Identification of Sleep Substances in the Brain Using Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

摘要 睡眠是一种基本的生理需求，充足的睡眠是身心健康所必需的。减少或 睡眠障碍会导致一系列代谢、炎症、心血管和神经系统疾病， 疾病，降低生产力和性能，并对公共安全构成威胁。了解控制 睡眠对人类健康和社会至关重要。睡眠压力（或嗜睡）长期以来一直是 被认为是由大脑中积累的物质驱动的，在那里它们通过作用于 控制睡眠的神经中枢脑内睡眠压力的分子介质的鉴定将 这是非常重要的，因为它将为改善日常睡眠提供明显的目标。这里我们 建议对符合促进睡眠物质标准的分子进行大规模无偏见的筛选 它驱动睡眠压力的日常变化，并促进睡眠后睡眠的自我平衡增加 剥夺为此，我们已经开发了区分分子的方法， 睡眠压力从那些直接响应于保持睡眠所必需的机械扰动而上升的压力中分离出来。 动物实验对象醒了。在这样做的时候，我们解决了一个根本性的挑战， 睡眠压力的分子相关性。使用基质辅助激光解吸/电离-飞行时间 质谱分析，我们将确定分子的丰度跟踪睡眠压力在中央 大脑是控制睡眠的地方。候选睡眠物质的鉴定将为新的睡眠物质奠定基础。 果蝇睡眠机制的研究，其目的是确定保守的分子机制 自我平衡的控制。考虑到动物王国中存在类似睡眠的状态， 即使在神经系统非常简单的动物中，也存在稳态睡眠控制，发现 可能揭示了调节睡眠的生理系统的保守特征。

Abstract Sleep is a basic biological need and sufficient sleep is required for physical and mental health. Reduced or disordered sleep contributes to an array of metabolic, inflammatory, cardiovascular, and neurological disorders, reduces productivity and performance, and is a threat to public safety. Understanding the control of sleep is therefore critical for human health and society. Sleep pressure (or sleepiness) has long been thought to be driven by substances that accumulate in the brain, where they promote sleep by acting on neural centers of sleep control. The identification of molecular mediators of sleep pressure in the brain would be highly significant, in that it would provide obvious targets for the improvement of daily sleep. Here we propose a large and unbiased screen for molecules that the fit the criteria for sleep promoting substances that drive daily changes in sleep pressure and promote homeostatic increases in sleep following sleep deprivation. To this end we have developed methods to differentiate molecules that rise with increasing sleep pressure from those that rise in direct response to the mechanical perturbation necessary to keep animal subjects awake. In doing so we have addressed a fundamental challenge to the identification of molecular correlates of sleep pressure. Using Matrix Assisted Laser Desorption/Ionization – Time of Flight Mass Spectrometry, we will identify molecules whose abundance tracks sleep pressure within the central brain, where sleep is controlled. The identification of candidate sleep substances will set the stage for new mechanistic investigations of sleep in Drosophila whose aim is to identify conserved molecular mechanisms of its homeostatic control. Given the existence of sleep-like states across the animal kingdom and the presence of homeostatic sleep control even in animals with extremely simple nervous systems, discoveries made in Drosophila are likely to reveal conserved features physiological systems regulating sleep.