Department of Cell and Systems Biology, University of Toronto, Toronto, Ontario, Canada.
Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
PLoS Biol. 2020 Jul 15;18(7):e3000710. doi: 10.1371/journal.pbio.3000710. eCollection 2020 Jul.
During gestation, uterine smooth muscle cells transition from a state of quiescence to one of contractility, but the molecular mechanisms underlying this transition at a genomic level are not well-known. To better understand these events, we evaluated the epigenetic landscape of the mouse myometrium during the pregnant, laboring, and postpartum stages. We generated gestational time point-specific enrichment profiles for histone H3 acetylation on lysine residue 27 (H3K27ac), histone H3 trimethylation of lysine residue 4 (H3K4me3), and RNA polymerase II (RNAPII) occupancy by chromatin immunoprecipitation with massively parallel sequencing (ChIP-seq), as well as gene expression profiles by total RNA-sequencing (RNA-seq). Our findings reveal that 533 genes, including known contractility-driving genes (Gap junction alpha 1 [Gja1], FBJ osteosarcoma oncogene [Fos], Fos-like antigen 2 [Fosl2], Oxytocin receptor [Oxtr], and Prostaglandin G/H synthase 2 (Ptgs2), for example), are up-regulated at day 19 during active labor because of an increase in transcription at gene bodies. Labor-associated promoters and putative intergenic enhancers, however, are epigenetically activated as early as day 15, by which point the majority of genome-wide H3K27ac or H3K4me3 peaks present in term laboring tissue is already established. Despite this early exhibited histone signature, increased noncoding enhancer RNA (eRNA) production at putative intergenic enhancers and recruitment of RNAPII to the gene bodies of labor-associated loci were detected only during labor. Our findings indicate that epigenetic activation of the myometrial genome precedes active labor by at least 4 days in the mouse model, suggesting that the myometrium is poised for rapid activation of contraction-associated genes in order to exit the state of quiescence.
在妊娠期间,子宫平滑肌细胞从静止状态转变为收缩状态,但在基因组水平上,这种转变的分子机制尚不清楚。为了更好地理解这些事件,我们评估了怀孕、分娩和产后阶段小鼠子宫的表观基因组图谱。我们通过大规模平行测序的染色质免疫沉淀(ChIP-seq)生成了妊娠时间点特异性的组蛋白 H3 赖氨酸 27 乙酰化(H3K27ac)、组蛋白 H3 赖氨酸 4 三甲基化(H3K4me3)和 RNA 聚合酶 II(RNAPII)占有率的富集图谱,以及总 RNA-seq(RNA-seq)的基因表达图谱。我们的研究结果表明,包括已知的收缩驱动基因(缝隙连接蛋白α 1 [Gja1]、成骨肉瘤癌基因[Fos]、Fos 样抗原 2 [Fosl2]、催产素受体[Oxtr]和前列腺素 G/H 合酶 2 [Ptgs2])在内的 533 个基因在活跃分娩的第 19 天被上调,因为基因体的转录增加。然而,与分娩相关的启动子和推定的基因间增强子在第 15 天就被表观遗传激活,到那时,在足月分娩组织中存在的大多数全基因组 H3K27ac 或 H3K4me3 峰已经建立。尽管表现出这种早期的组蛋白特征,但只有在分娩期间才能检测到推定的基因间增强子中非编码增强子 RNA(eRNA)的产生增加,以及 RNAPII 募集到与分娩相关基因座的基因体。我们的研究结果表明,在小鼠模型中,子宫基因组的表观遗传激活至少在活跃分娩前提前了 4 天,这表明子宫平滑肌为快速激活与收缩相关的基因做好了准备,以退出静止状态。