PMID: 8464873Apr 1, 1993Paper

Cholesterol deprivation affects the fluorescence properties of a ceramide analog at the Golgi apparatus of living cells

Proceedings of the National Academy of Sciences of the United States of America
O C MartinR E Pagano

Abstract

Previous studies have established that a fluorescent analog of ceramide, N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)] -6-aminohexanoyl-D-erythro-sphingosine (C6-NBD-Cer), is a vital stain for the Golgi apparatus and a useful tool for studying the sorting and transport of sphingolipids along the secretory pathway in animal cells. Here, we examine the effects of various culture conditions on labeling of the Golgi apparatus of human skin fibroblasts by C6-NBD-Cer and demonstrate that cholesterol deprivation affects the fluorescence properties of the probe at this organelle. Labeling of the Golgi apparatus by C6-NBD-Cer was dramatically reduced in cells grown in medium containing lipoprotein-deficient serum compared to cells grown in medium containing normal serum. Quantitative fluorescence microscopy showed that this apparent reduction in labeling resulted from accelerated photo-bleaching of the fluorescent analog. C6-NBD-Cer labeling of the Golgi apparatus was restored in cholesterol-deprived cells by stimulating endogenous cholesterol biosynthesis with mevalonic acid or by adding exogenous nonlipoprotein cholesterol or low density lipoprotein to the culture medium. In addition, when cells grown in medium containing normal serum were p...Continue Reading

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Citations

Aug 2, 2007·Bioscience Reports·Gerald Gimpl, Katja Gehrig-Burger
May 21, 2005·Methods : a Companion to Methods in Enzymology·David L MarksRichard E Pagano
Dec 24, 2002·Trends in Cell Biology·David L Marks, Richard E Pagano
Feb 5, 2003·The International Journal of Biochemistry & Cell Biology·Alexander D DergunovGerard Siest
Apr 27, 2004·Molecular Biology of the Cell·Deepak K SharmaRichard E Pagano
Aug 5, 2004·Molecular Biology of the Cell·Amit ChoudhuryRichard E Pagano
Jul 31, 2014·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Daniel WüstnerDaniel Sage
Apr 29, 1998·The Journal of Cell Biology·P Keller, K Simons
Jan 8, 2016·Progress in Lipid Research·Mélanie CarquinDonatienne Tyteca
Nov 16, 2010·Steroids·Gerald Gimpl, Katja Gehrig-Burger
Mar 20, 2016·Biochimica Et Biophysica Acta·Takuma KishimotoToshihide Kobayashi
Jan 30, 2008·Current Protocols in Cell Biology·Raman Deep SinghRichard E Pagano
Jul 24, 2007·American Journal of Physiology. Lung Cellular and Molecular Physiology·Sanda A PredescuAsrar B Malik
Jun 2, 1995·The Journal of Biological Chemistry·C S ChenR E Pagano
Aug 30, 1996·The Journal of Biological Chemistry·E B NeufeldE J Blanchette-Mackie
Jun 20, 2006·Molecular and Cellular Biology·Paola GiussaniSarah Spiegel
Jul 20, 1999·The Journal of Biological Chemistry·R LiS Ladisch
Mar 27, 2003·The Journal of Biological Chemistry·Vishwajeet PuriRichard E Pagano

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