Cellular and Molecular Mechanisms of REM Sleep

描述（由申请人提供）：本研究的长期目标是进一步了解脑干细胞、分子和网络机制对快速眼动睡眠的调节。具体来说，这项更新应用的目标是研究桥脚被（cc - ppt）和背蓝下核（SubCD）胆碱能细胞区内的分子机制，以及它们在快速眼动睡眠的稳态调节中的作用。本研究的中心假设是，脑源性神经营养因子（BDNF）受体（原肌球蛋白相关激酶B [TrkB]）和细胞外信号调节激酶1/2 （ERK1/2）信号在cc - ppt和SubCD中的相互作用积极调节REM睡眠和相脑桥波（p波）活动的稳态调节。为了系统地检验这一假设，设计了三个具体目标：验证在选择性快速眼动睡眠剥夺过程中，慢波睡眠的增加和CCC-PPT中BDNF表达的增加对快速眼动睡眠稳态驱动的发展至关重要，而在相同条件下，SubCD中BDNF表达的增加对p波活动的稳态调节至关重要。这一目标将通过测量在不同强度的动态眼动睡眠动态驱动下CCC-PPT、SubCD和一些控制区域BDNF水平的变化来实现。2. 验证CCC-PPT中BDNF TrkB受体与ERK1/2信号之间的相互作用对于REM睡眠的稳态调节至关重要，而SubCD中相同的相互作用对于p波活动的稳态调节至关重要的假设。为了验证这一假设，在选择性REM睡眠剥夺期开始时，我们将在大鼠的cc - ppt或亚cd中应用BDNF TrkB受体抑制剂，ERK1/2激活抑制剂或载体控制，以阻断快速眼动睡眠增加的稳态驱动。3. 在杂合BDNF敲除（BDNF+/-）大鼠中，CCC-PPT中BDNF产生的减少减弱了REM睡眠的稳态调节，而SubCD中BDNF产生的减少减弱了p波活性的稳态调节。为了验证这一假设，在选择性快速眼动睡眠剥夺期开始时，我们将BDNF应用于BDNF+/-大鼠的cc - ppt或SubCD。所有这些实验都将在自由活动的成年大鼠身上进行。我们相信这些研究的结果将扩展快速眼动睡眠调节的基本神经生物学机制的前沿知识。此外，这些结果将首次开启一个新的研究领域，旨在了解快速眼动睡眠个体生理体征的稳态调节的局部细胞和分子机制。我们相信，这些研究的结果将揭示在许多精神和神经疾病（如内源性抑郁症、精神分裂症、阿尔茨海默氏症、亨廷顿氏症、帕金森病和中风）中快速眼动睡眠稳态调节功能障碍的神经病理机制，并将帮助我们设计治疗干预措施来消除这些功能障碍。

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 molecular mechanisms within the cholinergic cell compartment of the pedunculopontine tegmentum (CCC-PPT) and dorsal subcoeruleus nucleus (SubCD) with respect to their roles in homeostatic regulation of REM sleep. The central hypothesis of this proposal is that homeostatic regulation of REM sleep and phasic pontine- wave (P-wave) activity are actively regulated by the interaction between brain-derived neurotrophic factor (BDNF) receptors (tropomyosin-related kinase B [TrkB]) and extracellular signal-regulated kinase1/2 (ERK1/2) signaling in the CCC-PPT and SubCD. Three specific aims have been designed to systematically test this hypothesis: 1. Test the hypothesis that during selective REM sleep deprivation, increased slow-wave sleep and increased expression of BDNF in the CCC-PPT are critical for the development of homeostatic drive for REM sleep, and under the same conditions, increased BDNF expression in the SubCD is critical for the homeostatic regulation of P-wave activity. This goal will be achieved by measuring homeostatic drive-associated changes in BDNF levels in the CCC-PPT, SubCD, and in a number of control areas at different intensities of increased REM sleep homeostatic drive. 2. Test the hypothesis that the interaction between BDNF TrkB receptors and ERK1/2 signaling in the CCC-PPT is critical for the homeostatic regulation of REM sleep, and that this same interaction in the SubCD is critical for the homeostatic regulation of P-wave activity. To test this hypothesis, at the beginning of a selective REM sleep deprivation period we will apply a BDNF TrkB receptor inhibitor, ERK1/2 activation inhibitor, or vehicle control into either the CCC-PPT or SubCD of rats, in order to block the increased homeostatic drive for REM sleep. 3. Test the hypothesis that, in heterozygous BDNF knockout (BDNF+/-) rats, decreased BDNF production in the CCC-PPT attenuates homeostatic regulation of REM sleep, and decreased BDNF production in the SubCD attenuates homeostatic regulation of P-wave activity. To test this hypothesis, at the beginning of the selective REM sleep deprivation period we will apply BDNF either into the CCC-PPT or SubCD of BDNF+/- rats. All of these experiments will be performed on adult, freely moving rats. We believe that the results of these studies will extend the leading edge of knowledge on the basic neurobiological mechanisms of REM sleep regulation. Also, for the first time, these results will launch a new area of investigation aimed at understanding the localized cellular and molecular mechanisms of homeostatic regulation of individual physiological signs of REM sleep. We believe that the results of these studies will shed light on the neuro-pathological mechanisms of REM sleep homeostatic regulatory dysfunctions in a number of psychiatric and neurological disorders (e.g., endogenous depression, schizophrenia, Alzheimer's, Huntington's, Parkinson's, and stroke), and will help us to design therapeutic interventions to eliminate these dysfunctions.