Fraser L, Parda A, Filipowicz K, Strzeżek J
Department of Animal Biochemistry and Biotechnology, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
Reprod Domest Anim. 2010 Oct;45(5):e155-60. doi: 10.1111/j.1439-0531.2009.01537.x.
In this study, we tested the hypothesis whether the neutral Comet assay (NCA) and the Sperm-Sus-Halomax (SSH) test kit could provide similar measurements of post-thaw DNA fragmentation of boar spermatozoa. Whole ejaculates or sperm-rich fractions of boar semen were frozen in an extender containing lactose, lipoprotein fractions isolated from ostrich egg yolk (LPFo), glycerol (lactose-LPFo-G) or in a standard boar semen extender (K3), without the addition of cryoprotective substances. In all boars, both the NCA and SSH test showed similar levels of post-thaw sperm DNA fragmentation in samples of the same ejaculates, regardless of the ejaculate collection procedure and extender. Yet, the levels of post-thaw sperm DNA damage, detected by the NCA and SSH test, were more accentuated in spermatozoa frozen in the absence of cryoprotective substances. Both the NCA and SSH detected variations among individual boars in terms of post-thaw sperm DNA fragmentation. Agreement between the measurements of the NCA and SSH was confirmed by scatter plots of differences, suggesting that the DNA integrity tests could detect the same sperm populations, which were susceptible to cryo-induced DNA damage. The findings of this study indicate that the NCA and the SSH test are effective in detecting similar levels of sperm DNA fragmentation and reinforce their importance in the assessment of frozen-thawed boar semen quality.
在本研究中，我们验证了以下假设：中性彗星试验（NCA）和精子悬浮-光晕最大法（SSH）检测试剂盒是否能够提供相似的公猪精子解冻后DNA碎片化测量结果。将公猪全份精液或富含精子的部分精液，在含有乳糖、从鸵鸟蛋黄中分离的脂蛋白组分（LPFo）、甘油的稀释液（乳糖-LPFo-G）中冷冻，或在不添加冷冻保护剂的标准公猪精液稀释液（K3）中冷冻。在所有公猪中，无论射精采集程序和稀释液如何，同一射精样本经解冻后，NCA和SSH检测均显示出相似水平的精子DNA碎片化。然而，在未添加冷冻保护剂的情况下冷冻的精子中，NCA和SSH检测到的解冻后精子DNA损伤水平更为明显。NCA和SSH均检测到不同公猪解冻后精子DNA碎片化存在差异。差异散点图证实了NCA和SSH测量结果之间的一致性，这表明DNA完整性检测能够检测到相同的易受冷冻诱导DNA损伤的精子群体。本研究结果表明，NCA和SSH检测在检测相似水平的精子DNA碎片化方面是有效的，并强化了它们在评估冷冻解冻公猪精液质量中的重要性。
