Differential distribution of vimentin and neurofilament protein immunoreactivity in NB2a/d1 neuroblastoma cells following neurite retraction distinguishes two separate intermediate filament systems.

Mouse NB2a/d1 cells assemble all 3 neurofilament protein subunits (NFPs) into the detergent-insoluble cytoskeleton and segregate phosphorylated forms of the 200-kDa subunit (NFP-H) within neurites when differentiation is induced with dibutyryl cyclic AMP (dbcAMP). Before and after differentiation, these cells also incorporate vimentin into both the perikaryal and neuritic cytoskeleton (Shea et al., 1988, Dev. Brain Res., submitted). To determine whether NFPs and vimentin constitute separate intermediate filament systems or exist as heteropolymers, we perturbed cytoskeletal architecture by inducing the retraction of neurites with colchicine. After cells were exposed to colchicine, vimentin immunoreactivity partitioned into perikarya in the form of fibrous whorls that did not cross-react with antisera to NFPs. By contrast, NFP immunoreactivity remained dispersed throughout the cell body following neurite retraction. We interpret these different responses to colchicine to indicate that NFPs and vimentin are assembled into separate intermediate filaments in NB2a/d1 cells.