摘要
旨在研究维生素C对藏猪精液4℃保存效果及其精子全基因组DNA甲基化的影响。用手握法采集健康藏猪精液,在改进的猪精液低温稀释液中添加不同浓度的维生素C(0.0、2.5、5.0、7.5、10.0g·L^(-1)),4℃保存到第5天时,以藏猪精子活力、精子畸形率、精子顶体完整率和质膜完整率为评价指标,探讨维生素C对藏猪精液4℃保存的效果;用甲基化荧光定量法检测藏猪新鲜精液组、保存到第5天时未添加维生素C组、添加维生素C组(添加维生素C最佳效果组5.0g·L^(-1))精液DNA甲基化的水平,以探讨维生素C对藏猪精液4℃保存所造成的精子全基因组DNA甲基化的影响。结果显示:添加5.0g·L^(-1)维生素C组其精子活力、畸形率、顶体完整率和质膜完整率均显著好于其它组(P<0.05);鲜精组、未添加维生素C组、添加维生素C组的DNA甲基化水平分别为(0.604 7±0.040 3)、(0.920 2±0.016 9)、(0.656 9±0.048 3)ng,鲜精组与添加组均显著低于未添加组(P<0.05),而鲜精组与添加组间差异不显著(P>0.05)。综上表明,在改进的猪精液稀释液中添加5.0g·L^(-1)维生素C可以明显改善藏猪精液4℃保存的效果,并且可以明显降低4℃保存对藏猪精液精子全基因组DNA甲基化的影响,这将为藏猪精液低温保存的生产应用奠定基础。
This research was conducted to study the effect of vitamin C on Tibetan boar semen preserved at 4℃and the subsequent influence on global genome DNA methylation. The semen was collected from Tibetan boar and preserved at 4 ℃ in modified medium supplemented with vi- tamin C at 0.0,2. 5,5. 0,7. 5,10. 0 g · L-1. Sperm motility, abnormality, acrosome integrity and plasma membrane integrity were evaluated 5 days after preservation. And the global methylation level of the fresh and preserved sperm was detected by using MethylFlashTM. The results showed that a significantly higher sperm motility and plasma membrane integrity, acrosome integrity and less abnormality was observed in group supplemented with 5.0 g· L-1 vitamin C(P〈0.05). DNA methylation levels of fresh sperm (0. 604 7±0. 040 3) ng and sperm preserved with vitamin C (0. 656 9 ±0. 048 3) ng were significantly lower than those preserved without vitamin C (0. 920 2±0. 016 9) ng (P〈0.05),but no significant difference was found between fresh semenand sperm preserved with vitamin C(P〈0.05). These results indicated that supplementation of vitamin C at 5.0 g ·L-1 semen extender could improve quality of Tibetan boar semen preservation at 4 ℃ and mitigate effect on DNA methylation of Tibetan boar sperm. The results in the current study would be helpful for further investigation of the freezing protocol for Tibetan boar semen.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2016年第5期1057-1061,共5页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
2014年畜牧资源平台建设
2013年畜牧体系研究团队
科技部支撑项目(2012BAD03B03)
西藏自治区自然科学基金资助项目(2015ZR-13-32)
关键词
维生素C
藏猪精液
4℃保存
DNA甲基化
vitamin C
Tibetan boar semen
preservation at 4 ℃
DNA methylation