同型半胱氨酸对小鼠卵母细胞成熟过程中组蛋白表观遗传修饰的影响

梁蓉* 陈曦 张育军 石程

doi:10.16098/j.issn.0529-1356.2017.05.015

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解剖学报 ›› 2017, Vol. 48 ›› Issue (5) : 585-589. DOI: 10.16098/j.issn.0529-1356.2017.05.015

组织学胚胎学发育生物学

同型半胱氨酸对小鼠卵母细胞成熟过程中组蛋白表观遗传修饰的影响

梁蓉^1* 陈曦¹ 张育军² 石程¹

作者信息 +

Effects of homocysteine on histone modification during oocyte development in mice

LIANG Rong¹^* CHEN Xi¹ ZHANG Yu-jun² SHI Cheng¹

Author information +

文章历史 +

摘要

目的探讨卵母细胞发育过程中同型半胱氨酸（HCY）对组蛋白表观遗传修饰的影响。方法首先使用10只2周ICR雌性小鼠建立完整卵泡的体外培养体系，在卵母细胞发育早期，利用免疫组织化学方法，观察HCY对甲基化组蛋白H3K4、H3K9以及乙酰化组蛋白H3K9分布的影响；其次使用10只4周ICR雌性小鼠，利用卵母细胞的体外成熟培养体系，观察HCY对卵母细胞成熟过程的影响，同时利用实时定量PCR方法，观察在此成熟过程中HCY对卵母细胞内组蛋白乙酰化水平的调控酶GCN5和HDAC表达的影响。结果 HCY明显抑制卵母细胞的体外成熟，在HCY作用下，甲基化组蛋白H3K4、H3K9以及乙酰化组蛋白H3K9的分布没有变化，但是表达强度降低，核呈现去浓缩的趋势。成熟过程中HCY并不改变基因GCN5的表达水平，却明显抑制卵母细胞内HDAC基因的表达。结论卵母细胞发育过程中高水平的同型半胱氨酸影响组蛋白的表观遗传修饰，HCY对卵母细胞核内组蛋白甲基化和乙酰化修饰的影响有可能是造成核染色体稳定性下降的重要原因。

Abstract

Objective To understand the effect of homocysteine（HCY）on histone modification during oocyte development. Methods In this paper the follicle culture system (10 female ICR mouse of 2 weeks) and method of oocyte maturated in vitro (10 female ICR mouse of 4weeks) were used. The processes of oocyte development and maturation were observed. Immunofluorescent staining was performed to show the influence of HCY on distribution of methylated histone H3K4, H3K9, and acetylated histone H3K9 during early stage of oocyte development. With the approach of Real-time PCR, the expression levels of two enzymes controlling histone acetylation, histone deacetylase(HDAC) and histone acetyltransferase (GCN5), in oocyte maturated in vitro were compared between the groups of coculture with homocysteine and control. Results The process of oocyte maturation was significantly inhibited by homocysteine. The distribution patterns of methylated histone H3K4, H3K9 and acetylated histone H3K9 and the staining intensity were decreased by homocysteine. The chromatin appeared and slightly decondensated. The influernce of HCY on GCN5 expression was small, but the expression of HDAC in oocyte was significantly inhibited.Conclusion The effect of homocysteine on histone methylation and acetylation is present during oocyte development. This may be the good reason for the decline of nuclear chromosome stability caused by homocysteine.

导出引用

梁蓉陈曦张育军石程. 同型半胱氨酸对小鼠卵母细胞成熟过程中组蛋白表观遗传修饰的影响[J]. 解剖学报. 2017, 48(5): 585-589 https://doi.org/10.16098/j.issn.0529-1356.2017.05.015

LIANG Rong CHEN Xi ZHANG Yu-jun SHI Cheng. Effects of homocysteine on histone modification during oocyte development in mice[J]. Acta Anatomica Sinica. 2017, 48(5): 585-589 https://doi.org/10.16098/j.issn.0529-1356.2017.05.015

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