Cellular heterogeneity in primary cultures of brain cells revealed by immunocytochemical localization of glutamine synthetase

Abstract

The co-localization of glutamine synthetase, glial fibrillary acidic protein, galactocerebroside and fibronectin was investigated by immunofluorescence double staining in primary cultures of dissociated brain cells from newborn mice. In cultures, grown in serum-free medium, containing dibutyryl-cAMP, glutamine synthetase was found in about half of the glial fibrillary acidic protein containing astroblasts. After addition of dexamethasone to the cultures, glutamine synthetase appeared also in another cell type, in which no glial fibrillary acidic protein, but fibronectin was detectable. This demonstrates that these cells, which are present in substantial amounts, are not of glial nature. In cultures treated with dibutyryl-cAMP no other cell type was found positive for fibronectin. For cultures grown in serum-containing medium lacking dibutyryl-cAMP, evidence was obtained that glutamine synthetase was induced by dexamethasone only in part of all cells staining for fibronectin. This suggests the presence of two different populations of fibronectin-positive cells. Oligodendrocytes, revealed by staining for galactocerebroside, never contained detectable amounts of glutamine synthetase, irrespective of the presence of dexamethasone. This also holds for cultures grown in serum-free medium containing dibutyryl-cAMP. However, under these conditions, oligodendroblasts are seen only very rarely. Our results demonstrate an unexpected heterogeneity in the cellular composition of primary cultures from newborn mouse brain.