Two actin isoforms, gamma and beta, are contained within neuroblastoma cells. However, the relative amount and distribution of both isoforms within the cells are differentially regulated during neurite extension. The proportion of gamma-actin isoform became about four times greater than that of beta actin during neuroblastoma cell differentiation. Additionally, whereas beta actin appears to be concentrated in the cell cortex, gamma actin is also present throughout the cell body. Upon differentiation, neuroblastoma cells reorganize their actin cytoskeleton and gamma actin is induced to polymerize whereas beta actin polymers are partially disassembled. Moreover, both actin isoforms are differentially distributed within differentiated cells. Thus, gamma actin polymers are located both in the soma and proximal regions of extended neurites, whereas beta actin is enriched in the terminal tip of the neurites. Our results strongly suggest that both actin isoforms are involved in a different way in neuroblastoma cell differentiation.