ICMAN-Institute of Marine Science of Andalusia, Spanish National Research Council, Av. Republica Saharaui 2, 11510 Puerto Real, Cádiz, Spain.
Anim Reprod Sci. 2011 May;125(1-4):189-95. doi: 10.1016/j.anireprosci.2011.03.003. Epub 2011 Mar 15.
During cryopreservation, dilution in the extender media reduces the seminal plasma constituents being cells more vulnerable to oxidative stress. Vitamins (C and E) and the amino acids taurine and hypotaurine are powerful antioxidants naturally present in seminal plasma. Whether their effect may improve sperm quality and reduce sperm DNA damage after cryopreservation in fish sperm still remains unclear. Thus, the aim of the present work was to analyse the effect of extender supplementation with several antioxidant components on post-thawed sperm motility, viability and DNA integrity of two commercial species, gilthead seabream (Sparus aurata) and European seabass (Dicentrarchus labrax). Sperm collected from ten to twelve individuals was cryopreserved in ten different extenders containing: taurine and hypotaurine (1 and 10mM), ascorbic acid (1 and 10mM), α-tocoferol (0.1 and 0.5 mM) or 1 ml/l of a commercial cell antioxidant supplement. Cell viability, motility and DNA fragmentation were determined in post-thawed samples. Addition of antioxidants (vitamins and amino acids) to D. labrax and S. aurata extenders did not significantly increase the parameters of motility (TM, PM, VCL, VSL and Lin) or viability, although 1mM taurine slightly increased the percentage of motile cells (TM) in S. aurata. DNA fragmentation (DNA in tail and Olive tail moment) in D. labrax sperm was higher in treatments containing vitamins than amino acids or control. However in S. aurata sperm, antioxidants especially taurine and hypotaurine, significantly reduced both DNA fragmentation parameters, protecting DNA against strand breaks. These results suggest a species-specific effect depending on the type of antioxidants used.
在冷冻保存过程中，稀释 extender 介质会降低精液中细胞成分对氧化应激的敏感性。维生素（C 和 E）以及氨基酸牛磺酸和次牛磺酸是精液中天然存在的强效抗氧化剂。它们的作用是否可以改善鱼类精子冷冻保存后的精子质量和减少 DNA 损伤仍不清楚。因此，本研究的目的是分析在几种抗氧化成分的添加对两种商业鱼类（真鲷 Sparus aurata 和欧洲鲈鱼 Dicentrarchus labrax）解冻后精子活力、活力和 DNA 完整性的影响。从 10 到 12 个个体中采集的精子在 10 种不同的 extender 中冷冻保存，这些 extender 含有：牛磺酸和次牛磺酸（1 和 10mM）、抗坏血酸（1 和 10mM）、α-生育酚（0.1 和 0.5mM）或 1 毫升/升商业细胞抗氧化补充剂。在解冻后的样品中测定细胞活力、活力和 DNA 片段化。在 D. labrax 和 S. aurata 的 extender 中添加抗氧化剂（维生素和氨基酸）并没有显著增加运动参数（TM、PM、VCL、VSL 和 Lin）或活力，尽管 1mM 牛磺酸略微增加了 S. aurata 中运动细胞的百分比（TM）。D. labrax 精子中的 DNA 片段化（尾部 DNA 和 Olive 尾部矩）在含有维生素的处理中高于含有氨基酸或对照的处理。然而，在 S. aurata 精子中，抗氧化剂特别是牛磺酸和次牛磺酸，显著降低了这两个 DNA 片段化参数，保护了 DNA 免受链断裂。这些结果表明，根据使用的抗氧化剂类型，存在特定于物种的效应。
