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）：我们如何以及为什么睡眠是医学中未解决的中心问题。据估计，在美国，将近4000万人患有慢性或间歇性睡眠障碍，例如睡病，睡眠呼吸暂停，不安的腿综合症和失眠症。传统方法已经确定了几个神经元人群，它们的相互作用对于产生睡眠和清醒很重要。如何建立相互作用，如何改变病理学及其细胞和分子后果，仍然对其进行了较低的了解。该建议的长期目标是确定与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，以了解控制睡眠产生的分子机制。基于我们上一个资金时期的发现，该应用集中在电压门控钙通道的身份和功能上，以控制中桥胆碱能神经元的活性，这些神经元被认为在产生的觉醒和REM睡眠中起着至关重要的作用。我们的中心假设是，通过不同的钙通道的钙涌入受胆碱能和单胺能输入的差异调节，从而在改变行为跨行为状态的间泊胆碱能神经元的整合性中起不同的作用。为了检验该假设，我们将使用全细胞斑块夹，单细胞贴片夹，单光子和两光子钙成像以及激光未整体衰老方法的脑切片，来自野生型，钙通道基因敲除和毒蕈碱受体敲除小鼠的脑切片。这些研究的结果将1）确定小鼠中胆碱能“自动”受体的CAV2.3是否被胆碱能“自动”受体抑制。 2）确定这些神经元表达的多种肌肉受体抑制了这些钙通道。 3）确定这些神经元的soma和树突中的甲肾上腺素，5-羟色胺和腺苷抑制了哪些Ca2+通道； 4）确定通过CAV2.3通道在调节树突状和体细胞兴奋性方面的作用。这些结果将有助于我们理解睡眠调节的分子基础，并继续推进小鼠作为未来睡眠研究的平台。