Specification of sleep-wake control neurons in the basal forebrain

慢性应激是引发精神和躯体疾病的危险因素。部署和作战 暴露经常被认为是服役人员的压力源，与压力相关的精神疾病是一种 VA Healthcare中导致永久性残疾和疾病的主要原因。在神经学层面上，压力会扰乱 兴奋性和抑制性神经传递之间的生理平衡。突触的这种失调 网络活动经常被认为是导致许多棘手的精神疾病的原因，包括 抑郁、精神分裂症和失眠。退伍军人事务部面临发病率急剧上升和 近年来对失眠等睡眠障碍的诊断，代表着医疗费用的增加 对于退伍军人来说，更重要的是，这是一个增加了退伍军人自杀风险的因素。 家长拨款的首要目标是促进对睡眠神经生物学的理解， 促进下一代疗法的发展。它将识别GABA能神经元的新群体 在基底前脑(BF)中，调节睡眠和注意力的大脑综合区域。在这些细胞类型中， 我们的初步数据表明，BF神经元PAS结构域1表达(NPAS1)发生了化学激活 细胞强烈促进清醒。最近的临床前文献已经确认了BF NPAS1细胞的作用 应激敏感性和影响中的激活(Morais-Silva等人，2021)，潜在地由Dense介导 应激激活促肾上腺皮质激素释放因子(CRF)神经元在杏仁中央核的投射 Al.，2018年)。这些发现支持一种假设，即压力上调BF NPAS1细胞的活性， 导致失眠和相关的精神疾病。这一假设将是 研究2个目标，探索应激对NPAS1神经元体外电特性的影响 1)以及由此产生的体内睡眠行为效应(目标2)。 本研究补充资料为受训者Timothy Troppoli博士提供了理想的培训机会。 特罗波利博士于9月获得马里兰大学分子医学专业博士学位 2021年，最近加入了哈佛医学院精神病学系(HMS)和退伍军人事务部 一位健康科学专家。被辅导者的主要研究兴趣是关于压力诱导的干扰 突触神经传递与抑郁障碍和精神疾病的发病。特罗波利医生有一个 该领域多产、高影响力的研究历史，发表了4篇论文(其中两篇是第一作者) 研究新型抗抑郁药类化合物的作用机制和靶向结合。 学员的研究背景和专业知识与本研究补充材料的目标相辅相成。 虽然拟议的实验将显著提高特罗波利博士作为 VA BHS的神经科学家使用最先进的全细胞膜片钳技术，先进的显微镜， 化学遗传学和睡眠/脑电分析。这些方法的培训将由导师里奇博士提供 布朗和共同导师詹姆斯·麦肯纳博士，退伍军人管理局BHS/HMS成员和睡眠领域的专家 研究。布朗博士已经成功地指导了8名初级研究人员，其中包括VA CDA2获得者。他会的 对学员进行电生理学和化学遗传学方面的培训，指导学员的职业发展。麦肯纳医生的 BF神经解剖学背景和应激性失眠的临床前模型将支持分析 这项工作的严谨性，有助于解释结果，并协助为未来的高炉工作进行实验设计。 成功完成拟议的目标将表征应激诱导的功能缺陷 BF的NPAS1细胞，提供了它们在应激诱导的失眠和精神疾病中作用的第一个证据 疾病。建议的实验扩大了家长资助的范围，将指导小说的发展 退伍军人治疗学，并为未来退伍军人职业发展奖提供初步数据 明确关注压力、睡眠和抑郁的共同机制。

Chronic stress is a risk factor for the onset of psychiatric and somatic disease. Deployment and combat exposure are routinely cited as stressors for Service members, and stress-related psychiatric conditions are a leading cause of permanent disability and illness within VA Healthcare. On a neurological level, stress disrupts the physiological balance between excitatory and inhibitory neurotransmission. This dysregulation of synaptic network activity is routinely cited as a cause for numerous intractable psychiatric conditions, including depression, schizophrenia, and insomnia. Veterans Affairs has faced a steep rise in the incidence and diagnosis of sleep disorders like insomnia in recent years, representing an increasing healthcare cost for the VA and, critically, an enhanced risk factor for suicide among Veterans. An overarching goal of the parent grant is to advance understanding of the neurobiology of sleep, facilitating development of next-generation therapeutics. It will identify novel populations of GABAergic neurons in the basal forebrain (BF), an integratory brain region that regulates sleep and attention. Of these cell types, our preliminary data indicate chemogenetic activation of BF neuronal PAS domain 1-expressing (NPAS1+) cells strongly promotes wakefulness. Recent preclinical literature has identified a role of BF NPAS1+ cell activation in stress susceptibility and affect (Morais-Silva et al., 2021), potentially mediated by dense projections of stress-activated corticotropin-releasing factor (CRF) neurons from the central amygdala (Hunt et al., 2018). These findings support a hypothesis that stress upregulates the activity of BF NPAS1+ cells, contributing to the onset of insomnia and related psychiatric conditions. This hypothesis will be investigated in 2 aims, exploring the effects of stress on in vitro electrical properties of NPAS1+ neurons (aim 1) and resultant in vivo effects on sleep behavior (aim 2). This research supplement provides an ideal training opportunity for Dr. Timothy Troppoli, the mentee. Dr. Troppoli received a Ph.D. from the University of Maryland’s Molecular Medicine program in September 2021 and has recently joined the Department of Psychiatry at Harvard Medical School (HMS) and VA BHS as a Health Science Specialist. The mentee’s primary research interest concerns stress-induced disruption of synaptic neurotransmission and the onset of depressive disorders and psychiatric disease. Dr. Troppoli has a history of productive, high-impact research in this field, publishing 4 papers (two as first author) investigating the mechanisms and target engagement of novel antidepressant-like compounds. The mentee’s research background and expertise compliment the goals of this research supplement, while the proposed experiments will significantly enhance Dr. Troppoli’s independence and productivity as a neuroscientist at VA BHS using state-of-the-art whole-cell patch-clamp techniques, advanced microscopy, chemogenetics, and sleep/EEG analysis. Training in these approaches will be provided by mentor Dr. Ritchie Brown and co-mentor Dr. James McKenna, members of VA BHS/HMS and experts in the field of sleep research. Dr. Brown has successfully mentored 8 junior researchers, including VA CDA2 recipients. He will train the mentee in electrophysiology and chemogenetics and guide his career development. Dr. McKenna’s background in BF neuroanatomy and preclinical models of stress-induced insomnia will bolster the analytical rigor of this work, aid in interpretation of results and assist in experimental design for future BF work. Successful completion of the proposed aims will characterize stress-induced functional deficits of NPAS1+ cells of the BF, providing the first evidence of their role in stress-induced insomnia and psychiatric disease. The proposed experiments expand the scope of the parent grant, will guide the development of novel therapeutics for Veterans, and provide preliminary data for a future VA Career Development Award with a distinct focus on convergent mechanisms of stress, sleep, and depression.