小鼠生发泡期卵母细胞成熟过程中转录沉默机制的探讨
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国家自然科学基金(30700275)


The explore of the mechanism of transcriptional silencing during germinal vesicle oocytes maturation in mice
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    摘要:

    目的:探讨小鼠生发泡卵母细胞成熟过程中发生大规模转录沉默的机制-方法:选用4~6周龄ICR雌性小鼠,分离生发泡(germinal vesicle, GV)期卵母细胞,运用RNA聚合酶Ⅱ抗体(anti-RNA PolⅡCTD, clone 4H8)及其羧基末端结构域(CTD)基团第2位丝氨酸磷酸化的抗体(anti-RNA PolⅡCTD Ser2-P, clone H5)-组蛋白H3K4/H3K9三甲基化的抗体(anti-H3K4me3-anti-H3K9me3)进行免疫荧光染色-观察伴随GV期小鼠卵母细胞染色质构型由非包围核仁(non-surround nucleolus,NSN)型转变为包围核仁(surround nucleolus, SN)型的过程,这两对标记物的表达及分布变化-结果:在NSN型的GV期卵母细胞中,RNA PolⅡCTD Ser2-P于核浆中呈特征性斑块状分布,同时H3K4me3在核浆中也显示强阳性信号;但在SN型卵母细胞中,RNA PolⅡCTD Ser2-P失去特征性分布,转变为核浆中弥漫性分布,H3K4me3的核浆中阳性信号也消失-结论:RNA聚合酶Ⅱ的活性降低及组蛋白修饰状态的改变在小鼠GV期卵母细胞成熟过程的大规模转录沉默中发挥了重要作用-

    Abstract:

    Objective: To explore the mechanism of large-scale transcriptional silence during germinal vesicle (GV) oocytes maturation in mice. Methods:The separated GV oocytes were obtained from 4~6-week-old ICR female mice. As for the observation about the expressional and distributional changes of both markers in the process of the chromatin configuration from non-surround nucleolus(NSN) type into surround nucleolus(SN) type in mice GV oocytes, these oocytes were immunofluorescence stained with RNA polymeraseⅡ antibody(anti-RNA PolⅡCTD, clone 4H8) and the second serine phosphorylated antibody at RNA polymeraseⅡCTD group (anti-RNA PolⅡCTD Ser2-P, clone H5); as well as histone H3K4/H3K9 tri-methylation antibodies (anti-H3K4me3, anti-H3K9me3). Results:In NSN type GV oocytes, RNA CTD PolⅡSer2-P was distributed with characteristic plaques shape in nucleus; meanwhile, H3K4me3 also presented the strong positive signals in nucleus. However, in SN type oocytes, RNA CTD PolⅡSer2-P lost the characteristic distribution and diffused in nucleus; moreover, the positive signal in nucleus showed by H3K4me3 disappeared. Conclusion:The reduction of RNA polymeraseⅡactivity and the change of histone modification status played an important role in the large-scale transcriptional silencing during germinal vesicle oocytes maturation of mice.

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夏 梦,刘明兮,周 涛,何 辉,万 茹,霍 然.小鼠生发泡期卵母细胞成熟过程中转录沉默机制的探讨[J].南京医科大学学报(自然科学版),2011,(7):956-961

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  • 收稿日期:2011-01-21
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