Pharmacogenomic research for mood disorder

情绪障碍的药物基因组学研究

• 批准号: 12558089
• 负责人: SAITO Naoaki
• 金额: $ 7.17万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12558089/
• 关键词: serotonin transporter; Genomic analysis; polymorphism; mode disorder; promoter; reporter assay

Three variants of routine serotonin transporter (5-HTT) mRNA, which consist of a different exon-one (exon-la, exon-1b or exon-1c) and the same exon-two to exon-five, were identified. Any promoter region, ligated to PGL-3 enhancer vector, had activities significantly higher than the empty vector in some cell lines tested, where as the activity for the exon-1c was significantly lower than the others. In COS-7 cell lines, dibutyryl-cyclic AMP (Dib-cAMP) treatment decreased the activity for the exon-lc and did not change the others. Human interferon-*** (ENF-***) treatment decreased the activity for the exon-la, did not change the one for the exon-lb and increased the one for exon-lc. In PC12 cell lines the promoter regions for exon-la and exon-lb had higher activities than the mock vector. The promoter activity for exon-l c did not differ from that of mock vector. Dib-cAMP treatment increased the activities of all of the constructs whereas human INF-α application did not change any of the … More m. In immortalized rat serotoneigic raphe neurons, RN46A, the insertion of the promoter region for exonl a, exonlb and exonlc increased the activity twelve, sixteen or three times respectively. Dib-cAMP slightly increased their activity where as murine interferon *** did not change them. These three promoter regions may play a role in transcription of 5-HTT and could offer a model of the regulation of 5-HTT production in human and further the pathogenesis of depression and other serotonin spectrum disorders.Enhancer/silencer activity of each allelic variant of the human serotonin transporter linked polymorphic region (5HTTLPR) including newly found ones was measured in several cell lines including raphe-nucleus-derived RN46A. 5-HTTLR variants ligated in pGL-3 promote1 vector increased hiciferase activity in COS-7 cells and PC12 cells, where no significant differences among the variants were observed. In RN46A cell lines, however, 5-HTTLPRs decreased hiciferase activity to eighty to thirty percent, acting as silencers not as enhancers. Some allelic variants (15, 19, 20, and 22) showed even significantly stronger silencer activities than others in RN46A. We also examined relationship between allelic frequencies, the enhancer/silencer activities and incidents of mood disorder. The categorized genotypic or allelic frequencies was not significantly different between the mood disorder and the control. No significant difference was detected either when analyzed by silencer activities of each allelic variant.Three variants ofmurine serotonin transporter (5-HTT) mRNA, which consist of a different exon-one (exon-la, exon-lb or exon-lc) and the same exon4wo to exon-five, were identified. Any promoter region, ligated to pGL-3 enhancer vector, had activities significantly higher than the empty vector in some cell lines tested, where as the activity for the exon-lc was significantly lower than the others. Less

常规5-羟色胺转运体（5-HTT）mRNA的三种变体，由不同的外显子1（外显子1a，外显子1b或外显子1c）和相同的外显子2至外显子5组成。任何启动子区域，连接到PGL-3增强子载体，在一些测试的细胞系中具有比空载体显著更高的活性，其中外显子-1c的活性显著低于其他。在COS-7细胞系中，二丁酰环腺苷酸（Dib-cAMP）处理降低了外显子-lc的活性，而没有改变其他。人干扰素-*（ENF-*）处理降低了外显子-Ia的活性，不改变外显子-Ib的活性，而增加了外显子-Ic的活性。在PC 12细胞系中，外显子-Ia和外显子-Ib的启动子区域具有比模拟载体更高的活性。外显子lc的启动子活性与模拟载体的启动子活性没有差异。Dib-cAMP处理增加了所有构建体的活性，而人INF-α的应用没有改变任何结构。 ...更多信息 M.在永生化的大鼠中缝核神经元RN 46 A中，插入exonla、exonlb和exonlc的启动子区域分别使活性增加12、16或3倍。Dib-cAMP略微增加了它们的活性，而鼠干扰素 * 没有改变它们。这三个启动子区可能在5-HTT的转录中起作用，并可能提供一个模型，调节5-HTT的生产在人类和进一步的发病机制，抑郁症和其他5-羟色胺spectrum disorders.Enhancer/silencer活性的人5-羟色胺转运蛋白连锁多态性区域（5-HTTLPR）的每个等位基因的变体，包括新发现的几个细胞系，包括中缝核-衍生RN 46 A。5-在COS-7细胞和PC 12细胞中，连接在pGL-3启动子1载体中的HTTLR变体增加了hiciferase活性，其中在变体之间没有观察到显著差异。然而，在RN 46 A细胞系中，5-HTTLPRs将hiciferase活性降低至80%至30%，充当沉默剂而不是增强剂。一些等位基因变体（15、19、20和22）显示出比RN 46 A中的其他等位基因变体更强的沉默物活性。我们还研究了等位基因频率，增强子/沉默子活动和情绪障碍事件之间的关系。分类基因型或等位基因频率之间的情绪障碍和控制没有显着差异。通过对每个等位基因变体的沉默子活性进行分析，也没有检测到显着差异。鉴定了小鼠5-羟色胺转运蛋白（5-HTT）mRNA的三种变体，它们由不同的第一外显子（第一外显子la、第一外显子lb或第一外显子lc）和相同的第四外显子wo至第五外显子组成。连接到pGL-3增强子载体的任何启动子区域在一些测试的细胞系中具有显著高于空载体的活性，其中外显子-Ic的活性显著低于其它。少

项目成果

Mayumi Shindo: "Diacylglycerol kinase γ is one of the specific receptors of tumorpromoting phorbol esters"Biochem. Biophys. Res. Commun.. 289. 451-456 (2001)

Mayumi Shindo：“二酰基甘油激酶 γ 是促进肿瘤的佛波酯的特异性受体之一”Biochem. Commun. 289. 451-456 (2001)

Lersen, E. C., DiGennaro, J. A., Saito, N., Mehta, S., Loegering, D. J., Mazurkiewicz, J. E., Lennartz, M. R.: "Differential requirement for classic and novel PKC isoforms in respiratoiy burst and pnagocyrosis in RAW 264.7 cells"J. Immunol.. 165. 2809-281

Lersen, E. C.、DiGennaro, J. A.、Saito, N.、Mehta, S.、Loegering, D. J.、Mazurkiewicz, J. E.、Lennartz, M. R.：“RAW 264.7 细胞呼吸爆发和 pnagocyrosis 中经典和新型 PKC 亚型的不同要求”J

Naoaki Saito: "The family of protein kinase C and membrane lipid mediators"J. Diabetes and its Complications. 16. 1-5 (2002)

Naoaki Saito：“蛋白激酶 C 家族和膜脂介质”J。

Ucyama,T.: "cDNA cloning of an alternative splicing variant of protein kinase Cδ(PKCδIII).a new truncated form of PKCδ,in rats."Biochem.Biophys.Res.Commun.. 269. 557-563 (2000)

Ucyama, T.：“蛋白激酶 Cδ (PKCδIII) 的替代剪接变体的 cDNA 克隆。一种新的 PKCδ 截短形式，在大鼠中。”Biochem.Biophys.Res.Commun.. 269. 557-563 (2000)

Foley,P.: "The L-DOPA story-an early Japanese contribution."Parkinsonism & Related Disorders. 6. 1-6 (2000)

Foley,P.：“左旋多巴的故事——日本早期的贡献。”帕金森病