Journal of Andrology
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Published-Ahead-of-Print August 21, 2008, DOI:10.2164/jandrol.107.004333

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Effects of Various Physical Stress Factors on Rat Sperm Motility, Acrosome and Plasma Membrane Integrity

Omer Varisli , Cevdet Uguz , Cansu Agca , and Yuksel Agca *

* To whom correspondence should be addressed. E-mail: agcay{at}missouri.edu.

The objective of this study was to determine the effects of various physical interventions such as centrifugation regimes, Percoll gradient separation, and repeated pipetting on various viability parameters of epididymal sperm of Fischer 344 (F-344) and Sprague Dawley (SD) rat strains. Three experiments were conducted. In experiment 1, sperm motility, acrosomal and membrane integrity were compared after exposing sperm samples to 200, 400, 600, and 800xg centrifugal forces for 5, 10 or 15 min. In experiment 2, sperm motility, acrosomal and membrane integrity were compared after passing them through a Percoll separation using centrifugal forces of 600, 800, 1000, and 1200xg for either 15 or 30 min. In experiment 3, effect of repeated pipetting (2, 4, 6, 8, and 10 times) on motility and membrane integrity of rat sperm was compared to that of mouse, ram, bull and boar sperm. The results revealed that both F-344 and SD rat sperm motility and membrane integrity were significantly affected by centrifugation (P<0.05). The acrosomal integrity of SD rat sperm was affected after using 800xg centrifugation force for 10 or 15 min (P<0.05), while F-344 rat sperm was not affected by any centrifugation regimes (P>0.05). However, sperm from SD rats had higher motility and membrane integrity loss than those F-344 rats after centrifugation and pipetting (P<0.05). Percoll gradient separation did not cause significant motility loss or acrosomal damage to either F-344 or SD sperm (P>0.05). Repeated pipetting had dramatic adverse affect on both rat and mouse sperm motility (P<0.05) as compared to sperm from bull, boar and ram, which were not affected at all (P>0.05). These data suggest that rat sperm have unique properties which need to be considered during centrifugation, Percoll gradient separation, and pipeting procedures.


Key words: Epididymis • Semen Analysis • Percoll gradient • centrifugation • pipeting • rat sperm






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