Effect of fetal exposure to multiple chemicals on the epigenetic regulation in the testis of mice

本研究的主要目的是研究胎儿暴露于多种化学物质(己烯雌酚)对小鼠睾丸表观遗传学改变的影响。然而，利用限制性地标基因组扫描(RLGS)方法对男性生殖道进行基因扫描并没有得到明确的结果。另一方面，胎儿的腭裂(CP)表现为母亲暴露于全反式维甲酸(AtRA)，这是一种先天性异常，并能够通过化学方法获得一些关于表观遗传学发生机制的发现。尽管过去已经进行了大量的遗传学研究，提出了各种候选基因，但病因仍不清楚。然而，近年来，参与细胞发育和分化的表观遗传学作为一种新的方法受到了人们的关注。在这份报告中，表观遗传学方法(具体地说，dna甲基化)被用于分析…。此外，孕妇暴露于全反式维甲酸中，可能是胎儿发生CP.CPs的机制之一。他们的次级味觉被切除以进行分析。用胞嘧啶延伸试验(CEA)和RLGS分析DNA甲基化状态。实时定量RT-PCR检测DNA甲基转移酶的表达水平。用CEA检测，在妊娠14.5天，atRA组CpG岛和整体DNA的甲基化状态显著低于对照组(p&lt；0.01和p&lt；0.05)。在对照组中，GD14.5的CpG岛内状态显著高于GD13.5和18.5，p&lt；0.01。实时荧光定量RT-PCR检测结果显示，除GD18.5外，DNMTs的表达无明显变化。用RLGS对GD18.5进行检测，发现对照组有5个斑点(0.49%)，atRA处理组有1个斑点(0.1%)，提示DNA甲基化的改变可能在母亲暴露于atRA等环境因素引起的CP的表现中起重要作用。较少

The main purpose of the study was to examine the effect of fetal exposure to multiple chemical (Diethlstilbestrol) on epigenetic alteration in mouse testis. However, we did not obtain a clear finding using Restriction Landmark Genomics Scanning (RLGS) method for male reproductive tract. On the other hand, cleft palate (CP) in fetuses were manifested by maternal exposure to all-trans retinoic acid (atRA) that was the congenital anomaly, and was able to obtain a few findings concerning the generation mechanism of epigenetic with the chemical.CP is caused by both genetic and environmental factors. Although numerous genetic studies have been performed in the past, with various candidate genes suggested, the etiology remains yet unclear. However, in recent years, epigenetics that participates in the development and the differentiation of cells has received the spotlight as a new approach. In this report, an epigenetic approach (in specific, DNA methylation) was utilized in order to analyze … More the mechanisms of CP.CPs in fetuses were manifested by maternal exposure to atRA. Their secondary palates were excised for analysis. The cytosine extension assay (CEA) and RLGS were performed to analyze DNA methylation status. The expression levels of the DNA methyltransferases were examined by real- time RT-PCR.Using CEA, on gestation day (GD) 14.5, the status of DNA methylation within CpG islands and in global DNA was significantly decreased in atRA-treated groups compared to the controls (p<0.01 and p<0.05). In the controls, the status within CpG islands on GD14.5 was significantly increased compared to GD13.5 and 18.5, p<0.01. Using real-time RT-PCR, there was no significant change in the expression of Dnmts, except for on GD 18.5. Using RLGS on GD 18.5, 5 spots (0.49%) in the controls and 1 spot (0.1%) in atRA-treated groups were specifically detected.These results indicate that changes in DNA methylation may play an important role in the manifestation of CP caused by environmental factors such as maternal exposure to atRA. Less