Previous studies indicated that a chemically-defined, differentiation medium (DM) induces neuroblastoma cells, especially IMR32K cells, to exhibit phenotypes of mature neurons (including neurite outgrowth and synthesis of neurofilament polypeptides) and develop certain attributes of the neurons which are affected by neurofibrillary degeneration in Alzheimer's disease, such as expression of tangle-associated epitopes and accumulation of paired helical filaments-(PHF-) like fibrils. Immunocytochemical staining suggested that this cytoskeletal abnormality most likely results from altered expression of tau proteins. In the current study, we addressed this issue by analyzing tau-enriched preparations of IMR32K cells that were previously exposed to different incubation media using a panel of antibodies specific to tau and related microtubule-associated proteins. These cultured cells exhibited three groups of tau immunoreactivities which differ in molecular weight. Among them the level of high molecular weight tau (MW 90-112 kDa) was selectively augmented after DM incubation. The tau proteins produced in these neuron-like cells shared phosphorylated sites with PHF-tau and fetal tau, but differed from PHF-tau in their lack of the N-terminal insert which characterizes adult isoforms.