Research on DNA methylation for two-generation pairs with schizophrenia (Anticipation and modulation of gene expression in schizophrenia)

精神分裂症两代人DNA甲基化研究（精神分裂症基因表达的预测和调节）

• 批准号: 13671016
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671016/
• 关键词: schizophrenia; anticpation; monozygotic twin; DNA methylation; methylation-sensitive restriction enzyme; HPLC; 精神分裂病(統合失調症); 遺伝子発現調整; RLGS法; M-PCR法; 精神分裂病

The aim of this study was to explore the molecular mechanisms including epigenetic DNA modification underlying genetic anticipation in schizophrenia. Twelve two-generation pairs with schizophrenia, which showed prominent anticipation., were examined.First, we investigated whether the promoter region of dopamine D2 receptor gene (DRD2) was methylated or not, using the methylation-sensitive restriction enzyme, Hap II, and the methylation-insensitive enzyme, MspI. After enzyme digestion, polymerase chain reaction (PCR) was performed, and. the. size of PCR products was determined with polyacrylamide gel electrophoresis. No difference was found between two generation in methylation of the promoter region of DRD2.Second, we measured 5-methyl-2'-deoxycytidine (mC) contents by high performance liquid chromatography (HPLC) in peripheral leukocytes DNA obtained from eight two-generation pairs with schizophrenia, to compare the global methylation levels between two generation. We obtained no significant results in this method.In summary, we found no evidence for the relationship between genetic anticipation and DNA methylation in schizophrenia. Therefore in future studies we will aim to collect more subjects and try other enzymes.

本研究的目的是探索精神分裂症遗传预期的分子机制，包括表观遗传DNA修饰。首先，我们利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，研究了多巴胺D2受体基因(DRD2)的启动子区是否发生甲基化。酶消化后，进行聚合酶链式反应(PCR)。这个。用聚丙烯酰胺凝胶电泳法测定扩增产物的大小。第二，用高效液相色谱法测定8对两代精神分裂症患者外周血白细胞DNA中5-甲基-2‘-脱氧胞苷(MC)的含量，比较两代精神分裂症患者外周血白细胞DNA的整体甲基化水平。我们在这种方法中没有得到显著的结果。总而言之，我们没有发现精神分裂症患者的遗传预期和DNA甲基化之间存在联系的证据。因此，在未来的研究中，我们将致力于收集更多的对象，并尝试其他酶。

项目成果

Matsumoto S., Sasaki T., Imamura A., et al.: "HLA classI distribution in Japanese patients with schizophrenia"Am J Med Genet. 114. 42-45 (2002)

Matsumoto S.、Sasaki T.、Imamura A. 等：“日本精神分裂症患者的 HLA I 类分布”Am J Med Genet。

The Japanese Schizophrenia Sib-pair Linkage Group (JSSLG): "Initial genome-wide scan for linkage with schizophrenia in the Japanese Schizophrenia Sib-pair Linkage Group (JSSLG) families"Am J Med Genet. 120B. 22-28 (2003)

日本精神分裂症同胞对连锁群 (JSSLG)：“对日本精神分裂症同胞对连锁群 (JSSLG) 家族中与精神分裂症的连锁进行初始全基因组扫描”Am J Med Genet。

Imamura A..Tsujita T., Kayashima T., et al.: "Lack of association between the hKCa3 gene and the Japanese schizophrenia patients"Psychiatric Genetics. 11. 227-229 (2001)

Imamura A..Tsujita T.、Kayashima T. 等人：“hKCa3 基因与日本精神分裂症患者之间缺乏关联”精神病遗传学。