The venom of the scorpion Leiurus quinquestriatus, well-known for its actions on voltage-sensitive sodium channels, has now been shown to have pronounced effects on the in vitro polymerization and stability of neuronal microtubules purified by temperature-dependent cycles of assembly and disassembly. The crude venom, at concentrations as low as 100 micrograms/ml, alters both the extent of tubulin polymerization, as monitored by turbidity, and the appearance of polymerized material under electron microscopic examination. Structures formed in the presence of the venom retain the temperature sensitivity characteristic of normal microtubules, but respond to calcium ions abnormally with a dispersal of ordered structures, as reflected by both increased light scattering and electron microscopic analysis. Fractionation of the crude venom suggested that the active component was the same as the polypeptide neurotoxin which interacts with voltage-sensitive sodium channels and this identity was subsequently verified. Thus, the effect on microtubules of highly purified L. quinquestriatus sodium channel toxin obtained from an independent source was indistinguishable from that of the crude venom. These results indicate that the sodium channel toxin from L. quinquestriatus is also a potent cytoskeletal agent in vitro. This finding may be related to the growing body of evidence suggesting that the neuronal cytoskeleton plays a functional role in the maintenance of membrane excitability.