Non-circadian Role for Clock Genes in Sleep Homeostasis

时钟基因在睡眠稳态中的非昼夜节律作用

• 批准号: 7263210
• 负责人: H Craig Heller
• 金额: $ 40.44万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-04 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7263210
• 关键词: Acute; Animals; Area; Arrhythmia; Behavior; Behavioral; Brain; Brain region; Cerebral cortex; Chronic; Circadian Rhythms; Condition; Coupled; Crying; Drosophila genus; Electroencephalography; Enzymes; Exhibits; Gene Expression; Gene Family; Generations; Genes; Homeostasis; Inbred Strains Mice; Individual; Knock-out; Knockout Mice; Lesion; Measures; Messenger RNA; Metabolism; Modeling; Molecular; Molecular Genetics; Monitor; Motor Activity; Mus; Mutant Strains Mice; Mutation; Neurons; Output; Oxidation-Reduction; Periodicity; Pharmaceutical Preparations; Phase; Phenotype; Play; Process; Prosencephalon; Proteins; Recording of previous events; Recovery; Regulation; Relative (related person); Reporting; Research Personnel; Role; Sleep; Sleep Deprivation; Sleep Disorders; Specificity; Structure; Study models; System; Testing; Time; Transcriptional Regulation; Translating; Wakefulness; Wild Type Mouse; awake; base; circadian pacemaker; cryptochrome; day; design; improved; insight; interest; lactate dehydrogenase A; non rapid eye movement; paralogous gene; pressure; response; sleep regulation

DESCRIPTION (provided by applicant): The mechanisms of the homeostatic regulation of sleep are largely unknown, although a role for gene expression seems likely based on genetic and molecular studies. In this proposal we suggest that several of the genes identified as critical components of the circadian pacemaker in the SCN also have a separate, non-circadian role in the cerebral cortex (and perhaps other brain regions) in sleep homeostasis. We found that mice lacking the circadian 'clock'-genes cryptochrome (cry1,2-/-), not only lack circadian rhythms but also exhibit all the electrocortical hallmarks of high sleep pressure. This unexpected phenotype was associated with elevated brain expression of the transcriptional regulators period (per)l, and per2, which are transcriptionally inhibited by CRY1,2. Wild-type mice following sleep deprivation also have elevated levels of per1,2 mRNA in the cortex specifically. Here we propose to confirm and further characterize the relationship between sleep-wake and these molecular changes at both mRNA and protein levels. Determining this relationship and the neuro-anatomical specificity of these changes is critical to test the hypothesis that a transcriptional network of interacting clock genes outside the SCN underlies the sleep homeostat. To separate homeostatic from circadian sleep regulatory aspects, SCN lesions will be performed. We expect that in the absence of circadian output provided by the SCN, the cortical oscillation in per expression will be driven by sleep and wake, as in intact animals. We will directly manipulate this clock-gene network by examining sleep regulation in mice lacking cry1, cry2, per1, per2, clock, and npas2. Mice lacking either cry1 or cry2 will be recorded to assess whether inactivation of only one cry gene can replicate the cry1,2-/-sleep phenotype. CLOCK and NPAS2 are positive transcriptional regulators of per1,2, cry1,2, and other genes, and should provide an important contrast to the cry1,2-/- phenotype. Given the partial redundancy in these gene families, we expect comparisons of mice lacking both per1,2, or both npas2,clock, to be especially informative. An improved understanding of the molecular substrate of sleep homeostasis could be helpful to a wide variety of individuals with acute or chronic sleep problems.

描述（由申请人提供）：尽管基于遗传和分子研究，基因表达的作用似乎很可能存在，但睡眠稳态调节的机制在很大程度上尚不清楚。在这项提议中，我们建议，被确定为 SCN 昼夜节律起搏器关键组成部分的几个基因在大脑皮层（可能还有其他大脑区域）的睡眠稳态中也具有独立的非昼夜节律作用。我们发现，缺乏昼夜节律“时钟”基因隐花色素（cry1,2-/-）的小鼠不仅缺乏昼夜节律，而且还表现出高睡眠压力的所有皮层电特征。这种意想不到的表型与转录调节因子period(per)1和per2的脑表达升高有关，它们在转录上受到CRY1,2的抑制。睡眠剥夺后的野生型小鼠皮质中 per1,2 mRNA 的水平也特别升高。在这里，我们建议在 mRNA 和蛋白质水平上确认并进一步表征睡眠-觉醒与这些分子变化之间的关系。确定这种关系以及这些变化的神经解剖学特异性对于检验 SCN 外部相互作用的时钟基因的转录网络是睡眠稳态的基础这一假设至关重要。为了将体内平衡与昼夜节律睡眠调节分开，将进行 SCN 损伤。我们预计，在缺乏 SCN 提供的昼夜节律输出的情况下，每个表达的皮质振荡将由睡眠和觉醒驱动，就像在完整的动物中一样。我们将通过检查缺乏cry1、cry2、per1、per2、时钟和npas2的小鼠的睡眠调节来直接操纵这个时钟基因网络。将记录缺乏cry1或cry2的小鼠，以评估仅一个cry基因失活是否可以复制cry1,2-/-睡眠表型。 CLOCK 和 NPAS2 是 per1,2、cry1,2 和其他基因的正转录调节因子，并且应该提供与cry1,2-/- 表型的重要对比。考虑到这些基因家族的部分冗余，我们预计对同时缺乏 per1,2 或同时缺乏 npas2,clock 的小鼠进行比较会特别提供信息。更好地了解睡眠稳态的分子基础可能对患有急性或慢性睡眠问题的各种个体有所帮助。

项目成果

A non-circadian role for clock-genes in sleep homeostasis: a strain comparison.

• DOI: 10.1186/1471-2202-8-87
• 发表时间: 2007-10-18
• 期刊: BMC neuroscience
• 影响因子: 2.4
• 作者: Franken P;Thomason R;Heller HC;O'Hara BF
• 通讯作者: O'Hara BF

Melanopsin as a sleep modulator: circadian gating of the direct effects of light on sleep and altered sleep homeostasis in Opn4(-/-) mice.

• DOI: 10.1371/journal.pbio.1000125
• 发表时间: 2009-06-09
• 期刊: PLoS biology
• 影响因子: 9.8
• 作者: Tsai JW;Hannibal J;Hagiwara G;Colas D;Ruppert E;Ruby NF;Heller HC;Franken P;Bourgin P
• 通讯作者: Bourgin P

cGMP-dependent protein kinase type I is implicated in the regulation of the timing and quality of sleep and wakefulness.

• DOI: 10.1371/journal.pone.0004238
• 发表时间: 2009
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Langmesser S;Franken P;Feil S;Emmenegger Y;Albrecht U;Feil R
• 通讯作者: Feil R