Cholinergic Phenotype in Murine Models of Sleep

小鼠睡眠模型中的胆碱能表型

• 批准号: 6774695
• 负责人: RALPH LYDIC
• 金额: $ 26.39万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 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 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6774695
• 关键词: G protein; SDS polyacrylamide gel electrophoresis; acetylcholine; biological models; brain electrical activity; brain mapping; electroencephalography; electromyography; high performance liquid chromatography; histochemistry /cytochemistry; laboratory mouse; model design /development; muscarinic receptor; neural transmission; prefrontal lobe /cortex; protein localization; purinergic receptor; sleep; sleep deprivation; western blottings

DESCRIPTION (provided by applicant): This research program was initiated in 1999 in response to RFA HL99001. The goal of the RFA was to stimulate improved molecular, cellular, and systems approaches to investigate sleep in mice. Every human gene has a mouse homologue. This remarkable homology means that the mouse genome may provide unique insights into human disease. Advances in sequencing the mouse genome now require complimentary data regarding normal and abnormal phenotypes. In accord with consensus statements published by The Jackson Laboratory, this application focuses on the C57BL/6J (B6) mouse strain. The long-term objectives are to advance scientific knowledge by providing data not presently available concerning molecules that regulate ACh release and alter electroencephalographic (EEG) excitability, sleep, and breathing. Aim 1 will test the hypothesis that microinjecting neostigmine into the pontine reticular formation of B6 mouse causes a REM sleep-like state and changes in breathing that are concentration-dependent, site-specific within the pons, and blocked by atropine. Aim 2 will use in vivo microdialysis and high performance liquid chromatography (HPLC) to test the hypothesis that dialysis delivery of an adenosine A1 receptor agonist into the prefrontal cortex of B6 mouse will decrease cortical ACh release and EEG power, and delay wake-up time from anesthesia. Aim 3 will use combined microdialysis and microinjection to test the hypothesis that ACh release in the pontine reticular formation of B6 mouse is altered by nitric oxide donors and by inhibitors of nitric oxide synthase. Aim 4 will use a quantitative Western assay to test the hypothesis that brain expression of M2 muscarinic receptor protein varies significantly as a function of mouse strain and brain region. These aims will take this research program in new directions by developing a pharmacological model of rapid eye movement sleep in mouse (Aim 1), quantifying the effects of endogenous neuromodulators on ACh release (Aims 2 and 3), and initiating strain comparisons of muscarinic receptor protein expression (Aim 4). The unifying conceptual scheme of this proposal is that higher level phenotypes such as sleep and breathing (Aim 1) emerge from the expression of lower level phenotypes such as ACh (Aims 2 and 3) and muscarinic cholinergic receptors (Aim 4).

描述（由申请人提供）：该研究计划于1999年启动了RFA HL99001。 RFA的目的是刺激改进的分子，细胞和系统方法来研究小鼠的睡眠。每个人类基因都有小鼠的同源物。这种非凡的同源性意味着小鼠基因组可以对人类疾病提供独特的见解。现在，对小鼠基因组进行测序的进展需要有关正常和异常表型的免费数据。根据杰克逊实验室发表的共识声明，该应用集中在C57BL/6J（B6）小鼠菌株上。长期目标是通过提供有关调节ACH释放并改变脑电图（EEG）兴奋性，睡眠和呼吸的分子的数据来提高科学知识。 AIM 1将检验以下假设：将新生氨氨酸微注射到B6小鼠的桥丁网状形成中会导致REM睡眠状状态和呼吸的变化，这些呼吸依赖性浓度依赖性，特定于PON的位点特异性，并被阿托品阻塞。 AIM 2将使用体内微透析和高性能液相色谱（HPLC）来检验以下假设：透析腺苷A1受体激动剂透析到B6小鼠的前额叶皮层中透析会降低皮质ACH释放和EEG释放和EEEG功率，并延迟从麻醉中唤醒时间。 AIM 3将使用合并的微透析和显微注射来检验以下假设，即一氧化氮供体和一氧化氮合酶的抑制剂改变了B6小鼠蓬蒂恩网状形成中的ACH释放。 AIM 4将使用定量的西部测定法来检验以下假设：M2毒蕈碱受体蛋白的大脑表达随小鼠菌株和脑区域的函数而变化很大。这些目标将通过开发小鼠快速眼动睡眠的药理学模型（AIM 1），量化内源性神经调节剂对ACH释放的影响（AIMS 2和3），并启动毒蕈碱受体蛋白表达的菌株比较（AI​​M 4），从而将该研究计划沿新方向进行了新的方向（AIM 1）。该提案的统一概念方案是，较高水平的表型，例如睡眠和呼吸（AIM 1），从较低水平表型（例如ACH（AIMS 2和3）（目标2和3）和毒蕈碱胆碱能受体（AIM 4）中出现。