The investigation of the histone proteins acetylation and the DNA methylation in monozygotic twins discordant for schizophrenia

精神分裂症不一致同卵双胞胎组蛋白乙酰化和 DNA 甲基化的研究

基本信息

批准号：
17591221
负责人：
IMAMURA Akira
金额：
$ 2.24万
依托单位：
Nagasaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

项目摘要

We tried to investigate the relationship between schizophrenia and the epigenetic events such as the methylation of DNA and the acetylation of histone proteins. First, we examined whether the promoter region of dopamine D2 receptor gene (DRD2) was methylated, using the methylation-sensitive restriction enzyme, Hap II, and the methylation-insensitive enzyme, MspI. The subjects were five pairs of monozygotic twins discordant for DSM-IV schizophrenia (all pairs were male, mean age 42.6 years; all twins had been discordant more than ten years). The genome DNA samples were extracted from lymphocytes by the phenol method. After enzyme digestion, polymerase chain reaction (PCR) was performed, and the size of PCR products was determined using polyacrylamide gel electrophoresis. In four out of five pairs, the level of methylation of the DRD2 promoter region was higher in the affected twin than the normal twin. In one pair, the level was nearly equal in both. In this study, we found some evidenc … More e for the relationship between phenotypic difference in monozygotic twins discordant for schizophrenia and DNA methylation. Therefore in future studies we will try to collect more subjects and use other enzymes.Next, we performed SNPs genotyping and gene expression analysis in monozygotic twins discordant for schizophrenia and compared between twins. The subjects were a pair of monozygotic male twins (age:51), one had DSM-IV schizophrenia, undifferentiated type, and the other had a job, was married, and was well-adapted to social life. The samples were genomic DNA and RNA from the subjects. The SNPs were typed using Affymetrix GeneChip Human Mapping 100K Set (50K Xba Array, 50K Hind Array) and the gene expressions were analyzed by Affymetrix Human Genome U133 Plus 2.0 Array. In SNPs genotyping, forty six SNPs had different signal intensity among all 116204 SNPs between twins. Also, we measured the expression level of 12857 genes. As a results, we detected 4 SNPs that had different signal intensity and located near the 5' end or intron of the genes that showed the different gene expression between twins. By sequencing, we confirmed that there were no alterations of DNA sequences in these regions. Therefore, it could be thought that some epigenetic events caused the change of gene expression. We will investigate the associations between the genes including these SNPs and schizophrenia. Less

我们试图研究精神分裂症与DNA甲基化和组蛋白乙酰化等表观遗传事件之间的关系，首先，我们利用甲基化敏感限制条件检查了多巴胺D2受体基因（DRD2）的启动子区域是否甲基化。酶 Hap II 和甲基化不敏感酶 MspI 受试者是五对 DSM-IV 精神分裂症不一致的同卵双胞胎。（所有配对均为男性，平均年龄42.6岁；所有双胞胎均已不一致十多年）通过苯酚法从淋巴细胞中提取基因组DNA样本，进行酶消化，进行聚合酶链式反应（PCR）。使用聚丙烯酰胺凝胶电泳测定 PCR 产物的大小，在五对中的四对中，受影响的双胞胎中的 DRD2 启动子区域的甲基化水平高于正常双胞胎中的水平。在这项研究中，我们发现了一些证据证明精神分裂症不一致的同卵双胞胎的表型差异与 DNA 甲基化之间的关系。因此，在未来的研究中，我们将尝试收集更多的受试者并使用其他酶。接下来，我们进行了 SNP 基因分型和分析。对精神分裂症不一致的同卵双胞胎进行基因表达分析，并在双胞胎之间进行比较。双胞胎（年龄：51），其中一个患有DSM-IV型精神分裂症，未分化型，另一个有工作，已婚，并且很好地适应了社会生活。样本是来自受试者的基因组DNA和RNA。使用 Affymetrix GeneChip Human Mapping 100K Set（50K Xba Array、50K Hind Array）分型，并通过 Affymetrix Human Genome 分析基因表达U133 Plus 2.0 Array 在双胞胎之间的所有 116204 个 SNP 中，有 46 个 SNP 具有不同的信号强度。此外，我们还测量了 12857 个基因的表达水平，结果，我们检测到了 4 个具有不同信号强度且位于附近的 SNP。显示双胞胎之间基因表达不同的基因的 5' 端或内含子通过测序，我们确认双胞胎中的 DNA 序列没有改变。因此，可以认为一些表观遗传事件导致了基因表达的变化，我们将研究包括这些SNP在内的基因与精神分裂症之间的关联。