Drobná Zuzana, Henriksen Anne D, Wolstenholme Jennifer T, Montiel Catalina, Lambeth Philip S, Shang Stephen, Harris Erin P, Zhou Changqing, Flaws Jodi A, Adli Mazhar, Rissman Emilie F
Center for Human Health and the Environment and Department of Biological Sciences, North Carolina State University, Raleigh, North Carolina.
Department of Integrated Science and Technology, MSC 4102, James Madison University, Harrisonburg, Virginia.
Endocrinology. 2018 Jan 1;159(1):132-144. doi: 10.1210/en.2017-00730.
Bisphenol A (BPA) is a ubiquitous man-made endocrine disrupting compound (EDC). Developmental exposure to BPA changes behavioral and reproductive phenotypes, and these effects can last for generations. We exposed embryos to BPA, producing two lineages: controls and BPA exposed. In the third filial generation (F3), brain tissues containing the preoptic area, the bed nucleus of the stria terminalis, and the anterior hypothalamus were collected. RNA sequencing (RNA-seq) and subsequent data analyses revealed 50 differentially regulated genes in the brains of F3 juveniles from BPA vs control lineages. BPA exposure can lead to loss of imprinting, and one of the two imprinted genes in our data set, maternally expressed gene 3 (Meg3), has been associated with EDCs and neurobehavioral phenotypes. We used quantitative polymerase chain reaction to examine the two imprinted genes in our data set, Meg3 and microRNA-containing gene Mirg (residing in the same loci). Confirming the RNA-seq, Meg3 messenger RNA was higher in F3 brains from the BPA lineage than in control brains. This was true in brains from mice produced with two different BPA paradigms. Next, we used pyrosequencing to probe differentially methylated regions of Meg3. We found transgenerational effects of BPA on imprinted genes in brain. Given these results, and data on Meg3 methylation in humans, we suggest this gene may be a biomarker indicative of early life environmental perturbation.
双酚A（BPA）是一种普遍存在的人造内分泌干扰化合物（EDC）。发育期接触BPA会改变行为和生殖表型，且这些影响可持续数代。我们将胚胎暴露于BPA中，产生了两个谱系：对照组和BPA暴露组。在第三代（F3）中，收集了包含视前区、终纹床核和下丘脑前部的脑组织。RNA测序（RNA-seq）及后续数据分析揭示了BPA谱系与对照谱系的F3幼体大脑中有50个差异调节基因。BPA暴露会导致印记丢失，我们数据集中的两个印记基因之一，母源表达基因3（Meg3），已与EDC和神经行为表型相关联。我们使用定量聚合酶链反应来检测数据集中的两个印记基因，即Meg3和含微小RNA的基因Mirg（位于同一基因座）。与RNA-seq结果一致，BPA谱系的F3大脑中Meg3信使核糖核酸高于对照大脑。在两种不同BPA模式产生的小鼠大脑中均是如此。接下来，我们使用焦磷酸测序来探测Meg3的差异甲基化区域。我们发现BPA对大脑中的印记基因有跨代影响。鉴于这些结果以及人类中Meg3甲基化的数据，我们认为该基因可能是早期生活环境扰动的生物标志物。
