Cholinergic Regulation of the Circadian Clock

描述(申请人提供)：乙酰胆碱(ACh)具有多种调节作用 在中枢神经系统中的作用。脑干和基底的胆碱能神经元 前脑结构将上升纤维发送到整个大脑，在那里它们可能 参与广泛的功能，包括调节学习， 记忆提取、情绪状态、中央自主神经控制和过程 调节睡眠/唤醒。胆碱能信号诱导的机制 神经元状态的长期变化及其对行为的影响， 都是对人类健康非常重要的研究兴趣领域。 我们建议研究通过m1mAChRs的胆碱能信号转导机制 (M1.Rs)与行为相关的下丘脑神经元和永生化细胞 来自大鼠的，以及转基因小鼠模型中的。M1.R分布的成像 在视交叉上核(SCN)内显示实质性受体 浓度。我们发现M1亚型类药理试剂 选择性地改变胆碱能反应。利用转基因技术获得的初步数据 小鼠和永生化细胞系支持药理学并指出这一点 脑部位作为评价M1.R信号的重要模型。我们 建议采用从行为分析到细胞外的各种技术 以及脑片上的膜片钳记录，伴随着免疫细胞化学， 生化和细胞生物学方法，共同旨在揭示 M1.R信号的基本机制。我们的具体目标包括：1) 充分评价M1.R基因缺失对胆碱能的影响 响应，2)评估下游可扩散信使的行为，以及3) 确定特定的PKG亚型在该M1.R信号级联中的作用。在……里面 除了对理解毒鼠强做出基本贡献外 中枢神经系统神经元的神经调节，这些研究将使我们能够评估 M1.R介导的神经传递在确定的行为轴中的作用。这 多学科方法将为中枢胆碱能提供新的见解 神经传递，毒鼠碱信号转导机制，以及 形成行为改变的神经基础的决策过程。 信号转导是一个细胞过程，通过识别 特定的受体，第二信使系统和靶点，我们将能够 了解调节神经元长期调节的因果机制 州政府。本研究具有战略应用上的现实意义。 胆碱能障碍的合理治疗，包括那些改变 睡眠/觉醒、自主神经功能、老年性痴呆、阿尔茨海默型(SDAT)、 帕金森氏病、亨廷顿舞蹈症等神经精神和运动 精神错乱。

DESCRIPTION (provided by applicant): Acetylcholine (ACh) has diverse regulatory roles in the central nervous system. Cholinergic neurons in brainstem and basal forebrain structures send ascending fibers throughout the brain where they may participate in a wide range of functions, including modulation of learning, memory retrieval, mood states, central autonomic control, and the processes regulating sleep/arousal. Mechanisms by which cholinergic signals induce long-lasting changes in neuronal state, and their consequences for behavior, are areas of intense research interest with great importance for human health. We propose to investigate mechanisms of cholinergic signaling via M1 mAChRs (M1.Rs) to behaviorally relevant hypothalamic neurons and immortalized cells from rat, as well as in transgenic mouse models. Imaging of M1.R distribution within the suprachiasmatic nucleus (SCN) reveals substantial receptor concentrations. We have found that M1 subtype-like pharmacological reagents selectively alter the cholinergic response. Preliminary data using transgenic mice and immortalized cell lines support the pharmacology and point to this brain site as an important model in which to evaluate M1.R signaling. We propose to employ techniques ranging from behavioral analyses to extracellular and patch-clamp recordings in brain slices, accompanied by immunocytochemical, biochemical and cell biological approaches, together aimed at uncovering fundamental mechanisms of M1.R signaling. Our specific aims include: 1) To fully evaluate the effect of genetic deletion of the M1.R on the cholinergic response, 2) To assess actions of downstream diffusible messengers, and 3) To define the role of specific PKG isoforms in this M1.R signaling cascade. In addition to making fundamental contributions to understanding muscarinic neuromodulation of CNS neurons, these studies will permit us to evaluate the roles of M1.R-mediated neurotransmission within a defined behavioral axis. This multidisciplinary approach will provide new insights into central cholinergic neurotransmission, muscarinic signal transduction mechanisms, and decision-making processes that form the neural substrates of behavioral change. Signal transduction is a cellular process, and by identifying the roles of specific receptors, second messenger systems and targets, we will be able to understand the causal mechanisms that mediate long-term adjustments in neuronal state. This research has applied relevance for strategies in developing rationally-based therapies for cholinergic disorders, including those altering sleep/arousal, autonomic function, senile dementia, Alzheimer's type (SDAT), Parkinson's disease, Huntingtons chorea and other neuropsychiatric and movement disorders.