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Reactive oxygen species and equine sperm function.

机译：活性氧和马精子的功能。

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摘要

The use of cryopreserved equine spermatozoa has increased; however, the fertility of frozen semen is not comparable to fresh spermatozoa and there is a significant variation between stallions. An improved understanding of equine sperm physiology and the cell changes occurring during the freeze-thaw process is required. This dissertation was directed towards understanding the impact of oxidative stress on equine spermatozoa. Previous studies indicate that equine spermatozoa generate reactive oxygen species (ROS). In this study I have demonstrated that equine neutrophils are an additional source of ROS. The addition of activated equine neutrophils to equine spermatozoa in vitro resulted in a significant (P 0.01) increase in hydrogen peroxide generation and a significant (P 0.001) decrease in sperm motility that could be counteracted by the addition of the antioxidant, catalase.; The generation of ROS by equine spermatozoa suggests a physiological role for ROS in sperm function. I report that low-level generation of ROS resulted in a significant increase in tyrosine phosphorylation (P 0.005) and capacitation (P 0.0001) in equine spermatozoa, prevented by the addition of catalase; implicating hydrogen peroxide as the causative ROS. Therefore, ROS generation by equine spermatozoa may be involved in the signaling pathways underlying sperm capacitation. In contrast to the physiological effect of low-level ROS generation, high levels of ROS generation were associated with a significant (P 0.005) increase in DNA fragmentation of equine spermatozoa; counteracted by the addition of catalase or reduced glutathione. Cryopreservation of equine spermatozoa was also associated with a significant (P 0.01) increase in DNA fragmentation.; Equine seminal plasma contains a relatively high activity of enzyme scavengers; however, cryopreservation is associated with the removal of seminal plasma and consequently the predominant source of antioxidants protection. Since, the addition of antioxidants in vitro counteracted the detrimental effects of oxidative stress on equine spermatozoa, I investigated whether antioxidant addition to the cryopreservation extender could improve post-thaw sperm parameters. The results of this study show no improvement in post-thaw equine sperm motility, DNA fragmentation, viability, acrosomal integrity or mitochondrial membrane potential with the addition of enzyme scavengers or antioxidants. The role of oxidative stress in cryopreservation-induced damage to equine spermatozoa requires further investigation.
机译：冷冻保存的马精子的使用有所增加。然而，冷冻精液的育性不能与新鲜精子相提并论，种马之间存在显着差异。需要更好地了解马精子生理和冻融过程中发生的细胞变化。本文旨在了解氧化应激对马精子的影响。先前的研究表明，马精子会产生活性氧（ROS）。在这项研究中，我证明了马中性粒细胞是ROS的另一个来源。在体外向马精子中添加活化的马中性粒细胞会导致过氧化氢生成量显着（P <0.01）的增加和精子活动性的显着（P <0.001）的降低，这可以通过添加抗氧化剂，过氧化氢酶来抵消。 ;马精子产生的R​​OS表明ROS在精子功能中的生理作用。我报告说，ROS水平的降低导致马精子中酪氨酸磷酸化（P <0.005）和获能（P <0.0001）的显着增加，这可以通过添加过氧化氢酶来预防。暗示过氧化氢是致病性ROS。因此，马精子产生的R​​OS可能参与精子获能的信号通路。与低水平的ROS产生的生理作用相反，高水平的ROS产生与马精子的DNA片段显着增加（P <0.005）有关；通过添加过氧化氢酶或减少的谷胱甘肽来抵消。马精子的冷冻保存还与DNA片段的显着增加（P <0.01）有关。马精浆含有相对较高的酶清除剂活性。然而，冷冻保存与精浆的清除有关，因此是抗氧化剂保护的主要来源。由于在体外添加抗氧化剂可以抵消氧化应激对马精子的有害影响，因此我研究了将抗氧化剂添加到冷冻保存扩展剂中是否可以改善解冻后的精子参数。这项研究的结果表明，添加酶清除剂或抗氧化剂后，解冻后马的精子活力，DNA片段化，生存力，顶体完整性或线粒体膜电位均无改善。氧化应激在冷冻保存诱导的马精子损伤中的作用尚需进一步研究。

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  • 作者

    Baumber, Julie.;

  • 作者单位

    University of California, Davis.;

  • 授予单位 University of California, Davis.;
  • 学科 Biology Veterinary Science.; Agriculture Animal Pathology.; Biology Animal Physiology.
  • 学位 Ph.D.
  • 年度 2003
  • 页码 p.3133
  • 总页数 199
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 动物学;
  • 关键词

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