A novel fluorescent ceramide analogue for studying membrane traffic in animal cells: accumulation at the Golgi apparatus results in altered spectral properties of the sphingolipid precursor.

摘要: A series of ceramide analogues bearing the fluorophore boron dipyrromethene difluoride (BODIPY) were synthesized and evaluated as vital stains for Golgi apparatus, tools studying lipid traffic between apparatus plasma membrane living cells. Studies spectral properties several BODIPY-labeled ceramides in vesicles demonstrated that fluorescence emission maxima strongly dependent upon molar density probes membrane. This was especially evident using N-[5-(5,7-dimethyl BODIPY)-1-pentanoyl]-D-erythro-sphingosine (C5-DMB-Cer), which exhibited a shift its maximum from green (integral 515 nm) to red 620 wavelengths with increasing concentrations. When C5-DMB-Cer used label cells, this property allowed us differentiate membranes containing high concentrations fluorescent metabolites (the corresponding sphingomyelin glucosylceramide) other regions cell where smaller amounts probe present. Using approach, prominent labeling apparatus-associated tubulovesicular processes, putative transport seen human skin fibroblasts, well types. Based on ratio imaging microscopy, we estimate present at up 5-10 mol %. In addition, concentration-dependent monitor C5-DMB-lipids surface 37 degrees C.