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Copyright E American Society of Andrology Comparative Study of Four Different Sperm Washing Methods Using Apoptotic Markers in Ram Spermatozoa

By Elena Martí, Rosaura Pérez-pé, Teresa Muiño-blanco and José A. Cebrián-pérez


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/swimup 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

Topics: Key words, Swim-up, caspase activity, annexin V. J Androl 2006, 27, 746–753
Year: 2013
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