Interleukin-1: A promoter of slow wave sleep

描述（由申请人提供）：许多与睡眠不足相关的症状，如疲劳、表现障碍、代谢综合征、慢性炎症、嗜睡等；可通过给予白细胞介素-1 β (IL1)或相关细胞因子引起，或可通过阻断它们来预防。然而，il - 1家族在大脑中的调节机制大多是不确定的。最近发现了一种新的脑特异性il - 1受体辅助蛋白（IL1 AcPb）。虽然其功能尚不清楚，但初步数据显示睡眠剥夺增强了其皮层表达。另一个IL1家族成员IL36（以前称为ILF7）的功能最近被确定（未发表）；IL36抑制包括IL1在内的几种促炎促睡细胞因子，同时促进抗炎促睡细胞因子的表达。在Aim 1中，我们确定了IL1 AcPb和IL36在睡眠调节中的作用。100多年来，人们已经知道长时间的清醒(W)会促进大脑产生和释放睡眠调节物质（srs）。然而，导致SRS活性增强的W的性质仍有待确定。在Aim 2中，我们研究了在神经传递过程中释放的ATP是一种信号，可以提供先前W活动的测量。具体来说，ATP通过嘌呤P2受体，通过神经胶质细胞释放il - 1等细胞因子，转化为更持久的先前大脑使用指数。初步数据表明，ATP激动剂促进睡眠，而ATP拮抗剂抑制睡眠，P2X7受体的表达随睡眠倾向而变化。在Aim 2中，我们通过使用缺乏关键模型成分基因（如P2X7受体）的小鼠来测试我们的模型，即atp -细胞因子-腺苷假说。例如，这些小鼠对睡眠不足和炎症刺激脂多糖的睡眠反应减弱。在Aim 3中，我们重点研究了SRS基因表达和EEG δ功率的活动依赖性。我们利用光敏通道视紫红质2 （ChR2）基因在细胞培养中表达，然后用不同的光模式激活细胞，确定SRS和模型基因的表达。我们还使用ChR2转基因小鼠在体内控制皮层神经元的激活和随后的脑电图表现。预期的结果将为炎症如何改变睡眠以及细胞活动如何转化为SRS机制的问题提供机制答案。研究结果将有实际应用于IL1相关的脑部病理，包括炎症相关的睡眠障碍，发生在睡眠呼吸暂停和代谢综合征。

DESCRIPTION (provided by applicant): Many of the symptoms associated with sleep loss, e.g. fatigue, performance impairments, metabolic syndrome, chronic inflammation, sleepiness, etc; can be elicited by administration of interleukin-1 beta (IL1) or related cytokines or can be prevented by blocking them. Nevertheless, the regulation of the IL1 family in the brain is mostly undetermined. Recently a new brain-specific IL1 receptor accessory protein (IL1 AcPb) was identified. Although its function remains unknown preliminary data show that sleep deprivation enhances its cortical expression. The function for another IL1 family member, IL36 (formerly ILF7) has very recently (unpublished) been identified; IL36 inhibits several pro-inflammatory somnogenic cytokines including IL1, while simultaneously promoting expression of anti-inflammatory anti-somnogenic cytokines. In Aim 1, we determine the roles that IL1 AcPb and IL36 have in sleep regulation. For over 100 years it has been known that prolonged wakefulness (W) enhances brain production and release of sleep regulatory substances (SRSs). Nevertheless, the property of W that is responsible for enhanced SRS activity remains to be identified. In Aim 2, we investigate the hypothesis that ATP, released during neurotransmission, is a signal that provides a measure of prior W activity. Specifically, ATP is translated, via purine P2 receptors, into a longer lasting index of prior brain usage through release of cytokines such as IL1 from glia. Preliminary data indicate that ATP agonists promote sleep while ATP antagonists inhibit sleep and P2X7 receptor expression varies with sleep propensity. In Aim 2, we test our model, the ATP-cytokine-adenosine hypothesis, by using mice lacking key model component genes such as the P2X7 receptor. These mice, for example have attenuated sleep responses to sleep loss and the inflammatory stimulus, lipopolysaccharide. In Aim 3, we focus on activity-dependency of SRS gene expression and EEG delta power. We make use of the light-sensitive channelrhodopsin 2 (ChR2) gene by expressing it in cell cultures, then activating the cells with various patterns of light and determine SRS and model gene expressions. We also use ChR2- transgenic mice for in vivo controlled activation of cortical neurons and subsequent manifestations on the EEG. Anticipated results will provide mechanistic answers to questions of how inflammation alters sleep and how cellular activity is translated into SRS mechanisms. Results will have practical application to IL1- associated brain pathologies including inflammation-associated sleep disturbances occurring in sleep apnea and metabolic syndrome.