The neurotoxin tetanospasmin causes tetanus when it reaches the central nervous system. In this autoradiographic study, 125-I-labeled tetanospasmin was injected into the leg muscles of rodents, and the nerves supplying these muscles were crushed. The labeled toxin accumulated within axons on the distal side of the crush. This study provides direct evidence for retrograde axonal transport of a macromolecular toxin that acts at synapses in the central nervous system.