Seminal plasma represents an important and active contributor to mammalian fertility. Specific components of seminal plasma are adsorbed onto the surface of ejaculated spermatozoa while they pass through the male reproductive tract and provide sperm cells with the ability to reach and fertilize the oocyte. Among the components of seminal plasma, there are enzymatic and non-enzymatic antioxidants that modulate the production of reactive oxygen species (ROS), which are by-products generated during mitochondrial electron transport. Oxidative stress, which is induced by an incorrect balance between antioxidants and ROS production, impairs sperm function. In this context, seminal plasma represents the most important defense system able to prevent the damages that these species exert upon sperm cells. In spite of this, removal of seminal plasma is usually recommended prior to sperm storage, cooled or cryopreserved, as the presence of this fluid may be detrimental for sperm longevity. Against this background, the main aim of this Dissertation was to determine the relevance of the antioxidant enzymes present in seminal plasma for equine reproduction strategies. On the one hand, and as a first objective of this work, we observed much higher activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GSR) in jackass seminal plasma than in that of stallions. Since equids are seasonal breeders, this study also investigated season influence on the antioxidant composition of seminal plasma. Interestingly, activities of SOD and GPX in seminal plasma showed seasonal variations in jackasses but not in stallions. On the other hand, because the demand of cryopreserved semen in the equine industry is increasing and no study had previously addressed whether the activities of these four enzymes are related to sperm cryotolerance, the second and third study evaluated the relationship between SOD, CAT, GPX and GSR activities in fresh seminal plasma and the sperm ability to withstand cryopreservation in these two species. It was found that the specific SOD activity in seminal plasma is related to sperm cryotolerance in both stallions and jackasses. Since seminal plasma has been reported to modulate ROS balance in several mammalian species, this Dissertation also investigated how the presence of seminal plasma modulates ROS generation by jackass spermatozoa under oxidative stress induced exogenously by hydrogen peroxide. With regard to these results, however, prolonged contact of seminal plasma was found to have limited effect against oxidative stress induced exogenously to jackass spermatozoa. Finally, sperm selection is getting prominent in equine assisted reproduction technologies and also leads to the removal of seminal plasma components. The last objective of this study aimed to investigate the effects of single layer centrifugation (SLC) with Equicoll on jackass sperm parameters and its interactions with the female reproductive tract. While SLC-Equicoll was found to increase the percentages of viable spermatozoa after selection, the impact on sperm motility parameters was marginal. In addition, removal of seminal plasma components through SLC-Equicoll increased sperm phagocytosis by polymorphonuclear neutrophils (PMNs), thereby confirming that seminal plasma modulates the interaction between sperm and PMNs. In conclusion, this Dissertation indicates that although the activities of seminal plasma antioxidants differ between stallions and jackasses, SOD appears to be a good predictor of sperm cryotolerance in these two species. In addition, this Dissertation has also demonstrated, for the first time, that the modulating role of seminal plasma with regard to the interaction between jackass spermatozoa and PMNs is more apparent than that observed when oxidative stress is induced by hydrogen peroxide.
|Date of Award||13 Dec 2019|
|Supervisor||Jordi Miro Roig (Director) & Marc Yeste Oliveras (Director)|