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。这些发现与睡眠障碍的诊断增加了6倍， 近年来，。最普遍的睡眠障碍，失眠，与以下风险增加有关： 自杀此外，睡眠中断和异常皮层活动是常见的严重神经精神病 影响退伍军人的疾病，如痴呆症和精神分裂症。因此，更好的机械理解 控制睡眠-觉醒周期和皮层振荡的大脑回路的发展是迫切需要的 为退伍军人和其他弱势群体提供新型治疗。因此，这一广泛目标 一项在小鼠身上进行的研究计划是确定新的治疗靶点，以纠正睡眠和皮层的异常。 电活动。我们将在小鼠中使用一种创新的方法，该方法基于 并确定了控制其在成年人中活性的转录因子， 允许靶向治疗重新校准它们的活动以恢复正常的睡眠和皮质节律。我们 重点放在基底前脑，一个涉及睡眠-觉醒活动、注意力和奖励的区域， 在痴呆症中，以及在这个区域中释放抑制性神经递质的最大神经元群上， γ-氨基丁酸（GABA）。我们靶向表达三种转录因子的神经元， 失眠症和其他神经精神疾病在退伍军人中常见的遗传和死后研究。 帮助退伍军人的路线图是：我们将根据他们的神经元来识别新的神经元组。 发展起源（Aim 1）。我们将使用转基因小鼠，允许操纵他们的活动， 确定它们如何影响睡眠-觉醒行为和皮层振荡（目标2）。最后，我们将最先进的 基因技术，以确定转录因子控制其活动在成人（目标3），使我们能够 纠正神经精神疾病中的异常睡眠-觉醒行为和皮层电活动， 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能神经元产生于发育中的大脑的一个区域， 与影响退伍军人的各种神经精神疾病的风险有关。内 在腭板下，尾侧、内侧和外侧的祖细胞产生不同的GABA能神经元群， 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 指定唯一神经元组。我们将使用转基因小鼠，它们表达一种酶， 重组酶（Cre），在这些转录因子的控制下，以鉴定它们并操纵它们的转录。 活动将这些小鼠品系与另一种表达红色荧光蛋白的小鼠品系杂交， Cre存在与免疫染色相结合将允许我们绘制它们的位置和表型 (Aim 1）。在目标2中，我们将通过注射病毒增强它们的活性或消除它们来研究它们的功能。 在Cre存在下表达蛋白质的载体。这些实验将揭示这些神经元的作用 在睡眠-觉醒行为和控制皮层电活动的第一次，并设置了阶段， 翻译研究来改变它们的活性。最后，在目标3中，我们将开始迈向翻译的第一步。 通过使用最先进的遗传技术来调节这些转录之一的活性 因子Lhx6，其表达在涉及精神分裂症的GABA能细胞类型中改变。我们将使用一个 最先进的基因编辑技术，称为重复的规则间隔短回文重复序列 （CRISPR），目前正在各种疾病的临床试验中进行测试。 如果成功，正如我们强大的初步数据所表明的那样，该项目将允许新的翻译 研究和纠正退伍军人疾病相关异常的方法， 前脑神经元和基因调节睡眠-觉醒行为和皮质电活动。

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.