Neuronal sensitivity to tetanus toxin requires gangliosides.
Tetanus toxin produces spastic paralysis in situ by blocking inhibitory neurotransmitter release in the spinal cord. Although di- and trisialogangliosides bind tetanus toxin, their role as productive toxin receptors remains unclear. We examined toxin binding and action in spinal cord cell cultures grown in the presence of fumonisin B(1), an inhibitor of ganglioside synthesis. Mouse spinal cord neurons grown for 3 weeks in culture in 20 microM fumonisin B(1) develop dendrites, axons, and synaptic terminals similar to untreated neurons, even though thin layer chromatography shows a greater than 90% inhibition of ganglioside synthesis. Absence of tetanus and cholera toxin binding by toxin-horseradish peroxidase conjugates or immunofluorescence further indicates loss of mono- and polysialogangliosides. In contrast to control cultures, tetanus toxin added to fumonisin B(1)-treated cultures does not block potassium-stimulated glycine release, inhibit activity-dependent uptake of FM1-43, or abolish immunoreactivity for vesicle-associated membrane protein, the toxin substrate. Supplementing fumonisin B(1)-treated cultures with mixed brain gangliosides completely restores the ability of tetanus toxin to bind to the neuronal surface and to block neurotransmitter release. These data demonstrate that fumonisin B(1) protects against toxin-induced synaptic blockade and that gangliosides are a necessary component of the receptor mechanism for tetanus toxin.
Williamson
L C
LC
Laboratory of Developmental Neurobiology, NICHHD, National Institutes of Health, Bethesda, Maryland 20892-4480, USA.
Bateman
K E
KE
Clifford
J C
JC
Neale
E A
EA
D000818:Animals; D002264:Carboxylic Acids / Q000494:pharmacology*; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D037341:Fumonisins; D005732:Gangliosides / Q000494:pharmacology*; D005998:Glycine / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008858:Microscopy, Phase-Contrast; D009474:Neurons / Q000187:drug effects*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors; D010446:Peptide Fragments; D011485:Protein Binding; D011726:Pyridinium Compounds / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D050683:R-SNARE Proteins; D013116:Spinal Cord / Q000187:drug effects*; D013744:Tetanus Toxin / Q000494:pharmacology*
D002264:Carboxylic Acids; D004791:Enzyme Inhibitors; C073804:FM1 43; D037341:Fumonisins; D005732:Gangliosides; D008565:Membrane Proteins; D010446:Peptide Fragments; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D050683:R-SNARE Proteins; D013744:Tetanus Toxin; C056933:fumonisin B1; D010088:Oxidoreductases; C108138:dihydroceramide desaturase; D005998:Glycine