The Effect of mWAKE in the Subfornical Organ on the Circadian Regulation of Water Consumption

穹窿下器官 mWAKE 对耗水量昼夜节律调节的影响

• 批准号: 10464743
• 负责人: Elijah Blank
• 金额: $ 4.68万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-23 至 2025-05-22
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10464743
• 关键词: Address; Animals; Area; Arousal; Behavior; Biological Clocks; Biological Rhythm; Biomedical Research; Brain; Brain region; Cells; Circadian Dysregulation; Circadian Rhythms; Darkness; Data; Dependovirus; Development; Drosophila genus; Exhibits; Feedback; Fluid Balance; Future; Gene Expression; Genes; Genetic Screening; Genetic Transcription; Glutamates; Health; Human; Impairment; Invertebrates; Kidney; Knock-out; Label; Lead; Light; Liver; Luciferases; Mammals; Mediating; Mentors; Molecular; Mus; Nature; Neurons; Organ; Organism; Orthologous Gene; Outcome Study; Output; Pathway interactions; Pattern; Periodicity; Physiological Processes; Physiology; Positioning Attribute; Process; Research Personnel; Sleep; Slice; Solid; Students; Subfornical Organ; System; Technical Expertise; Thirst; Time; Tissues; Training; Transgenic Mice; Virus; Visceral; Water; Water consumption; Western Blotting; Work; awake; career; cell type; circadian; circadian pacemaker; circadian regulation; designer receptors exclusively activated by designer drugs; drinking; drinking behavior; experimental study; fly; gamma-Aminobutyric Acid; insight; molecular clock; motivated behavior; mutant; patch clamp; receptor; shift work; single cell sequencing; success; suprachiasmatic nucleus

Project Summary/Abstract Circadian rhythms are important for human health. At a molecular level, they are controlled by a cell-autonomous molecular clock which is generally synchronized across different tissues. In mammals, these local clocks are coordinated by the suprachiasmatic nucleus (SCN), the circadian pacemaker. Most previous work in this field has focused on how the SCN regulates neuronal physiology and behavior, and the function of local clocks in these processes is relatively understudied. In particular, the functions of extra-SCN oscillators in the brain are poorly understood. Our lab previously identified a molecule Wide Awake (WAKE) in Drosophila, which acts downstream of the clock to reduce arousal at night by inhibiting neuronal activity in a cell-autonomous manner. WAKE is conserved from flies to mammals, including mice and humans. In mice, our lab has recently found that WAKE in mice is enriched in the subfornical organ (SFO). This brain region is known to regulate behaviors controlling fluid balance and water consumption. Moreover, the SFO has been shown to exhibit robust cycling of local clock oscillators, and furthermore water consumption is under circadian control. My preliminary data suggest that loss of mWAKE results in an increase in drinking, specifically during the night. Taken together, these findings lead me to hypothesize that mWAKE normally inhibits thirst-promoting SFO neurons at night. Because SCN-specific mechanisms have also been shown to regulate circadian aspects of drinking, this system may allow for characterization of the interplay between local and central clock mechanisms in regulating a motivated behavior. In Aim 1, I propose to examine whether mWAKE expression in the SFO is rhythmic and under clock control and to characterize the nature of mWAKE+ cells in the SFO. In Aim 2, I will investigate the effects of mWAKE on SFO neuronal activity and the function of mWAKE and mWAKE+ cells in regulating rhythmic water consumption. These studies should provide new insights into how local clocks help tune neuronal activity to modulate rhythmic behaviors. Because desynchrony between local body clocks and the SCN can be produced by common activities such as shift work and is likely associated with poor health outcomes, these studies may one day inform future therapies related to dysregulation of circadian clocks. Finally, the proposed experiments will provide a solid platform for me to strengthen my technical skills and conceptual background and, combined with my proposed training in career and professional development, will position me for future success as an independent investigator and a mentor for diverse students in biomedical research.

项目摘要/摘要 昼夜节律对人类健康很重要。在分子水平上，它们由细胞自主控制 分子时钟通常在不同组织中同步。在哺乳动物中，这些本地时钟是 由昼夜节奏起搏器上核（SCN）协调。该领域的大多数工作 专注于SCN如何调节神经元的生理和行为以及本地时钟的功能 这些过程相对研究。特别是，大脑中SCN振荡器的功能是 理解不佳。我们的实验室以前在果蝇中识别出分子宽（唤醒），它起作用 时钟下游通过以细胞自主方式抑制神经元活性来减少唤醒。 唤醒是从苍蝇到哺乳动物的保守，包括小鼠和人类。在老鼠中，我们的实验室最近发现 小鼠的唤醒富集在亚长质器官（SFO）中。已知这个大脑区域可以调节行为 控制流体平衡和用水量。此外，已显示SFO表现出强大的循环 当地时钟振荡器，以及进一步的水消耗在昼夜节律的控制之下。我的初步数据 表明MWAKE的损失会导致饮酒增加，特别是在夜间。在一起， 这些发现使我假设Mwake通常在晚上抑制口渴的SFO神经元。 由于SCN特异性机制也已被证明可以调节饮酒的昼夜节律方面，因此该系统 可以允许在调节局部和中央时钟机制之间的相互作用表征 动机行为。在AIM 1中，我建议检查Mwake在SFO中的表达是否有节奏和 在时钟控制下并表征SFO中MWAKE+细胞的性质。在AIM 2中，我将调查 MWAKE对SFO神经元活性以及MWAKE和MWAKE+细胞的功能在调节中的影响 有节奏的用水量。这些研究应提供有关本地时钟如何帮助调整神经元的新见解 活动以调节节奏行为。因为当地身体时钟和SCN之间的偏斜可能是 由转变工作等常见活动产生的，可能与健康状况不佳有关，这些 研究可能有一天会为与昼夜节律失调有关的未来疗法提供信息。最后，提议 实验将为我提供一个坚实的平台，以增强我的技术技能和概念背景 而且，结合我在职业和专业发展方面的培训，将为我定位 作为独立研究者和生物医学研究学生的导师的成功。