J. Lipid Res.
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Originally published In Press as doi:10.1194/jlr.M800159-JLR200 on June 24, 2008

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Journal of Lipid Research, Vol. 49, 2365-2378, November 2008
Copyright © 2008 by American Society for Biochemistry and Molecular Biology

Lipid levels in sperm, eggs, and during fertilization in Xenopus laevis

Douglas W. Petcoff{dagger}, William L. Holland* and Bradley J. Stith1,*

* Department of Biology, University of Colorado Denver, Denver, CO
{dagger} Metropolitan State College, Denver, CO

Published, JLR Papers in Press, June 24, 2008.

This work has been supported by National Science Foundation Grant IBN-0110609, and University of Colorado Denver Downtown Campus Undergraduate Research Opportunity Program grants.

1 To whom correspondence should be addressed. e-mail: brad.stith{at}cudenver.edu

Critical developmental periods, such as fertilization, involve metabolic activation, membrane fusion events such as sperm-egg or plasma membrane-cortical granule merger, and production and hydrolysis of phospholipids. However, there has been no large-scale quantification of phospholipid changes during fertilization. Using an enzymatic assay, traditional FA analysis by TLC and gas chromatography, along with a new method of phospholipid measurement involving HPLC separation and evaporative light-scattering detection, we report lipid levels in eggs, sperm, and during fertilization in Xenopus laevis. Sperm were found to contain different amounts of phospholipids as compared with eggs. During fertilization, total phosphatidylinositol, lysophosphatidylcholine, sphingomyelin, and phosphatidylserine decreased, and ceramide increased, whereas there was no change in phosphatidylcholine, cardiolipin, or phosphatidylethanolamine. FA analysis of phospholipids found numerous changes during fertilization. Because there is an increase in sn-1,2-diacylglycerol at fertilization, the FAs associated with this increase and the source of the increase in this neutral lipid were examined. Finally, activation of phospholipase C, phospholipase D, phospholipase A2, autotoxin, and sphingomyelinase at fertilization is discussed.

Supplementary key words diacylglycerol • phospholipase C • phospholipase D • phospholipase A2 • autotaxin • sphingomyelinase

Abbreviations: CL, cardiolipin; DAG, diacylglycerol; DHA, docosahexaenoic acid; ELSD, evaporative light-scattering detection; EPA, eicosapentaenoic acid; IP3, inositol 1,4,5-trisphosphate; LPC, lysophosphatidylcholine; MBS, modified Barth's medium; MUFA, monounsaturated fatty acid; PA, phosphatidic acid; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PI, phosphatidylinositol; PIP2, phosphatidylinositol 4,5-bisphosphate; PKC, protein kinase C; PLD, phospholipase D; PS, phosphatidylserine; SM, sphingomyelin


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