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

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/PSl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/PSl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21d,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4d.两种繁育方式产仔数量阳性生小鼠数量有显著差异（P＜0.05）,阳性生率无显著性差异（P＞0.05）. 12周龄、24周龄、36周龄的APP/PSl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.

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

To study the techniques of in vitro fertilization （IVF） and embryo transfer applied for breeding APP/PSI double transgenic mice. Methods The fertilization rates and birth rates were analyzed in both natural mating group and IVF group for breeding APP/PS 1 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/PS 1 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/PS 1 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 ofAPP/PS 1 double transgenic mice.