遗传 ›› 2008, Vol. 30 ›› Issue (11): 1421-1426.doi: 10.3724/SP.J.1005.2008.01421

• 研究报告 • 上一篇    下一篇

外源基因对精子的影响及其在山羊早期胚胎中的表达

叶华虎1;董罡1;袁菊芳1;隋丽华1;胡娟峰1;李瑞生1;刘彦2;马啸3;陈振文3;曾林1   

  1. 1. 军事医学科学院实验动物中心, 北京 100071;
    2. 北京市农林科学院畜牧兽医研究所, 北京 100097;
    3. 首都医科大学实验动物部, 北京 100069
  • 收稿日期:2008-02-29 修回日期:2008-06-10 出版日期:2008-11-10 发布日期:2008-11-10
  • 通讯作者: 曾林

Exogenous DNA influencing fertilization of goat sperm cells and ex-pression in early embryos

YE Hua-Hu1;DONG Gang1;YUAN Ju-Fang1;SUI Li-Hua1;HU Juan-Feng1;LI Rui-Sheng1;LIU Yan2;MA Xiao3;CHEN Zhen-Wen3;ZENG Lin1   

  1. 1. Laboratory Animal Center, Academy of Military Medical Sciences, Beijing 100071, China;
    2. Institute of Husbandry and Veterinary Science, Beijing Academy of Agriculture and Forestry, Beijing 100097, China;
    3. Laboratory Animal Center, Capital University of Medical Sciences, Beijing 100069, China
  • Received:2008-02-29 Revised:2008-06-10 Online:2008-11-10 Published:2008-11-10
  • Contact: ZENG Lin

摘要: 摘要: 在前期实验中发现, 山羊精子可自发结合外源DNA, 但结合能力在不同动物个体之间差异显著。挑选结合能力明显不同的3只公羊, 进一步探讨了外源DNA对精子的影响及其在早期胚胎中的表达, 结果发现: 外源基因与精子共同孵育后, 精子的活率、顶体反应发生率和受精能力均呈下降态势, 其降幅与精子的结合能力密切相关。利用与DNA共育后的精子进行体外受精, 外源基因可被导入卵母细胞并在早期胚胎中获得表达, 但胚胎阳性率因精子供体不同而差异显著(P<0.05); 其中来源于高、中结合能力供体生产的胚胎, 分别有16.2%(25/154)和5.3%(4/76)可检测到外源基因存在, 但表达仅见于高结合能力供体生产的早期胚胎, 表达率为6.5%(10/154); 低结合能力供体生产的胚胎无外源基因。研究表明, 在以精子载体方法生产转基因动物的实验过程中, 筛选对DNA结合能力较强的精子供体是提高转基因效率的前提, 但需要考虑外源DNA对精子受精能力的影响。

关键词: 精子介导基因转移, 精子, 胚胎, 山羊

Abstract: Abstract: Our early study found that goat spermatozoa could spontaneously take up foreign DNA and vary in capabilities of spermatozoa from different donors to bind and internalize exogenous DNA. In this study, three goats with considerable differences of capability were used to investigate the effect of exogenous DNA on goat spermatozoa, and feasibility and efficiency of transgenic embryo production by sperm-mediated gene transfer method. The viability, acrosomal reaction fre-quencies and cleavages were decreased in the groups co-cultured with exogenous DNA, compared with the control groups, and the range of decrease was correlated with the capability of sperm cells up-take foreign DNA. After fertilizing with co-cultured spermatozoa, GFP gene was introduced into oocytes and expressed in early embryos. However, different effi-ciencies of transgenic embryos appeared in sperm donors (P<0.05). GFP gene was detected in 16.2% (25/154), 5.3% (4/76), and 0% (0/36) embryos, respectively, when high, middle and low capability of sperm donors were used. But only 6.5% (10/154) embryos from high capability sperm donor expressed GFP. Our results demonstrate that selecting high capability of sperm donor is a key step for improving efficiency of sperm mediated-gene transfer method. However, the adverse influ-ence of foreign DNA on spermatozoa needs to be further studied.