Biosynthesis of gangliosides in cultured retina from chick embryos.

Abstract

Retina tissue from 7-day chick embryos was maintained in culture for up to 10 days. After 5 days in culture the incorporation of [3H]leucine into proteins and of [3H]glucosamine into gangliosides was similar to that found in retinas from 12-day embryos. The incorporation of [3H]thymidine into DNA decreased steadily with time in culture; after 5 days it was about 20% of the initial value and approximately twice that determined in retinas from 12-day embryos. The radioactivity pattern of gangliosides labeled with [3H]glucosamine showed a predominance of the label in disialosyllactosylceramide (GD3); up to the 3rd day of culture. From then on, there was a progressive increase in the labeling of disialosylgangliotetraosylceramide (GD1a); by day 7 of culture, labeling of GD1a predominated and the labeling pattern was indistinguishable from that found in retinas from 12-day-old embryos. The specific activities of the CMP-NeuAc:GM3 sialosyl- and UDP-GalNAc:GM3 N-acetylgalactosaminyl-transferases decreased to 15% and increased to 400%, respectively, of the values determined in the retinas of 7-day embryos. The cultured retinas progressed in their organization into layers with culture time. The labeling transition from GD3 to GD1a was also detected after inhibition of the histotypic organization by addition of 5-bromo 2-deoxyuridine to the culture medium. Results suggest that high activity of GM3:sialosyl transferase and high labeling of GD3 are associated with the proliferative state of retina cells, while high activity of GM3:N-acetylgalactosaminyltransferase and high labeling of GD1a are associated with the non-proliferative, differentiated state of these cells.