Maintenance of neurons isolated in bulk from rat brain: incorporation of radiolabeled substrates.

Neurons isolated in bulk from 10- to 15-day-old rat brain can be maintained for 18--24 h. Phase microscopy shows that the cells remain morphologically intact during this time. As a criterion of viability, the incorporation of radiolabeled amino acids and uridine into trichloroacetic acid-insoluble material was selected. The cells are capable of the incorporation both after isolation and after maintenance. The uptake is constant with time, is proportional to substrate concentration, and is inhibited by puromycin and cycloheximide, but not by chloramphenicol. Thus the cells remain viable during this maintenance period. This system may provide a model for the study of the synthesis of specific neuronal components.