Role of Shaker Channel Function in the Regulation of Sleep in Drosophila

Shaker Channel 功能在果蝇睡眠调节中的作用

• 批准号: 8201103
• 负责人: Terry Dean
• 金额: $ 1.69万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8201103
• 关键词: Alleles; American; Antihypertensive Agents; Area; Biological; Biological Assay; Brain; Cell membrane; Cells; Cities; Complement; Data; Development; Diabetes Mellitus; Dissection; Dominant-Negative Mutation; Drosophila genus; Equilibrium; Exhibits; Foundations; Future; Gene Expression; Gene Mutation; Genes; Genetic; Health; Heart Diseases; Homeostasis; Hour; Human; In Vitro; Intervention; Ion Channel; Knowledge; Laboratories; Life; Link; Malignant Neoplasms; Mammals; Modeling; Molecular; Mutation; Nature; Neuroanatomy; Neuromuscular Junction; Neurons; Phenotype; Play; Population; Potassium Channel; Proteins; Regulation; Role; Severities; Sleep; Sleep Deprivation; Subcellular Anatomy; Synapses; Synaptic Transmission; System; Time; Wakefulness; awake; drug mechanism; fly; in vivo; insight; mutant; neurophysiology; novel; pressure; public health relevance; research study; sleep regulation; voltage

DESCRIPTION (provided by applicant): Sleep is ubiquitous[1] and vital for life[2], while sleep loss leads to several detrimental effects[3]. However, as demonstrated by the absence of sleep-specific drugs, the mechanism of sleep regulation is unknown. Studies in Drosophila (i.e. fruit flies) show that mutations in genes encoding the Shaker K+ channel[4] and two Shaker channel modulators (Hyperkinetic[5] and Sleepless[6]) dramatically reduce daily sleep time, which is significant because K+ channels seem to have a similar role in the regulation of mammalian sleep as well[7-8]. This proposal includes three lines of studies to investigate the relationships between Shaker, its modulators, and sleep. The first aim is to identify the neuronal populations responsible for the Shaker-dependent regulation of sleep in Drosophila. This will be accomplished by ectopically expressing dominant negative Shaker subunits in select brain areas (via the bipartite GAL4 system) and assaying for abnormal sleep phenotypes. The anticipated results will contribute to our understanding of the neuroanatomy of sleep in Drosophila and allow for future molecular dissection of the sleep regulatory mechanism(s) in these candidate cells. The second aim is to explore the possible cellular mechanism by which the aforementioned sleep- reducing alleles are responsible for their abnormal sleep phenotypes. This will be accomplished by correlating the effects of each of the alleles on synaptic activity and cellular ionic currents recorded at the Drosophila neuromuscular junction with the severities of sleep loss that they induce. The third aim is to determine whether the mechanism by which Sleepless modulates Shaker channel activity is through a direct interaction at the plasma membrane. Identification of such a mechanism would not only provide insight into sleep regulation, but, because this would be a completely novel means by which to specifically enhance Shaker channel activity, could have significant non-sleep implications (e.g. the development of new anti-hypertensive or antiarrythmic agents). PUBLIC HEALTH RELEVANCE: The majority of Americans do not sleep for the recommended "8 hours a night"[9], resulting in sleep deprivation that incurs an estimated ~$15-75 billion per year strain[10-11] on the US economy and is linked to several major health problems (e.g. diabetes, heart disease, cancer)[12-14]. However, we know very little about the biological mechanism of sleep regulation, which could be useful to develop interventions to manipulate our need for sleep. The proposed studies will investigate the brain anatomy and cellular activity that play a role in regulating sleep in the fruit fly, laying the foundation for future studies in mammals.

描述（由申请人提供）：睡眠无处不在[1]，对生命至关重要[2]，而睡眠损失会导致几种有害影响[3]。然而，正如缺乏睡眠特异性药物所证明的那样，睡眠调​​节的机制尚不清楚。果蝇（即水果蝇）的研究表明，编码编码振动筛K+通道[4]和两个振动摇通道调节剂的基因突变（Hyperionic [5]和无睡[6]）大大缩短了每日睡眠时间，这很重要，因为K+通道似乎在哺乳动物睡眠的调节中也具有相似的作用。该提案包括三条研究，以调查振动器，其调节剂和睡眠之间的关系。 第一个目的是确定负责果蝇中振荡器依赖性睡眠调节的神经元种群。这将通过异位表达在某些大脑区域（通过双分式GAL4系统）中的显性负振动筛子亚基并进行异常睡眠表型的测定来实现。预期的结果将有助于我们理解果蝇中睡眠的神经解剖学，并允许对这些候选细胞中睡眠调节机制的未来分子解剖。 第二个目的是探索上述睡眠降低等位基因的可能的细胞机制，导致其异常的睡眠表型。这将通过将每个等位基因对果蝇神经肌肉连接处记录的突触活性和细胞离子电流的影响与它们诱导的睡眠损失严重性相关联。 第三个目的是确定失眠调节振动弹通道活动的机制是否是通过质膜直接相互作用的。这种机制的识别不仅可以洞悉睡眠调节，而且因为这将是一种完全新颖的手段，可以专门增强振荡器通道活性，可能具有重要的非结论含义（例如，新的抗高血压或抗抑制剂的发展）。 公共卫生相关性：大多数美国人在建议的“每晚8个小时” [9]中不睡觉，导致睡眠不足，估计每年约15-7.5亿美元在美国经济上造成约15-7.5亿美元的压力[10-11]，与几个主要的健康问题有关（例如，糖尿病，心脏病，心脏病，癌症）[12-14] [12-14]。但是，我们对睡眠调节的生物学机制知之甚少，这对于制定干预措施以操纵我们的睡眠需求可能是有用的。拟议的研究将研究大脑的解剖结构和细胞活性，在调节果蝇中的睡眠中起作用，为哺乳动物的未来研究奠定了基础。