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A more recent version of this article appeared on February 1, 2007

Papers In Press, published online ahead of print November 8, 2006
J. Lipid Res., doi:10.1194/jlr.M600344-JLR200
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Submitted on July 26, 2006
Revised on November 6, 2006
Accepted on November 7, 2006

Ceramide and sphingomyelin species of fibroblasts and neurons in culture

Manuela Valsecchi, Laura Mauri, Riccardo Casellato, Simona Prioni, Nicoletta Loberto, Alessandro Prinetti, Vanna Chigorno, and Sandro Sonnino

Medical Chemistry, Biochemistry and Biotechnology, University of Milan, Segrate, Milano 20090

Corresponding Author: sandro.sonnino{at}unimi.it

The ceramide and sphingomyelin species of cultured differentiated rat cerebellar granule cells and human fibroblasts were characterised by electron spray ionization mass spectrometry (ESI-MS). We identified 35 different species of ceramide and 18 of sphingomyelin in human fibroblasts and 35 different species of ceramide and 9 of sphingomyelin were characterized in rat neurons . The main ceramide species of rat cerebellar granule cells contained d18:1 sphingosine linked with palmitic, stearic or nervonic fatty acid and the two main sphingomyelin species were d18:1,16:0 and d18:1,18:0. Both sphingolipids were enriched in detergent resistant membranes (DRMs or lipid rafts), and significant differences were found in the sphingolipid patterns of DRMs and of detergent soluble fraction (DSF) from these cells. In human fibroblasts the main ceramide species were d18:1,16:0, d18:2,16:0, d18:1,24:0, d18:2,24:0, d18:1,24:1 and d18:2,24:1; and the most represented species of sphingomyelin were d18:1,16:0, d18:1,24:0 and d18:1,24:1. In these cells, sphingomyelin was highly enriched in DRMs, instead ceramide was mainly associated with the detergent soluble fraction and the species found in DRMs were markedly different from those of DSF.


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