Inhibition of GCS-derived ganglioside biosynthesis results in increased insulin sensitivity in hypothalamic cells. Inhibition of GCS-derived ganglioside.

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Volume 68, Issue 4, Pages (October 2005)

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Inhibition of GCS-derived ganglioside biosynthesis results in increased insulin sensitivity in hypothalamic cells. Inhibition of GCS-derived ganglioside biosynthesis results in increased insulin sensitivity in hypothalamic cells. A: In a hypothalamic cell line, GCS is inhibited by treatment with the ceramide analog Genz. Genz treatment inhibits subsequent ganglioside biosynthesis. B: Thin-layer chromatography shows that the adult hypothalamic cell line (A2–12 cells) expresses the more complex neuronal gangliosides GM1 and GD1a, besides GM3. Treatment with Genz (5 µmol/L, 7 days) inhibits ganglioside biosynthesis. C and D: Western blot from A2–12 cell lysates indicates that IR levels are increased upon Genz treatment (n = 4). E and F: Hypothalamic cells were stimulated with 10 nmol/L insulin for the indicated time points. Western blot analysis for tyrosine phosphorylation of immunoprecipitated IR indicates increased IR phosphorylation in Genz-treated neurons (n = 5). The blot showing IR was obtained after stripping the membrane from p-Tyr antibody. G: When normalized to precipitated IRs, tyrosine phosphorylation is still significantly increased in Genz-treated neurons 3 min after stimulation (n = 5). Data are reported as the mean ± SEM. *P ≤ 0.05, **P ≤ 0.01. p-Tyr, tyrosine phosphorylation; UDP, uridine diphosphate. Silke Herzer et al. Diabetes 2015;64:3363-3376 ©2015 by American Diabetes Association