In the present work, taurine and hypotaurine were evaluated as potential additives to improve European sea bass (Dicentrarchus labrax) sperm quality after cryopreservation. For cryopreservation, three different extenders were used: control extender (NAM), supplemented with 1mM taurine or supplemented with 1mM hypotaurine, all of them containing 10% Me₂SO as cryoprotectant. To evaluate sperm quality of fresh and thawed sperm, motility (CASA: computer assisted sperm analysis), viability (SYBR Green/propidium iodide), lipid peroxidation (malondialdehyde level), protein oxidation (carbonyl content), glutathione peroxidase, glutathione reductase and superoxide dismutase activities and DNA fragmentation (comet assay) were quantified. The result demonstrated that 1 mM hypotaurine supplemented extender increased total motility (30.1 ± 3.2%), and that 1 mM taurine extender produced higher velocity (18.1 ± 2.6 μm/s) and linearity (46.0 ± 4.8%) than the control extender (21.8 ± 3.2%, 15.5 ± 1.3 μm/s, 41.8 ± 2.4%, respectively). Cell viability, lipid peroxidation and protein oxidation were not statistically different between treatments. Similar results were obtained for glutathione peroxidase and superoxide dismutase activities. Only glutathione reductase showed differential activity before and after freezing, increasing its activity in thawed sperm. Regarding the comet assay results, taurine and hypotaurine significantly reduced DNA fragmentation (52.8 ± 0.9% and 51.8 ± 0.9%, respectively) in comparison to the control (55.7 ± 0.8%). In conclusion, for European sea bass sperm cryopreservation, extenders supplemented with 1 mM taurine and 1 mM hypotaurine improved some parameters of sperm quality after thawing, resulting in better motility and lower DNA damage than the control, two very important factors related to fertilization success.

译文

在目前的工作中,牛磺酸和次牛磺酸被评估为潜在的添加剂,可在冷冻保存后提高欧洲鲈鱼 (Dicentrarchus labrax) 的精子质量。对于冷冻保存,使用三种不同的增量剂: 补充1mm牛磺酸或补充1mm次牛磺酸的对照增量剂 (NAM),所有这些都含有10% me2 so作为冷冻保护剂。为了评估新鲜和解冻的精子质量,运动能力 (CASA: 计算机辅助精子分析),活力 (SYBR Green/碘化丙啶),脂质过氧化 (丙二醛水平),蛋白质氧化 (羰基含量),谷胱甘肽过氧化物酶,定量了谷胱甘肽还原酶和超氧化物歧化酶活性和DNA片段化 (彗星试验)。结果表明,添加1毫米的次牛磺酸增量剂可提高总运动性 (30.1 ± 3.2%),而1毫米的牛磺酸增量剂产生的速度 (18.1 ± 2.6 μ m/s) 和线性度 (46.0 ± 4.8%) 高于对照增量剂 (21.8 ± 3.2%,15.5 ± 1.3 μ m/s,分别为41.8 ± 2.4%)。处理之间的细胞活力,脂质过氧化和蛋白质氧化无统计学差异。谷胱甘肽过氧化物酶和超氧化物歧化酶活性获得了相似的结果。冷冻前后只有谷胱甘肽还原酶表现出不同的活性,从而增加了其在解冻精子中的活性。关于彗星测定结果,与对照 (55.7 ± 0.8%) 相比,牛磺酸和次牛磺酸显着减少了DNA片段化 (分别为52.8 ± 0.9% 和51.8 ± 0.9%)。综上所述,对于欧洲鲈鱼精子冷冻保存,补充1毫米牛磺酸和1毫米次牛磺酸的增量剂可改善解冻后精子质量的某些参数,从而比对照组具有更好的运动性和更低的DNA损伤,这两个非常重要的因素与受精成功有关。

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