ME491/CD63对小鼠胚胎发育及着床的影响

Effect of tumor metastasis inhibitory factor ME491/CD63 gene on embryo development and implantation in mice

目的:研究ME491/CD63对小鼠胚胎体外发育及着床的影响,以及ME491/CD63mRNA及蛋白在假孕D1～8小鼠子宫中的表达规律.方法:①将小鼠8-细胞胚胎培养于含不同浓度ME491/CD63抗体的培养液中,观察胚胎发育,记录囊胚形成及脱透明带的情况.②妊娠D4小鼠宫角注射ME491/CD63抗体,于妊娠D8观察小鼠胚胎植入数量.③用RT-PCR及免疫组织化学技术观察ME491/CD63mRNA及蛋白在假孕小鼠子宫的表达规律.结果:①ME491/CD63抗体可影响胚胎在体外的发育.与对照组相比,在ME491/CD63抗体浓度为1∶400及以上时,囊胚的形成率明显降低(P<0.05);在抗体浓度为1∶800及以上时,胚胎脱带率显著下降(P<0.05),ME491/CD63抗体对胚胎发育的影响具有浓度依赖性.②宫角注射一定量的ME491/CD63抗体可以显著提高小鼠胚胎的着床数[(5.8±2.3)vs(4.1±1.8),P<0.05].③ME491/CD63mRNA在假孕D1～8小鼠子宫中均有表达,于D6表达最丰富.ME491/CD63蛋白在假孕D1～8的子宫腺上皮细胞均有表达,假孕D1～5腔上皮细胞无表达,假孕D6～8,子宫腔上皮细胞阳性表达.结论:①ME491/CD63在小鼠胚胎发育和着床过程中发挥重要作用.②ME491/CD63在妊娠小鼠子宫的表达是非胚胎依赖性的,其表达量的变化可能与胚胎存在与否有一定关系.

AIM： To study the effect of tumor metastasis suppressor gene ME491/CD63 on embryo development in vitro and implantation, and identify expression pattern of ME491/CD63 mRNA and protein in pseudopregnant mice. METHODS：① 8-cell embryos were cultured in medium containing ME491/CD63 antibody at various concentrations, then the rates of formation and hatching of blastocysts were observed. ② The number of implantation sites was examined in uterine horns on D8, which were injected with ME491/CD63 antibody on D4. ③ Protein and mRNA expressions of ME491/CD63 in uterus of pseudopregnant mice were detected by immunohistochemistry and RT-PCR. RESULTS： ① ME491/CD63 ployclonal antibody significantly inhibited the rate of formation of blastocysts （ 1 ： 400, P 〈 0.05 ） and prevented hatching of blastocysts from zona pellucida （ 1 ： 800, P 〈 0. 05 ） in a dose-dependent manner.② The number of implanted embryos markedly increased in uterine horns injected with ME491/CD63 antibody on D4 of pregnancy[ （5.8 ±2.3） vs （4.1 ± 1.8）, P 〈 0.05 ]. ③ In uterus of pseudopregnant mice, expression of ME491/CD63 mRNA was detected during D1-8, and it was dramatically abundant on D6. Expression of ME491/ CD63 protein was detected mainly in glandular epithelial cells during D1-8, however, no expression was detected in luminal epithelial cells during D1-5, and positive expression was observed during D6-8. CONCLUSION： ① MEA91/CD63 might play an important role in embryo development and implantation in mice. ② Expression of ME491/CD63 in uterus was independent of embryos in pregnant mice, its quantity was related to whether embryo existed or not.