Oct4对鼠-猪异种核移植胚胎早期发育的影响  

Influence of oct4 on Early Embryonic Development of Mouse-Porcine Interspecies Nuclear Transfer

作　　者：阎新龙[1] 林艳丽[1] 周艳荣[1] 温叶飞[1] 叶华虎[1] 陈红星[1] 邓继先[1] 

机构地区：[1]军事医学科学院生物工程研究所一室,北京100071

出　　处：《中国实验动物学报》2007年第1期15-20,I0002,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家高技术发展计划(863)重大专项(编号:2002AA206621)基金资助

摘　　要：目的探讨Oct4转录因子能否促进鼠-猪异种核移植胚胎的早期发育。方法RT-PCR获得小鼠Oct4基因,构建pEGFP-N1-Oct4-EGFP融合质粒及pEGFP-N1-Oct4-EGFP终止质粒,pEGFP-N1质粒为阴性对照,脂质体法转染小鼠NIH3T3细胞,阳性克隆经RT-PCR,荧光显微镜验证正确后,移入去核的猪卵母细胞,观察并记录发育率。结果未转染的NIH3T3阴性对照组、转染Oct4-EGFP的小鼠NIH3T3细胞和转染pEGFP-N1组均能够支持猪异种核移植胚胎的早期发育,但转染pEGFP-N1-Oct4-EGFP实验组重构胚的卵裂率和8细胞发育率与转染pEGFP-N1组和未转染的NIH3T3组的重构胚发育率差异不显著。结论NIH3T3细胞能够支持鼠-猪异种核移植胚胎早期发育,Oct4对鼠猪异种核移植胚胎的发育并没有表现出促进作用,可能也受到NIH3T3来源的异种核移植胚胎本身发育率低以及本实验室核移植显微操作水平的限制,具体的机制尚需进一步探讨。Objective To explore whether the Oct4 transcription factor can improve the mouse-porcine interspecies nuclear transfer development or not. Methods Oct4 cDNA was amplified by BT-PCB. Two recombinant expression plasmids were constructed as the pEGFP-N1-Oct4-EGFP fusion vector and pEGFP- N1-Oct4-EGFP termination vector, and pEGFP-N1 as the negative control, then transfect the NIH3T3 cell with lipofectamineTM2000. RT-PCR assay showed that oct4 DNA fragment could be amplified, and confirmed by fluorescence microscopy. Then the cell was inserted into the perivitellinc space of the enucleated porcine oocyte to investigate the development of interspecies embryo. Results All the non-transgenic NIH3T3 group, transgenie Oct4-EGFP NIH3T3 and pEGFP-N1 groups could support the early development of cloned interspecies embryo, and the cleavage rate and development rate into the eight-cell stage were not significantly different. Conclusion It is clear that NIH3T3 can support the early embryonic development of mouse-porcine interspecies nuclear transfer. But maybe due to the low development of NIH3T3 derived nuclear transfer embryo and our limited condition of micromanipulation, the Oct4 gone does not have an improving effect on interspecies nuclear transfer. Further studies are still needed to elucidate its concrete mechanism.

关 键 词：猪 体细胞核移植 Oct4-EGFP NIH3T3 胚胎发育 

分 类 号：R321.33[医药卫生—人体解剖和组织胚胎学]