These authors contributed equally to this article and share senior coauthorship.
Comparative Study of Four Different Sperm Washing Methods Using Apoptotic Markers in Ram Spermatozoa
Article first published online: 2 JAN 2013
2006 American Society of Andrology
Journal of Andrology
Volume 27, Issue 6, pages 746–753, November-December 2006
How to Cite
Martí, E., Pérez-Pé, R., Muiño-Blanco, T. and Cebrián-Pérez, J. A. (2006), Comparative Study of Four Different Sperm Washing Methods Using Apoptotic Markers in Ram Spermatozoa. Journal of Andrology, 27: 746–753. doi: 10.2164/jandrol.106.000109
Supported by grants CICYT-FEDER AGL 2004–02882, INIA RZ03–035, CICYT-FEDER AGL 2005–02614, and DGA A-26/2005.
- Issue published online: 2 JAN 2013
- Article first published online: 2 JAN 2013
- Received for Publication March 13, 2006; accepted for Publication June 18, 2006
- caspase activity;
- annexin V
ABSTRACT: 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.