DNA methylation changes during cleft palate formation induced by retinoic acid in mice
DNA methylation changes during cleft palate formation induced by retinoic acid in mice
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DOI：
10.1597/07-134.1
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发表时间：
2008-09-01
期刊：
影响因子：
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通讯作者：
Mori, Chisato
中科院分区：
文献类型：
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作者：
Kuriyama, Motone;Udagawa, Akikazu;Mori, Chisato
目的：本研究的目的是分析表观遗传（特别是DNA甲基化）参与小鼠全反式维甲酸暴露诱导的腭裂机制。设计：胎儿腭裂仅由母体接触全反式维甲酸诱发。切除第二腭进行分析。胞嘧啶延伸试验和限制性标记基因组扫描分析DNA甲基化状态。实时逆转录-聚合酶链反应检测DNA甲基转移酶的表达水平。结果：胞嘧啶延长试验显示，妊娠第14.5天，与对照组相比，全反式维甲酸处理组CpG岛内和全DNA甲基化水平显著降低（p < 0.01和p < 0.05）。对照组妊娠第14.5天CpG岛内状态较妊娠第13.5和18.5天显著升高（p < 0.01）。实时逆转录-聚合酶链反应显示，除妊娠第18.5天外，DNA甲基转移酶的表达无显著变化。妊娠第18.5天限制性限制标记基因组扫描，在对照组中特异性检测到5个位点（0.49%），在全反式维甲酸处理组中特异性检测到1个位点（0.1%）。结论：上述结果提示，DNA甲基化的改变可能仅在母亲全反式维甲酸暴露等环境因素引起的腭裂表现中起重要作用。
Objectives: The aim of this study was to analyze epigenetic (specifically, DNA methylation) participation in the mechanisms of cleft palate only induced by maternal exposure to all-trans retinoic acid in mice.Design: Cleft palate only was induced in fetuses by maternal exposure to all-trans retinoic acid. Their secondary palates were excised for analysis. Cytosine extension assay and restriction landmark genomic scanning were performed to analyze DNA methylation status. The expression levels of the DNA methyltransferases were examined by real-time reverse transcriptase-polymerase chain reaction.Results: Using cytosine extension assay, on gestation day 14.5, the status of DNA methylation within CpG islands and in global DNA was decreased significantly in all-trans retinoic acid-treated groups compared with the controls (p < .01 and p < .05). In the controls, the status within CpG islands on gestation day 14.5 was significantly increased compared with gestation days 13.5 and 18.5 (p < .01). Using real-time reverse transcriptase-polymerase chain reaction, there was no significant change in the expression of DNA methyltransferases, except on gestation day 18.5. Using restriction landmark genomic scanning on gestation day 18.5, five spots (0.49%) in the controls and one spot (0.1%) in all-trans retinoic acid-treated groups were specifically detected.Conclusions: These results indicate that changes in DNA methylation may play an important role in the manifestation of cleft palate only caused by environmental factors such as maternal exposure to all-trans retinoic acid.