The Impact of Ovarian Stimulation on the Expression of Candidate Reprogramming Genes in Mouse Preimplantation Embryos
The Impact of Ovarian Stimulation on the Expression of Candidate Reprogramming Genes in Mouse Preimplantation Embryos
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DOI：
10.1159/000343755
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发表时间：
2012-11
影响因子：
1.7
通讯作者：
Matthias Linke;A. May;Kurt Reifenberg;T. Haaf;Ulrich Zechner
中科院分区：
文献类型：
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作者：
Matthias Linke;A. May;Kurt Reifenberg;T. Haaf;Ulrich Zechner
促性腺激素刺激卵巢是辅助生殖技术治疗人类生育力低下/不孕症的一个组成部分。最近的研究结果表明，卵巢刺激与人类印记疾病的发病率增加以及全基因组甲基化重编程缺陷有关，特别是小鼠的印记。在这里，我们提出了第一项研究，确定卵巢刺激对发育重要的重编程基因（Apex 1，Lig 1，Lig 3，Mbd 2，Mbd 3，Mbd 4和Polb）在单个早期小鼠桑椹胚（16细胞期）的表达的影响。通过实时定量PCR对mRNA进行绝对定量，我们观察到卵巢刺激与单个桑椹胚中编码碱基切除修复蛋白APEX 1和POLB以及5-甲基-CpG结合结构域蛋白MBD 3的mRNA下调相关。用抗APEX 1抗体对早期和晚期桑椹胚进行全胚免疫荧光染色，发现卵巢刺激后单个胚胎的蛋白表达水平较低，mRNA表达与蛋白丰度相关。我们的数据表明，在女性配子发生和/或早期胚胎发育过程中，卵巢刺激会对候选重编程因子的表达产生负面影响。
Ovarian stimulation with gonadotrophins is an integral part of assisted reproductive technologies in human subfertility/infertility treatment. Recent findings have associated ovarian stimulation with the increased incidence of imprinting disorders in humans as well as defects in genome-wide methylation reprogramming and, in particular, imprinting in mice. Here, we present the first study that determined the impact of ovarian stimulation on the expression of developmentally important reprogramming genes (Apex1, Lig1, Lig3, Mbd2, Mbd3, Mbd4, and Polb) in single early mouse morula embryos (16-cell stage). Using absolute quantification of mRNA by quantitative real-time PCR, we observed an association of ovarian stimulation with a downregulation of mRNAs encoding the base excision repair proteins APEX1 and POLB as well as the 5-methyl-CpG-binding domain protein MBD3 in individual morula embryos. Whole mount immunofluorescence staining of early and late morula embryos with an antibody against APEX1 revealed individual embryos with lower protein expression levels after ovarian stimulation and a correlation of mRNA expression with protein abundance. Our data argue for a negative impact of ovarian stimulation during female gametogenesis and/or early embryo development affecting the expression of candidate reprogramming factors.