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Changes in histone modifications during in vitro maturation of porcine oocytes

机译:猪卵母细胞体外成熟过程中组蛋白修饰的变化

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Nuclear core histone modifications influence chromosome structures and functions. Recently, the involvement of histone acetylations in the cell memory of gene expression has been suggested in mouse oocyte maturation. At present, there is little available data on histone modifications in mammalian oocyte maturation. In the present study, we examined changes in the acetylation of histone H3 lysines 9 (H3K9) and 14 (H3K14), and histone H4 lysines 5 (H4K5), 8 (H4K8) and 12 (H4K12), and trimethylation of H3K9 during in vitro maturation of porcine oocytes. Immunocytochemical analyses revealed that the all of the lysines examined were highly acetylated in the germinal vesicle stage, and this level of acetylation was maintained until the first prometaphase. In the first metaphase, the lysines near the N-terminal end, H3K9 and H4K5, were completely deacetylated. The acetylation of the lysines far from the N-terminal end, H3K14, H4K8, and H4K12, was markedly decreased but still present. The acetylations were increased transiently at the first anaphase and telophase, and then decreased again at the second metaphase to the same level as the first metaphase. Since effective concentrations of trichostatin A (TSA) to inhibit the deacetylation were different in various lysine residues, multiple histone deacetylases (HDACs) were suggested to function during meiotic maturation. The trimethylation of H3K9 was maintained in a high level throughout maturation. These results suggest that the histone acetylation during porcine oocyte maturation is precisely controlled by the cell cycle.
机译:核核心组蛋白修饰影响染色体结构和功能。最近,在小鼠卵母细胞成熟中已经提出组蛋白乙酰化参与基因表达的细胞记忆。目前,关于哺乳动物卵母细胞成熟中的组蛋白修饰的可用数据很少。在本研究中,我们研究了在肝癌发生期间,组蛋白H3赖氨酸9(H3K9)和14(H3K14),组蛋白H4赖氨酸5(H4K5),8(H4K8)和12(H4K12)的乙酰化以及H3K9的三甲基化猪卵母细胞的体外成熟。免疫细胞化学分析显示,所检查的所有赖氨酸在生小泡阶段均高度乙酰化,这种乙酰化水平一直维持到第一个前中期。在第一个中期,N末端附近的赖氨酸H3K9和H4K5被完全脱乙酰。远离N末端的H3K14,H4K8和H4K12赖氨酸的乙酰化程度明显降低,但仍然存在。乙酰化在第一后期和末期短暂增加,然后在第二中期再次降低到与第一中期相同的水平。由于抑制各种不同赖氨酸残基的曲古抑菌素A(TSA)的有效浓度不同,因此建议多种组蛋白脱乙酰基酶(HDAC)在减数分裂成熟过程中发挥作用。在整个成熟过程中,H3K9的三甲基化水平保持较高水平。这些结果表明,猪卵母细胞成熟过程中的组蛋白乙酰化受到细胞周期的精确控制。

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