Effects of UV irradiation and hydrogen peroxide on DNA fragmentation, motility and fertilizing ability of rainbow trout (Oncorhynchus mykiss) spermatozoa
Effects of UV irradiation and hydrogen peroxide on DNA fragmentation, motility and fertilizing ability of rainbow trout (Oncorhynchus mykiss) spermatozoa
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DOI：
10.1016/j.theriogenology.2005.04.010
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发表时间：
2005-11-01
期刊：
影响因子：
2.8
通讯作者：
Ciereszko, A
中科院分区：
文献类型：
--
作者：
Dietrich, GJ;Szpyrka, A;Ciereszko, A
保持 DNA 完整性对于保护精子质量至关重要。本研究使用彗星测定法检查了虹鳟鱼（Oncorhynchus mykiss）kiss）精子在受到紫外线照射（2075 mu W/cm(2)，0-15 分钟）或过氧化氢（0-20 mM）诱导的氧化应激时的 DNA 碎片。还测量了精子活力和受精能力。紫外线照射 5 分钟后，DNA 碎片显着增加，但此时没有观察到精子活力的显着变化。较长时间的照射导致运动参数降低，DNA 碎片进一步增加。紫外线照射导致有眼胚胎的百分比明显下降，并且大多数胚胎没有孵化。当高度稀释的精子悬浮液（50,000 倍）暴露于 0.1 mM H2O2 时，观察到 DNA 碎片明显增加。另一方面，当使用更浓缩的精子悬浮液（仅稀释 40 倍）时（为了同时进行活力和受精测量）1-20 mM H2O2 仅导致 DNA 碎片适度增加以及精子活力和受精能力的剂量依赖性下降。这表明 H2O2 的毒性作用主要与抑制精子活力有关。我们的结果表明，彗星试验可用于监测紫外线照射对鱼精子 DNA 灭活的有效性。因此，彗星试验与精子运动分析一起可以应用于鱼类雌核发育过程的优化工作。 H2O2 在诱导主要 DNA 片段化方面缺乏有效性，表明虹鳟鱼精子中存在抗氧化防御机制。 (c) 2005 Elsevier Inc. 保留所有权利。
Preservation of DNA integrity is essential for protection of sperm quality. This study examined, with the use of comet assay, DNA fragmentation of rainbow trout (Oncorhynchus mykiss)kiss) spermatozoa subjected to UV irradiation (2075 mu W/cm(2), 0-15 min) or oxidative stress induced by hydrogen peroxide (0-20 mM). Sperm motility and fertilizing ability were also measured. A dramatic increase in DNA fragmentation was recorded after 5 min UV irradiation but no significant changes in sperm motility were observed at this time. Longer irradiation resulted in a decrease in motility parameters and further increase of DNA fragmentation. UV irradiation caused a clear decrease in the percentage of eyed embryos and most of the embryos did not hatch. When highly diluted sperm suspensions (50,000-fold) were exposured to 0.1 mM H2O2 evident increase in DNA fragmentation was observed. On the other hand, when more concentrated sperm suspensions (diluted only 40-fold) were employed (in order to conduct motility and fertilization measurements at the same time) 1-20 mM H2O2 caused only moderate increase in DNA fragmentation and dose-dependent decline in sperm motility and fertilizing ability. This suggests that toxic effects of H2O2 were primarily related to inhibition of sperm motility. Our results demonstrate that comet assay can be used for monitoring the effectiveness of fish sperm DNA inactivation by UV irradiation. Therefore, the comet assay together with sperm motility analysis can be applied in optimization works of gynogenetic procedures in fish. Lack of effectiveness of H2O2 in inducing major DNA fragmentation suggests presence of mechanisms of antioxidative defense in rainbow trout spermatozoa. (c) 2005 Elsevier Inc. All rights reserved.