Characterization and molecular cloning of loci controlling sleep in Drosophila

描述(申请人提供)：尽管经过几十年的密集研究，睡眠的功能仍不清楚。同样，我们对控制睡眠和清醒的分子和机制的理解也是不完整的。最近在黑腹果蝇中发现了一种睡眠状态，这为使用强大的正向遗传学方法在更容易获得的模式生物中剖析睡眠的功能和调节提供了可能性。这项建议的具体目标集中在果蝇睡眠突变体的正向遗传筛查中分离的两类突变体。其中一类突变体表现出明显的睡眠时相改变和睡眠持续时间的增加。第二节课表现出睡眠减少。将使用包括杂交、减数分裂图谱和互补测试在内的遗传方法来确定这些突变体的性质，包括它们的遗传模式、染色体位置以及与已经确定的调节睡眠和生物钟的基因的关系。这些突变体的其他行为和分子特征将决定它们的唤醒阈值、昼夜节律和对睡眠剥夺的反应。这项建议的最终目标是通过缺乏非互补、转基因挽救和基因测序等手段，对影响这些睡眠表型的基因进行分子克隆。这些基因的分子克隆将使编码产物的结构、分布和活性能够在未来的实验中进行研究和操纵。归根结底，这项研究的广泛目标是更好地了解人类正常和功能失调睡眠的基因和机制。与公共健康相关：睡眠的功能尚不清楚，控制正常和功能失调睡眠的机制也不完全清楚。了解睡眠的重要性体现在睡眠相关疾病的成本上，据估计，美国每年的睡眠相关疾病成本高达数百亿至数千亿美元。这项研究将使用一种强大的遗传学方法来扩大我们对控制睡眠的基因的理解。

DESCRIPTION (provided by applicant): Despite decades of intensive study, the function of sleep is still not known. Similarly, our understanding of the molecules and mechanisms that govern sleep and wakefulness is incomplete. The recent identification of a sleep state in Drosophila melanogaster has opened up the possibility of using powerful forward genetic methods to dissect the function and regulation of sleep in a more accessible model organism. The specific aims of this proposal focus on two classes of mutants isolated in a forward genetic screen for Drosophila sleep mutants. One class of mutants exhibits a striking alteration of sleep phase and an increase in sleep duration. The second class exhibits reduced sleep. Genetic methods including crosses, meiotic mapping, and complementation testing will be employed to determine the properties of these mutants, including their mode of inheritance, chromosomal position, and relationship to already characterized genes that regulate sleep and the circadian clock. Additional behavioral and molecular characterization of these mutants will determine their arousal threshold, circadian period, and response to sleep deprivation. The final aim of this proposal is the molecular cloning of the affected genes underlying these sleep phenotypes, by means of deficiency noncomplementation, transgenic rescue, and gene sequencing. The molecular cloning of these genes will enable the structure, distribution, and activity of the encoded products to be studied and manipulated in future experiments. Ultimately, the broad objective of this research is to better understand the genes and mechanisms that underlie normal and dysfunctional sleep in humans. Public health relevance: The function of sleep is not known, and the mechanisms that control normal and dysfunctional sleep are not fully understood. The importance of understanding sleep is indicated by the cost of sleep related disorders, which is estimated at tens to hundreds of billions of dollars in the United States each year. This research will use a powerful genetic approach to broaden our understanding of genes that control sleep.