SUMMARY: Binding of purified type A, C and E toxins to cultured cells was studied by an immunocytochemical method. Type A and C toxins bound strongly to neuron cultures prepared from brains of foetal mice, but binding of type E toxin was weak. None of the toxin types bound to the feeder layer, composed of non-neuronal cells. The heavy-chain component of the type C toxin bound to neurons, but the light chain component did not. Type C toxin also bound only to cell lines of neuronal origin. When type C toxin [final concentration 4 × 10 LD (10 ng) per well] was added to primary neuron cultures in 96-well plates, degeneration of neuronal processes and rounding of neuronal somas were observed, but type A and E toxins did not produce such changes. The binding and cytotoxic activities of type C toxin were blocked by heat treatment (80 °C for 30 min) or by preincubation of the toxin with polyclonal anti-C IgG and some of the monoclonal antibodies which neutralized the toxin activity in mice. In the neuronal processes treated with C toxin, many degenerated mitochondria, membranous dense bodies and vesicles were observed by electron microscopy; these ultrastructural changes were similar to those of Wallerian degeneration .


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