Epigenetic Transgenerational Actions of Vinclozolin on Promoter Regions of the Sperm Epigenome
Epigenetic Transgenerational Actions of Vinclozolin on Promoter Regions of the Sperm Epigenome
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DOI：
10.1371/journal.pone.0013100
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发表时间：
2010-09-30
期刊：
影响因子：
3.7
通讯作者：
Skinner, Michael K.
中科院分区：
文献类型：
--
作者：
Guerrero-Bosagna, Carlos;Settles, Matthew;Skinner, Michael K.
先前的观察表明，胚胎在性腺性别决定过程中暴露于内分泌干扰物长春花碱会促进跨代成人发病疾病，如男性不育症、肾脏疾病、前列腺疾病、免疫异常和肿瘤的发展。目前的研究调查了F3代大鼠精子中全基因组启动子DNA甲基化的变化，这些大鼠的F0代母亲暴露于长春花碱。用甲基胞嘧啶抗体甲基化DNA免疫沉淀和启动子切割微阵列(MeDIP-Chip)程序鉴定了精子启动子组中52个甲基化显著改变的不同区域。用质谱仪对CpG DNA甲基化进行分析，确定了16个差异DNA甲基化区域，其余区域由于亚硫酸氢盐技术的限制而无法分析。对这些有效区域的分析确定了一个与75%的启动子相关的共识DNA序列(基序)。有趣的是，在随机选取的125个启动子中，只有16.8%包含这个基序。一个候选启动子(Fam111a)被发现是由于拷贝数变异(CNV)而不是甲基化改变，这表明种系表观基因组的初始改变可能促进遗传异常，如在后代中诱导CNV。这项研究确定了最初暴露三代后启动子区域的差异DNA甲基化位点，并确定了这些区域存在的共同基因组特征。除了初级表观突变外，还发现了一种潜在的间接遗传异常，两者都被认为与所观察到的表观遗传跨代遗传有关。这项研究证实，环境因素有能力诱导精子表观基因组的表观遗传跨代变化。
Previous observations have demonstrated that embryonic exposure to the endocrine disruptor vinclozolin during gonadal sex determination promotes transgenerational adult onset disease such as male infertility, kidney disease, prostate disease, immune abnormalities and tumor development. The current study investigates genome-wide promoter DNA methylation alterations in the sperm of F3 generation rats whose F0 generation mother was exposed to vinclozolin. A methylated DNA immunoprecipitation with methyl-cytosine antibody followed by a promoter tilling microarray (MeDIP-Chip) procedure was used to identify 52 different regions with statistically significant altered methylation in the sperm promoter epigenome. Mass spectrometry bisulfite analysis was used to map the CpG DNA methylation and 16 differential DNA methylation regions were confirmed, while the remainder could not be analyzed due to bisulfite technical limitations. Analysis of these validated regions identified a consensus DNA sequence (motif) that associated with 75% of the promoters. Interestingly, only 16.8% of a random set of 125 promoters contained this motif. One candidate promoter (Fam111a) was found to be due to a copy number variation (CNV) and not a methylation change, suggesting initial alterations in the germline epigenome may promote genetic abnormalities such as induced CNV in later generations. This study identifies differential DNA methylation sites in promoter regions three generations after the initial exposure and identifies common genome features present in these regions. In addition to primary epimutations, a potential indirect genetic abnormality was identified, and both are postulated to be involved in the epigenetic transgenerational inheritance observed. This study confirms that an environmental agent has the ability to induce epigenetic transgenerational changes in the sperm epigenome.