Astroglial Orexin in Sleep Disorders

睡眠障碍中的星形胶质细胞食欲素

• 批准号: 8585663
• 负责人: Priyattam J. Shiromani
• 金额: $ 22.43万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8585663
• 关键词: Affect; Animal Disease Models; Animal Model; Area; Astrocytes; Axon; Behavior; Brain; Brain region; Calcium; Cataplexy; Cations; Chemicals; Complex; Corpus striatum structure; Data; Development; Disease; Distal; Gene Delivery; Gene Transfer; Genes; Genetic; Genetic Engineering; Glial Fibrillary Acidic Protein; Hour; Image; In Vitro; Knock-out; Leg; Length; Ligands; Light; Link; MJD1 protein; Measures; Methods; Modeling; Motor; Movement; Mus; Narcolepsy; Nerve Degeneration; Neurobiology; Neuroglia; Neurons; Obstructive Sleep Apnea; Optics; Peptides; Pharmacologic Substance; Pharmacology; Pontine structure; Publications; Research; Role; Site; Sleep; Sleep Disorders; Specificity; Symptoms; Testing; Transgenic Mice; behavior change; cost; cost effective; hypocretin; improved; mammilloinfundibular nucleus structure; mouse model; novel strategies; optogenetics; promoter; public health relevance; receptor; research study; synaptic function; tool

DESCRIPTION (provided by applicant): We have demonstrated the first successful use of neuronal-orexin gene transfer to ameliorate symptoms of narcolepsy in two established animal models of the disease. The effects were site specific and depended on the connectivity of the surrogate neurons. Since neuronal orexin can be secreted at multiple distal sites, some of which may regulate narcoleptic behavior whereas others do not, we propose a localized gene delivery method by expressing orexin in astroglia. Astroglial-orexin will re-establish the ligand-receptor link in a localized area that already contains the orexin receptors. Astroglial-orexin gene deliver will serve as a cost-effective neurobiological tool to understand the networking underlying sleep. In aim 1 the gene for orexin will be inserted into astroglia (GFAP promoter driven) in three specific brain regions, two of which are implicated in network sleep models (TMN, pons) and the third, striatum, will serve as control. Our preliminary results in the orexin-ataxin-3 mice model indicate that in the TMN glial-orexin (rAAV-GFAP-orexin) completely rescues waking but not cataplexy. In the pons it produces a 78% reduction in cataplexy but not waking. Thus, there is site- specificity of the glial-orexin. In aim 2 astroglial-orexin will be driven by optogenetic stimulation to determine whether such stimulation further stimulates waking or blocks cataplexy, especially in ineffective sites in aim 1. Experiments with appropriate controls, including orexin receptor antagonist are proposed to strengthen the conclusions. In-vitro calcium imaging study will determine functionality of the genetically inserted ChR2 receptors. To the best of our knowledge this is the first use of astroglial-orexin in sleep disorders. Gene transfer is extremely cost-efficient, it is used clinically and it will demonstrate that genetic pharmacology can be used to elucidate the network in sleep disorders.

描述（由申请人提供）：我们已经证明了神经元 - 蛋白质基因基因转移到了两种已建立的疾病动物模型中的改善症状。效果是特定于位点的，取决于替代神经元的连通性。由于可以在多个远端部位分泌神经元蛋白，其中一些可能调节麻醉性行为，而其他一些则不能调节固有的基因递送方法，通过在星形胶质细胞中表达局部基因递送方法。星形胶质细胞蛋白将在已经包含Orexin受体的局部区域中重新建立配体受体链接。星形胶质细胞 - 蛋白质基因的传递将成为一种具有成本效益的神经生物学工具，以了解睡眠的网络。在AIM 1中，Orexin的基因将在三个特定的大脑区域中插入星形胶质细胞（GFAP启动子驱动）中，其中两个与网络睡眠模型（TMN，PON）有关，第三个纹状体将作为对照。我们的初步结果在OREXIN-ATAXIN-3小鼠模型中表明，在TMN神经胶质蛋白（RAAV-GFAP-蛋白质）中，完全挽救了醒来，但没有弹出。在PON中，它会降低78％，但不会醒来。因此，神经胶质蛋白的位点特异性。在AIM 2中，星形胶质细胞 - 蛋白质将由光遗传学刺激驱动，以确定这种刺激是否进一步刺激醒来或阻塞弹药，尤其是在AIM 1中无效的位置。提议具有适当控制的实验，包括针对选择性对照的实验，包括Orexin受体拮抗剂，以增强结论。体外钙成像研究将确定遗传插入的CHR2受体的功能。据我们所知，这是星形胶质细胞在睡眠障碍中首次使用。基因转移极为 具有成本效益，在临床上使用，它将证明可以使用遗传药理学 阐明睡眠障碍中的网络。