程序化冷冻前后小鼠孵化囊胚和休眠胚胎的Hba-α蛋白分布和差异表达

摘要

Objective This study is aimed to investigate the distribution and differential expression of Hba⁃αpro⁃tein in mouse normal hatched blastocysts and dormant embryos before and after cryopreservation, and to provide theoretical basis of the development and use of new mammalian antifreeze proteins. Methods Normal hatched blastocysts were ob⁃tained from d5 pregnant mice while dormant embryos were collected from mouse delayed implantation models. Confocal mi⁃croscopy and western blot were used to detect the differential expression of Hba⁃α protein among those groups. Results Hba⁃αprotein expressed in mouse normal hatched blastocysts and dormant embryos before and after cryopreservation. There were no significant differences of expression of Hba⁃α protein between normal hatched blastocysts before and after cryopr⁃eservation (P>0�05). Compared with the normal hatched blastocysts before and after cryopreservation, the expression of Hba⁃αprotein in dormant embryos was significantly lower (P<0�05). The expression of Hba⁃αprotein in dormant embryos after cryopreservation was significantly lower than the other groups (P<0�05). Conclusions The effect of cryopreserva⁃tion on the expression of Hba⁃α protein in mouse dormant embryos is significantly higher than normal hatched blastocysts. Hba⁃α protein has a potential to be used as a novel mammalian⁃derived antifreeze protein.%目的：利用程序化冷冻的方法，探究冷冻前、后小鼠孵化囊胚和休眠胚胎中Hba⁃α蛋白的分布和差异表达的变化，为今后开发和利用新型哺乳动物源抗冻蛋白提供理论依据。方法从妊娠d5小鼠体内获取孵化囊胚；从小鼠延迟着床模型获取休眠胚胎，利用Confocal显微镜和Western Blot技术对程序化冷冻前、后的各组胚胎进行Hba⁃α蛋白的分布和表达检测。结果孵化囊胚和休眠胚胎在程序化冷冻前、后均有Hba⁃α蛋白的表达；孵化囊胚经程序化冷冻后Hba⁃α蛋白的表达与冷冻前相比差异无显著性（ P＞0�05）；冷冻前休眠胚胎与程序化冷冻前后的孵化囊胚相比，Hba⁃α蛋白的表达量显著下调（P＜0�05）；冷冻后休眠胚胎Hba⁃α蛋白的表达量显著低于其他各组（P＜0�05）。结论程序化冷冻处理对小鼠休眠胚胎Hba⁃α蛋白的表达影响明显大于孵化囊胚；Hba⁃α基因具有作为新型哺乳动物源抗冻蛋白的潜力。