H3K4和H3K9三甲基化在猪体外成熟卵母细胞和克隆胚胎中的表达分析

发布时间：2018-06-21 10:25

本文选题：HK三甲基化 + HK三甲基化　；参考：《西南大学学报(自然科学版)》2017年05期

【摘要】：该研究主要运用单细胞免疫荧光技术分析猪卵母细胞和克隆胚胎发育过程中,H3K4和H3K9三甲基化(H3K4me3,H3K9me3)的表达情况.收集体外成熟0h,6h,12h,24h,30h,36h,42h的猪卵母细胞;同时通过体细胞核移植操作获得一细胞、四细胞、桑葚胚和囊胚期克隆胚胎,经过免疫荧光染色,在激光共聚焦显微镜下观察H3K4me3和H3K9me3修饰在卵母细胞和胚胎中的变化情况.免疫荧光结果显示,在不同减数分裂阶段的猪卵母细胞内均能观察到H3K4me3的表达,并且呈现出逐渐下降的趋势;在生发泡期(GV),生发泡晚期(L-GV)和生发泡破裂(GVBD)阶段的卵母细胞中能发现明显的H3K9me3表达,随着减数分裂的进行,H3K9me3表达消失,在第一次减数分裂中期前阶段(Pre-MI)到第二次减数分裂中期(MⅡ)阶段卵母细胞中未发现H3K9me3荧光信号.H3K4me3和H3K9me3在不同发育阶段的猪克隆胚胎中都有表达,其中H3K4me3在不同阶段胚胎中荧光信号强度相近,H3K9me3在四细胞胚胎中荧光强度降低,在桑葚胚和囊胚中荧光强度出现升高的趋势.以上结果说明,H3K4me3参与调控猪卵母细胞体外成熟的整个过程,H3K4me3和H3K9me3在猪体细胞核移植胚胎早期发育过程中可能具有调控作用.
[Abstract]:In this study, the expression of H3K4 and H3K9 trimethylated H3K4 and H3K9 in porcine oocytes and cloned embryos were analyzed by single cell immunofluorescence technique. Porcine oocytes were collected from mature porcine oocytes in vitro from 0 h to 6 h ~ (12) h ~ (24) h ~ (30) h ~ (36) h ~ (-1) and 1 cell, 4 cell, morula and blastocyst stage cloned embryos were obtained by somatic cell nuclear transfer, and were stained by immunofluorescence. The changes of H3K4me3 and H3K9me3 modification in oocytes and embryos were observed under confocal laser microscope. The results of immunofluorescence showed that the expression of H3K4me3 was observed in porcine oocytes at different meiosis stages, and the expression of H3K4me3 decreased gradually. The expression of H3K9me3 was found in the oocytes at the stage of foaming stage, late foaming stage (L-GV) and foaming rupture stage (GVBDD), and disappeared with the meiosis of meiosis, and the expression of H3K9me3 disappeared with the meiosis, and the expression of H3K9me3 disappeared with the development of meiosis. No H3K9me3 fluorescence signal. H3K4me3 and H3K9me3 were expressed in porcine cloned embryos at different stages of development. The fluorescence intensity of H3K4me3 was similar in different stages of embryos. The fluorescence intensity of H3K9me3 decreased in four-cell embryos and increased in morula and blastocyst. These results suggest that H3K4me3 is involved in the regulation of porcine oocyte maturation in vitro. H3K4me3 and H3K9me3 may play a regulatory role in the early development of porcine somatic nuclear transfer embryos.
【作者单位】：广西大学亚热带农业生物资源保护与利用国家重点实验室;柳州市人民医院;
【基金】：国家自然科学基金(31560632) 广西自然科学基金(2014GXNSFAA118084,2014GXNSFAA118099,2016GXNSFDA380030)
【分类号】：S828

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