FGF10对小鼠植入前胚体外发育及2-细胞胚内G蛋白偶联受体30表达的影响

The effect of FGF10 on the in vitro development of mouse preimplantation embryo and G protein-coupled receptor 30 in 2-cell embryos

目的观察成纤维细胞生长因子10(FGF10)对小鼠受精卵体外发育和2-细胞胚内G蛋白偶联受体30(GPR30)水平的影响,为进一步探讨FGF10在小鼠植入前胚体外发育中的作用提供依据。方法以M16培养液为对照组,M16中分别添加不同浓度的FGF10为实验组,收集昆明(KM)雌性小鼠与KM雄性小鼠交配所得的受精卵,分别观察各组受精卵体外发育情况。收集KM小鼠受精卵,分别置于M16和含不同浓度FGF10的M16培养液中培养,获取体外发育的2-细胞胚,采用免疫荧光染色结合激光共聚焦扫描显微镜检测细胞内GPR30水平。结果添加FGF10实验组发育到4-细胞胚、桑葚胚和囊胚的比率明显高于对照组,其中以添加浓度为10ng/ml的FGF10实验组的效果最明显。免疫荧光染色结果显示,10ng/ml的FGF10实验组体外发育2-细胞胚内GPR30免疫荧光染色强度明显高于对照组。结论 FGF10可促进KM小鼠植入前胚的体外发育,其作用可能与上调2-细胞胚内GPR30有关。

Objective To investigate the effect of fibroblast growth factor 10 （FGF10） on mouse zygote in vitro development, and on G protein-coupled receptor 30 （GPR30） in 2-cell embryos, and lay the foundation for further studies regarding the roles of FGF10 in mouse preimplantation embryo development. Methods Zygotes were collected from female KM mice that had been mated with male KM mice, and cultured in Ml6 medium （control group） and Ml6 medium supplemented with FGF10 （2. 5ng/ml,5ng/ml, 10ng/ ml, 20ng/ml, 40ng/ml, respectively） . The development potential of each group was recorded. Furthermore, 2-cell embryos were col-lected from M16 medium and M16 medium supplemented with 10ng/ml FGF10 respectively, and the level of GPR30 was detected by laser confocal scanning microscopy combined with immunofluorescent staining. Results The ratios of 4-cell embryos, morulae, and blas-tocysts in the groups supplemented with FGF10 were significantly higher than in the control group. The maximum effect was observed in the lOng/ml group. Immunostaining results showed that the immunofluorescence staining intensity of GPR30 in the 2-cell embryos was obviously elevated after 10ng/ml FGF10 treatment. Conclusion FGF10 promotes the in vitro development of mouse preimplantation em-bryos, possibly through the up-regulation of GPR30 in 2-cell embryos.