《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (9): 2502-2507.doi: 10.16431/j.cnki.1671-7236.2016.09.041

• 经验交流 • 上一篇    下一篇

猪精子质量的流式细胞检测分析

吴梦1,2, 丁玉春1,3, 刘作华1,2,4, 刘文1,3, 罗林1, 葛良鹏1,2,4   

  1. 1. 重庆市畜牧科学院, 重庆 402460;
    2. 农业部养猪科学重点实验室, 重庆 402460;
    3. 农业部西南设施养殖观测试验站, 重庆 402460;
    4. 养猪科学重庆市市级重点实验室, 重庆 402460
  • 收稿日期:2016-01-28 出版日期:2016-09-20 发布日期:2016-09-20
  • 通讯作者: 葛良鹏 E-mail:geliangpeng1982@163.com
  • 作者简介:吴梦(1987-),男,重庆人,硕士,助理研究员,研究方向:生物工程,E-mail:404264245@qq.com
  • 基金资助:

    863项目(2014AA021602);重庆市国际合作项目(CSTC2013gjhz80002);重庆市基础与前沿研究重大项目(cstc2013jcyjC80001);重庆市农发资金项目(12402);重庆市基本科研业务费项目(15447)

Detection and Analysis of the Pig Sperm Quality by Flow Cytometry

WU Meng1,2, DING Yu-chun1,3, LIU Zuo-hua1,2,4, LIU Wen1,3, LUO Lin1, GE Liang-peng1,2,4   

  1. 1. Chongqing Academy of Animal Sciences, Chongqing 402460, China;
    2. Key Laboratory of Pig Industry Sciences, Ministry of Agriculture, Chongqing 402460, China;
    3. Scientific Observing and Experimental Station of Animal Housing System in Southwest Areas, Ministry of Agriculture, Chongqing 402460, China;
    4. Chongqing Key Laboratory of Pig Industry Sciences, Chongqing 402460, China
  • Received:2016-01-28 Online:2016-09-20 Published:2016-09-20

摘要:

本试验将流式细胞术应用于猪精子分析,同时检测精子功能的多个特性,以保证精子具备多种特性且功能完整,顺利完成授精过程。分别采用SYBR-14/PI(Propidium Iodide)、YO-PRO-1/PI、PNA/PI及Mito Tracker/YO-PRO-1 4种染色方法对荣昌猪精子的质膜完整性、细胞凋亡、顶体完整性及线粒体功能进行了检测和分析;以SYBR-14+/PI-表示质膜完整的活精子,YO-PRO-1-/PI-表示活精子,PNA-/PI-表示未发生顶体反应的活精子,Mito Tracker+/YO-PRO-1-表示线粒体活性高的活精子,以上述检测指标的综合结果判定精子功能的完整性。本试验通过流式细胞术对荣昌猪精子进行质量检测和分析,结果表明,该方法可为临床诊断种猪不孕症提供理论依据,对有效治疗不孕症有重要意义。

关键词: 荣昌猪; 精子; 流式细胞术; 检测

Abstract:

The flow cytometry was used in this trail to determine the pig sperm quality,in order to ensure the sperm had complete functions during the fertilization process.Four stain methods of SYBR-14/PI (Propidium Iodide),YO-PRO-1/PI,PNA/PI,Mito tracker/YO-PRO-1 were used to detect the plasma membrane integrity,apoptosis,top body ntegrity and mitochondrial function of Rongchang pig sperm.SYBR-14+/PI- expressed live sperm with complete plasma membrane,YO-PRO-1-/PI- expressed live sperm,PNA-/PI- expressed live sperm had no top body reaction,and Mito Tracker+/YO-PRO-1-expressed live sperm with high mitochondrial activity.The functional integrity of the tested sperm was evaluated according to the above comprehensive results.Through the study of Rongchang pig sperm quality detection by flow cytometry,it suggested that the method could provide theoretical basis for clinical diagnosis,and was very important for swine infertility treatments.

Key words: Rongchang pig; sperm; flow cytometry; detection

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