维生素C对藏猪精液4 ℃保存及其精子全基因组DNA甲基化的影响

doi:10.11843/j.issn.0366-6964.2016.05.025

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (5): 1057-1061.doi: 10.11843/j.issn.0366-6964.2016.05.025

维生素C对藏猪精液4 ℃保存及其精子全基因组DNA甲基化的影响

任子利，赵彦玲，王建洲，李瑜鑫，商鹏，强巴央宗^*

（西藏农牧学院动物科学学院，林芝 860000）

收稿日期:2015-11-25 出版日期:2016-05-23 发布日期:2016-05-23
通讯作者: 强巴央宗，教授，E-mail：qbyz628@126.com
作者简介:任子利（1969-），男，河南卫辉人，副教授，博士，主要从事动物繁殖研究，E-mail：zlren0925@163.com, Tel：0894-5822924
基金资助:
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）

Effect of Vitamin C on Tibetan Boar Semen Preserved at 4 ℃ and Levels of the Sperm Global Genome DNA Methylation

REN Zi-li，ZHAO Yan-ling，WANG Jian-zhou，LI Yu-xin，SHANG Peng，CHAMBA Yangzom^*

（Faculty of Animal Science，Tibet Agricultural and Animal Husbandry College，Linzhi 860000，China）

Received:2015-11-25 Online:2016-05-23 Published:2016-05-23

摘要/Abstract

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、10.0 g•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5.0 g•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5.0 g•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5.0 g•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。

Abstract:

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 vitamin 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 MethylFlash^ＴＭ.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 semen and 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.

中图分类号:

任子利，赵彦玲，王建洲，李瑜鑫，商鹏，强巴央宗. 维生素C对藏猪精液4 ℃保存及其精子全基因组DNA甲基化的影响[J]. 畜牧兽医学报, 2016, 47(5): 1057-1061.

REN Zi-li，ZHAO Yan-ling，WANG Jian-zhou，LI Yu-xin，SHANG Peng，CHAMBA Yangzom. Effect of Vitamin C on Tibetan Boar Semen Preserved at 4 ℃ and Levels of the Sperm Global Genome DNA Methylation[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(5): 1057-1061.

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维生素C对藏猪精液4 ℃保存及其精子全基因组DNA甲基化的影响

Effect of Vitamin C on Tibetan Boar Semen Preserved at 4 ℃ and Levels of the Sperm Global Genome DNA Methylation

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