目的:研究14-3-3ε蛋白调节小鼠受精卵纺锤体组装的作用。方法:采用间接免疫荧光实验检测1-细胞期受精卵14-3-3ε和仪.微管蛋白的亚细胞定位;利用显微注射方法将14.3—3εsiRNA注射入1-细胞期受精卵,观察小鼠受精卵纺锤体形态及检测Polo样激酶l(Plkl)的活性。结果:14-3-3ε和α-微管蛋白在G1期、S期和G2期卵中主要共定位于细胞质,M期卵14-3-3ε主要位于细胞质皮质。小鼠受精卵注射14-3-3ε siRNA后导致异常形态纺锤体形成及Plkl活性下降。结论:14-3-3ε蛋白在调节小鼠受精卵纺锤体组装中发挥重要作用。
Objective: To study the effect of 14-3-3ε protein on the spindle assembly in mouse fertilized eggs. Methods: Indirect immunofluorescence was used to detect the localization of 14-3-3ε and tx-tubulin in the mouse fertilized eggs. 14-3-3ε siRNA was microinjected into 1-cell fertilized eggs to study morphology of spindle and polo like kinase 1 (Plkl) activity. Results: In all phases of fertilized eggs investigated, 14-3-3ε had a mainly cytoplasmic distribution and co-localized with α-tubulin in the cytoplasm at the Gx, S and G2 phases. Knockdown of 14-3-3ε resulted in abnormal morphology of spindle and decreased the Plkl activity. Conclusion: 14-3-3ε plays critical roles in spindle assembly in mouse fertilized eggs.