BLR&D Research Career Scientist Award Application

BLR

• 批准号: 9899097
• 负责人: Priyattam J. Shiromani
• 金额: --
• 依托单位: RALPH H JOHNSON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9899097
• 关键词: 3-Dimensional; Aging; American; Anxiety; Area; Arousal; Atypical depressive disorder; Autopsy; Award; Behavior; Behavioral Symptoms; Books; Brain; Brain Mapping; Brain imaging; Complex; Data; Defect; Disease; Drowsiness; Emotional; Environment; FDA approved; Fluorescence; Funding; Gene Transfer; General Population; Generations; Genetic Engineering; Genetically Engineered Mouse; Goals; Grant; Grant Review; Growth and Development function; Heart; Hypothalamic structure; Image; Individual; Injury; International; Jet Lag Syndrome; Journals; Kidney; Laboratories; Laboratory Research; Life; Limbic System; Link; Liver; Magnetic Resonance Imaging; Malignant Neoplasms; Mammals; Maps; Measures; Memory; Mental Depression; Mentors; Methodology; Methods; Microscope; Microtomy; Modern Medicine; Molecular; Moods; Mus; Narcolepsy; Nerve Degeneration; Neurobiology; Neurodegenerative Disorders; Neurons; Neurosciences; Obstructive Sleep Apnea; Pathology; Patients; Peer Review; Pharmacogenetics; Phenotype; Philosophy; Post-Traumatic Stress Disorders; Prosthesis; Proteins; Publishing; Rattus; Research; Research Personnel; Resolution; Review Committee; Rodent Model; Science; Scientist; Series; Services; Signal Transduction; Sleep; Sleep Deprivation; Sleep Disorders; Sleep disturbances; Sleeplessness; Slice; Spinal Cord; Stroke; Students; Substance abuse problem; Testing; Time; Tissues; Training; Trauma; Traumatic Brain Injury; United States National Institutes of Health; Veterans; Visual; Wakefulness; Work; awake; brain tissue; calcium indicator; career; cell type; conditioned fear; emotional behavior; experience; experimental study; falls; hypnotic; hypocretin; imaging modality; interest; liver imaging; microendoscope; mild traumatic brain injury; mouse model; neural circuit; neuronal circuitry; non-alcoholic fatty liver disease; optogenetics; post-traumatic symptoms; programs; reconstruction; skills; tool; wound healing

70 million Americans suffer from some sort of sleep disorder. Behavior, mood and memory deteriorate with sleep loss and it gets worse with continuing sleep deprivation. Sleep disturbance is a frequent and common complaint among our Veterans. Lack of sleep due to hyperarousal is one symptom of PTSD, but it is not known why Veterans with PTSD cannot fall asleep. My research focuses on identifying and mapping the brain neurons that induce sleep. The overall impact of my research is that it will provide the first direct evidence linking specific phenotypes of neurons and their circuits responsible for inducing sleep. This will make it possible to induce sleep in conditions where the arousal drive is very strong, such as in the insomnia of PTSD, or to maintain wakefulness when there is excessive sleepiness, such as in patients with obstructive sleep apnea or atypical depression. One series of experiments uses optogenetics and pharmacogenetics to identify functional circuits in the brain. The brain contains many different types of cells and through optogenetics and pharmacogenetics it is now possible to disassemble the brain to identify the culprit neurons responsible for complex behaviors, such as sleep. My lab was the first in the area of sleep neurobiology to use optogenetics to induce sleep (Konadhode et al., 2013, attached). We activated a specific phenotype of neurons in the hypothalamus and discovered that it induced sleep at a time of day when the mouse should have been awake. We have now shown the same effect in rats, indicating that activating these neurons drives sleep across mammals. We now want to test the sleep-inducing effect in conditions of high arousal, such as fear-conditioning (PTSD) or anxiety. In conjunction with optogenetics and pharmacogenetics, I am using the deep-brain imaging method to image the activity of phenotype-specific neurons in the brains. This method measures changes in fluorescence of a genetically encoded calcium indicator in individual neurons. The fluorescence signal is captured via a microendoscope attached to a miniature microscope (2g). The microendoscope can be placed anywhere in the brains of mice providing unprecedented record of neuronal activity. I am using it to obtain visual evidence of the activity of specific neuronal circuits during sleep and waking. Another series of studies uses the CLARITY method to map the circuit activated by optogenetics. CLARITY is a new neuroanatomical method that makes postmortem tissue, such as the brain, transparent. The PI collaborated with RHJVAMC researchers to acquire a Zeiss Lightsheet microscope and used it to produce a 3D reconstruction of brain neuronal circuits (see Shiromani and Peever, 2017 attached). My intent is to use CLARITY to visualize postmortem brains in rodent models of TBI, and also image a transparent heart, liver and kidney. The goal is to provide a visual record of the break in a circuit in diseased tissue. The fourth series of experiments use the gene transfer approach to fix defective circuits and restore behavior. I have used it to successfully to correct behavioral symptoms in a mouse model of the neurodegenerative sleep disorder, narcolepsy. We are now using the gene transfer method to block triggering of abnormal behavior, for example in fear-conditioning. Overall, these neuroscience methods and tools aid the collective research effort at RHJ VAMC.

7000 万美国人患有某种睡眠障碍。行为、情绪和记忆力恶化 随着睡眠不足，情况会随着持续的睡眠不足而变得更糟。睡眠障碍是一种常见且 我们退伍军人的共同抱怨。过度警觉导致的睡眠不足是 PTSD 的症状之一，但 不知道为什么患有创伤后应激障碍的退伍军人无法入睡。我的研究重点是识别和绘制地图 诱导睡眠的大脑神经元。我的研究的总体影响是它将提供第一个直接的 将神经元的特定表型及其负责诱导睡眠的回路联系起来的证据。这将使 在唤醒驱动力非常强的情况下，例如在失眠的情况下，可以诱导睡眠 PTSD，或在过度困倦时保持清醒，例如患有阻塞性睡眠障碍的患者 睡眠呼吸暂停或非典型抑郁症。 一系列实验利用光遗传学和药物遗传学来识别大脑中的功能回路。 脑。大脑包含许多不同类型的细胞，通过光遗传学和药物遗传学，它是 现在可以分解大脑来识别负责复杂行为的罪魁祸首神经元，例如 作为睡眠。我的实验室是睡眠神经生物学领域第一个利用光遗传学诱导睡眠的实验室 （Konadhode 等人，2013 年，附后）。我们激活了下丘脑神经元的特定表型 发现它可以在一天中本应清醒的时间小鼠入睡。我们现在有 在大鼠中显示出相同的效果，表明激活这些神经元可以促进哺乳动物的睡眠。我们现在 想要测试在高度唤醒的情况下的睡眠诱导效果，例如恐惧调节（PTSD）或焦虑。 结合光遗传学和药物遗传学，我使用深部脑成像方法 对大脑中表型特异性神经元的活动进行成像。该方法测量的变化 单个神经元中基因编码的钙指示剂的荧光。荧光信号为 通过连接到微型显微镜（2g）的显微内窥镜捕获。可放置显微内窥镜 小鼠大脑中的任何地方都提供了前所未有的神经元活动记录。我用它来获取 睡眠和清醒期间特定神经元回路活动的视觉证据。 另一系列研究使用 CLARITY 方法来绘制光遗传学激活的电路图。 CLARITY 是一种新的神经解剖学方法，可以使死后组织（例如大脑）变得透明。 该 PI 与 RHJVAMC 研究人员合作购买了一台蔡司 Lightsheet 显微镜，并用它来 生成大脑神经元回路的 3D 重建（参见 Shiromani 和 Peever，2017 年附件）。我的意图是 使用 CLARITY 可视化 TBI 啮齿动物模型中的死后大脑，并对透明心脏进行成像， 肝脏和肾脏。目标是提供病变组织中电路断裂的视觉记录。 第四系列实验使用基因转移方法修复有缺陷的电路并恢复 行为。我已经用它成功地纠正了小鼠模型的行为症状 神经退行性睡眠障碍，发作性睡病。我们现在使用基因转移方法来阻止触发 异常行为，例如恐惧条件反射。总的来说，这些神经科学方法和工具有助于 RHJ VAMC 的集体研究工作。