文章基本信息

标题：Effects of ceramide-1-phosphate on cultured cells: dependence on dodecane in the vehicle
本地全文：下载
作者：Tauzin, Loic ; Graf, Christine ; Sun, Mei 等
期刊名称：JLR Papers In Press
印刷版ISSN：0022-2275
电子版ISSN：1539-7262
出版年度：2007
卷号：48
期号：1
页码：66-76
DOI：10.1194/jlr.M600399-JLR200
出版社：American Society for Biochemistry and Molecular Biology
摘要：Ceramide-1-phosphate (C1P), the product of ceramide kinase,is a sphingophospholipid with recently recognized signalingproperties. In particular, it was reported to be mitogenic andcapable of direct stimulation of cytosolic phospholipase A₂.Much of the present knowledge has relied on the use of C1P ofvarious acyl chain lengths, together with diverse protocolsto 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 ofC1P has not been documented. Here, we show that addition ofC1P in ethanol-dodecane to culture medium leads to irreversiblecytotoxic effects. These culminate in mitochondrial swelling,vacuole formation, and cell death. Not only the toxicity ofC1P, 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 toC1P. They result from the capacity of dodecane to interact withphospholipids; hence, they go undetected with a vehicle control.This study should raise awareness about the use of dodecanefor phospholipid delivery and, in turn, help in unraveling C1Psignaling, which is still poorly understood. Supplementary key words ceramide kinase • sphingolipid • mitochondria • viability • phosphatidic acid • phospholipase A₂ • prostaglandin E₂ Abbreviations: ATCC, American Type Culture Collection; C1P, ceramide-1-phosphate; cPLA₂, cytosolic phospholipase A₂; ER, endoplasmic reticulum; LAMP2, lysosome-associated membrane protein 2; NBD-Cer, nitrobenzo-2-oxa-1,3-diazole-labeled C6-ceramide; PA, phosphatidic acid; PG, phosphatidylglycerol; PGE₂, prostaglandin E₂; YFP, yellow fluorescent protein