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Published-Ahead-of-Print June 28, 2006, DOI:10.2164/jandrol.106.000109
Journal of Andrology, Vol. 27, No. 6, November/December 2006
Copyright © American Society of Andrology
DOI: 10.2164/jandrol.106.000109

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Comparative Study of Four Different Sperm Washing Methods Using Apoptotic Markers in Ram Spermatozoa

ELENA MARTÍ, ROSAURA PÉREZ-PÉ, TERESA MUIñO-BLANCO* AND JOSÉ A. CEBRIÁN-PÉREZ*

From the Department of Biochemistry and Molecular and Cell Biology, School of Veterinary Medicine, University of Zaragoza, Zaragoza, Spain.

Correspondence to: Dr J.A. Cebrián-Pérez, Departamento de Bioquímica y Biología Molecular y Celular, Facultad de Veterinaria, C/Miguel Servet, 177. 50013-Zaragoza. Spain (e-mail: pcebrian{at}unizar.es).


The accurate measurement of semen fertilizing potential is of great importance in determining the acceptability of processed semen for breeding purposes. A good sperm preparation technique results in a sample with high viability and motility and also takes into account other parameters such as the capacitation and apoptotic state which could compromise the ability to fertilize an oocyte. In this study, we investigate the effects of 4 sperm preparation techniques (a dextran/swim-up procedure, discontinuous Percoll density gradient centrifugation, sucrose washing, and filtration) on ram sperm quality parameters. Besides the evaluation of viability and the capacitation state, we also analyzed the apoptotic status of the sperm samples by assessing the phosphatidylserine translocation and caspase-3 and -7 activities. This is the first report, to our knowledge, that evidences the presence of active caspases in ram sperm. The results confirm the better ability of the dextran/swim-up procedure to select nonapoptotic spermatozoa, in addition to viable and noncapacitated sperm, compared with other sperm preparation methods. This should be considered to enhance results of artificial insemination techniques in ovine reproduction.

     Key words: Swim-up, caspase activity, annexin V




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