Abstract
Cerebellar granule cells in culture were subjected to a pulse (0.5–4 h)-chase (0–4 h) of 10 −6 M [ 3H]ganglioside GM1 carrying the radioactive label at the level of NeuAc ([ 3H-NeuAc]GM1), Sph ([ 3H-Sph]GM1) or Gal ([ 3H-GallGMl) and the formed [ 3H]metabolites were determined. With all forms of [ 3H]GM1, there was formation of [ 3H]catabolites, including [ 3H]H 2O and [ 3H]biosynthetic products obtained by recycling of [ 3H]NeuAc, [ 3H]Sph and [ 3H]Gal released during intralysosomal ganglioside degradation (salvage processes). Much higher amounts of [ 3MH 2O were produced from [ 3H-Gal]GM1 than [ 3H-Sph]GM1 and [ 3H-NeuAc]GM1; conversely, more products from salvage processes (polysialogangliosides GD1a, GDlb, GTlb, O-acetylated GT1b, protein-bound radioactivity) were obtained with [ 3H-NeuAc]GM1; than the two other forms of [ 3H]GM1, Liberated [ 3H]NeuAc produced 10-fold less tritiated water and 10-fold higher salvage products than [ 3H]Gal. Using [ 3H-NeuAc]GM1; granule cells appeared to metabolize 7.7% of membrane-incorporated exogenous GM1 per hour with a high degree of NeuAc recycling and the calculated metabolic half-life was 6.5 h.
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