Summary
Local tetanus was elicited in rats and cats by intramuscular injection of 125I-tetanus toxin. After different times spinal radioactivity was extracted with either non-ionic (Lubrol PX) or ionic (sodium dodecyl sulfate, SDS) detergents and compared with native or 125I-toxin by gel filtration, SDS-gel electrophoresis, immunological procedures, and toxicity tests. In double-isotope experiments, 131I-toxin was added to the extracts as standard.
In rats, the bulk of extracted material was indistinguishable from native toxin. However, there was a slight shift of the extracted material towards smaller molecular weights in gel filtration with Lubrol. In gel filtration with SDS, the toxin peak was followed by some tailing of 125I radioactivity. Accordingly a small part of extracted radioactivity moves faster than the standard in SDS disc gel electrophoresis. These findings taken together indicate some degradation in vivo.
Adsorption to solid-phase antibodies indicated that more than 80% of the radioactivity extracted from rats was still immunoreactive. It yielded a zone confluent with extrinsic toxin in immunodiffusion.
The spinal cord Lubrol extract from rats was still toxic in the expected range. Due to the very small amounts of toxin present, no precise toxicity data could be given.
In cats, there was also some evidence for radioactive split products in both SDS gel filtration and disc gel electrophoresis. The patterns closely resembled those obtained with extracts from rat spinal cord.
SDS extracts from rat and cat spinal cords, poisoned with 125I tetanus toxin in vivo, were also subjected to SDS disc gel electrophoresis followign reduction with dithioerythritol (DTE). They yielded large and small chains of the same size as did native toxin.
In vitro, extensive degradation with brain homogenate from rats took place at pH 3.65, but not at pH 7.5. This indicates that lysosomal degradation is not a major metabolic pathway of tetanus toxin in vivo, although it is possible in principle.
It is concluded that a) unlike other toxins, tetanus toxin is not necessarily degraded during its cellular uptake, b) the bulk of radioactive material is indistinguishable, following its neuronal ascent, from native or labeled toxin, c) a part of the radioactivity is recovered as split products.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bizzini, B., Turpin, A., Raynaud, M.: Immunochimie et mécanisme d'action de la toxine tétanique. Bull. Inst. Pasteur 72, 177–219 (1974)
Cheng-Raude, D., Treloar, M., Habermann, E.: Preparation and pharmacokinetics of labeled derivatives of apamin. Toxicon 14, 467–476 (1976)
Dimpfel, W., Habermann, E.: Histoautoradiographic localisation of 125I-labeled tetanus toxin in rat spinal cord. Naunyn-Schmiedebergs Arch. Pharmacol. 280, 177–182 (1973)
Dimpfel, W., Neale, J. H., Habermann, E.: 125I-labelled tetanus toxin as a neuronal marker in tissue cultures derived from embryonic CNS. Naunyn-Schmiedeberg's Arch. Pharmacol. 290, 329–333 (1975)
Erdmann, G., Wiegand, H., Wellhöner, H. H.: Intraaxonal and extraaxonal transport of 125I-tetanus toxin in early local tetanus. Naunyn-Schmiedeberg's Arch. Pharmacol. 290, 357–373 (1975)
Fairbanks, G., Steck, T. L., Wallach, D. F. H.: Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10, 2606–2617 (1971)
Fedinec, A. A.: Tetanospasmin spreading, metabolism and possibilities of neutralization. Proc. of 4th International Conf. on Tetanus, pp. 123–144. Lyon: Fondation Mérieux, 1975
Fildes, P., Ledingham, J. C. G.: A system of bacteriology in rerelation to medicine. Vol. III, pp. 298–372. London: His Majesty's Stationary Office 1929
Fillenz, M., Gagnon, C., Stoeckel, K., Thoenen, H.: Selective uptake and retrograde axonal transport of dopamine β-hydroxylase antibodies in peripheral adrenergic neurons. Brain Res. 114, 293–303 (1976)
Gill, D. M.: Protein toxins that act within cells. Bull. Inst. Pasteur 74, 65–84 (1976)
Habermann, E.: Ein neues Prinzip zur Bestimmung hochmolekularer Antigene (Verknüpfungstest) und seine Anwendung auf Tetanustoxin, Serumalbumin und Ovalbumin. Z. Klin. Chem. Klin. Biochem. 8, 51–55 (1970a)
Habermann, E.: Pharmakokinetische Besonderheiten des Tetanustoxins und ihre Beziehungen zur Pathogenese des lokalen bzw. generalisierten Tetanus. Naunyn-Schmiedeberg's Arch. Pharmakol. 267, 1–19 (1970b)
Habermann, E.: Interaction of labelled tetanus toxin and toxoid with substructures of rat brain and spinal cord in vitro. Naunyn-Schmiedeberg's Arch. Pharmacol. 276, 341–359 (1973)
Habermann, E.: Affinity chromatography of tetanus toxin, tetanus toxoid, and botulinum A toxin on synaptosomes, and differentiation of their acceptors. Naunyn-Schmiedeberg's Arch. Pharmacol. 293, 1–9 (1976)
Habermann, E., Wellhöner, H. H.: Advances in tetanus research. Klin. Wschr. 52, 255–265 (1974)
Habermann, E., Dimpfel, W., Räker, K. O.: Interaction of labelled tetanus toxin with substructures of rat spinal cord in vivo. Naunyn-Schmiedeberg's Arch. Pharmacol. 276, 361–373 (1973)
Jaenike, R., Knof, S.: Molecular weight and quaternary structure of lactic dehydrogenase. 3. Comparative determination by sedimentation analysis, light scattering and osmosis. Eur. J. Biochem. 4, 157–163 (1968)
Neuhoff, V.: Micromethods in molecular biology. Berlin-Heidelberg-New York: Springer 1973
Pagé, M., Godin, C.: On the determination of molecular weight of protein subunits on Sephadex G-200 in the presence of detergent. Glutamate dehydrogenase. Can. J. Biochem. 47, 401–403 (1969)
Price, D. L., Griffin, J., Young, A., Peck, K., Stocks, A.: Tetanus toxin: Direct evidence for retrograde axonal transport. Science 188, 945–947 (1975)
Putnam, F. W.: Structure and function of plasma proteins. In: The proteins, vol. 3, pp. 154–267 (H. Neurath, ed.). New York-London: Academic Press 1965
Reynolds, J. A., Tanford, C.: Binding of dodecyl sulfate to proteins at high binding ratios. Possible implications for the state of proteins in biological membranes. Proc. Natl. Acad. Sci. U.S.A. 66, 1002–1007 (1970)
Robbins, J. B., Haimovich, J., Sela, M.: Purification of antibodies with immunoadsorbents prepared using bromacetyl cellulose. Immunochemistry 4, 11–22 (1967)
Schwab, M. E., Thoenen, H.: Electron microscopic evidence for a transsynaptic migration of tetanus toxin in spinal cord motoneurons: an autoradiographic and morphometric study. Brain Res. 105, 213–227 (1976a)
Schwab, M. E., Thoenen, H.: Selective trans-synaptic migration of tetanus toxin in rat sympathetic ganglia after retrograde axonal transport. Experientia 32, 811 (1976b)
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. 99, 1–16 (1975)
Van Heyningen, S.: Binding of ganglioside by the chains of tetanus toxin. FEBS Lett. 68, 5–7 (1976)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Habermann, E., Wellhöner, H.H. & Räker, K.O. Metabolic fate of 125I-tetanus toxin in the spinal cord of rats and cats with early local tetanus. Naunyn-Schmiedeberg's Arch. Pharmacol. 299, 187–196 (1977). https://doi.org/10.1007/BF00498561
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00498561