Transient increase in vimentin in axonal cytoskeletons during differentiation in NB2a/d1 cells

The localization of vimentin (Vm) within the Triton-insoluble cytoskeleton was characterized during differentiation of mouse NB2a/dl neuroblastoma cells. Vm staining increased within neurites during the first day of differentiation, and then rapidly declined in both perikarya and neurites. By contrast, immunoreactivity against extensively phosphorylated forms of the high molecular weight neurofilament subunit (NF-H) was absent until the third day after differentiation. Immunoblot analyses confirmed that these alterations reflected specific changes in Vm and NF-H steady-state levels. Metabolic labeling demonstrated a decrease in the rate of Vm synthesis by the third day of differentiation. We conclude that changes in incorporation of intermediate filament species into the axonal cytoskeleton reflect distinct stages in neurite outgrowth and maturation; i.e., the Vm filament system may participate in initial stages of neuritogenesis during which outgrowth is most rapid, while NFPs may subsequently function in the establishment of a stabilized axonal cytoskeleton.