冻融后小鼠休眠胚胎超微结构的研究.doc

研究报告 冻融后小鼠休眠胚胎超微结构的研究 顾美超，卢天罡，刘云海，倪和民，张劭俣，翟椿东，邢书涵，郭勇* (北京农学院动物科学技术学院，北京 102206) 【摘要】 目的 观察小鼠休眠胚胎与正常孵化期胚胎在细胞连接和各细胞器形态与分布上的差异，以及冻融培养后的变化，从亚细胞超微结构的角度揭示其抗冻能力优于正常孵化胚胎的原因。方法 利用透射电子显微镜观察各类小鼠胚胎的超微结构，并进行相关比较分析。结果 通过亚细胞结构对比分析发现：冷冻前小鼠休眠胚胎为紧缩状，处于能量代谢较低的“基态”，通过冻融后培养，细胞器结构恢复与正常孵化胚胎冷冻前相似。异染色质减少，甚至消失；而正常孵化胚胎经过冻融后，线粒体数量减少，细胞核松散甚至碎裂，异染色质增多。结论 小鼠休眠胚胎与正常孵化胚胎冻融后相比，其细胞状态更有利于物质储存及能量代谢，表明小鼠休眠胚胎从亚细胞超微结构的角度比其正常孵化胚胎更具抗冻性。 【关键词】小鼠；休眠胚胎；超微结构；冻融 Study on the ultra-structure of mouse dormant embryos Cultured in vitro after frozen-thawn GU Mei-chao, LU Tian-gang , LIU Yun-hai, NI He-min, ZHANG Shao-yu, ZhAI Chun-dong, XING Shu-han , GUO Yong* (College of the Animal Science and Technology, Beijing University of Agriculture, Beijing 102206, China) *: correspondence author: GUO Yong [Abstract] Objective In this study, it was aimed to investigate the differences of the cell junction, cellular organelles morphology and distribution, as well as being cultured in vitro after frozen-thawn between normal mouse hatched blastocysts and their dormant ones, and then, to reveal the reason from the ultra-structure about how their dormant embryos initiated better anti-freezing shock effect than those normal hatched ones did. Methods By transmission electron microscope, the ultra-structure of these two different types of murine embryos was observed and analyzed respectively. Results By the comparative analysis of their ultra-structures, the results showed that the dormant embryos be austerity and lower energy metabolism at ‘ground state’. Its cellular structure seemed to be similar to the normal embryos cultured in vitro after frozen-thawn. The heterochromatin decreased or disappeared, as well as the number of mitochondria decreased, and the nucleus of those normal hatched blastocysts after frozen-thawn, were loose and fractured. In addition, their heterochromatin also increased. Conclusion Collectively, it can be concluded that the satisfied cellular ultra-structure of mouse delayed embryos, can improve their e