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(P<0.05);休眠胚胎冷冻后C3mRNA相对表达量与冷冻前相比显著下调(P<0.05);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(P<0.05);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(P<0.05)。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。
The study was aimed to investigate the protein location and the transcriptional expression of C3 on mouse normal hatched blastocysts and dormant embryos before and after cryopreservation,and to provide theoretical basis for elucidating the regulation mechanism of mammalian embryos during antifreeze.The ICR female mouse were selected,normal hatched blastocysts were collected from 5th pregnant mouse and dormant embryos were obtained from mouse delayed implantation model,then the confocal and Real-time PCR were used to detect ICC and relative expression of C3 mRNA on each group.The results showed that C3 were both expressed on transcriptional and translational levels in mouse normal hatched blastocysts and dormant embryos before and after cryopreservation;The relative expression of C3 mRNA in normal hatched blastocysts was significantly up-regulated after cryopreservation（P〈0.05）;The relative expression of C3 mRNA in dormant embryos was significantly down-regulated after cryopreservation（P〈0.05）;The relative expression of C3 mRNA in dormant embryos before cryopreservation was significantly higher thanthe normal hatched blastocysts before cryopreservation（P〈0.05）;The relative expression of C3 mRNA in dormant embryos after cryopreservation was significantly lower than the normal hatched blastocysts after cryopreservation（P〈0.05）.The results showed that the transcriptional down-regulated expression of C3 in embryos had a positive effect on the antifreeze of embryos,and C3 gene might be involved in the regulation of embryo freezing-thawing process.
出处
《中国畜牧兽医》
CAS
北大核心
2017年第2期514-520,共7页
China Animal Husbandry & Veterinary Medicine
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)
