An immunohistochemical study of two myelin-specific proteins in enriched oligodendroglial cell cultures combined with an autoradiographic investigation using [3H]thymidine.

Abstract

The aim of the present work was to examine the possible relationship between proliferation and expression of 2 myelin specific proteins in cultured oligodendroglial cells. Mixed cultures of glial cells, from newborn rat brain, containing astroglia and oligodendroglia were grown in 2 different culture media, minimum Eagle's medium and Waymouth's medium both supplemented with 10% calf serum in presence or absence of adult rat brain soluble extract. The proliferative activity of the cells was followed over a 28-day period by autoradiography after radioactive thymidine incorporation. It was found that in cultures grown in Waymouth's medium the proportion of oligodendroglial cells was higher and that proliferation was more active than in minimum Eagle's medium. Addition of brain extract elicited a stimulation of the proliferation of the cells in the 2 basal media. Under all conditions W1 protein appeared earlier than MBP by immunofluorescent visualization. Some oligodendroglial cells synthesizing W1 protein were still able to proliferate. MBP appears to be a marker of a later stage of cell maturation since very few MBP-positive cells incorporated tritiated thymidine. More cells contained MBP in the presence of brain extract. These results suggest that oligodendroglial cell maturation proceeds by steps, the step of W1 protein expression is compatible with proliferation while that of MBP expression appears at the end of the proliferation phase.