DIFFERENCES IN GLYCOGEN-METABOLISM IN ASTROGLIA-RICH PRIMARY CULTURES AND SORBITOL-SELECTED ASTROGLIAL CULTURES DERIVED FROM MOUSE-BRAIN

Recently it has become possible by chemical selection using sorbitol instead of glucose in the culture medium to produce pure astroglial cultures from astroglia-rich primary cultures from mouse brain. The glycogen-degrading enzyme glycogen phosphorylase in brain is localized in astrocytes and ependymal cells. In view of this fact it appeared necessary to study the influence of glucose and other hexoses on the glycogen metabolism in these cultures lacking the influence of other cell types in comparison to the astroglia-rich primary cultures containing several types of cells. The sorbitol-fed selected cultures and the glucose-deprived astroglia-rich primary cultures contain less than 10% of the glycogen encountered in glucose-fed primary cultures. During incubation with glucose the glycogen content of the selected cultures and the glucose-deprived primary cultures increases by more than one order of magnitude. Nevertheless, not all cells are found to have accumulated glycogen. The time course of the replenishment of glycogen is similar in both types of culture, although maximal levels reached in the selected cultures are 3 times those in the astroglia-rich primary cultures. This difference might be explained by the fact that the ratio of the maximal activities of glycogen synthase and glycogen phosphorylase in selected cultures was found to be twice that in the unselected cultures. During glucose deprivation the glycogen content is reduced in both culture systems with half-maximal contents being reached at 15 min (primary culture) and 45 min (selected culture). Both types of culture can also utilize mannose for the synthesis of glycogen and the production of lactate. In contrast, fructose or galactose are poor substrates, or none at all, for the generation of glycogen. In conclusion, the absence of non-astroglial cells and the high glycogen content after glucose feeding makes pure astroglial cultures an even better system for investigating the regulation and the function of astrocytic glycogen than astroglia-rich primary cultures.