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A more recent version of this article appeared on January 1, 2007
Papers In Press, published online ahead of print October 3, 2006
J. Lipid Res., doi:10.1194/jlr.M600399-JLR200
Submitted on September 6, 2006
Revised on September 29, 2006
Accepted on October 3, 2006
Effects of ceramide-1-phosphate on cultured cells: dependence on dodecane in the vehicle
Loic Tauzin, Christine Graf, Mei Sun, Philipp Rovina, Nicolas Bouveyron, Markus Jaritz, Anthony Winiski, Nicole Hartmann, Frank Staedtler, Andreas Billich, Thomas Baumruker, Mei Zhang, and Frederic Bornancin
Novartis Institutes for BioMedical Research, Vienna
Corresponding Author: frederic.bornancin{at}novartis.com
Ceramide-1-phosphate (C1P), the product of ceramide kinase, is a sphingophospholipid with recently recognized signaling properties. In particular, it was reported to be mitogenic and capable of direct stimulation of cytosolic phospholipase A2 . Much of the present knowledge has relied on the use of C1P of various acyl chain length, together with diverse protocols to deliver it to cultured cells. A mixture of ethanol (or methanol) with dodecane, as the vehicle, has become popular. However, the contribution of this solvent to the observed effects of C1P has not been documented. Here we show that, addition of C1P in ethanol/dodecane to culture medium, leads to irreversible cytotoxic effects. These culminate in mitochondrial swelling, vacuole formation and cell death. Not only the toxicity of C1P, but also its ability to trigger PGE2 release, are fully dependent upon addition of a pre-made C1P-dodecane mixture. Furthermore, we show that these effects are not restricted to C1P. They result from the capacity of dodecane to interact with phospholipids; hence, they go undetected with a vehicle control. This study should raise awareness about the use of dodecane for phospholipid delivery and in turn, help unraveling C1P signaling, which is still poorly understood.

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Copyright © 2006 by the American Society for Biochemistry and Molecular Biology.
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