamine secretory vesicles from PC12 cells. J Neurochem 1985; 45: 1244.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1993

Authors and Affiliations

  • Philip Lazarovici
    • 1
  • Alexander Fedinec
    • 2
  • Bernard Bizzini
    • 3
  1. 1.Department of Pharmacology and Experimental Therapeutics, School of Pharmacy, Faculty of MedicineThe Hebrew University of JerusalemJerusalemIsrael
  2. 2.Department of Anatomy and NeurobiologyUniversity of TennesseeMemphisUSA
  3. 3.Department of Molecular ToxinologyInstitut PasteurParisFrance

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