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 12, 2003.
Biol Reprod 2003, 10.1095/biolreprod.103.022996
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Submitted September 5, 2003
Returned for revision September 17, 2003
Accepted October 27, 2003

Gamete Biology


Low-Density Lipoprotein Fraction from Hen's Egg Yolk Decreases the Binding of the Major Proteins of Bovine Seminal Plasma to Sperm and Prevents Lipid Efflux from the Sperm Membrane

Annick Bergeron , Marie-Hélène Crête , Yves Brindle , and Puttaswamy Manjunath *

* To whom correspondence should be addressed. E-mail: manjunap{at}medclin.umontreal.ca.

Abstract
For sperm preservation, semen is generally diluted with extender containing egg yolk (EY) but the mechanisms of sperm protection by EY are unclear. The major proteins of bull seminal plasma (BSP proteins: BSP-A1/A2, BSP-A3 and BSP-30kDa) bind to sperm surface at ejaculation and stimulate cholesterol and phospholipid efflux from the sperm membrane. Since EY low-density lipoprotein fraction (LDF) interacts specifically with BSP proteins, it is proposed that the sequestration of BSP proteins in seminal plasma by EY-LDF represents the major mechanism of sperm protection by EY. In order to gain further insight into this mechanism, we investigated the effect of seminal plasma, EY and EY-LDF on the binding of BSP proteins to sperm and the lipid efflux from the sperm membrane. As shown by immunodetection, radioimmunoassays and lipid analysis, when semen was incubated undiluted or diluted with control extender (without EY or EY-LDF), BSP proteins bound to sperm in a time-dependent manner and there is a continuous cholesterol and phospholipid efflux from the sperm membrane. In contrast, when semen was diluted with extender containing EY or EY-LDF, there was 50-80% less BSP proteins associated with sperm and a significant amount of lipid added to sperm membrane during incubation. In addition, sperm functions analysis show that the presence of EY or EY-LDF in the extender preserved sperm motility. These results show that LDF is the constituent of EY that prevents binding of the BSP proteins to sperm, lipid efflux from the sperm membrane and is beneficial to sperm functions during sperm preservation.

Key words: Gamete Biology • Male Reproductive Tract • Assisted Reproductive Technology • Seminal vesicles • Sperm


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