Uptake and metabolism of fluorescent ceramide analogs by rat oligodendrocytes in culture

FEBS Letters

Vos, Jan P.²  Marchesini, Sergio¹  Lopes-Cardozo, Matthijs²  Van Golde, Lambert M.G.²  Giudici, Maria L.¹ 

[1] Department of Biomedical Sciences and Biotechnology, University of Brescia, Italy;Laboratory of Veterinary Biochemistry, Utrecht University, Utrecht, The Netherlands

关键词: Ceramide metabolism;    Sphingomyclin;    Glycolipids of myelin;    N-lissamine rhodamine;    NBD;    Rat oligodendrocyte;    BSA;    Bovine serum albumin;    Cer;    Ceramides;    FA;    Fatty acid;    FITC;    Fluorescein isothiocyanate;    GalCer;    Galactosylcer-amides (galactocerebrosides);    GlcCer;    Glucosylceramides (glucocerebrosides);    HBSS;    Hanks;    balanced salt solution;    HPTLC;    High-performance thin-layer chromatography;    LRh;    N-lissamine rhodamine;    NBD;    N-(7-nitrobenz-2-oxa-1;    3-diazol-4-yl)-;    SGalCer;    Sulfogalactosylceramide (sulphatide);    SM;    Sphingomyelin;   

DOI  :  10.1016/0014-5793(92)81529-U

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We studied the metabolism of sphingolipids by oligodendrocytes derived from rat spinal cord by providing lipid vesicles with either N-lissamine-rhodaminyl-ceramide (LRh-Cer) or N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-ceramide (NBD-Cer) to the cells cultured in a chemically-defined medium. With both probes them the fluorescent product turned out to be sphingomyelin (SM). Most or LRh-SM was not cell-associated but recovered from the culture medium, probably due to back-exchange to the lipid vesicles. The accumulation of LRh-SM, both in the cells and in the medium, was inhibited in the presence of monensin or brefeldin A, whereas the production of NBD-SM was much less affected by these Golgi perturbing drugs. With LRh-Cer as substrate, LRh-labelled fatty acid (FA), galactosyl- and sulfogalactosyl-ceramides (GalCer and SGalCer) were also formed. NBD-Cer, however, was metabolized to glucosylecramide (GlcCer) and GalCer but not to SGalCer or NBD-FA. These data demonstrate that chemical modifications of ceramide alter its metabolism in oligodendrocytes and that the metabolites of LRh-Cer reflect the glycolipid composition of myelin more closely than those of NBD-Cer.

【 授权许可】

Unknown