Sleep Neurobiology and Circuitry

描述(由申请人提供)： 7000万美国人患有某种睡眠障碍。大脑如何在意识状态之间切换尚不清楚，但研究人员现在已经确定，一个分布的神经元网络负责清醒、非快速眼动和快速眼动睡眠。一阶电路模型已经出现，并使用传统的损伤和基因缺失方法进行了测试。我们现在建议使用基因转移方法修复网络。这种方法已被证明是治疗许多神经退行性疾病的有效神经生物学工具，但令人惊讶的是，还没有人用它来纠正睡眠障碍。在这个项目中，重组腺相关病毒载体(rAAV-血清型8)将被用来将觉醒肽下克汀素(也称为增食欲素)的基因转移到特定的神经元表型。一个由四个目标组成的综合系列将在两个靶向缺失下丘脑肌素基因或下丘脑神经元靶向缺失的小鼠模型中测试特定的假说。为了加强这一结论，我们建议进行适当控制的实验。这些目标得到了我们非常强大的初步数据的支持，这些数据显示，在两个小鼠模型中，特定神经元中的下丘脑泌素基因产物强劲表达，同时发作性睡病行为明显下降。这些研究将证明遗传药理学可以作为修复网络的一种有价值的神经生物学工具。

DESCRIPTION (provided by applicant): 70 million Americans suffer from some sort of sleep disorder. How the brain switches between states of consciousness is not known, but researchers have now determined that a distributed network of neurons are responsible for wake, non-REM and REM sleep. First-order circuit models have emerged, and these have been tested using traditional lesion and gene deletion methods. We now propose to repair the network using the gene transfer method. This approach has proven to be an effective neurobiological tool in a number of neurodegenerative diseases but surprisingly no one has used it to correct a sleep disorder. In this project recombinant adeno-associated viral vector (rAAV-serotype 8) will be used to transfer the gene for the arousal peptide hypocretin (also known as orexin) into a specific neuronal phenotype. An integrated series of four aims will test specific hypotheses in two murine models with targeted deletions of the hypocretin gene or of the hypocretin neurons. Experiments with appropriate controls are proposed to strengthen the conclusions. These aims are supported by our very strong preliminary data that shows robust expression of the hypocretin gene product in specific neurons along with unambiguous decline of narcoleptic behavior in the two murine models. These studies will demonstrate that genetic pharmacology can serve as a valuable neurobiological tool to repair the network.