Effect of tumor metastasis inhibitory factor MFA491/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. ②The number of implantation sites was examined in uterine horns on D8, which were injected with ME491/CD63 antibody on D4. ③Protein and mRNA expressions of ME491/CD63 in uterus of pseudopregnant mice were detected by immunohistochemistry and RT-PCR. RESULTS: ①ME491/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. ②The 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]. ③In 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: ①ME491/CD63 might play an important role in embryo development and implantation in mice. ②Expression of ME491/CD63 in uterus was independent of embryos in pregnant mice, its quantity was related to whether embryo existed or not.
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NSTL
万方数据
维普
