Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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BOR - Papers in Press, published online ahead of print November 3, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.036095
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Submitted September 7, 2004
Returned for revision September 30, 2004
Accepted October 15, 2004

Gamete Biology


Percoll-Gradient-Centrifuged Capacitated Mouse Sperm Have Increased Fertilizing Ability and Higher Contents of Sulfogalactosylglycerolipid and Docosahexaenoic Acid-Containing Phosphatidylcholine Compared to Washed Capacitated Mouse Sperm

Anna Furimsky , Ngoc Vuong , Hongbin Xu , Premkumari Kumarathasan , Min Xu , Wattana Weerachatyanukul , Maroun Bou Khalil , Morris Kates , and Nongnuj Tanphaichitr *

* To whom correspondence should be addressed. E-mail: ntanphaichitr{at}ohri.ca.

Abstract
Although Percoll gradient centrifugation has been routinely used to prepare motile human sperm, its use in preparing motile mouse sperm is limited. Here, we reported that Percoll gradient centrifuged (PGC) capacitated mouse sperm had markedly higher fertilizing ability (sperm-zona pellucida (ZP) binding and in vitro fertilization) than washed capacitated mouse sperm. We also showed that the lipid profiles of PGC capacitated sperm and washed capacitated sperm differed significantly. PGC sperm had much lower contents of cholesterol and phospholipids. This resulted in relative enrichment of male germ cell specific sulfogalactosylglycerolipid (SGG), a ZP binding ligand, in PGC capacitated sperm, and this would partly explain their increased ZP binding ability, compared with washed capacitated sperm. Analyses of phospholipid fatty acyl chains revealed that PGC capacitated sperm were enriched in phosphatidylcholine (PC) molecular species containing highly unsaturated fatty acids (HUFAs) with docosahexaenoic acid (DHA, C22:6n-3) being the predominant HUFA (42% of total hydrocarbon chains of PC). In contrast, the level of PC-HUFAs (comprising arachidonic acid (20:4n-6), docosapentaenoic acid (C22:5n-6) and DHA in washed capacitated sperm was only 27%. Having the highest unsaturation degree among all HUFAs in PC, DHA would enhance membrane fluidity to the uppermost. Therefore, membranes of PCG capacitated sperm would undergo fertilization-related fusion events at higher rates than washed capacitated sperm. These results suggested that PGC mouse sperm should be used in fertilization experiments and SGG and DHA should be considered as important biomarkers for sperm fertilizing ability.

Key words: Gamete Biology • Fertilization • In vitro fertilization • Sperm • Sperm capacitation


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