Zhang Zixiao, Chen Changchao, Cui Panpan, Liao Yaya, Yao Lingyun, Zhang Yue, Rui Rong, Ju Shiqiang
College of Veterinary Medicine, Nanjing Agricultural University, Jiangsu, 210095, China.
J Assist Reprod Genet. 2017 Mar;34(3):399-407. doi: 10.1007/s10815-016-0864-4. Epub 2017 Jan 10.
This study was conducted to examine the dynamic distribution of polo-like 1 kinase (Plk1) and the possible role it plays in first mitotic division during early porcine embryo development.
Indirect immunofluorescence and confocal microscopy imaging techniques combined with western blot analyses were used to study the dynamic expression and subcellular localization of Plk1 protein in pig parthenogenetic embryos. Finally, a selective Plk1 inhibitor, GSK461364, was used to evaluate the potential role of Plk1 during this special stage.
The results showed that Plk1 upon expression exhibited specific dynamic intracellular localization, which closely correlated with the α-tubulin distribution during the first mitotic division. GSK461364 treatment resulted in cleavage failure, with majority of the GSK461364-treated embryos being arrested in prometaphase. Further results of the subcellular structure examination showed that GSK461364 treatment led to a significantly higher proportion of the treated embryos having abnormal spindles and misarranged chromosomes at the prometaphase stage.
Thus, these results indicated that Plk1 is essential for porcine embryos to complete the first mitotic division. Furthermore, Plk1 regulation was associated with effects on spindle assembly and chromosome arrangement.
本研究旨在检测polo样激酶1(Plk1)在猪早期胚胎发育过程中的动态分布及其在首次有丝分裂中可能发挥的作用。
采用间接免疫荧光和共聚焦显微镜成像技术,并结合蛋白质免疫印迹分析,研究Plk1蛋白在猪孤雌胚胎中的动态表达及亚细胞定位。最后,使用选择性Plk1抑制剂GSK461364评估Plk1在此特殊阶段的潜在作用。
结果显示,Plk1表达时呈现出特定的细胞内动态定位,这与首次有丝分裂期间α-微管蛋白的分布密切相关。GSK461364处理导致卵裂失败,大多数经GSK461364处理的胚胎停滞在前中期。亚细胞结构检查的进一步结果表明,GSK461364处理导致在前中期阶段,处理后的胚胎中纺锤体异常和染色体排列紊乱的比例显著更高。
因此,这些结果表明Plk1对猪胚胎完成首次有丝分裂至关重要。此外,Plk1的调控与对纺锤体组装和染色体排列的影响有关。