Differential developmental requirements for individual histone H3K9 methyltransferases in cleavage-stage porcine embryos
Differential developmental requirements for individual histone H3K9 methyltransferases in cleavage-stage porcine embryos
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DOI：
10.1071/rd10280
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发表时间：
2011-01-01
影响因子：
1.9
通讯作者：
Cabot, Ryan A.
中科院分区：
文献类型：
--
作者：
Park, Ki-Eun;Johnson, Christine M.;Cabot, Ryan A.
二甲基H3K9是一种可遗传的表观遗传标记，与转录沉默密切相关，并在卵裂发育过程中经历全球重塑。哺乳动物中有5种组蛋白甲基转移酶(HMTase)，即SUV39H1、Suv39H2、SetDB1、EHMT1和EHMT2，它们介导H3K9的甲基化。本研究的目的是确定这些HMTase在猪胚胎卵裂发育过程中的发育需求。我们假设上述HMTase的敲除将对猪卵裂发育产生不同的影响。为了验证这一假设，将猪体外受精和体外受精的卵母细胞分为三个处理组，包括未注射对照组、注射针对HMTase之一的双链干扰RNA分子的卵母细胞和注射相应突变(对照)双链RNA分子的卵母细胞。受精后6天，对所有胚胎进行核计数。虽然注射EHMT1和EHMT2干扰RNA的胚胎细胞总数与各自的对照组相比没有显著差异，但注射针对SUV39H1、Suv39H2和SetDB1的干扰RNA的胚胎的细胞总数显著低于各自的对照组(P
Dimethylated H3K9 is a heritable epigenetic mark that is closely linked with transcriptional silencing and known to undergo global remodelling during cleavage development. Five mammalian histone methyltransferases (HMTases), namely Suv39H1, Suv39H2, SetDB1, EHMT1 and EHMT2, have been shown to mediate the methylation of H3K9. The aim of the present study was to determine the developmental requirements of these HMTases during cleavage development in porcine embryos. We hypothesised that knockdown of the above mentioned HMTases would differentially affect porcine cleavage development. To test this hypothesis, IVM and IVF porcine oocytes were divided into one of three treatment groups, including non-injected controls, oocytes injected with a double-stranded interfering RNA molecule specific for one of the HMTases and oocytes injected with a corresponding mutated (control) double-stranded RNA molecule. Nuclei were counted in all embryos 6 days after fertilisation. Although no significant difference in total cell number was detected in embryos injected with EHMT1 and EHMT2 interfering RNAs (compared with their respective control groups), embryos injected with interfering RNAs that targeted Suv39H1, Suv39H2 and SetDB1 had significantly lower cell numbers than their respective control groups (P