Glycogen accumulation in rat cerebral cortex in dissociated cell culture

Incorporation of [3H]glucose into [3H]glycogen was demonstrated in individual coverslip cultures of dissociated rat cerebral cortex. The time course of incorporation of [3H] glucose into [3H]glycogen was investigated, and it was found that [3H]glycogen accumulation monotonically increased during at least the first hour of incubation with [3H]glucose. In order to identify which cells accumulate glycogen in these cultures we attempted to demonstrate cytochemically the localization of glycogen. We found, however, that conventional aqueous methods of glycogen cytochemistry did not reliably or consistently stain the cultures. By labelling glycogen using [3H]glucose, we were able to show that the entire [3H]glycogen compartment was extractable by water after ethanol fixation. Therefore we developed a non-aqueous technique which preserves tissue glycogen by exploiting its solubility properties. Using this technique we were able to cytochemically demonstrate glycogen in at least two different cell types in the cultures.