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)克隆了人BMAL1基因的BAC克隆，并测定了其序列。结果表明，人类BMAL1编码区由17个外显子组成，几个BMAL1变异体是剪接变异体的产物。2)我们克隆了小鼠BMAL1的BAC克隆。3)对小鼠BMAL1启动子区进行了序列测定。我们确定了BMAL1的启动子/增强子区域。4)确定了BMAL1的转录起始点。5)寻找了睡眠障碍患者BMAL1基因的多态性。6)建立了检测Clock基因产物相互作用的双杂交体系。7)建立了检测mPer1启动子激活能力的荧光素酶检测体系。8)制备了Clock基因融合蛋白，用于检测Clock基因产物之间的相互作用。9)克隆了一个诺贝尔bHLH-PAS因子，命名为BMAL2。10)检测了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》。