ME491／CD63对小鼠胚胎发育及着床的影响

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／CI）63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／CI）63mRNA在假孕D1～8小鼠子宫中均有表达，于D6表达最丰富．MFA91／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．

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

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.