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Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm

机译：冷冻保存对牛精子活力，活性氧合成和DNA损伤的影响

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The objective was to examine if there are relationships between alterations in sperm viability, reactive oxygen species (ROS) synthesis, and DNA integrity induced by cryopreservation of bovine sperm. Four ejaculates were collected from each of six bulls. Each ejaculate was diluted and divided into two aliquots; one was incubated for 24 hours at 37 degrees C, and the other frozen, thawed, and incubated for 24 hours at 37 degrees C. Analyses of quality of sperm were performed after 0, 3, 6, 12, and 24 hours of incubation. Progressive motile sperm was determined with computer assisted sperm analysis. Percentages of plasma membrane- and acrosome-intact sperm, sperm with a high mitochondrial membrane potential, sperm showing a high degree of DNA fragmentation (%DFI), and their reactive oxygen species content were assessed with dichlorofluorescein-diacetate, dihydrorhodamine, diaminofluorescein diacetate, and mitochondrial superoxide indicator using flow cytometry. Although all other sperm parameters showed alterations (P 0.05) during the 24-hour incubation time, %DFI stayed constant (P > 0.05, 0.91 +/- 0.23) in nonfrozen sperm. Cryopreservation induced changes of all sperm parameters (P 0.05). In contrast to all other sperm parameters, dichlorofluorescein-diacetate-fluoroescence indicating the synthesis of H2O2 showed a similar exponential rise (P 0.05) like the %DFI values in frozen sperm. In conclusion, changes of DNA integrity in frozen sperm seem to be related to synthesis of H2O2 but not to sperm viability and synthesis of other reactive oxygen species. (c) 2016 Elsevier Inc. All rights reserved.
机译：目的是检查在精子的活力，活性氧（ROS）合成和冷冻保存的牛精子诱导的DNA完整性之间是否存在关系。从六头公牛的每只中收集了四个射精。将每个射精稀释并分成两等分;将一个在37℃下孵育24小时，将另一个冷冻，解冻并在37℃下孵育24小时。在0、3、6、12和24小时的孵育之后进行精子质量的分析。用计算机辅助精子分析确定进行性运动精子。用二氯荧光素-二乙酸盐，二氢若丹明，二氨基荧光素-二乙酸盐，流式细胞仪检测线粒体超氧化物指示剂。尽管所有其他精子参数在24小时温育时间内均显示出变化（P <0.05），但非冷冻精子中的％DFI保持恒定（P> 0.05，0.91 +/- 0.23）。冷冻保存引起所有精子参数的变化（P <0.05）。与所有其他精子参数相反，表明过氧化氢合成的二氯荧光素-二乙酸酯-荧光显示了相似的指数上升（P <0.05），类似于冷冻精子中的％DFI值。总之，冷冻精子中DNA完整性的变化似乎与H2O2的合成有关，但与精子的活力和其他活性氧的合成无关。 （c）2016 Elsevier Inc.保留所有权利。

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