BLR&D Research Career Scientist Award Application

BLR

• 批准号: 10265393
• 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/10265393
• 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 由于睡眠损失，由于持续的睡眠剥夺而变得更糟。睡眠障碍经常 在我们的退伍军人中普遍投诉。缺乏由于超阳性引起的睡眠是PTSD的一种症状，但这是 不知道为什么具有PTSD的退伍军人无法入睡。我的研究重点是识别和映射 诱导睡眠的脑神经元。我的研究的总体影响是它将提供第一个直接 与神经元的特定表型及其电路有关的证据，导致睡眠。这会做到 在唤醒驱动非常强的情况下，可以诱导睡眠，例如 PTSD，或在过度嗜睡时保持清醒，例如阻塞性患者 睡眠呼吸暂停或非典型抑郁症。 一系列实验使用光遗传学和药物遗传学来识别功能电路 脑。大脑包含许多不同类型的细胞，并且通过光遗传学和药物遗传学是 现在可以拆卸大脑以识别负责复杂行为的罪魁祸首 睡觉。我的实验室是睡眠神经生物学领域的第一个使用光遗传学诱导睡眠的实验室 （Konadhode等人，2013年，附件）。我们激活了下丘脑中神经元的特定表型， 发现它在一天中应该醒着的时候引起睡眠。我们现在有 在大鼠中显示出相同的作用，表明激活这些神经元在跨哺乳动物中驱动睡眠。我们现在 想要在高唤醒状况（例如恐惧条件（PTSD）或焦虑症）的情况下测试诱导睡眠的效果。 与光遗传学和药物遗传学结合使用，我使用了深脑成像方法 图像大脑中表型特异性神经元的活性。此方法衡量变化 单个神经元中遗传编码的钙指标的荧光。荧光信号为 通过连接到微型显微镜（2G）的微型内镜捕获。可以放置微镜 小鼠大脑中的任何地方都提供前所未有的神经元活动记录。我正在用它来获得 在睡眠和醒来过程中特定神经元电路活性的视觉证据。 另一系列研究使用清晰度方法来绘制光遗传学激活的电路。 清晰度是一种新的神经解剖学方法，它使术后组织（例如大脑）透明。 PI与RHJVAMC研究人员合作获取了Zeiss Lightsheet显微镜，并将其用于 产生脑神经元电路的3D重建（请参见Shiromani and Peever，2017年附带）。我的意图是 要使用清晰度可视化TBI啮齿动物模型中的尸体大脑，并形象透明的心脏， 肝脏和肾脏。目的是提供患病组织中电路中断裂的视觉记录。 第四系列实验使用基因转移方法来固定有缺陷的电路并恢复 行为。我已经用它成功地纠正了鼠标模型中的行为症状 神经退行性睡眠障碍，发作性睡病。我们现在正在使用基因转移方法阻止触发 异常行为，例如恐惧条件。总体而言，这些神经科学方法和工具有助于 RHJ VAMC的集体研究工作。