摘要
本试验旨在研究比较猪体细胞克隆胚胎体外发育与凋亡变化规律,探讨其体外发育潜能与细胞凋亡间的关系。将体外成熟培养获得的猪MⅡ期卵母细胞随机分为2组,分别进行体细胞核移植(SNCT)与体外受精(IVF)处理,采用彗星试验(Comet assay)和Real-time RT-PCR技术对体外发育过程中各时期胚胎的细胞凋亡事件及抗凋亡基因Bcl-2 mRNA的表达水平进行研究。结果表明,两组在卵裂率上无显著差异(77.6%vs 80.3%,P>0.05),在囊胚发育率上,SCNT组胚胎明显低于IVF组,但无统计学差异(18.9%vs 24.5%,P>0.05);随着胚胎发育,SCNT与IVF组胚胎的凋亡率均呈现不断增加的趋势,Bcl-2 mRNA表达量则逐渐下降;到16-细胞以后,SCNT组胚胎的凋亡率均显著高于IVF组,同时Bcl-2 mRNA表达量均显著低于IVF组(P<0.05)。本试验结果表明,猪克隆胚胎生产效率低下可能与胚胎发育过程中的细胞凋亡有关。
The purpose of this experiment was to determine the relationship between embryonic development and apoptosis in porcine nuclear transfer （SCNT） embryos during development in vitro. The apoptotic incidence was evaluated via Comet assay, and the mRNA expression of gene implicated in anti-apoptosis （Bcl-2） was also examined using Real-time RT-PCR. The results showed that the SCNT embryos had a similar cleavage rate （77.6% vs 80.3%, P0.05） but a lower developmental rate of blastocysts （18.9% vs 24.5%,P0.05） compared to in vitro fertilization （IVF） embryos. The results of Comet assay demonstrated that the apoptotic rate both of the two types of embryos maintained increased tends along embryonic developmental stages, but the SCNT embryos exhibited higher apoptotic rate from 16-cell to morula stage compared to IVF embryos. In the whole developmental process, the relative abundance of Bcl-2 mRNA was gradually reduced both in SCNT and IVF embryos, and the expression of Bcl-2 mRNA in SCNT embryos was significantly lower than that of IVF embryos at 16-cell and 32-cell stages. It was concluded that the higher degree of apoptotic incidence may be associated with the low efficiency of porcine SCNT in vitro production.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2010年第6期678-684,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家高技术研究与发展计划(2008AA101003)
农业部"十一五"科技支撑计划(2008BADB2B11)
国家转基因生物新品种培育重大专项(2009ZX08009-1438)
关键词
猪
体细胞核移植
发育
凋亡
基因表达
swine
somatic cell nuclear transfer
development
apoptosis
gene expression
作者简介
作者简介：剧世强（1980-），男，山西朔州人，博士生，主要从事动物生殖生物学研究，E-mail：jsqiang2005@yahoo．com.cn
通讯作者：芮荣，E—mail：rrui@njau．edu．cn
