Nitric Oxide and the Regulation of Behavioral State

一氧化氮与行为状态的调节

• 批准号: 6716897
• 负责人: PAUL ALLEN ROSENBERG
• 金额: $ 22.63万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-08 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6716897
• 关键词: REM sleep; behavior; behavior test; biological signal transduction; electroencephalography; enzyme inhibitors; guanosine monophosphate; guanylate cyclase; in situ hybridization; laboratory rat; microdialysis; molecular psychobiology; neuroanatomy; neuropsychological tests; neuropsychology; nitric oxide; nitric oxide synthase; phosphodiesterases; polysomnography; prosencephalon; protein kinase; protein localization; sleep; sleep disorders

Although much has been learned about the transmitter systems regulating behavioral state, we know little about the actual molecular mechanisms causing the brain to sleep and to wake up. This project is directed at defining these mechanisms. Cholinergic neurons of the laterodorsal tegrnental/pedunculopontine tegmental nucleus, which project to the basal forebrain (BF), as well as intrinsic cholinergic neurons of the BF, both contain nitric oxide synthase (NOS), and are active during waking. We found that: 1) NO evokes the release of adenosine from forebrain neurons in culture by a cGMP-independent mechanism; 2) A NOS inhibitor dialyzed into the BF suppressed NREM sleep following sleep deprivation in rats, and a nitric oxide donor dialyzed into the BF increased NREM sleep. These observations are the basis for one of two major hypothesis motivating this project, which is that NO, released during wakefulness in the BF by cholinergic neurons, is a major stimulus to the release of adenosine during waking, and thus is a key factor regulating the extracellular level of this important somnogen. In addition, we found that: 1) The high affinity cGMP degrading enzyme cyclic nucleotide phosphodiesterase (PDE) 9A is expressed in large neurons of the BF; 2) An inhibitor of cGMP degrading PDEs dialyzed into the BF increased NREM sleep. On the basis of these observations, we hypothesize that NO also has important effects relevant to the regulation of behavioral state that are mediated by the NO/cGMP signaling pathway, and are independent of the adenosine-releasing effects of NO. The specific aims of this project are to: 1) Use mierodialysis with behavioral and electroeneephalographie monitoring to test for the role of NO in homeostatic sleep regulation in the rat using NOS inhibitors and NO donors. 2) Determine the anatomic and cellular localization of the components of the NO/cGMP signal transduction system (cGMP hydrolyzing cyclic nucleotide phosphodiesterases, soluble guanylyl cyclase, NO synthase, protein kinase G) in regions of the brain relevant for sleep/wake regulation. 3) Characterize the electrophysiological effects of NO on neurons of the BF and ventrolateral preoptic nucleus (VLPO) using a basal forebrain/preoptic area BF/POA) slice preparation.

尽管有关调节行为状态的发射机系统的了解很多，但我们对导致大脑睡眠和醒来的实际分子机制知之甚少。该项目旨在定义这些机制。后期dodor虫的胆碱能神经元/花梗细分核的核元，它们的基础前脑（BF）以及BF的固有性胆碱能神经元都包含一氧化氮合酶（NOS），并且在饮食过程中都活跃。我们发现：1）不唤起腺苷的释放 培养中的前脑神经元是通过CGMP独立的机制； 2）在大鼠睡眠剥夺后抑制NREM睡眠的NOS抑制剂，一氧化氮供体透析到BF中增加了NREM睡眠。这些观察结果是激励该项目的两个主要假设之一的基础，即胆碱能神经元在BF中释放的NO是对醒来过程中腺苷释放的主要刺激，因此是调节这一重要脾脏外细胞外水平的关键因素。此外，我们发现：1）高亲和力CGMP降解酶环状核苷酸磷酸二酯酶 （PDE）9a在BF的大神经元中表达； 2）CGMP降解PDE的抑制剂透析到BF中增加了NREM睡眠。根据这些观察结果，我们假设NO还具有与NO/CGMP信号通路介导的行为状态相关的重要作用，并且与NO的腺苷释放效应无关。该项目的具体目的是： 1）使用MierodiaiSIS进行行为和电气仪监控 测试使用NOS抑制剂和NO供体在大鼠中调节NO在稳态睡眠中的作用。 2）确定NO/CGMP信号转导系统组件的解剖学和细胞定位（CGMP水解环核苷酸磷酸二酯酶，可溶性鸟叶叶尼糖环酶，NO合酶，蛋白质激酶G）在大脑与睡眠/唤醒相关的区域中的蛋白质激酶G）。 3）使用基础前脑/前脑/前bf/pOA的NO对NO对BF神经元和腹外侧前核（VLPO）的电生理影响。