Incubation of nerve with Ca2+ and the divalent cation ionophores A23187 or ionomycin, causes a prompt vesiculation of the myelin at the paranodes and Schmidt-Lanterman incisures. The vesiculation appears to be dependent upon a rise in the intracellular calcium concentration of the affected Schwann cells. To determine whether a similar vesiculation might occur in vivo, and to examine the long term consequences of ionophore exposure, the sciatic nerves of rats and mice were injected with ionomycin and examined histologically after intervals of 1 h to 75 days. A prompt myelin vesiculation was again observed, and this spread from the paranodes and incisures to invade regions of formerly compact myelin: segmentally demyelinated axons were common 7 days post-injection. The lesions were large, and involved up to 95% of the fibres in the mouse. There was little evidence of either Schwann cell necrosis, or axonal degeneration in either species. The appearance and spatio-temporal progression of the lesion was strikingly similar to that seen after the intraneural injection of lysophosphatidylcholine or phospholipase A2. However, in the ionophore lesion, not all the demyelination was effected by myelin vesiculation, for up to 30% of the affected fibres showed evidence of macrophage-mediated stripping. Macrophage stripping was confined to internodes already affected by vesiculation. Most of the myelin debris was removed by macrophages, and myelin debris was only rarely observed within Schwann cells. Remyelination was initiated in nearly all the affected fibres by 21 days, and by 2 months all affected fibres were invested with thin, new myelin sheaths. These findings may be relevant to immune-mediated demyelination in the peripheral nervous system.