Molecular mechanisms of circadian rhythm system and sleep disorders

昼夜节律系统与睡眠障碍的分子机制

• 批准号: 10670914
• 负责人: IKEDA Masaaki
• 金额: $ 2.05万
• 依托单位: Saitama Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670914/
• 关键词: circadian rhythm; BMAL1; clock gene; suprachiasmatic nuclei; sleep disorder; BMAL2; gene analysis; transcription factor; サーカディアンリズム; ヒトゲノム; 細胞内局在; 睡眠覚醒・リズム障害; ノックアウト; ヌプライスバリアント

1) We have cloned a BAC clone for human BMAL1 gene and determined its sequence. It revealed that human BMAL1 coding region consists from 17 exons and several BMAL1 variants are products from splicing variants.2) We have cloned a BAC clone for mouse BMAL1. Its coding region splits over 30kb.3) Mouse BMAL1 promoter region was sequenced. We identified the promoter/enhancer region of BMAL1.4) The transcription start sites for BMAL1 have been identified.5) Polymorphism of BMAL1 gene from sleep disorders were searched. No polymorphism were identified in the BMAL1 coding region.6) Two-hybrid system to test the interaction of clock gene products have been established.7) Luciferase assay system to test the ability to activate mPer1 promoter have been established.8) Fusion proteins of clock gene have been prepared for examining the interaction of clock gene products.9) A nobel bHLH-PAS factor have been cloned and designated BMAL2.10) The expression, chromosomal localization and cellular localization of BMAL2 were examined.

1）我们已将BAC克隆克隆用于人BMAL1基因并确定其序列。它揭示了人类BMAL1编码区域由17个外显子组成，几个BMAL1变体是剪接变体的产物。2）我们已经克隆了一个BAC克隆以用于小鼠BMAL1。其编码区域分裂超过30KB.3）小鼠BMAL1启动子区域进行了测序。我们确定了BMAL1.4的启动子/增强子区域，已经鉴定出BMAL1的转录起始位点。5）搜索了来自睡眠障碍的BMAL1基因的多态性。在BMAL1编码区域中未发现多态性。6）已经建立了两种杂交系统来测试时钟基因产物的相互作用。7）7）萤光素酶测定酶测定系统测试激活MPER1启动子的能力。88）时钟基因的融合蛋白已准备好检查了Clock Gene and clone and clone and clone and nebel fincon and。 BMAL2.10）检查了BMAL2的表达，染色体定位和细胞定位。

项目成果

池田正明: "BMAL1と生体リズム"日本神経精神薬理学雑誌(印刷中). 20. 189-198 (2000)

Masaaki Ikeda：“BMAL1 和生物节律”日本神经精神药理学杂志（出版中）20. 189-198（2000）。

Ebisawa T, Uchiyama M et al: "Genetic polymorphisms of human melatonin 1b receptor gene in circadian rhythm sleep disorders"Neurosci Lett,. 280. 29-32 (2000)

Ebisawa T、Uchiyama M 等人：“人褪黑素 1b 受体基因在昼夜节律睡眠障碍中的遗传多态性”Neurosci Lett，。

Ikeda,M,Yu,W et al: "cDNA Cloning of a Novel bHLH-PAS Transcription Factor Superfamily Gene, BMAL2 ; Its mRNA Expression, Subcellular Distribution, and Chromosomal Localizatio. "Biochem Biophys Res Commun. 275. 493-502 (2000)

Ikeda,M,Yu,W 等人：“新型 bHLH-PAS 转录因子超家族基因 BMAL2 的 cDNA 克隆；其 mRNA 表达、亚细胞分布和染色体定位。”Biochem Biophys Res Commun。

Abe H, Honma S, Namihira M, Masubuchi S, Ikeda M, Ebihara S, and Honma K: "Clock gene expressions in the suprachiasmatic mucleus and other areas of the brain during thythm splitting in CS mice"Mol.Brain Res.. 87. 92-99 (2001)

Abe H、Honma S、Namihira M、Masubuchi S、Ikeda M、Ebihara S 和 Honma K：“CS 小鼠胸腺分裂期间视交叉上粘核和大脑其他区域的时钟基因表达”Mol.Brain Res.. 87

Masubuchi S,Honma S et al: "Clock genes outside the suprachiasmatic nucleus involved in manifestation of locomotor activity rhythm in rats "Eur J of Neurosci. 12. 4206-4214 (2000)

Masubuchi S、Honma S 等人：“视交叉上核外的时钟基因参与大鼠运动活动节律的表现”《Eur J of Neurosci》。