Abstract
Early events of in vitro neuronal development were studied by inducing neuron formation in a neuroectodermal cell line, NE-4C/A3, derived from the embryonic forebrain vesicles of p53-deficient mice. Neuronal differentiation was initiated by treating the cells with all-trans retinoic acid (RA). By the second day of RA treatment compact cell aggregates were formed. The first signs of neuronal cell fate decision were revealed inside the aggregates. To elucidate the process of aggregate formation, the dynamics of cell clustering and the migration of individual cells were investigated by a novel computer-controlled videomicroscopic system. Besides real-time observation of cell motility, the system allowed statistical analysis of large sets of data providing quantitative evaluation of cell locomotion during an early, critical phase of RA induced neuron formation. The results showed that chemoattractants did not play a principal role in cell aggregation. Retinoic acid, on the other hand, was found to cause a rapid decrease in the average migratory velocity without changing the randomness of migratory routes. The data indicated that aggregation was facilitated by increased cohesion upon incident collision of randomly encountering cells. The resulting compact cell clusters provided the structural conditions for contact communication apparently needed for the neuronal differentiation of NE-4C/A3 cells.
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