In vivo characterization of a novel ion channel

这项研究的目的是表征一个独特的离子通道基因，α1U(A1U)，在昼夜节律中的作用。虽然这个基因在动物中高度保守，但之前没有任何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反转的活动模式，在恒定的黑暗中表现出心律失常行为。果蝇的基因技术使其成为研究Alu功能的理想系统。GAL4-UAS系统和RNA干扰将被用来确定Dma1U在昼夜节律中的组织特异性作用(S)。免疫荧光将被用来评估Dma1U突变体中时钟成分的振荡。鉴于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未来对哺乳动物节律的研究。具体目的：(1)验证Dma1U调节果蝇脑内时钟成分昼夜振荡的假说。(2)验证Dma1U在果蝇起搏细胞内发挥作用以调节其对昼夜节律的影响的假说。(3)检验昼夜节律起搏器内的1U功能足以促进节律性行为的假设

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