Pulse-labeled cytoplasmic proteins from cultured fetal calf-muscle cells at various stages of development were analyzed by one- and two-dimensional gel electrophoresis. The high resolution of the two-dimensional technique allows the determination of those protein species that begin to be synthesized after cell fusion. In addition, actin has been found to exist in three forms possessing similar biochemical properties and identical molecular weights but having slightly different isoelectric points. Two of the forms are found in prefusion dividing myoblasts and also in cultured kidney cells. The third form is the only one found in fetal muscle tissue and is predominant in cultures of fused muscle cells. Thus, it would seem that actin can exist in several isozymic forms of which one is specific to fused muscle tissue.