Transformation of fast-twitch into slow-twitch skeletal muscle was induced in adult rabbits by chronic low-frequency stimulation and studied at the ultrastructural level. With the use of stereological techniques, a time course was established for changes in mitochondrial volume, sarcotubular system, and Z-band thickness for periods of stimulation ranging from 6 h to 24 weeks. T-tubules, terminal cisternae, and sarcoplasmic reticulum decreased at an early stage and reached levels typical of slow muscle after only 2 weeks of stimulation. Transformation of Z-band structure took place between 1 1/2 and 3 weeks after the onset of stimulation. Mitochondrial volume increased several fold over the first 3 weeks of stimulation, and fell rapidly after 7 weeks, although it still remained above the levels typical of slow muscle. Although there was no sign of degradation and regeneration of the muscle fibers themselves, considerable structural reorganization was evident at the subcellular level after 1 week of stimulation. The fibers passed through a less well organized transitional stage in which fibers could not be assigned to a normal ultrastructural category. After 3 weeks all of the stimulated fibers could be assigned to the normal slow-twitch category although some subcellular irregularities persisted even after 24 weeks. The ultrastructural alterations are discussed in relation to functional and biochemical changes in the whole muscle.