牦牛鲜精与冻精HSP70表达差异比较研究

doi:10.11843/j.issn.0366-6964.2018.11.024

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (11): 2514-2520.doi: 10.11843/j.issn.0366-6964.2018.11.024

牦牛鲜精与冻精HSP70表达差异比较研究

唐朋^1,2, 凌笑笑¹, 贾聪俊¹, 梁春年¹, 吴晓云¹, 王宏博¹, 褚敏¹, 阎萍^1*

1. 中国农业科学院兰州畜牧与兽药研究所, 甘肃省牦牛繁育重点实验室, 兰州 730050;
2. 甘肃农业大学动物科技学院, 兰州 730050

收稿日期:2018-01-25 出版日期:2018-11-23 发布日期:2018-11-23
通讯作者: 阎萍,研究员,主要从事动物遗传育种与繁殖研究,E-mail:pingyanlz@163.com.
作者简介:唐朋(1988-),男,陕西宝鸡人,硕士生,主要从事动物遗传育种与繁殖研究,E-mail:844768241@qq.com
基金资助:
现代肉牛牦牛产业技术体系（CARS-37）；中国农业科学院创新工程项目（CAAS-ASTIP-2014-LIHPS-01）

Study on the Differences of HSP70 Expression in Fresh Sperm and Frozen-thawed Sperm in Yak

TANG Peng^1,2, LING Xiao-xiao¹, JIA Cong-jun¹, LIANG Chun-nian¹, WU Xiao-yun¹, WANG Hong-bo¹, CHU Min¹, YAN Ping^1*

1. Key Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China;
2. College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730050, China

Received:2018-01-25 Online:2018-11-23 Published:2018-11-23

摘要/Abstract

摘要：

旨在研究冷冻前后牦牛精子质量和HSP70表达变化。采集健康且繁殖能力正常的6头大通牦牛精液，将试验分为鲜精组和冻精组，以精子活力和质膜完整率作为评价精子质量的指标。采用实时荧光定量PCR、Western blot和免疫荧光技术检测HSP70在精子冷冻前后的表达差异。结果表明，与鲜精相比，冻融后精子活力和质膜完整率均极显著下降（P<0.01），HSP70 mRNA转录水平显著降低（P<0.05），蛋白水平极显著下降（P<0.01）。鲜精HSP70蛋白定位于精子顶体区、顶体赤道段、后顶体区和精子中段，而冻融精子HSP70蛋白主要位于后顶体区和精子中段，且平均光密度值极显著降低（P<0.01）。综上表明，冻融后牦牛精子质量与HSP70表达量均降低，推测两者之间存在一定关联。

Abstract:

The aims of this study were to investigate changes of sperm quality and heat shock protein(HSP)70 expression before and after freezing yak semen. The semen of 6 healthy Datong yaks with normal reproductive ability was collected and assigned to 2 groups:fresh group and frozen-thawed group. The quality of sperm was evaluated by assessing sperm motility and plasma membrane integrity. The expression of HSP70 in both groups was detected by the real-time quantitative polymerase chain reaction,Western blot and immunofluorescence techniques. The results showed that,compared with fresh sperm,sperm motility and plasma membrane integrity of frozen-thawed sperm were significantly decreased (P<0.01); In addition,the expression of HSP70 was significantly lower at both the transcriptional (P<0.05) and protein (P<0.01) levels. HSP70 protein was distributed over the sperm acrosome,acrosome equatorial segment,post-acrosomal and middle sections of sperm in the fresh group. In contrast,after freezing and thawing,HSP70 protein was mainly located in post-acrosomal and middle sections of sperm,and the average optical density was significantly decreased(P<0.01). In conclusion,yak sperm quality and the expression of HSP70 declined simultaneously after freezing and thawing,suggesting that they may be associated.

中图分类号:

S823.8⁺5.2

唐朋, 凌笑笑, 贾聪俊, 梁春年, 吴晓云, 王宏博, 褚敏, 阎萍. 牦牛鲜精与冻精HSP70表达差异比较研究[J]. 畜牧兽医学报, 2018, 49(11): 2514-2520.

TANG Peng, LING Xiao-xiao, JIA Cong-jun, LIANG Chun-nian, WU Xiao-yun, WANG Hong-bo, CHU Min, YAN Ping. Study on the Differences of HSP70 Expression in Fresh Sperm and Frozen-thawed Sperm in Yak[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(11): 2514-2520.

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牦牛鲜精与冻精HSP70表达差异比较研究

Study on the Differences of HSP70 Expression in Fresh Sperm and Frozen-thawed Sperm in Yak

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