In vivo characterization of a novel ion channel

新型离子通道的体内表征

• 批准号: 6740596
• 负责人: BRIDGET C LEAR
• 金额: $ 4.73万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6740596
• 关键词: Drosophilidae; RNA interference; antibody; circadian rhythms; fluorescent in situ hybridization; gene expression; genetic regulation; immunofluorescence technique; ionophores; membrane channels; mutant; neurogenetics; neurons; postdoctoral investigator; protein localization; protein structure function; time resolved data

The goal of this research is to characterize the role of a unique ion channel gene, alpha1U (a1U), in circadian rhythms. Although this gene is highly conserved in animals, no functional data existed previously for any a1U ortholog. Drosophila a1U (Dma1U) mutants exhibit inverted activity patterns in light-dark conditions and arrhythmic behavior in constant darkness. The genetic techniques available in Drosophila make this an ideal system in which to study alU function. The GAL4-UAS system and RNA interference will be used to determine the tissue-specific role(s) of Dma1U in circadian rhythms. Immunofluorescence will be used to assess the oscillation of clock components in Dma1U mutants. Given the conservation of circadian components between Drosophila and mammals, these analyses could directly impact future research in mammalian rhythms. Specific Aims: (1) To test the hypothesis that Dma1U regulates the circadian oscillation of clock components in the Drosophila brain. (2) To test the hypothesis that Dma1U functions within the Drosophila pacemaker cells to mediate its effects on circadian rhythms. (3) To test the hypothesis that a1U function within the circadian pacemaker is sufficient to promote rhythmic behavior

本研究的目的是描述一个独特的离子通道基因alpha 1U（a1 U）在昼夜节律中的作用。虽然该基因在动物中高度保守，但以前没有任何a1 U直系同源物的功能数据。果蝇a1 U（Dma 1U）突变体在光暗条件下表现出倒置的活动模式，在恒定黑暗中表现出行为学行为。果蝇的遗传技术使其成为研究alU功能的理想系统。GAL 4-UAS系统和RNA干扰将用于确定Dma 1U在昼夜节律中的组织特异性作用。免疫荧光将用于评估Dma 1U突变体中时钟成分的振荡。考虑到果蝇和哺乳动物之间昼夜节律成分的保守性，这些分析可能会直接影响哺乳动物节律的未来研究。具体目的：（1）验证Dma 1U调控果蝇脑内生物钟成分昼夜节律振荡的假说。(2)验证Dma 1U在果蝇起搏细胞中介导其对昼夜节律的影响的假设。(3)检验昼夜节律起搏器内的a1 U功能足以促进节律行为的假设