Journal of Andrology
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Published-Ahead-of-Print October 31, 2007, DOI:10.2164/jandrol.107.003640

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Detection of Apoptosis-Like Changes During the Cryopreservation Process in Equine Sperm

C. Ortega-Ferrusola , Y. Sotillo-Galán , E. Varela-Fernández , J. M. Gallardo-Bolaños , A. Muriel , L. González-Fernández , J. A. Tapia , and F. J. Peña *

* To whom correspondence should be addressed. E-mail: fjuanpvega{at}unex.es.

The kinematic of the apparition of apoptotic markers was studied flow cytometrically and by inmunoblot assays in equine sperm subjected to freezing and thawing. Caspase activity, low mitochondrial membrane potential and increases in sperm membrane permeability were observed in all the phases of the cryopreservation procedure. Freezing and thawing caused an increase in membrane permeability, and changes in the pattern of caspase activity; while decreases in mitochondrial membrane potential were observed after centrifugation and cooling to 4°C and after freezing and thawing. It is proposed that the sperm mitochondria may be directly involved is the subtle damage that is present in most of the spermatozoa surviving freezing and thawing.


Key words: Semen • Semen Analysis • Sperm • flow cytometry






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