Neural Gene Expression in Sleep Deprivation and Recovery

睡眠剥夺和恢复中的神经基因表达

• 批准号: 7655840
• 负责人: Thomas S Kilduff
• 金额: $ 55万
• 依托单位: SRI INTERNATIONAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7655840
• 关键词: Adenosine; Affect; Anxiety; Brain Stem; Cells; Cerebral cortex; Circadian Rhythms; Classification; Complement; Corticotropin-Releasing Hormone; Electrophysiology (science); Enzymes; Epilepsy; Family; Family member; Funding; Gene Expression; Gene Targeting; Genetic Transcription; Hypothalamic structure; Immunohistochemistry; In Vitro; Influentials; Knockout Mice; Label; Link; Mental disorders; Modeling; Mood Disorders; Mus; Napping; Neurobiology; Neurologic; Neuromodulator; Neurons; Neurotransmitters; Nitric Oxide Synthase Type I; Population; Process; Prosencephalon; Proteins; Rattus; Recovery; Regulation; Role; Schizophrenia; Sleep; Sleep Deprivation; Sleep Disorders; Sleeplessness; Somatotropin-Releasing Hormone; System; Testing; Time; Transcription Factor AP-1; Transgenic Mice; Wakefulness; base; cholinergic; cholinergic neuron; cortistatin; cytokine; in vivo; locus ceruleus structure; member; neural circuit; noradrenergic; patch clamp; public health relevance; relating to nervous system; research study; response; sleep regulation; suprachiasmatic nucleus; transcription factor

Description (provided by applicant): The Two Process model of homeostatic sleep regulation posits that the timing of sleep is due to the interaction between a circadian process (Process C) and a homeostatically regulated sleep process (Process S). Whereas Process C is linked to the suprachiasmatic nucleus, the neural basis of Process S is currently unknown. During the previous funding period, we identified a population of cells in the cerebral cortex that express the neuronal nitric oxide synthase (nNos) within which the transcription factor Fos is expressed specifically during recovery sleep (RS) after sleep deprivation (SD). Expression of Fos in cortical nNos neurons also occurs in conjunction with the natural occurrence of sleep. Based on these results, the hypothesis that underlies this proposal is that cortical Fos+/nNos neurons are fundamentally related to the homeostatic regulation of sleep and are a neuroanatomical substrate of Process S. To test this hypothesis, we will systematically vary the duration of SD and RS and determine Fos expression in cortical nNos neurons by double-label immunohistochemistry. To determine whether nNos per se is related to homeostatic sleep regulation, we will evaluate homeostatic sleep regulation in the nNos knockout mouse and use a co-localized phenotypic marker to determine whether Fos is induced during RS in cortical neurons that normally express nNos. To determine the afferent inputs that influence cortical nNos neurons, we will use a combination of in vitro and in vivo approaches. We will use patch clamp electrophysiology of cortical nNos/EGFP neurons to determine the neurotransmitters and neuromodulators that excite or inhibit these cells, particularly substances implicated in the control of sleep and wakefulness. Lastly, we will determine the role(s) of cholinergic, noradrenergic, histaminergic, and hypocretinergic inputs in state-specific cortical Fos+/nNos expression by elimination of these putative afferent populations using neurochemically-specific saporin conjugates. Information from these in vivo experiments will complement the in vitro studies and is essential to understand the neural circuitry that results in state-specific activation of nNos neurons. These experiments will elucidate the neurobiology of cortical nNos neurons and allow us to determine whether the activity of these cells is indeed related to Process S. The results will not only enhance our understanding of sleep/wake regulation, but may also have implications for understanding sleep disorders and neurological and psychiatric diseases involving the cerebral cortex such as epilepsy, anxiety, and schizophrenia. PUBLIC HEALTH RELEVANCE: During the previous funding period, we identified a population of cells in the cerebral cortex that express the neuronal nitric oxide synthase (nNos) that appear to be activated during sleep. This is the first neuronal population in the cerebral cortex known to be active during sleep. We will determine whether nNos itself is important for sleep or whether nNos is simply a marker for these cells. We will also identify the neurotransmitters and neuromodulators that activate or inhibit nNos neurons and which may therefore activate these cells during sleep and inhibit nNos neurons during wakefulness. The results will not only enhance our understanding of sleep/wake regulation, but may also have implications for understanding sleep disorders and neurological and psychiatric diseases involving the cerebral cortex such as epilepsy, anxiety and schizophrenia.

描述（由申请人提供）：体内稳态睡眠调节的两个过程模型认为，睡眠时间是由于昼夜节律过程（过程C）与体内稳态调节的睡眠过程（过程S）之间的相互作用所致。尽管过程C与上核核链接相关，但过程S的神经基础目前尚不清楚。在上一个资金期间，我们确定了表达神经元一氧化氮合酶（NNOS）中大脑皮层中的细胞群，其中转录因子FOS在睡眠剥夺后的恢复睡眠（SD）（SD）后在恢复睡眠（RS）期间特别表达。皮质NNOS神经元中FOS的表达也与睡眠的自然发生结合。基于这些结果，该建议的基础是皮质+/NNOS神经元与睡眠的稳态调节根本性相关，并且是过程S的神经解剖学基材料。为了检验该假设，我们将系统地在Corortical nnos Neurons中系统地表达SD和RS的持续时间，并确定FOS的表达方式。为了确定NNOS本身是否与稳态睡眠调节有关，我们将评估NNOS基因敲除小鼠中的稳态睡眠调节，并使用共定位的表型标记来确定是否在通常表达NNOS的皮质神经元中诱导FOS。为了确定影响皮质NNOS神经元的传入输入，我们将使用体外和体内方法的组合。我们将使用皮质NNOS/EGFP神经元的斑块夹电生理学来确定激发或抑制这些细胞的神经递质和神经调节剂，尤其是与睡眠和唤醒控制有关的物质。最后，我们将通过消除这些假定的传入群体使用神经化学特异性的saporin conjugates来确定胆碱能，去甲肾上腺素能，组胺能和低载蛋白能输入的作用。来自这些体内实验的信息将补充体外研究，对于了解导致州特异性NNOS神经元激活的神经回路至关重要。 These experiments will elucidate the neurobiology of cortical nNos neurons and allow us to determine whether the activity of these cells is indeed related to Process S. The results will not only enhance our understanding of sleep/wake regulation, but may also have implications for understanding sleep disorders and neurological and psychiatric diseases involving the cerebral cortex such as epilepsy, anxiety, and schizophrenia.公共卫生相关性：在上一个资金期间，我们确定了表达神经元氧化物合酶（NNOS）的大脑皮层中的细胞群，这些细胞似乎在睡眠期间被激活。这是已知在睡眠期间活跃的大脑皮层中的第一个神经元种群。我们将确定NNOS本身对于睡眠还是NNOS是否仅仅是这些细胞的标记。我们还将确定激活或抑制NNOS神经元的神经递质和神经调节剂，因此在睡眠过程中可能会激活这些细胞并在清醒期间抑制NNOS神经元。结果不仅会增强我们对睡眠/唤醒调节的理解，而且还可能对了解睡眠障碍以及涉及脑皮质（例如癫痫，焦虑症和精神分裂症）的神经系统和精神疾病有影响。