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J. Lipid Res.
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A more recent version of this article appeared on September 1, 2007

Papers In Press, published online ahead of print June 7, 2007
J. Lipid Res., doi:10.1194/jlr.M700069-JLR200
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Submitted on February 7, 2007
Revised on June 7, 2007
Accepted on June 7, 2007

Neutral sphingomyelinase-induced ceramide triggers germinal vesicle breakdown and oxidant-dependent apoptosis in Xenopus laevis oocytes

Olga Coll, Albert Morales, José C. Fernández-Checa, and Carmen Garcia-Ruiz

Liver Unit, Hospital Clinic, IDIBAPS-CSIC, Barcelona 08036

Corresponding Author: checa229{at}yahoo.com

Ceramide regulates many cellular processes, including cell growth, differentiation and apoptosis. Although the effects of exogenous bacterial neutral sphingomyelinase in Xenopus laevis oocytes have been investigated, its microinjection into oocytes has not been previously reported. Thus, we compared the incubation versus microinjection of the neutral Bacillus cereus sphingomyelinase (bSMase) to examine if the topology of ceramide generation determines its effects on the fate of oocytes. In agreement with previous findings, incubation of mature stage VI oocytes with bSMase increased ceramide levels in oocyte extracts over time causing the germinal vesicle breakdown indicative of maturation, without evidence of cytotoxicity. In contrast, bSMase microinjection which increased ceramide levels in a time and dose-dependent fashion resulted in oocyte apoptosis characterized by reactive oxygen species (ROS) generation, glutathione (GSH) depletion in cytosol and mitochondria, release of cytochrome c and Smac/Diablo from mitochondria, and caspase-3 activation. Microinjection of acidic SMase from human placenta recapitulated the apoptotic effects of bSMase microinjection. Preincubation of oocytes with reduced GSH-ethyl ester prior to bSMase microinjection, prevented ROS generation and mitochondrial dowstream events, thus protecting oocytes from bSMase-induced death. These findings show a divergent action of bSMase-induced ceramide on oocyte maturation or apoptosis depending on the intracellular site where ceramide is generated.


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D. W. Petcoff, W. L. Holland, and B. J. Stith
Lipid levels in sperm, eggs, and during fertilization in Xenopus laevis
J. Lipid Res., November 1, 2008; 49(11): 2365 - 2378.
[Abstract] [Full Text] [PDF]


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