A Study of the Mechanism of Internalisation of Tetanus Toxin by Primary Mouse Spinal Cord Cultures

A Study of the Mechanism of Internalisation of Tetanus Toxin by Primary Mouse Spinal Cord Cultures Abstract: The fate of tetanus toxin bound to neuronal cells at 0°C was followed using an anti‐toxin 125I‐protein A assay. About 50%; of surface‐bound toxin disappeared within 5 min of warming cells to 37°C. Experiments with 125I‐toxin showed that much of this loss was due to dissociation of bound toxin into the medium. Some toxin was however rapidly internalised, and could be detected only by permeabilising cells with Triton X‐100 prior to assay. To investigate the mechanism of internalisation, tetanus toxin was adsorbed to colloidal gold. Toxin‐gold was shown to be stable, and to recognise the same receptor(s) as free toxin. Quantitation of the distribution of toxin‐gold particles bound to the cell body at 4°C showed that it was concentrated in coated pits. After 5 min at 37°C, toxin‐gold appeared in coated vesicles, endosomes, and tubules. After 15 min, it was found largely in endosomes, and at 30 min in multivesicular bodies. The involvement of coated pits in internalisation of tetanus toxin, but not cholera toxin, was confirmed using the free toxins, anti‐toxins, and protein A‐gold. Toxin‐gold also entered nerve terminals and axons via coated pits, accumulating in synaptic vesicles and in‐traaxonal uncoated vesicles, respectively. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Neurochemistry Wiley

A Study of the Mechanism of Internalisation of Tetanus Toxin by Primary Mouse Spinal Cord Cultures

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Publisher
Wiley
Copyright
Copyright © 1987 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0022-3042
eISSN
1471-4159
DOI
10.1111/j.1471-4159.1987.tb09994.x
Publisher site
See Article on Publisher Site

Abstract

Abstract: The fate of tetanus toxin bound to neuronal cells at 0°C was followed using an anti‐toxin 125I‐protein A assay. About 50%; of surface‐bound toxin disappeared within 5 min of warming cells to 37°C. Experiments with 125I‐toxin showed that much of this loss was due to dissociation of bound toxin into the medium. Some toxin was however rapidly internalised, and could be detected only by permeabilising cells with Triton X‐100 prior to assay. To investigate the mechanism of internalisation, tetanus toxin was adsorbed to colloidal gold. Toxin‐gold was shown to be stable, and to recognise the same receptor(s) as free toxin. Quantitation of the distribution of toxin‐gold particles bound to the cell body at 4°C showed that it was concentrated in coated pits. After 5 min at 37°C, toxin‐gold appeared in coated vesicles, endosomes, and tubules. After 15 min, it was found largely in endosomes, and at 30 min in multivesicular bodies. The involvement of coated pits in internalisation of tetanus toxin, but not cholera toxin, was confirmed using the free toxins, anti‐toxins, and protein A‐gold. Toxin‐gold also entered nerve terminals and axons via coated pits, accumulating in synaptic vesicles and in‐traaxonal uncoated vesicles, respectively.

Journal

Journal of NeurochemistryWiley

Published: Oct 1, 1987

References

  • Fate of tetanus toxin bound to the surface of primary neurones in culture: evidence for rapid internalization
    Critchley, Critchley; Nelson, Nelson; Habig, Habig; Fishman, Fishman
  • Reevaluation of the role of gangliosides as receptors for tetanus toxin
    Critchley, Critchley; Habig, Habig; Fishman, Fishman
  • Internalization and degradation of cholera toxin by cultured cells: relationship to toxin action
    Fishman, Fishman
  • Endocytosis of cholera toxin in GERL‐like structures of murine neuroblastoma cells pretreated with GM1 ganglioside
    Joseph, Joseph; Stieber, Stieber; Gonatas, Gonatas
  • Localization of the epidermal growth factor (EGF) receptor within the endosome of EGF‐stimulated epidermal carcinoma (A431) cells
    Miller, Miller; Beardmore, Beardmore; Kanety, Kanety; Schlessinger, Schlessinger; Hopkins, Hopkins
  • Selective binding, uptake and retrograde transport of tetanus toxin by nerve terminals in rat iris: an electron microscope study using colloidal gold as a tracer
    Schwab, Schwab; Thoenen, Thoenen
  • Selective retrograde transsynaptic transfer of a protein, tetanus toxin, subsequent to its retrograde axonal transport
    Schwab, Schwab; Suda, Suda; Thoenen, Thoenen
  • Transport of macrophage Fc receptors and Fc receptor‐bound ligands to lysosomes
    Ukkonen, Ukkonen; Lewis, Lewis; Marsh, Marsh; Helenius, Helenius; Mellman, Mellman

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