程序化冷冻前后小鼠正常孵化囊胚和休眠胚胎中C3基因表达的研究

发布时间：2018-11-05 17:07

【摘要】：试验旨在研究程序化冷冻前后小鼠正常孵化囊胚和休眠胚胎中C3蛋白的分布及转录水平的差异表达情况,为阐明哺乳动物胚胎在抗冻过程中的相关调控机制提供理论依据。试验选用ICR系雌性小鼠,从妊娠第5天小鼠子宫回收正常孵化囊胚;构建延迟着床模型获取小鼠休眠胚胎,利用激光共聚焦和实时荧光定量PCR技术对各组胚胎进行细胞免疫荧光和C3mRNA相对表达量的检测。结果发现,程序化冷冻前后的正常孵化囊胚和休眠胚胎中C3在转录和翻译水平均有表达;正常孵化囊胚经冷冻处理后C3mRNA相对表达量显著上调(P0.05);休眠胚胎冷冻后C3mRNA相对表达量与冷冻前相比显著下调(P0.05);冷冻前休眠胚胎C3mRNA相对表达量显著高于冷冻前正常孵化囊胚(P0.05);冷冻后休眠胚胎C3mRNA相对表达量显著低于冷冻后正常孵化囊胚(P0.05)。结果表明,胚胎中C3基因在转录水平的下调表达对胚胎抗冻具有一定的积极作用,C3基因很可能参与了胚胎抗冻过程的相关调控。
[Abstract]:The purpose of this study was to study the distribution of C3 protein and the differential expression of transcription level in normal hatched blastocysts and dormant embryos before and after programmed freezing, and to provide a theoretical basis for elucidating the regulation mechanism of mammalian embryos in the process of freezing resistance. The normal hatched blastocysts were recovered from the uterus of the ICR female mice on the 5th day of gestation. Mouse dormant embryos were obtained by using delayed implantation model. The relative expression of C3mRNA and cellular immunofluorescence were detected by laser confocal and real-time fluorescence quantitative PCR. The results showed that C3 was expressed in normal blastocysts and dormant embryos before and after programmed freezing, and the relative expression of C3mRNA in normal hatched blastocysts was up-regulated after freezing (P0.05). The relative expression of C3mRNA in dormant embryos after freezing was significantly lower than that before freezing (P0.05), and the relative expression of C3mRNA in dormant embryos before freezing was significantly higher than that in normal hatched blastocysts before freezing (P0.05). The relative expression of C3mRNA in frozen dormant embryos was significantly lower than that in normal blastocysts after freezing (P0.05). The results showed that the down-regulation of C3 gene expression at the transcriptional level in embryos had a positive effect on embryo antifreeze, and C3 gene might be involved in the regulation of embryo antifreeze process.
【作者单位】：北京农学院动物科学技术学院兽医学(中医药)北京市重点实验室;
【基金】：国家自然基金面上项目(31272526) 2013年度北京市教委北京市属高等学校创新团队建设与教师职业发展计划项目(PXM2013_014207_000067)
【分类号】：S814.8

【参考文献】