Effects of ceramide-1-phosphate on cultured cells: dependence on dodecane in the vehicle.
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 A(2alpha). Much of the present knowledge has relied on the use of C1P of various acyl chain lengths, 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 prostaglandin E2 release, is fully dependent upon addition of a premade 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 in unraveling C1P signaling, which is still poorly understood.
Tauzin
Loïc
L
Novartis Institutes for BioMedical Research, A-1235 Vienna, Austria.
Graf
Christine
C
Sun
Mei
M
Rovina
Philipp
P
Bouveyron
Nicolas
N
Jaritz
Markus
M
Winiski
Anthony
A
Hartmann
Nicole
N
Staedtler
Frank
F
Billich
Andreas
A
Baumruker
Thomas
T
Zhang
Mei
M
Bornancin
Frédéric
F
D000473:Alkanes / Q000494:pharmacology*; D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D002518:Ceramides / Q000494:pharmacology*; D002522:Chlorocebus aethiops; D017931:DNA Primers; D000431:Ethanol / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D008933:Mitochondrial Swelling / Q000187:drug effects*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
D000473:Alkanes; D002518:Ceramides; D017931:DNA Primers; C065576:ceramide 1-phosphate; C007548:n-dodecane; D000431:Ethanol