模拟微重力条件下昆明小鼠早期胚胎体外发育及其G6PD基因表达  

Development of Kunming Mouse Early Embryos in Vitro and Expression of G6PD under Simulated Microgravity

作　　者：吴嫦丽[1] 高淑静[1] 江青艳[1] 冯定远[1] 田秀春 杨向中 张守全[1] 

机构地区：[1]华南农业大学动物科学学院 [2]Center for Regenerative Biology,University of Connecticut,Storrs,CT06269,USA

出　　处：《航天医学与医学工程》2006年第5期340-344,共5页Space Medicine & Medical Engineering

基　　金：广东省自然科学基金项目(04020611);广东省教育厅自然科学基金资助(Z03005)。

摘　　要：目的研究模拟微重力条件下培养的小鼠早期胚胎体外发育与1g重力条件下体外发育的差异。方法检测葡萄糖代谢关键酶6-磷酸葡萄糖脱氢酶(G6PD)在昆明小鼠早期胚胎体外不同重力条件下培养各个时期基因的转录与表达。结果模拟微重力培养条件下,胚胎体外发育受到明显的抑制,模拟微重力不利于小鼠胚胎的体外发育。用CZB+10%FCS进行培养,2-细胞期培养的囊胚率为零,显著低于常规培养的23.92%;4-细胞期培养的囊胚率为18.44%,显著低于常规培养的80.12%。利用G6PD的内、外两对引物,检测昆明小鼠早期胚胎体外发育各个阶段的G6PD基因的转录与表达。从2-细胞期开始体外常规培养的胚胎,发育至4-、8-细胞胚胎、桑椹胚、囊胚及退化囊胚都有G6PD基因的转录与表达。在模拟微重力条件下,从2-细胞期开始培养的胚胎,发育至4-细胞期、桑椹期、囊胚期胚胎和退化桑椹胚皆有G6PD基因的转录和表达,而退化的2-细胞胚胎没有检测到G6PD基因的转录与表达。结论两种培养方式之间,G6PD基因的转录与表达不存在差异,说明早期胚胎都存在磷酸戊糖途径。Objective To detect gene expression of G6PD in Kunming mouse preimplantation embryos which were cultured in vitro under 1 g gravity and simulated microgravity. Method Two sets of inner and outer primers were designed and synthesized according to cDNA sequences of G6PD. This gene expression was detected in Kunming mouse preimplantation embryos by nested RT-PCR （reverse transcription-polymerase chain reaction）. Under simulated microgravity condition, the percentages of passing 2-cell development block embryos and blastocysts in CZB ＋ 10%FCS were lower than those under lg gravity. Result It is suggested that the frequency of early embryonic lethality is possibly increased by simulated microgravity. With two pairs of inner and outer primers of G6PD gene, G6PD gene mRNA expression were observed in Kunming mouse 4-, 8-cell embryos, morula, blastocyst and degenerated blastocyst in vitro under 1 g gravity condition. In Embryos cultured under the simulated microgravity condition, G6PD gene mRNA expression were observed in 4-cell embryos, morula, blastocyst, and degenerated morula embryos, but not in 2-cell embryos collected from oviduct and degenerated in vitro. Conclusion It is concluded that there is no difference in gene expression of G6PD between 1g.qravitv and simulated microaravitv in vitro.

关 键 词：模拟微重力 早期胚胎 G6PD基因 基因表达 

分 类 号：R854[医药卫生—航空、航天与航海医学]