组蛋白H3S10及H3S28磷酸化对小鼠卵母细胞成熟的调控

Phosphorylation of histone H3S10 and H3S28 regulates oocyte maturation in mouse

[目的]进一步明确小鼠卵母细胞中组蛋白H3的磷酸化模式及其对卵母细胞成熟过程的调控。[方法]采用免疫荧光技术,对不同减数分裂阶段小鼠卵母细胞中组蛋白H3S10/H3S28的磷酸化状态进行了检测,在利用Aurora激酶广谱抑制剂ZM447439处理并检测卵母细胞成熟质量的基础上,进一步检测了处理后卵母细胞H3S10/H3S28磷酸化状态、染色体排列形态及Aurora激酶自身磷酸化水平的变化情况。[结果]小鼠卵母细胞减数分裂期间,组蛋白H3S10/H3S28持续磷酸化;ZM447439处理浓度依赖性地损害了卵母细胞的成熟能力,并相应导致H3S10/H3S28磷酸化减弱直至消失、染色体排列异常及Aurora激酶A/B/C自身磷酸化水平的改变。[结论]小鼠卵母细胞减数分裂过程中组蛋白H3S10/H3S28呈磷酸化状态,ZM447439处理引起组蛋白磷酸化减弱和染色体排列异常及Aurora激酶磷酸化水平的变化并导致了卵母细胞成熟能力损害。

[Objectives]The purpose of this experiment was to examine the phosphorylation patterns of histone H3 in mouse oocyte meiosis and their effects on the events related to oocyte maturation. [Methods]Phosphorylation patterns of histone H3S10/H3S28 at different stages of oocyte meiosis were detected by immunofluorescence technique. Then,Oocytes were treated with ZM447439 and the rates of maturation following treatments were checked. Finally,phosphorylation patterns of histone H3S10/H3S28,arrangement of chromosomes and phosphorylation levels of Aurora kinase family in oocytes treated with ZM447339 were evaluated. [Results]Results indicated that histone H3S10/H3S28 was phosphorylated during mouse oocyte meiosis. After ZM447439 treatment,the maturation capacity of mouse oocytes was damaged,the phosphorylation of histone H3S10/H3S28 decreased significantly and finally completely disappeared,the arrangement of chromosomes was abnormal and phosphorylation levels of Aurora kinase family were changed in a concentration dependent manner. [Conclusions]Histone H3S10/H3S28 were phosphorylated during mouse oocytes meiosis,and the decrease in histone phosphorylation and abnormality of chromosomes arrangement together with change in phosphorylation levels of Aurora kinase family led to the damage of oocytes maturation capacity following ZM447439 treatment.