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Lipid peroxidation, assessed with BODIPY-C₁₁, increases after cryopreservation of stallion spermatozoa, is stallion-dependent and is related to apoptotic-like changes

¹ Section of Reproduction and Obstetrics, Laboratory of Spermatology, Department of Herd Health and Medicine, Faculty of Veterinary Medicine, Veterinary Teaching Hospital² Department of Physiology, Faculty of Veterinary Medicine, University of Extremadura, Avda de la Universidad s/n, 10071 Cáceres, Spain³ Division of Reproduction, Faculty of Veterinary Medicine and Animal Sciences, Swedish University of Agricultural Sciences, SE-750 07 Uppsala, Sweden

Correspondence should be addressed to F J Peña; Email: fjuanpvega{at}unex.es

Lipid peroxidation (LPO) of stallion spermatozoa was assessed in fresh semen and in samples of the same ejaculates after freezing and thawing. Particular attention was paid to individual differences in the susceptibility to LPO and its possible relationship with freezability. Innate levels of LPO were very low in fresh spermatozoa but increased after thawing, a change that was largely stallion-dependent. The level of LPO in fresh spermatozoa was not correlated with that of the thawed spermatozoa. Negative correlations existed between LPO and intact membranes post-thaw (r=–0.789, P<0.001), and also between LPO and spermatozoa with high mitochondrial membrane potential (m) post-thaw (r=–0.689, P<0.001). LPO was also highly and significantly correlated with caspase activity. The correlation between caspase activity in ethidium positive cells and LPO was r=0.772, P<0.001. This LPO is unlikely to represent, per se, a sign of cryopreservation-induced injury, but it is apparently capable of triggering ‘apoptotic-like changes’ that could result in the sub-lethal cryodamage often seen among surviving spermatozoa.