Journal of Andrology
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Published-Ahead-of-Print June 28, 2006, DOI:10.2164/jandrol.106.000539

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Immunolocalization and Possible Functional Role of PSP-I/PSP-II Heterodimer in Highly-Extended Boar Spermatozoa

Ignacio Caballero , Juan M Vazquez *, Eva M García , Jordi Roca , Emilio Martinez , Juan J Calvete , Libia Sanz , Hans Ekwall , and Heriberto Rodriguez-Martínez

* To whom correspondence should be addressed. E-mail: vazquez{at}um.es.

PSP-I/PSP-II heterodimer is a major protein of boar seminal plasma which is able to preserve, in vitro, the viability, motility and mitochondrial activity of highly-extended boar spermatozoa for at least 5 hours. However, little is known about the binding pattern of the heterodimer to the sperm plasma membrane and its eventual relation with the maintenance of the sperm functionality. The present study investigated the effect of exposing highly-extended boar spermatozoa (1 million/mL) to 1.5 mg/mL of PSP-I/PSP-II for 0.5, 5 and 10 hours at 38°C on sperm characteristics and the changes in PSP-I/PSP-II localization as a result of both the addition of PSP-I/PSP-II to the extender and the incubation time. Exposure of the spermatozoa to PSP-I/PSP-II preserved sperm viability, motility and mitochondrial activity when compared to non-exposed spermatozoa. This protective effect lasted for 10 hours (p<0.05). After immunolabelling of highly-extended semen with rabbit monospecific polyclonal antibody against PSP-I/PSP-II, the percentage of immunopositive spermatozoa declines over time from 71% (0.5 h) to 49% (10 h). However, more than 80% of spermatozoa remained labelled during the 10h-incubation period if PSP-I/PSP-II was added. Scanning electron microscopy revealed 4 different binding patterns. The heterodimer was mainly localized to the acrosomal area, being redistributed to the post-acrosomal area or lost during in vitro incubation. In conclusion, the protective effect of the heterodimer appears to be related to its adhesion to the acrosomal area, and the loss of this protective effect coincides with a stepwise redistribution of PSP-I/PSP-II during incubation.


Key words: Reproductive Tract • Semen • Sperm • seminal plasma • sperm capacitation






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Copyright © 2006 by The American Society of Andrology.