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Published-Ahead-of-Print August 21, 2008, DOI:10.2164/jandrol.107.004333
Journal of Andrology, Vol. 30, No. 1, January/February 2009
Copyright © American Society of Andrology
DOI: 10.2164/jandrol.107.004333

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

OMER VARISLI*, CEVDET UGUZ{dagger}, CANSU AGCA{ddagger} AND YUKSEL AGCA{ddagger}

From the * Department of Animal Reproduction and Artificial Insemination, School of Veterinary Medicine, University of Ankara, Ankara, Turkey; the {dagger} Department of Medical Biology and Genetics, School of Veterinary Medicine, Afyon Kocatepe University, Afyon, Turkey; and the {ddagger} Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri, Columbia, Missouri.

Correspondence to: Dr Yuksel Agca, College of Veterinary Medicine, University of Missouri, 1600 East Rollins Road, Room W191, Columbia, MO 65211 (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 and acrosomal and membrane integrity were compared after exposing sperm samples to 200, 400, 600, and 800 x g centrifugal forces for 5, 10, or 15 minutes. In experiment 2, sperm motility and acrosomal and membrane integrity were compared after passing them through a Percoll separation using centrifugal forces of 600, 800, 1000, and 1200 x g for either 15 or 30 minutes. In experiment 3, the effect of repeated pipetting (2, 4, 6, 8, and 10 times) on motility and membrane integrity of rat sperm was compared with that on 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 < .05). The acrosomal integrity of SD rat sperm was affected after using 800 x g centrifugation force for 10 or 15 minutes (P < .05), whereas F-344 rat sperm acrosomal integrity was not affected by any centrifugation regimes (P > .05). Sperm from SD rats also had higher motility and membrane integrity loss than did sperm from F-344 rats after centrifugation and pipetting (P < .05). Percoll gradient separation did not cause significant motility loss or acrosomal damage to either F-344 or SD sperm (P > .05). Repeated pipetting had a dramatic adverse effect on both rat and mouse sperm motility (P < .05) as compared with sperm from bull, boar, and ram, which were not affected at all (P > .05). These data suggest that rat sperm have unique properties that need to be considered during centrifugation, Percoll gradient separation, and pipetting procedures.

     Key words: Centrifugation, Percoll gradient, pipetting






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