Regulation of cerebroside and sulfatide metabolism in glia cells.

Mouse oligodendroglioma cells, G-26 clone 20 and 24, contain galactosylceramide (cerebroside) and sulfogalactosylceramide (sulfatide) as determined by an HPLC technique. The synthesis of both these lipids was stimulated by 10(-6) M hydrocortisone (cortisol) and also by the removal of serum from the culture medium. Forty-eight hours after the addition of cortisol the incorporation of H235SO4 into sulfatide, the level of sulfatide and the specific activity of the enzyme 3'-phosphoadenosine 5'-phosphosulfate:galactosylceramide sulfotransferase in the cells increased three- to fourfold.

Biosynthesis of glycosphingolipids in cultured mouse neuroblastoma cells. Precursor-product relationships among sialoglycosphingolipids.

The reaction sequence for the biosynthesis of gangliosides by mouse neuroblastoma cells has been investigated by studying the pattern of incorporation of labeled precursors into sialoglycosphingolipids. Cultured NB41A cells incorporated N-[3H]acetylmannosamine into the sialic acid moiety of GM3 in less than 10 min. Labeled GM2 was not detected in cells incubated for less than 30 min, while measurable radioactivity did not appear in GM1 until after 60 to 90 min.

N-acetyl-beta-hexosaminidase: role in the degradation of glycosaminoglycans.

Extracts of cultured normal human skin fibroblasts released radioactivity from a (14)C-labeled heptasaccharide prepared by addition of [(14)C]N-acetylgalactosamine to the nonreducing terminus of a hexasaccharide derived from chondroitin 4-sulfate whereas fibroblast extracts from patients with Tay-Sachs and Sandhoff-Jatzkewitz diseases did not. The results suggest that N-acetyl-beta-hexosaminidase A is responsible for degradation of the oligosaccharide substrate.

Control of chondrogenic expression in mesodermal cells of embryonic chick limb.

Chick-limb mesodermal cells have been maintained in tissue culture under conditions that permit development of muscle and cartilage. 3-Acetylpyridine, a nicotinamide-antagonized muscle teratogen, potentiates chondrogenic expression in cell cultures, as evidenced by histological and biochemical criteria, including the production of chondromucoprotein. Xylosyltransferase and N-acetylgalactosaminyltransferase are two enzymes required for chondromucoprotein synthesis; the specific activities of these enzymes were measured in differentiating mesodermal cells cultured for various periods of time in the presence and absence of 3-acetylpyridine and in intact limb tissue.