The lipid composition of sperm membranes is crucial for fertilization and differs among species. As the evolution of internal fertilization modes in fishes is not understood, a comparative study of the sperm lipid composition in freshwater representatives of externally and internally fertilizing fishes is needed for a better understanding of taxa-specific relationships between the lipid composition of the sperm membrane and the sperm physiology. The lipidomes of spermatozoa from stingray, a representative of cartilaginous fishes possessing internal fertilization, and sterlet, a representative of chondrostean fishes with external fertilization, have been studied by means of nuclear magnetic resonance (NMR), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), electrospray MS, gas chromatography-(GC) MS, and thin-layer chromatography (TLC). NMR experiments revealed higher cholesterol content and the presence of phosphatidylserine in stingray compared to sterlet sperm. Unknown MS signals could be assigned to different glycosphingolipids in sterlet (neutral glycosphingolipid Gal-Cer(d18:1/16:0)) and stingray (acidic glycosphingolipid sulpho-Gal-Cer(d18:1/16:0)). Free fatty acids in sterlet sperm indicate internal energy storage. GC-MS experiments indicated a significant amount of adrenic acid, but only a low amount of docosahexaenoic acid in stingray sperm. In a nutshell, this study provides novel data on sperm lipid composition for freshwater stingray and sterlet possessing different modes of fertilization.

Department of Biology and Zootechny Ilha Solteira Faculty of Engineering São Paulo State University Neotropical Ichthyology Laboratory LINEO Monção Street 226 15385 000 Ilha Solteira SP Brazil

Faculty of Fisheries and Protection of Waters University of South Bohemia in Ceske Budejovice South Bohemian Research Centre for Aquaculture and Biodiversity of Hydrocenoses Zátiší 728 2 38925 Vodňany Czech Republic

Institute for Medical Physics and Biophysics Medical Faculty University of Leipzig Härtelstr 16 18 04107 Leipzig Germany

Leibniz Institute for Farm Animal Biology Institute of Muscle Biology and Growth Lipid Metabolism and Muscular Adaptation Workgroup Wilhelm Stahl Allee 2 18196 Dummerstorf Germany

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