In vivo characterization of a novel ion channel

本研究的目的是表征一个独特的离子通道基因α 1u （a1U）在昼夜节律中的作用。尽管该基因在动物中高度保守，但此前没有任何a1U同源基因的功能数据。果蝇a1U （Dma1U）突变体在明暗条件下表现出倒置的活动模式，在持续的黑暗条件下表现出不规律的行为。在果蝇中可用的遗传技术使其成为研究所有u功能的理想系统。GAL4-UAS系统和RNA干扰将用于确定Dma1U在昼夜节律中的组织特异性作用。免疫荧光将用于评估Dma1U突变体中时钟组分的振荡。考虑到果蝇和哺乳动物之间昼夜节律成分的保守性，这些分析可能直接影响哺乳动物节律的未来研究。具体目的：(1)验证Dma1U调节果蝇大脑生物钟成分昼夜节律振荡的假说。(2)验证Dma1U在果蝇起搏器细胞内调节其对昼夜节律影响的假说。(3)验证昼夜节律起搏器内a1U功能足以促进节律行为的假设

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