Developmental changes in gangliosides in cultured cerebellar granule neurons.

Abstract

The content and composition of gangliosides in cultures enriched in granule neurones and in astrocytes from rat cerebellum (P6-8) showed marked differences: astrocytes contained less than 10% of the amount of granule neurones and the profile was dominated by simple gangliosides with lactosyl ceramide backbone, while gangliosides of the 'b' series, which constitute about 40% in nerve cells, were virtually undetectable. Granule cell maturation was accompanied by a 16-fold increase in the ganglioside content during the initial 8 days in a serum-supplemented medium (S+), reaching a plateau much earlier and at a higher level than observed in the cerebellum in vivo. Developmental changes were characterized, as in vivo, by a pronounced decrease in the GD3 proportion and an increase in the 'b' series of gangliosides. Compared with S+, adhesion among cells and fibres is different in a serum-free medium (S-), in which the rise in cellular ganglioside content was less (30%), but the developmental changes in ganglioside profile were similar. However, in cultures in S- only, GM3 was not detectable, while the distribution of GM1 and GD3 indicated that maturation is retarded relative to cells in S+. Surface exposure of gangliosides (studied by the periodate/[3H]borohydride method) was similar under the two culture conditions. There was an initial delay, especially in S-, in the insertion of gangliosides into the plasma membrane, while the labelling of GD3 (the dominant ganglioside of immature granule cells) was very low compared with all the other species throughout the whole cultivation time.