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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

我们试图探讨精神分裂症与DNA甲基化和组蛋白乙酰化等表观遗传事件的关系。首先，我们研究了多巴胺D2受体基因（DRD 2）的启动子区域是否被甲基化，使用甲基化敏感的限制性内切酶，Hap II，和甲基化不敏感的酶，MspI。受试者是五对DSM-IV精神分裂症不一致的单卵双胞胎（所有对都是男性，平均年龄42.6岁;所有双胞胎都不一致超过十年）。采用酚法提取淋巴细胞基因组DNA。酶切后，进行聚合酶链反应（PCR），并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，DRD 2启动子区域的甲基化水平在受影响的双胞胎中高于正常双胞胎。在一对中，两者的水平几乎相等。在这项研究中，我们发现了一些证据， ...更多信息精神分裂症不一致的单卵双生子表型差异与DNA甲基化之间的关系。因此，在今后的研究中，我们将尝试收集更多的受试者，并使用其他酶。接下来，我们对精神分裂症不一致的单卵双生子进行SNPs基因分型和基因表达分析，并在双生子之间进行比较。受试者是一对同卵双胞胎（年龄：51岁），一个患有DSM-IV精神分裂症，未分化型，另一个有工作，已婚，适应社会生活。样本是来自受试者的基因组DNA和RNA。使用Affyscore GeneChip Human Mapping 100 K Set（50 K Xba Array，50 K Hind Array）对SNP进行分型，并使用Affyscore Human Genome U133 Plus 2.0 Array分析基因表达。在SNPs基因分型中，在双生子间的116204个SNPs中，有46个SNPs具有不同的信号强度。此外，我们测量了12857个基因的表达水平。结果发现4个信号强度不同的SNPs，它们位于基因的5'端或内含子附近，表明双生子之间存在基因表达差异。通过测序，我们证实在这些区域中没有DNA序列的改变。因此，可以认为某些表观遗传事件引起了基因表达的改变。我们将研究包括这些SNPs的基因与精神分裂症之间的关联。少