组蛋白3在小鼠嵌合胚泡中的表达与甲基化
Expression and Methylation of Histone 3 in Mice Chimeric Blastocysts.
作者信息
Salimi Maryam, Shirazi Abolfazl, Norouzian Mohsen, Jafari Ameneh, Edalatkhah Haleh, Mehravar Maryam, Majidi Mohammad, Mehrazar Mohammad Mahdi
机构信息
Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
出版信息
Rep Biochem Mol Biol. 2020 Oct;9(3):357-365. doi: 10.29252/rbmb.9.3.357.
BACKGROUND
Currently, the efficient production of chimeric mice and their survival are still challenging. Recent researches have indicated that preimplantation embryo culture media and manipulation lead to abnormal methylation of histone in the promotor region and consequently alter their gene expression pattern. This investigation was designed to evaluate the relationship between the methylation state of histone H3 and expression in mice chimeric blastocysts.
METHODS
Mouse 129/Sv embryonic stem cells (mESCs) expressing the green fluorescent protein (mESCs-GFP) were injected into the perivitelline space of 2.5 days post-coitis (dpc) embryos (C57BL/6) using a micromanipulator. H3K4 and H3K9 methylation, and H19 and expression was measured by immunocytochemistry and q-PCR, respectively, in blastocysts.
RESULTS
Histone H3 trimethylation in H3K4 and H3K9 in chimeric blastocysts was significantly less and greater, respectively (p< 0.05), than in controls. expression was significantly less (p< 0.05), while expression was less, but not significantly so, in chimeric than in control blastocysts.
CONCLUSION
Our results showed, that the alteration ofH3K4me3 and H3K9me3 methylation, change expression in chimeric blastocysts.
背景
目前，嵌合小鼠的高效生产及其存活仍然具有挑战性。最近的研究表明，胚胎植入前培养基和操作会导致启动子区域组蛋白异常甲基化，从而改变其基因表达模式。本研究旨在评估组蛋白H3甲基化状态与小鼠嵌合囊胚中基因表达之间的关系。
方法
使用显微操作器将表达绿色荧光蛋白的小鼠129/Sv胚胎干细胞（mESCs-GFP）注射到交配后2.5天（dpc）的胚胎（C57BL/6）的卵周隙中。分别通过免疫细胞化学和q-PCR检测囊胚中H3K4和H3K9甲基化以及H19基因表达。
结果
嵌合囊胚中H3K4和H3K9的组蛋白H3三甲基化分别显著低于和高于对照组（p<0.05）。嵌合囊胚中的基因表达显著降低（p<0.05），而基因表达降低，但差异不显著。
结论
我们的结果表明，H3K4me3和H3K9me3甲基化的改变会改变嵌合囊胚中的基因表达。
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