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

摘要：项目3 许多精神疾病都会影响睡眠。有些是与有压力相关障碍的人(例如，后 创伤应激障碍[创伤后应激障碍])睡眠比正常少，而其他人睡得更多，或有支离破碎的症状 睡眠模式反映了更频繁的睡眠和清醒。有大量证据表明 压力会扰乱睡眠，我们已经证明，慢性压力会导致小鼠睡眠和睡眠的深刻变化 昼夜节律。相比之下，睡眠失调对调节压力效应的大脑系统的影响是 不完全理解的。睡眠不佳或受限的时期可能会产生分子变化， 从而影响睡眠。在啮齿动物身上的研究显示了慢性肾功能衰竭和慢性肾功能衰竭的重要相似和不同之处 促肾上腺皮质激素释放因子和垂体腺苷环化酶激活多肽对血管紧张素转换酶活性的影响 行为。尽管这两种多肽都增加了声音惊吓，但这是一种警觉措施，通常用于临床前和 对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 迅速解决问题。这些类型的发现，以及CRF和PACAP系统中已知的基因变化 在易受压力相关疾病影响的人群中，为比较和对比神经生物学提供了理论基础 这些多肽中的一个。CRF和PACAP对睡眠的影响还没有被直接比较，特别是在 延长的时间段。我们的前提是，这些肽系统会影响睡眠，也会受到睡眠的影响。我们的 假设这些多肽与睡眠之间存在一种相互作用的关系，从而激活 CRF和/或PACAP系统将影响睡眠，反过来，睡眠限制将影响CRF和/或PACAP 但这些影响的持久性将有所不同。测试将在雄性和雌性小鼠身上进行 植入无线发射器，能够持续收集脑电、肌电、体温和 几周的活动数据。分子分析将集中在涉及情绪方面的大脑区域。 应激，包括杏仁核(AMG)、终纹床核(BNST)和前额叶皮质(PFC)，AS 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 对调节情绪行为的区域和这些区域之间的神经联系提供洞察的研究 来调节睡眠。在目标1中，我们将比较CRF和PACAP系统功能改变对睡眠的影响 和生物(昼夜和昼夜)节律。我们还将研究PAC1R或CRFR1消融的效果 在AMG中，追踪飞行员数据显示，应激增加了该区域PAC1R的表达。在目标2中，我们 将直接比较急性和反复睡眠限制对PACAP和CRF系统的影响 RNAScope可以量化任何变化的大小，并识别发生变化的细胞类型。在AIM 3，我们将使用跨突触病毒标记来绘制与睡眠和情绪有关的回路之间的联系 行为。项目3实验是基于当前知识设计的，但与Conte Center的预期一样， 我们将利用所有项目的新发现，通过管理核心来提炼和优化我们的目标。

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.