Roles of CRF-PACAP systems in sleep in mice (Carlezon)

概要：项目3 睡眠在许多精神疾病中受到影响。有些人患有与压力有关的疾病（例如，后 创伤性应激障碍[PTSD]）睡眠少于正常人，而其他人睡眠更多，或有碎片 反映更频繁的睡眠和觉醒的睡眠模式。有相当多的证据表明 压力会扰乱睡眠，我们已经证明，老鼠的慢性压力会导致睡眠的深刻改变， 昼夜节律相比之下，睡眠失调对介导压力效应的大脑系统的影响， 没有被彻底理解。睡眠不足或睡眠受限的时期可能会产生分子改变， 随后影响睡眠。啮齿类动物的研究表明，CRF之间存在重要的相似性和差异 （促肾上腺皮质激素释放因子）和PACAP（垂体腺苷酸环化酶激活多肽）对 行为虽然这两种肽都增加了声惊吓，但这是临床前和临床试验中经常使用的一种警惕措施。 焦虑和恐惧的临床研究，PACAP效应往往是持久的（持续>1周），而CRF效应 迅速解决。这些类型的发现，以及CRF和PACAP系统中已知的遗传改变， 在易受压力相关疾病影响的人群中，提供了比较和对比神经生物学的基本原理， 这些肽。CRF和PACAP对睡眠的影响还没有直接比较，特别是在 很长时间我们的前提是这些肽系统影响睡眠，也受睡眠的影响。我们 假设这些肽和睡眠之间存在相互关系， CRF和/或PACAP系统将影响睡眠，相反，睡眠限制将影响CRF和/或PACAP 系统，但这些影响的持久性将有所不同。试验将在雄性和雌性小鼠中进行 植入无线发射器，能够连续收集EEG、EMG、体温， 数周的活动数据。分子分析将集中在与情感方面有关的大脑区域 包括杏仁核（AMG），终纹床核（BNST）和前额叶皮层（PFC）， 以及传统上与睡眠和生物节律有关的区域。我们还将执行电路映射 研究提供了关于调节情绪行为的区域和调节情绪行为的区域之间的神经连接的见解。 调节睡眠目的1：比较改变CRF和PACAP系统功能对睡眠的影响 和生物（昼夜和昼夜）节律。我们还将检查PAC 1 R或CRFR 1消融的效果 在AMG中，追踪显示应激增加该区域中PAC 1 R表达的试验数据。在目标2中， 将直接比较急性和重复睡眠限制对PACAP和CRF系统的影响， RNAscope定量任何变化的幅度，并确定发生变化的细胞类型。在Aim中 3，我们将使用跨突触病毒标记来映射涉及睡眠和情绪的回路之间的连接， 行为项目3实验是根据现有知识设计的，但正如Conte中心所预期的那样， 我们将利用所有项目的新发现，通过行政核心来完善和优化我们的目标。

SUMMARY: PROJECT 3 Sleep is affected in many psychiatric illnesses. Some with people with stress-related disorders (e.g., Post- Traumatic Stress Disorder [PTSD]) sleep less than normal, whereas others sleep more, or have fragmented sleep patterns that reflect more frequent bouts of sleep and wakefulness. There is considerable evidence that stress disrupts sleep, and we have shown that chronic stress in mice causes profound alterations in sleep and diurnal rhythms. In contrast, the effects of sleep dysregulation on brain systems that mediate stress effects are not thoroughly understood. Periods of poor or restricted sleep may produce molecular alterations that subsequently affect sleep. Work in rodents demonstrates important similarities and differences between CRF (corticotropin-releasing factor) and PACAP (pituitary adenylate cyclase-activating polypeptide) effects on behavior. Although both peptides increase acoustic startle, a measure of vigilance often used in preclinical and clinical studies of anxiety and fear, PACAP effects tend to be persistent (lasting >1 week), whereas CRF effects resolve quickly. These types of findings, together with known genetic alterations in CRF and PACAP systems in people vulnerable to stress-related illness, provide a rationale for comparing and contrasting the neurobiology of these peptides. The effects of CRF and PACAP on sleep have not been directly compared, particularly over extended periods of time. Our premise is that these peptide systems affect—and are affected by—sleep. Our hypothesis is that there is a reciprocal relationship between these peptides and sleep, such that activation of CRF and/or PACAP systems will affect sleep and, conversely, sleep restriction will affect CRF and/or PACAP systems, but that the persistence of these effects will differ. Tests will be conducted in male and female mice implanted with wireless transmitters that enable continuous collection of EEG, EMG, body temperature, and activity data for several weeks. Molecular analyses will focus on brain areas implicated in the emotional aspects of stress, including amygdala (AMG), bed nucleus of the stria terminalis (BNST), and prefrontal cortex (PFC), as well as areas more traditionally implicated in sleep and biological rhythms. We will also perform circuit mapping studies to provide insights on neural connections between regions that regulate emotional behavior and those that regulate sleep. In Aim 1, we will compare the effects of altering CRF and PACAP system function on sleep and biological (diurnal and circadian) rhythms. We will also examine the effects of PAC1R or CRFR1 ablation in AMG, following up on pilot data showing that stress increases PAC1R expression in this region. In Aim 2, we will directly compare the effects of acute and repeated sleep restriction on PACAP and CRF systems using RNAscope to quantify the magnitude of any changes and identify the cell-types in which changes occur. In Aim 3, we will use trans-synaptic viral labeling to map connections between circuits implicated in sleep and emotional behavior. Project 3 experiments are designed based on current knowledge, but as expected with Conte Centers, we will use new discoveries from all projects to refine and optimize our objectives via the Administrative Core.