Aurora-A抑制对猪卵母细胞MⅠ期纺锤体组装的影响

[目的] 本试验旨在探究Aurora-A抑制对猪卵母细胞成熟和纺锤体组装的影响及潜在机制。[方法] 猪GV期卵母细胞分组进行体外成熟培养,在各组中分别添加0、1、3和5μmol？L-1 Aurora-A特异性抑制剂MLN8054,观察其对卵母细胞发育与成熟的影响及纺锤体结构的变化;免疫印迹检测磷酸化Aurora-A和TACC3的表达。[结果] Aurora-A抑制导致猪卵母细胞成熟明显受阻(P<0.05),且主要阻滞在Pro-MⅠ期及MⅠ期;MⅠ期纺锤体形态明显异常,出现单极或多极纺锤体,异常率显著上升(P<0.05)。Aurora-A抑制后,猪卵母细胞磷酸化Aurora-A和酸性卷曲转化相关蛋白3(TACC3)表达水平显著下降(P<0.05)。[结论] MLN8054处理后,猪卵母细胞减数分裂进程和纺锤体组装受阻,并抑制了磷酸化Aurora-A和其下游蛋白磷酸化TACC3的表达。
[Objectives] The purpose of this study was to investigate the effects of Aurora-A inhibition on maturation,spindle configuration of porcine oocytes and potential roles.[Methods] Porcine GV oocytes were randomly assigned into groups to culture in vitro. Each group was added with 0,1,3 and 5 μmol？L-1 MLN8054,an Aurora-A inhibitor,respectively. Maturation of oocytes and spindle structural alteration were observed. Phosphorylated Aurora-A and TACC3 (transforming acidic coiled-coil proteins) expression were analyzed by Western blot.[Results] The maturation rate of porcine oocytes was significantly inhibited when treated with the Aurora-A inhibitor (P<0.05),mainly arresting at Pro-MⅠand MⅠ-stage. Monopolar or multipolar spindles were observed at MⅠ-stage. The percentages of abnormal spindle morphology for MLN8054-treated groups were significantly increased as compared to the control (P<0.05). The pAurora-A and pTACC3 expressions of oocytes after Aurora-A inhibition were significantly decreased (P<0.05).[Conclusions] Maturation of oocytes and spindle configuration were affected and pAurora-A and its downstream protein pTACC3 expressions were inhibited by MLN8054 treatment