Specification of sleep-wake control neurons in the basal forebrain

部署和战斗暴露大大增加了失眠和其他睡眠障碍的风险 在军事人员中。这些发现与睡眠障碍的诊断增加了6倍相吻合 弗吉尼亚州在最近几年。最普遍的睡眠障碍，失眠，与增加的风险有关 自杀。此外，睡眠障碍和皮质活动异常在严重的神经精神病患者中也很常见。 影响退伍军人的疾病，如痴呆症和精神分裂症。因此，更好的机械性理解 控制睡眠-觉醒周期和大脑皮层振荡的大脑回路亟待发展 为退伍军人和其他弱势群体提供新的治疗方法。因此，这一行动的广泛目标是 在老鼠身上的研究计划是寻找新的治疗靶点来纠正睡眠和皮质的异常 电活动。我们将在小鼠身上使用一种创新的方法，它描述了基于 关于它们的发育起源，并确定在成人中控制它们活动的转录因子， 允许有针对性的治疗，重新校准他们的活动，以恢复正常的睡眠和皮质节律。我们 专注于基础前脑，这是一个参与睡眠-觉醒活动、注意力和奖赏退化的区域 在痴呆症中，在这一区域释放抑制性神经递质的最大神经元群上， γ-氨基丁酸(GABA)。我们的目标是表达三种转录因子的神经元 通过基因和尸检研究，失眠和其他神经精神障碍在退伍军人中很常见。 帮助退伍军人的路线图是：我们将根据他们的神经元识别新的神经元组 发展起源(目标1)。我们将使用转基因小鼠来控制它们的活动 确定它们如何影响睡眠-觉醒行为和大脑皮层振荡(目标2)。最后，我们将以最先进的技术 利用基因技术识别控制成人转录因子活性的转录因子(目标3)，使我们 通过以下方法纠正神经精神障碍患者的异常睡眠-觉醒行为和皮质电活动 设计作用于这些转录因子的基于病毒载体的疗法(长期目标)。 前脑GABA能神经元产生于大脑发育中的大脑皮层下区域 牵涉到影响退伍军人的各种神经精神障碍的风险。在 在大脑皮层下，不同组的GABA能神经元由祖细胞在尾侧、内侧和 外侧神经节隆起和胚胎视前区。在每个区域，不同的转录因子 指定独特的神经元组。我们将使用表达Cre酶的转基因小鼠 重组酶(Cre)，在这些转录因子的控制下，识别它们并操纵它们 活动。将这些小鼠品系与另一个表达红色荧光蛋白的小鼠品系进行杂交 Cre的存在，结合免疫染色，将使我们能够绘制出它们的位置和表型 (目标1)。在目标2中，我们将通过增加它们的活性或通过注射病毒来消融它们来研究它们的功能。 在Cre存在的情况下表达蛋白质的载体。这些实验将揭示这些神经元的作用 首次在睡眠-觉醒行为和大脑皮层电活动的控制上，为 翻译研究，以改变他们的活动。最后，在目标3中，我们将开始朝着翻译的第一步 通过使用最先进的基因技术来调节其中一种转录的活性 因子Lhx6，其表达在一种与精神分裂症有关的GABA能细胞类型中发生变化。我们将使用 最先进的基因编辑技术，称为有规律地散布的短回文重复 (CRISPR)，目前正在进行各种疾病的临床试验。 如果成功，正如我们强有力的初步数据所表明的那样，这个项目将允许新的翻译 通过调控基底细胞活性研究和纠正退伍军人疾病相关异常的方法 调节睡眠觉醒行为和大脑皮层电活动的前脑神经元和基因。

Deployment and combat exposure substantially increase the risk for insomnia and other sleep disorders in military personnel. These findings match with the 6-fold increase in the diagnosis of sleep disorders in the VA in recent years. The most prevalent sleep disorder, insomnia, is associated with an increased risk for suicide. Furthermore, disrupted sleep and abnormal cortical activity are common in severe neuropsychiatric conditions affecting veterans such as dementia and schizophrenia. Thus, a better mechanistic understanding of the brain circuitry controlling sleep-wake cycles and cortical oscillations are urgently needed to develop novel treatments for veterans and other vulnerable populations. Accordingly, the broad objective of this research program in mice is to identify new therapeutic targets to correct abnormalities of sleep and cortical electrical activity. We will use an innovative approach in mice which characterizes subgroups of neurons based on their developmental origin and identifies the transcription factors which control their activity in adults, allowing targeted therapies which recalibrate their activity to restore normal sleep and cortical rhythms. We focus on the basal forebrain, a region involved in sleep-wake activity, attention and reward which degenerates in dementia, and on the largest group of neurons in this region which release the inhibitory neurotransmitter, gamma-amino-butyric acid (GABA). We target neurons which express three transcription factors linked to insomnia and other neuropsychiatric disorders common in veterans by genetic and postmortem studies. The roadmap to helping veterans is: We will identify new groups of neurons based on their developmental origin (Aim 1). We will use transgenic mice which allow manipulation of their activity to determine how they affect sleep-wake behavior and cortical oscillations (Aim 2). Finally, we will state-of-the-art genetic techniques to identify the transcription factors which control their activity in adults (Aim 3), allowing us to correct abnormal sleep-wake behavior and cortical electrical activity in neuropsychiatric disorders by designing viral vector based therapies which act on those transcription factors (long-term goal). Forebrain GABAergic neurons are generated in the subpallium, an area of the developing brain implicated in the risk for developing diverse neuropsychiatric disorders affecting veterans. Within the subpallium, different groups of GABAergic neurons are generated by progenitors in the caudal, medial and lateral ganglionic eminences and embryonic preoptic regions. In each region, different transcription factors specify unique groups of neurons. We will use genetically modified mice which express an enzyme, Cre recombinase (Cre), under the control of these transcription factors to identify them and manipulate their activity. Crossing these mouse strains with another mouse strain which expresses a red fluorescent protein in the presence of Cre, in combination with immunostaining will allow us to map their location and phenotype (Aim 1). In Aim 2 we will study their function by increasing their activity or ablating them using injections of viral vectors which express proteins in the presence of Cre. These experiments will reveal the role of these neurons in sleep-wake behavior and control of cortical electrical activity for the first time and set the stage for translational studies to alter their activity. Finally, in Aim 3, we will begin the first step towards a translational approach by using state-of-the art genetic techniques to modulate the activity of one of these transcription factors, Lhx6, whose expression is altered in a GABAergic cell-type implicated in schizophrenia. We will use a state-of-the-art gene editing technique called Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR), which is currently being tested in clinical trials for various disorders. If successful, as suggested by our strong preliminary data, this project will allow novel translational approaches to study and correct disease-related abnormalities in veterans by manipulating the activity of basal forebrain neurons and genes which regulate sleep-wake behavior and cortical electrical activity.