Faculty of Pharmaceutical Sciences, International University of Health and Welfare, 2600-1 Ootawara, Tochigi 324-8501, Japan.
Biochem Biophys Res Commun. 2010 Mar 26;394(1):173-7. doi: 10.1016/j.bbrc.2010.02.139. Epub 2010 Feb 25.
To understand epigenetic regulation of neurotrophins in Neuro-2a mouse neuroblastoma cells, we investigated the alteration of CpG methylation of brain-derived neurotrophic factor (BDNF) promoter I and neurotrophin-3 (NT-3) promoter IB and that of histone modification in Neuro-2a cells. Bisulfite genomic sequencing showed that the CpG sites of BDNF promoter I were methylated in non-treated Neuro-2a cells and demethylated following 5-aza-2'-deoxycytidine (5-aza-dC) treatment. In contrast, methylation status of the NT-3 promoter IB did not change by 5-aza-dC treatment in Neuro-2a cells. Furthermore, we demonstrated that BDNF exon I-IX mRNA was induced by trichostatin A (TSA) treatment. However, NT-3 exon IB-II mRNA was not induced by TSA treatment. Chromatin immunoprecipitation assays showed that the levels of acetylated histones H3 and H4 on BDNF promoter I were increased by TSA. These results demonstrate that DNA methylation and/or histone modification regulate BDNF gene expression, but do not regulate NT-3 gene expression in Neuro-2a cells.
为了了解神经滋养因子在Neuro-2a 鼠神经母细胞瘤细胞中的表观遗传调控，我们研究了脑源性神经营养因子（BDNF）启动子 I 和神经营养因子-3（NT-3）启动子 IB 的 CpG 甲基化改变以及Neuro-2a 细胞中的组蛋白修饰。亚硫酸氢盐基因组测序显示，未经处理的 Neuro-2a 细胞中 BDNF 启动子 I 的 CpG 位点被甲基化，而经 5-氮杂-2'-脱氧胞苷（5-aza-dC）处理后则去甲基化。相比之下，5-aza-dC 处理并未改变 Neuro-2a 细胞中 NT-3 启动子 IB 的甲基化状态。此外，我们证明 Trichostatin A（TSA）处理可诱导 BDNF exon I-IX mRNA 的表达。然而，TSA 处理并未诱导 NT-3 exon IB-II mRNA 的表达。染色质免疫沉淀分析显示，TSA 增加了 BDNF 启动子 I 上乙酰化组蛋白 H3 和 H4 的水平。这些结果表明，DNA 甲基化和/或组蛋白修饰调节 Neuro-2a 细胞中 BDNF 基因的表达，但不调节 NT-3 基因的表达。
