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.

项目总结/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 分子钟，其通常在不同组织之间同步。在哺乳动物中，这些本地时钟 由视交叉上核（SCN）协调，昼夜节律起搏器。该领域以前的大部分工作 专注于SCN如何调节神经元的生理和行为，以及局部时钟的功能， 这些过程的研究相对不足。特别是，大脑中的额外SCN振荡器的功能是 不太了解。我们的实验室以前在果蝇中发现了一种分子Wide Awake（WAKE）， 在生物钟的下游，通过以细胞自主的方式抑制神经元活动来减少夜间觉醒。 WAKE从苍蝇到哺乳动物，包括小鼠和人类都是保守的。我们的实验室最近发现， 小鼠的WAKE在穹窿下器官（SFO）中富集。这个大脑区域被认为是调节行为的 控制液体平衡和水消耗。此外，SFO已被证明表现出稳健的循环， 本地时钟振荡器，而且水的消耗是在昼夜节律控制下。我的初步数据 这表明mWAKE的丧失导致饮酒增加，特别是在夜间。综合起来看， 这些发现使我假设mWAKE通常在夜间抑制促进口渴的SFO神经元。 由于SCN特异性机制也被证明可以调节饮酒的昼夜节律方面， 可以允许表征在调节生物钟时本地和中央时钟机制之间的相互作用， 动机行为在目标1中，我建议检查SFO中的mWAKE表达是否是有节奏的， 并表征SFO中mWAKE+单元的性质。在目标2中，我将研究 mWAKE对SFO神经元活动的影响以及mWAKE和mWAKE+细胞调节SFO神经元活动的功能 有节奏的用水这些研究应该为局部时钟如何帮助调节神经元提供新的见解。 调节节律行为的活动。因为局部生物钟和SCN之间的时间差可能是 由轮班工作等常见活动产生，可能与健康状况不佳有关，这些 研究可能有一天会告知未来与生物钟失调有关的治疗方法。最后，建议 实验将为我提供一个坚实的平台，以加强我的技术技能和概念背景 并且，结合我在职业和专业发展方面的培训，将使我在未来的工作中处于有利地位。 作为一个独立的研究者和导师，在生物医学研究的不同学生的成功。