他莫昔芬处理雄性大鼠胚胎中多个印记基因的异常甲基化。

Aberrant methylation of multiple imprinted genes in embryos of tamoxifen-treated male rats.

机构信息

Neuroendocrinology Division, National Institute for Research in Reproductive Health (ICMR), Parel, Mumbai 400 012, India.

出版信息

Reproduction. 2013 Jul 3;146(2):155-68. doi: 10.1530/REP-12-0439. Print 2013 Aug.

Abstract

Genomic imprinting is an epigenetic phenomenon known to regulate fetal growth and development. Studies from our laboratory have demonstrated that treatment of adult male rats with tamoxifen increased postimplantation loss around mid gestation. Further studies demonstrated the aberrant expression of transcripts of several imprinted genes in the resorbing embryos at days 11 and 13 of gestation including IGF2. In addition, decreased methylation at the Igf2-H19 imprint control region was observed in spermatozoa and in resorbing embryos sired by tamoxifen-treated males. In this study, methylation analysis of the imprinted genes, which were found to be differentially expressed, was done using EpiTYPER in the spermatozoa of tamoxifen-treated rats and in postimplantation embryos sired by tamoxifen-treated rats. Differentially methylated regions (DMRs) for most imprinted genes have not been identified in the rats. Hence, initial experiments were performed to identify the putative DMRs in the genes selected for the study. Increased methylation at CpG islands present in the putative DMRs of a number of imprinted genes was observed in the resorbing embryos sired by tamoxifen-treated male rats. This increase in methylation is associated with the downregulation of most of these genes at the transcript level in resorbing embryos. No change in the methylation status of these genes was observed in spermatozoa. These observations suggest that a deregulation of mechanisms protecting unmethylated alleles from a wave of de novo methylation occurs following implantation.

摘要

基因组印记是一种已知调节胎儿生长和发育的表观遗传现象。我们实验室的研究表明,用他莫昔芬治疗成年雄性大鼠会增加妊娠中期的胚胎着床后损失。进一步的研究表明,在妊娠第 11 和 13 天吸收的胚胎中,几个印记基因的转录本表达异常,包括 IGF2。此外,在他莫昔芬处理过的雄性大鼠的精子和吸收的胚胎中观察到 Igf2-H19 印记控制区的甲基化减少。在这项研究中,使用 EpiTYPER 对差异表达的印记基因进行了甲基化分析,这些基因存在于他莫昔芬处理过的大鼠的精子和他莫昔芬处理过的大鼠的着床后胚胎中。在大鼠中,大多数印记基因的差异甲基化区域(DMR)尚未被鉴定。因此,最初的实验是在研究中选择的基因中识别假定的 DMR。在他莫昔芬处理过的雄性大鼠的吸收胚胎中,在一些印记基因的假定 DMR 中存在的 CpG 岛的甲基化增加。这种甲基化的增加与这些基因在吸收胚胎中的转录水平的大部分下调有关。在精子中未观察到这些基因的甲基化状态发生变化。这些观察结果表明,在植入后,保护未甲基化等位基因免受新甲基化波影响的机制失调。

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