Glycosphingolipids (GSLs), consisting of hydrophilic oligosaccharides and hydrophobic ceramide, are crucial components of plasma membranes and play key roles in fundamental biological processes. In particular, sialo glycosphingolipids (gangliosides containing one or more sialic acid residues) are involved in cell-cell recognition and adhesion and signal transduction. For example, gangliosides are involved in LPS induced macrophage activation, which further involved in inflammation and production of cytokines. Gangliosides are mostly within specific cell surface microdomains, lipid rafts, and glycosphingolipid-enriched microdomains. In this report, we profiled major gangliosides (GM1, GM3, GD3) expression in cell surface, cell membrane, lipid rafts and their changes in THP-1 macrophages upon LPS stimulation using HPLC-fluorescence method. Briefly, we found that GM3, GM1 and GD3 decreased upon LPS stimulation overall. In cell lysate, GM3 and GM1 shown maximum decrease at 24 hrs, GD3 shown maximum decrease at 6 hrs than other time intervals and shows slight increase at 48 with LPS stimulation. In cell membrane, GM3, GD3 shown decrease maximum with LPS stimulation at 6hr than other time interval, while GM1 decrease was significant at 24 hr with LPS stimulation. In lipid rafts, the maximum decrease was observed in GM1, then GM3. Least decease was observed in GD3 with LPS stimulation for 24 hrs. On cell surface using confocal microscopy, there was decrease in ganglioside GM1, GM3, while least decrease was seen in GD3 with LPS stimulation. Overall, we have seen that ganglioside profiles were changed with LPS stimulation at different time interval and different cellular locations, indicating they may play different roles in receptor signaling regulation and pathogenesis.

Profiling ganglioside expression and change in THP-1 macrophages upon LPS stimulation

ACS Spring Meeting 2021

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