Glycosphingolipids are key components of eukaryotic cellular membranes. Through their propensity to form lipid rafts, they are important in membrane transport and signaling. At the cell surface, they are required for caveolar-mediated endocytosis, a process required for the action of many glycosphingolipid-binding toxins. Glycosphingolipids also exist intracellularly, on both leaflets of organelle membranes. It is expected that dissecting the mechanisms of cell pathology seen in the glycosphingolipid storage diseases, where lysosomal glycosphingolipid degradation is defective, will reveal their functions. Disrupted cation gradients in Mucolipidosis type IV disease are interlinked with glycosphingolipid storage, defective rab 7 function, and the activation of autophagy. Relationships between drug translocators and glycosphingolipid synthesis are also discussed. Mass spectrometry of cell lines defective in drug transporters reveal clear differences in glycosphingolipid mass and fatty acid composition. The potential roles of glycosphingolipids in lipid raft formation, endocytosis, and cationic gradients are discussed.