Uptake and metabolism of a fluorescent sulfatide analogue in cultured skin fibroblasts

Monti, Eugenio; Preti, Augusto; Novati, Alberto; Aleo, Maria Francesca; Clemente, Maria Luisa; Marchesini, Sergio

doi:10.1016/0005-2760(92)90129-j

Cited by 24 publications

(32 citation statements)

References 23 publications

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“…We compared LRh and ETBD as fluorescent labels. LRh-sulphatide was used recently in metabolic studies with fibroblasts [19] and with oligodendrocytes [13], while NBD has been used extensively to study the metabolism, transport and sorting of (glyco)sphingolipids in a variety of cell types [l&20 -22] . Specifically, NBDCer has been shown to label the Golgi compartment of the cell [23].…”

Section: Results and Dlscussionmentioning

confidence: 99%

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

Giudici¹,

Vos²,

Marchesini³

et al. 1992

FEBS Letters

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We studied tbc metabolism of sphingolipids by oligodendrocytes derived from rat spinal cord by providing lipid vesicles with cithcr N-lissamina rhodaminyl-cerumidc (LRh-Cer) or N-(7-nitrobcnz-2-oxo-I,3-diazol-4-yl)-ccramide (NBD-Cer) to the cells cultured in a chemically-defined mcdium. With both probes the major fluorescent product turned out to be sphingamyelin (SM). Most of LRh-SM WPS not cell-associated but recovered from lhe 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 brefcldin 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). galxtosyl-and sulfogalactosyl-ceramidcs (GalCer and SGalCer) were also formed. NBD-Cer, however, was metabolized to glucosylccramide (GlcCcr) and GalCcr but not to SGalCcr or NBD-FA. These data demonstrate that chemical mudiIicaliull> ofcclntilide alter its metabolism in oligcdcndrocytcs and that the mctsbo!itef of LRh-Cer reflect thc&~caligidcomposition of myclin more closely than those of NBD-Cer.

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Section: Results and Dlscussionmentioning

confidence: 99%

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

Giudici¹,

Vos²,

Marchesini³

et al. 1992

FEBS Letters

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“…C~2-Lrh-Cer or C6-NBD-Cer were incorporated into small unilamellar egg-PC vesicles (SUV) [15]. Alternatively, labelled ceramides were complexed with BSA immediately before use [16]. Ethanol solutions of C~2-LRh-Cer were prepared by dissolving the required amount of dried C,2-LRh-Cer in 70% (v/v) ethanol to a final concentration of 100 raM.…”

Section: Introductionmentioning

confidence: 99%

Sphingomyelin is synthesized at the plasma membrane of oligodendrocytes and by purified myelin membranes: a study with fluorescent‐ and radio‐labelled ceramide analogues

et al. 1995

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In most cell types sphingomyelin is synthesized predominantly in the cis-medial compartments of the Golgi stacks whereas the contribution of the plasma membrane is much lower. The aim of this study was to assess the contribution of both compartments to the synthesis of sphingomyelin in myelinating cells. Therefore, oligodendroeytes from rat spinal cord were incubated in culture with fluorescently-or radiolabelled ceramides, and the effects of a block in the vesicular flow (monensin, brefeldin A, low temperature) on surface synthesis of sphingomyelin were evaluated. The results indicate that = 50% of the sphingomyelin synthase is present at the plasma and myelin membranes of oligodendrocytes.

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“…BSA complexes of LR-CTH were prepared by a modifi cation of the method of Monte ( 1 ). For use in culture, 100 nmoles of LR-CTH in chloroform-methanol was dried under nitrogen in a glass vial and resuspended in 25 l of DMSO or ethanol.…”

Section: Preparation Of Lr-cth/bsa Complexesmentioning

confidence: 99%

“…LR-CTH (MW = 1502) was custom synthesized by Matreya, LLC (Pleasant Gap, PA) according to the method of Monti ( 1 ). It was provided in chloroform-methanol solution at a concentration of 1 mg/ml.…”

Section: Preparation Of Substratementioning

confidence: 99%

Use of lissamine rhodamine ceramide trihexoside as a functional assay for alpha-galactosidase A in intact cells

Kaneski

Schiffmann²,

Brady

et al. 2010

Journal of Lipid Research

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Uptake and metabolism of a fluorescent sulfatide analogue in cultured skin fibroblasts

Cited by 24 publications

References 23 publications

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

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

Sphingomyelin is synthesized at the plasma membrane of oligodendrocytes and by purified myelin membranes: a study with fluorescent‐ and radio‐labelled ceramide analogues

Use of lissamine rhodamine ceramide trihexoside as a functional assay for alpha-galactosidase A in intact cells

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