Cellular and Neurochemical Mechanisms of REM Sleep

快速眼动睡眠的细胞和神经化学机制

• 批准号: 8247816
• 负责人: Subimal Datta
• 金额: $ 34.19万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8247816
• 关键词: Address; Adenylate Cyclase; Adult; Alzheimer' s Disease; Animals; Antibodies; Appearance; Architecture; Behavioral; Behavioral Mechanisms; Biological Assay; Brain Stem; Calcium/calmodulin-dependent protein kinase; Catalytic Domain; Cataplexy; Cells; Choline O-Acetyltransferase; Clinical; Computer software; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Disease; Dose; Drops; Endogenous depression; Enzyme-Linked Immunosorbent Assay; Equilibrium; Event; FOS gene; Future Generations; GABA Receptor; GABA-B Receptor; Generations; Glutamate Receptor; Glutamates; Goals; Health; Homeostasis; Huntington Disease; Image; Image Analysis; Kainic Acid Receptors; Label; Laboratories; Lead; Light; Link; Malignant Neoplasms; Measures; Mediating; Medical; Methods; Microinjections; Mitogen-Activated Protein Kinase Inhibitor; Mitogen-Activated Protein Kinases; Molecular; Monitor; N-Methylaspartate; Narcolepsy; Nature; Neurobiology; Neurodegenerative Disorders; Neurons; Neurotransmitters; Outcome; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Phenotype; Physiological; Physiological Processes; Platelet aggregation; Process; Protein Kinase A Inhibitor; REM Sleep; Radioactive; Rattus; Receptor Activation; Regulation; Research; Research Design; Restless Legs Syndrome; Role; Schizophrenia; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Sleep Apnea Syndromes; Sleep Architecture; Sleep Deprivation; Sleep Disorders; Sleep Wake Cycle; Sleep disturbances; Slow-Wave Sleep; Staging; System; Techniques; Testing; Therapeutic; Time; U-0126; Wakefulness; Western Blotting; awake; base; calmodulin-dependent protein kinase II; cancer therapy; cell type; cholinergic; cholinergic neuron; design; experience; gamma-Aminobutyric Acid; inhibitor/antagonist; interdisciplinary approach; kainate; natural hypothermia; neurobiological mechanism; neurochemistry; novel; pedunculopontine tegmentum; rapid eye movement; receptor; receptor sensitivity; research study; sleep regulation

DESCRIPTION (provided by applicant): The long-term objective of this research is to further our understanding of brainstem cellular, molecular, and network mechanisms of REM sleep regulation. Specifically, the goal of this renewal application is to investigate the involvement of pedunculopontine tegmentum (PPT) intracellular signaling mechanisms in the regulation of REM sleep. The central hypothesis of this proposal is that REM sleep is regulated by a balance of cellular mechanisms in the PPT; specifically, in the PPT, REM sleep is induced by the activation of cAMP- dependent protein kinase A (PKA), and REM sleep is suppressed by the induction of wakefulness caused by activation of Ca2????dependent protein kinase II (CaMKII) and/or mitogen-activated protein kinase (MAPK). Three specific aims have been designed to systematically test this hypothesis: 1. Identify the neurotransmitter phenotype(s) of cells in the PPT whose activation induces REM sleep. To achieve this goal, state-dependently activated PPT cells will be identified by c-fos and pCREB expression, and their neurochemical identity will be determined by the immunocytochemical labeling of choline acetyltransferase (ChAT) and GABA. 2. Test the hypothesis that the activation of PKA in the PPT induces REM sleep. This goal will be achieved by quantifying the levels of PPT PKA activity after different amounts of REM sleep and by microinjecting a selective cAMP-PKA activation inhibitor into the PPT to block the homeostatic drive for REM sleep. 3. Test the hypothesis that CaMKII and MAPK activation in the PPT terminates REM sleep by inducing wakefulness. This goal will be achieved by measuring the levels of PPT CaMKII and MAPK activity at varying amounts of wakefulness and REM sleep and by applying inhibitors of CaMKII and MAPK activation into the PPT while quantifying their effects on the architecture of spontaneous as well as rebound REM sleep. All of these experiments will be performed on adult, freely moving rats. Preliminary data provide a rationale for each specific aim and demonstrate feasibility. This proposal addresses, at the mechanistic level, the general question in basic neurobiology: how is REM sleep regulated? Importantly, identification of the intracellular signaling molecules involved in the regulation of REM sleep may lead to the design of a future generation of drugs to treat REM sleep disorders as well as a variety of serious medical conditions that are exacerbated by disrupted REM sleep, such as narcolepsy/cataplexy, restless legs syndrome, endogenous depression, schizophrenia, Alzheimer's, and Huntington's disease. PUBLIC HEALTH RELEVANCE The goal of this research is to identify the intracellular signaling molecules involved in the normal regulation of rapid eye movement (REM) sleep. Recent evidence indicates that novel compounds designed to modify intracellular transduction pathways have therapeutic potential for endogenous depression, cancer, hypothermia, and pathological aggregation of platelets. Similarly, identification of the intracellular molecules involved in normal regulation of REM sleep may lead to the design of a future generation of drugs to treat REM sleep disorders as well as a variety of serious medical conditions that are exacerbated by disrupted REM sleep, such as narcolepsy/cataplexy, restless legs syndrome, endogenous depression, schizophrenia, Alzheimer's, and Huntington's disease.

描述(申请人提供)：这项研究的长期目标是进一步了解脑干细胞、分子和网络机制的快速眼动睡眠调节。具体地说，这种更新应用的目标是研究脚桥脑被盖(PPT)细胞内信号机制参与REM睡眠的调节。这一建议的中心假设是，REM睡眠受PPT中细胞机制的平衡调节；具体地说，在PPT中，REM睡眠是由cAMP依赖的蛋白激酶A(PKA)的激活诱导的，而REM睡眠是通过激活钙依赖的蛋白激酶II(CaMKII)和/或丝裂原激活的蛋白激酶(MAPK)而被抑制的。为了系统地验证这一假说，我们设计了三个特定的目标：1.确定PPT中激活诱导快速眼动睡眠的细胞的神经递质表型(S)。为了实现这一目标，状态依赖激活的PPT细胞将通过c-fos和pCREB的表达来鉴定，其神经化学鉴定将通过胆碱乙酰转移酶(ChAT)和GABA的免疫细胞化学标记来确定。2.验证PPT中PKA激活导致快速眼动睡眠的假设。这一目标将通过量化不同数量的REM睡眠后PPT PKA的活性水平以及通过向PPT中微量注射选择性cAMP-PKA激活抑制剂来阻断REM睡眠的动态平衡驱动来实现。3.验证PPT中CaMKII和MAPK激活通过诱导觉醒来终止REM睡眠的假设。这一目标将通过测量不同觉醒和REM睡眠时PPT、CaMKII和MAPK活性的水平，以及在PPT中应用CaMKII和MAPK激活的抑制剂，同时量化它们对自发和反弹REM睡眠结构的影响来实现。所有这些实验都将在成年、自由活动的大鼠身上进行。初步数据为每个具体目标提供了理由，并证明了可行性。这一建议在机械层面上解决了基础神经生物学中的一个普遍问题：快速眼动睡眠是如何调节的？重要的是，识别参与调节REM睡眠的细胞内信号分子可能会导致设计下一代药物来治疗REM睡眠障碍以及各种严重的医疗条件，这些疾病因REM睡眠中断而加剧，如发作性睡病/猝倒、不宁腿综合症、内源性抑郁症、精神分裂症、阿尔茨海默氏症和亨廷顿病。公共卫生相关性这项研究的目标是确定参与快速眼动(REM)睡眠正常调节的细胞内信号分子。最近的证据表明，设计用于修饰细胞内转导通路的新型化合物具有治疗内源性抑郁、癌症、低温和血小板病理性聚集的潜力。同样，识别参与快速眼动睡眠正常调节的细胞内分子可能会导致设计下一代药物来治疗快速眼动睡眠障碍以及各种因快速眼动睡眠中断而加剧的严重医疗状况，如发作性睡病/猝倒、不宁腿综合症、内源性抑郁症、精神分裂症、阿尔茨海默氏症和亨廷顿病。