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Journal of Lipid Research, Vol. 48, 417-424, February 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

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 Sonnino¹

Department of Medical Chemistry, Biochemistry, and Biotechnology, Center of Excellence on Neurodegenerative Diseases, University of Milan, 20090 Segrate, Italy

Published, JLR Papers in Press, November 8, 2006.

¹ To whom correspondence should be addressed. e-mail: sandro.sonnino{at}unimi.it

The ceramide (Cer) and sphingomyelin (SM) species of cultured differentiated rat cerebellar granule cells and human fibroblasts were characterized by electrospray ionization-mass spectrometry. We identified 35 different species of Cer and 18 species of SM in human fibroblasts, and 35 different species of Cer and 9 species of SM were characterized in rat neurons. The main Cer species of rat cerebellar granule cells contained d18:1 sphingosine linked with palmitic, stearic, or nervonic fatty acid, and the two main SM 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 fractions (DSF) from these cells. In human fibroblasts, the main Cer 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; the most represented species of SM were d18:1,16:0, d18:1,24:0, and d18:1,24:1. In these cells, SM was highly enriched in DRMs and Cer was mainly associated with DSF, and the species found in DRMs were markedly different from those found in DSF.

Supplementary key words molecular species • lipidomics • mass spectrometry • lipid rafts

Abbreviations: Cer, ceramide (N-acyl-sphingosine); DRM, detergent-resistant membrane; DSF, detergent-soluble fraction; ESI, electrospray ionization; SM, sphingomyelin (ceramide phosphocholine)

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