Enucleation of the rat pheochromocytoma clonal cell line, PC12: effect on neurite outgrowth

The effect of removal of PC12 cell nuclei on neurite outgrowth was studied. Enucleation (80-90%) was accomplished in the presence of cytochalasin B using a centrifugation technique that exploited the very tight adhesivity of PC12 cells for a substratum composed of an extracellular matrix secreted by bovine corneal endothelial cells in response to epidermal growth factor treatment. Neither nucleated nor enucleated PC12 cells showed significant neurite outgrowth on this particular matrix in the absence of nerve growth factor. In the presence of nerve growth factor both PC12 cell types initiated neurite outgrowth, but whereas neurites from nucleated cells grew continuously for two days, those from enucleated cells reached a maximum length after one day. The results suggest that neurite initiation but not continued neurite growth or stabilization can occur in the absence of transcription.