摘要
目的通过对比小鼠孵化囊胚以及其休眠胚胎程序化冷冻并解冻处理前后Mgst1基因的相对表达情况,以及Mgst1蛋白的差异表达分布情况,分析Mgst1基因在小鼠胚胎程序化冷冻过程中发挥的作用,为胚胎抗冻机理研究提供新的理论基础。方法使用激光共聚焦扫描显微镜采集Mgst1蛋白在胚胎中表达分布的免疫荧光图像;使用qRT-PCR技术鉴定Mgst1基因在胚胎中的相对表达量。结果通过免疫荧光图像发现,Mgst1蛋白在小鼠冷冻前后的孵化囊胚与休眠胚胎中均有表达;通过qRT-PCR发现,冷冻前的孵化囊胚与休眠胚胎的Mgst1基因相对表达无显著差异(P>0.05);冷冻后的孵化囊胚Mgst1基因相对表达量较冷冻之前显著上调(P<0.05);冷冻后的休眠胚胎Mgst1基因相对表达量较冷冻之前极显著上调(P<0.01);冷冻后的休眠胚胎Mgst1基因相对表达量较冷冻后的孵化囊胚显著上调(P<0.05)。结论Mgst1基因可能在小鼠胚胎抗冻机制中发挥着重要的正调控作用。
Objective To compare the relative expression of the Mgst1 gene and the expression distribution of Mgst1 protein in mouse hatching blastocysts and dormant embryos before and after cryopreservation.The role of the Mgst1 gene during the programmed freezing of mouse embryos was analyzed to provide a new theoretical basis for the study of embryo-related antifreeze mechanisms.Methods Immunofluorescence images of the expression distribution of Mgst1 protein in embryos were detected by confocal microscopy.By qRT-PCR,the relative gene expression of Mgst1 was determined.Results Mgst1 protein was expressed in mouse normal hatched blastocysts and dormant embryos before and after cryopreservation.There was no significant difference in the relative expressions of Mgst1 gene between hatched blastocysts and dormant embryos before cryopreservation(P>0.05).Compared with the normal hatched blastocysts before and after cryopreservation,the relative gene expression of Mgst1 in normal hatched blastocysts was significantly upregulated after cryopreservation(P<0.05),and its relative expression in dormant embryos was markedly upregulated after cryopreservation(P<0.01).Compared with the normal hatched blastocysts after cryopreservation,the relative gene expression of Mgst1 in dormant embryos was significantly upregulated(P<0.05).Conclusions The Mgst1 gene may play an important role in the antifreeze mechanism of mouse embryos.
作者
胡新宇
邢凯
李征
赵志显
常雪蕊
齐晓龙
盛熙晖
王相国
倪和民
郭勇
HU Xinyu;XING Kai;LI Zheng;ZHAO Zhixian;CHANG Xuerui;QI Xiaolong;SHENG Xihui;WANG Xiangguo;NI Hemin;GUO Yong(College of Animal Science and Technology,Beijing University of Agriculture,Beijing 102206,China;Beijing Institute of Feed Control,Beijing 100107)
出处
《中国实验动物学报》
CAS
CSCD
北大核心
2020年第2期175-179,共5页
Acta Laboratorium Animalis Scientia Sinica
基金
2016年度科技创新服务能力建设-科技计划重点项目(KZ201610020018)。
