Ion channels regulating REM-related cholinergic neurons

调节 REM 相关胆碱能神经元的离子通道

• 批准号: 7789543
• 负责人: CHRISTOPHER S LEONARD
• 金额: $ 35.78万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7789543
• 关键词: Action Potentials; Adenosine; Agonist; Arousal; Behavioral; Brain; Calcium; Calcium Channel; Carbachol; Cells; Chronic; Data; Dendrites; Doctor of Philosophy; Funding; Future; Generations; Glutamates; Growth; Image; Ion Channel; Knock-out; Knockout Mice; Lasers; Measures; Mediating; Medicine; Methods; Microscopy; Molecular; Mus; Muscarinic Acetylcholine Receptor; Muscarinics; Narcolepsy; Neurons; Neurotransmitter Receptor; Neurotransmitters; Nitric Oxide; Output; Pathology; Photons; Play; Population; Process; Property; REM Sleep; Regulation; Relative (related person); Research; Research Personnel; Restless Legs Syndrome; Role; Serotonin; Sleep; Sleep Apnea Syndromes; Sleep Disorders; Sleeplessness; Slice; Speed; Synapses; Synaptic plasticity; System; Testing; United States; Ursidae Family; Voltage-Gated Potassium Channel; Wakefulness; Whole-Cell Recordings; Width; base; cholinergic; cholinergic neuron; experience; imaging modality; neuronal cell body; novel; patch clamp; programs; receptor; research study; sleep regulation; tool; two-photon; voltage

DESCRIPTION (provided by applicant): How and why we sleep are central unsolved questions in medicine. Nearly 40 million people in the United States are estimated to experience chronic or intermittent sleep disorders such as narcolepsy, sleep apnea, restless leg syndrome and insomnia. Traditional approaches have identified several neuronal populations whose interplay is important in generating sleep and wakefulness. How that interplay is established, how it is altered in pathology and its cellular and molecular consequences, remain poorly understood. The long-term objective of this proposal is to determine the molecular identity and function of ion channels and receptors expressed by sleep-related neurons in order to understand the molecular mechanisms controlling sleep generation. Building on our findings from the previous funding period, this application focuses on the identity and function of voltage-gated calcium channels in controlling activity of mesopontine cholinergic neurons which are believed to play a pivotal role in the generation of wakefulness and REM sleep. Our central hypothesis is that the calcium influx through distinct calcium channels are differentially regulated by cholinergic and monoaminergic inputs and thereby play different roles in altering the integrative properties of mesopontine cholinergic neurons across behavioral state. To test this hypothesis we will use pharmacological methods with whole-cell patch clamp and single- and two-photon calcium imaging and laser uncaging methods in brain slices from wild-type, calcium channel knockout and muscarinic receptor knock-out mice. The results from these studies will 1) determine if Cav2.3 containing calcium channels are inhibited by cholinergic "auto" receptors in the soma and dendrites of important arousal-related neurons in mouse; 2) determine which of the multiple muscarinc receptors expressed by these neurons inhibit these calcium channels. 3) determine which Ca2+ channels are inhibited by noradrenalin, serotonin and adenosine in the soma and dendrites of these neurons; 4) Determine the role of calcium influx through Cav2.3 channels in regulating dendritic and somatic excitability. These results will contribute to our understanding of the molecular basis of sleep regulation as well as continue to advance the mouse as a platform for future sleep research.

描述（由申请人提供）：我们如何以及为何睡眠是医学上未解决的核心问题。据估计，美国有近 4000 万人患有慢性或间歇性睡眠障碍，如发作性睡病、睡眠呼吸暂停、不宁腿综合症和失眠。传统方法已经确定了几个神经元群，它们的相互作用对于产生睡眠和觉醒很重要。这种相互作用是如何建立的，它是如何在病理学中改变的，以及它的细胞和分子后果，仍然知之甚少。该提案的长期目标是确定睡眠相关神经元表达的离子通道和受体的分子身份和功能，以了解控制睡眠生成的分子机制。基于我们之前资助期的发现，该申请重点关注电压门控钙通道在控制中脑桥胆碱能神经元活动中的特性和功能，这些神经元被认为在觉醒和快速眼动睡眠的产生中发挥着关键作用。我们的中心假设是，通过不同钙通道的钙流入受到胆碱能和单胺能输入的差异调节，从而在改变中脑桥胆碱能神经元跨行为状态的整合特性方面发挥不同的作用。为了检验这一假设，我们将在野生型、钙通道敲除和毒蕈碱受体敲除小鼠的脑切片中使用全细胞膜片钳、单光子和双光子钙成像以及激光解笼方法的药理学方法。这些研究的结果将 1) 确定含有钙通道的 Cav2.3 是否被小鼠重要的唤醒相关神经元的体细胞和树突中的胆碱能“自身”受体抑制； 2)确定这些神经元表达的多种毒蕈碱受体中的哪一个抑制这些钙通道。 3）确定这些神经元的体细胞和树突中哪些Ca2+通道被去甲肾上腺素、血清素和腺苷抑制； 4)确定通过Cav2.3通道的钙流入在调节树突和体细胞兴奋性中的作用。这些结果将有助于我们了解睡眠调节的分子基础，并继续推动小鼠作为未来睡眠研究的平台。