MECHANISMS OF SLEEP RESPONSES TO VIRAL INFECTIONS

描述 (根据申请者的摘要改编)疲倦、嗜睡、 几乎所有人都出现了过度睡眠和睡眠障碍的症状。 传染病。这项拨款提案的广泛目标是 描述导致睡眠变化的分子机制 由流感病毒引起的。这些调查人员假设病毒 双链RNA诱导生理性睡眠素的上调 细胞因子与放大的其他细胞因子的从头合成 生理睡眠机制，包括激活一氧化氮 合酶(NOS)。大量的初步数据支持这一宽泛的假设。 用于这些睡眠研究的模型是A/PR/8/34-H1N1流感病毒 小鼠的感染。PR8引起肺炎伴发早发性肺炎 睡眠反应。在第一个特定目的中，诱导的细胞因子谱 流感病毒或由合成的dsRNA，聚I：C会在肺中 比较一下。预计病毒dsrna的出现将先于 细胞因子mRNA的上调和Poly I：C将诱导类似的 细胞因子的表达模式。此外，抗体的作用 病毒dsRNA的中和和细胞因子启动的作用将是 下定决心。具体目标2专注于大脑中细胞因子的产生和 它参与了病毒诱导的睡眠反应。三个基因敲除菌株 每只缺乏一种或多种细胞因子受体的小鼠将被感染和 睡眠反应已确定。这些小鼠的应对能力 传染性挑战预计会受到影响，他们的睡眠反应 迟钝的。同样感染的对照小鼠将接受细胞因子mRNA的检测 在大脑的不同区域均有表达。脑内细胞因子的表达是 预计在睡眠反应之前，并定位于下丘脑和 海马体。在特定目标#3中，一氧化氮合酶在病毒诱导中所起的作用 睡眠反应将被确定。三种不同的NOS基因敲除菌株 小鼠(分别缺乏NOS-1、-2和-3)将接种 将确定流感病毒和睡眠反应。的本土化 类似感染对照组小鼠脑内一氧化氮合酶mRNAs的表达 也要有决心。预计NOS-2基因敲除将会更多 严重损害小鼠，病毒诱导的睡眠反应将是 在NOS-1和NOS-2基因敲除小鼠中均减弱。病毒引起的变化 一氧化氮合酶的表达预计在脑中定位，在肺中明显。 组织。预期的结果将极大地帮助我们理解 病毒诱导睡眠反应的分子机制以及 生理睡眠机制。

DESCRIPTION (Adapted from the applicant's abstract) Fatigue, excessive sleepiness, excess sleep and sleep disturbances are presenting symptoms in nearly all infectious diseases. The broad objective of this grant proposal is to characterize the molecular mechanisms responsible for changes in sleep induced by influenza virus. These investigators hypothesize that viral double-stranded (ds)RNA induces an upregulation of physiological somnogenic cytokines and de novo synthesis of other cytokines that amplify physiological sleep mechanisms, which include activation of nitric oxide synthase (NOS). Substantial preliminary data support this broad hypothesis. The model used for these sleep studies is A/PR/8/34-H1N1 influenza virus infection in the mouse. PR8 causes a pneumonitis accompanied by early onset of sleep responses. In the first Specific Aim, cytokine profiles induced by influenza virus or by the synthetic dsRNA,poly I:C in lungs will be compared. It is expected that the appearance of viral dsRNA will precede an upregulation of cytokine mRNA and that poly I:C will induce a similar pattern of cytokine expression. Further, the effects of antibody neutralization of viral dsRNA and the role of cytokine priming will be determined. Specific Aim #2 focuses on cytokine production in the brain and its involvement in viral-induced sleep responses. Three knockout strains of mice, each lacking receptors of one or more cytokines, will be infected and sleep responses determined. The ability of these mice to cope with the infectious challenge is expected to be compromised and their sleep responses blunted. Control mice similarly infected will be examined for cytokine mRNA expression in various area of brain. Brain expression of cytokines is expected to precede sleep responses and be localized to the hypothalamus and hippocampus. In Specific Aim #3 the role that NOS plays in viral-induced sleep responses will be determined. Three different NOS-knockout strains of mice (lacking NOS-1, -2 and -3, respectively) will be inoculated with influenza virus and sleep responses will be determined. The localization of the expression of NOS mRNAs in brain of similarly infected control mice will also be determined. It is anticipated that NOS-2 knockouts will be the more severely impaired mice and that viral-induced sleep responses will be attenuated in both NOS-1 and NOS-2 knockout mice. Viral-induced changes in NOS expression are expected to be localized in brain and be evident in lung tissue. The anticipated results will greatly aid our understanding of the molecular mechanisms involved in viral-induced sleep responses as well as physiological sleep mechanisms.