Chigorno Vanna, Sciannamblo Mariateresa, Mikulak Joanna, Prinetti Alessandro, Sonnino Sandro
Center of Excellence on Neurodegenerative Diseases, Department of Medical Chemistry, Biochemistry and Biotechnology, University of Milan, 20090, Segrate, Italy.
Glycoconj J. 2006 May;23(3-4):159-65. doi: 10.1007/s10719-006-7921-7.
The membrane complex lipids of human fibroblasts and differentiated rat cerebellar granule cells in culture were metabolically radiolabeled with [1-(3)H]sphingosine, L-[3-(3)H]serine and [9,10-(3)H]palmitic acid. A relevant efflux of radioactive sphingolipids and phosphatidylcholine was observed from cells to the culture medium in the presence of fetal calf serum. This event was independent of the concentration and structure of the metabolic precursor administered to cells, and it was linearly time-dependent. The radioactive lipid patterns present in the medium were different from those present in the cells. Radioactive sphingomyelin and ganglioside GM3 containing short acyl chains were the main species present in the medium from human fibroblasts, while sphingomyelin and GD3 ganglioside in that from neuronal cells. In the absence of proteins in the culture medium, the efflux of complex lipids was much lower than in the presence of serum, and the patterns of released molecules were again different from those of cells.
用[1-(³H)]鞘氨醇、L-[3-(³H)]丝氨酸和[9,10-(³H)]棕榈酸对培养的人成纤维细胞和分化的大鼠小脑颗粒细胞的膜复合脂质进行代谢性放射性标记。在胎牛血清存在的情况下,观察到放射性鞘脂和磷脂酰胆碱从细胞向培养基的相关外流。该事件与给予细胞的代谢前体的浓度和结构无关,且呈线性时间依赖性。培养基中存在的放射性脂质模式与细胞中的不同。含短酰基链的放射性鞘磷脂和神经节苷脂GM3是来自人成纤维细胞的培养基中的主要成分,而来自神经元细胞的培养基中的主要成分是鞘磷脂和GD3神经节苷脂。在培养基中不存在蛋白质的情况下,复合脂质的外流比在血清存在时低得多,并且释放分子的模式也再次与细胞的不同。