The effect of 2,5-hexanedione (2,5-HD) on the distribution of the neurofilamental (NF) proteins and vimentin was examined in human neuroblastoma cell line SK-N-SH with immunocytochemical methods. Retinoic acid (10 mu M) induced differentiation into neuronal cells resulting in the outgrowth of processes and synthesis of NF proteins in the majority of the cells. A minority (4%) differentiated as large fibroblasts. Cells were exposed to 0-10 mM 2,5-HD for 3 days. In neuronal cells a concentration-dependent accumulation of NF proteins was detected as a spherical structure in the perikaryon. Neurofilaments in differentiated SK-N-SH cells were more susceptible to 2,5-HD than NF in undifferentiated cells, as the effects were observed at much lower 2,5-HD concentrations. In contrast, no accumulation of vimentin was detected in the fibroblastic cells.