实验动物与比较医学 ›› 2014, Vol. 34 ›› Issue (2): 140-144.DOI: 10.3969/j.issn.1674-5817.2014.02.012

• 论著 • 上一篇    下一篇

体外受精-胚胎移植技术在APP/PS1双转基因小鼠繁育中的应用

吕相川, 张婀娜, 周生来, 王惟, 于洋, 郑志红   

  1. 中国医科大学实验动物部辽宁省转基因研究重点实验室, 沈阳 110001
  • 收稿日期:2013-10-24 出版日期:2014-04-25 发布日期:2014-04-25
  • 作者简介:吕相川(1984-), 男, E-mail: dream04lv@163.com
  • 基金资助:
    辽宁省科学技术计划项目(2010232006)

Application of in Vitro Fertilization and Embryo Transfer in Breeding of APP/PSEN1 Double Transgenic Mice

LÜ Xiang-chuan, ZHANG E-nuo, ZHOU Sheng-lai, YU Yang, WANG Wei, ZHANG Mei-ying, ZHENG Zhi-hong   

  1. Laboratory Animal Center, China Medical University, Shenyang 110001, China
  • Received:2013-10-24 Online:2014-04-25 Published:2014-04-25

摘要: 目的 研究体外受精-胚胎移植(IVF-ET)技术在APP/PS1双转基因小鼠繁育中的应用。方法 分别应用自然交配与体外受精方法进行APP/PS1双转基因繁育,比较受精率及产仔数量; 腹腔注射PMSG-HCG超排雌性C57BL/6小鼠,分别与12周龄、24周龄、36周龄、52周龄、72周龄的成年雄性APP/PS1双转基因小鼠进行体外受精,将获得的2-细胞胚胎移植到假孕雌鼠的输卵管,通过PCR方法对仔代小鼠进行鉴定。结果 6只雄性转基因小鼠与18只野生型雌性小鼠自然方式交配,实验周期为21 d,获得胚胎的受精率为73.3%。应用体外受精的方法用1只雄性转基因小鼠的精子与18只雌性野生型小鼠的胚胎进行体外受精, 受精率达到95%, 实验周期为4 d。两种繁育方式产仔数量阳性小鼠数量有显著差异(P<0.05), 阳性率无显著性差异(P>0.05)。12周龄、24周龄、36周龄的APP/PS1转基因雄性小鼠的体外受精结果显示,三种不同周龄的雄性APP/PS1转基因小鼠受精率、产仔数量及阳性率均无显著性差异(P>0.05)。对应用常规繁育方法难以获得后代的老龄APP/PS1双转基因雄性小鼠(52周龄、72周龄)体外受精率分别为85.6%、84.1%,获得阳性后代18只和14只。结论 APP/PS1双转基因小鼠可以采用IVF-ET技术进行繁育,为APP/PS1双转基因小鼠快速扩大繁育提供一种新方式。

关键词: 小鼠, 体外受精-胚胎移植(IVF-ET), 阿尔兹海默病, 扩大繁育

Abstract: Objective To study the techniques of in vitro fertilization (IVF) and embryo transfer applied for breeding APP/PS1 double transgenic mice. Methods The fertilization rates and birth rates were analyzed in both natural mating group and IVF group for breeding APP/PS1 double transgenic mice. The female C57BL/6 mice were given intraperitoneal injection of appropriate dose of PMSG-hCG. The super ovulation oocytes were collected for IVF with sperms which were taken from 12 weeks, 24 weeks, 36 weeks, 52 weeks and 72 weeks old APP/PS1 double transgenic male mice respectively. The 2-cells embryos were than transfer to the fallopian of pseudopregnant mice. The genotypes of newborn mice were identified by PCR. Results The fertilization rates were 73.3% in group of natural mating, but 95% in group of IVF. The statistic differentiation was significant in newborn mice number (P<0.05) between natural crossing group to IVF group, but was not marked in genotype positive rate (P>0.05). The fertilization rates, birth rates and genotype positive rate were evaluated in 12 weeks, 24 weeks, and 36 weeks old APP/PS1 double transgenic male mice by statistic analysis which showed that there were no significant difference among them (P>0.05 ). The IVF rates were 85.6%, 84.1% in 52 weeks, and 72 weeks old APP/PS1 double transgenic male mice those were so aging enough that could not breed the filial generation by usual method. Conclusion The application the techniques of IVF-ET can enlarge rapidly the breeding population of APP/PS1 double transgenic mice.

Key words: Mouse, IVF-ET, Alzheimer's disease, Expanded breeding

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