Figure 3. Cellular gangliosides treatment.

CGCs were incubated with different gangliosides (GM3, GM1 or GD1a) and correspondent radiolabelled gangliosides ([³H]GM3, [³H]GM1 or [³H]GD1a), at a final concentration of 2×10⁻⁶ M at 37°C for 4 h. At the end of incubation, the ganglioside solution was removed and cells were washed 3 times with Locke's solution and maintained at 37°C for 20 min in 3 mL of FBS-BME. The lipids extract, from cell homogenates, were analysed to determine the ganglioside incorporation (panel A) and metabolism by HPTLC following radioactivity imaging (panel B). Lane 1: granule cell ganglioside pattern; lane 2: granule cells ganglioside extracted after incubation with GM3/[³H]GM3 2×10⁻⁶ M at 37°C for 4 h; lane 3: granule cells ganglioside extracted after incubation with GM1/[³H]GM1 2×10⁻⁶ M at 37°C for 4 h; lane 4: granule cells ganglioside extracted after incubation with GD1a/[³H]GD1a 2×10⁻⁶ M at 37°C for 4 h; lane 5: [³H]GM3 standard; lane 6. [³H]GM1 and [³H]GD1a standards. * Gangliosides endogenous content as reported by Palestini et al., 1991. [40]