Optogenetic dissection of basal forebrain neurons involved in sleep homeostasis

参与睡眠稳态的基底前脑神经元的光遗传学解剖

• 批准号: 8353608
• 负责人: RADHIKA BASHEER
• 金额: $ 18.57万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8353608
• 关键词: Acetylcholine; Adenosine; Alzheimer' s Disease; Attention; Attenuated; Bilateral; Biological; Brain region; Cells; Choline O-Acetyltransferase; Cognition; Cognitive; Depressive disorder; Dissection; Electroencephalography; Ensure; Evaluation; Frequencies; Glutamates; Goals; Health; Homeostasis; Hour; Human; Impaired cognition; Injection of therapeutic agent; Label; Lesion; Light; Measures; Mediating; Methods; Microdialysis; Mission; Modeling; Monitor; Mus; National Institute of Mental Health; Neurons; Neurotransmitters; Nitric Oxide; Optics; Parkinson Disease; Parvalbumins; Performance; Personal Satisfaction; Polysomnography; Population; Post-Traumatic Stress Disorders; Protocols documentation; REM Sleep; Recovery; Research; Role; Sleep; Sleep Deprivation; Technology; Testing; Time; Transgenic Mice; Viral Vector; Wakefulness; alertness; awake; basal forebrain; cell type; cholinergic; cholinergic neuron; combinatorial; design; extracellular; gamma-Aminobutyric Acid; human NOS2A protein; in vivo; mouse model; neurochemistry; neurotoxic; non rapid eye movement; novel; optogenetics; prevent; recombinase; research study; response; sleep regulation; therapy design; treatment planning

DESCRIPTION (provided by applicant): The biological underpinnings of homeostatic sleep regulation are an important aspect of sleep research given the relevance of sleep deprivation (SD) to human health, well-being, and cognitive performance. More and more people either are forced to, due to vocational demands, or by choice, stay awake for long periods or at biologically non-optimal times of the day. Moreover, sleep loss-associated cognitive impairments are often observed in conditions such as depressive disorders, post-traumatic stress disorder, Alzheimer's and Parkinson's diseases. Central mechanisms that mediate the effects of SD causing attention and cognitive impairment as well as induction of homeostatic sleep response are critical to be understood to design treatment paradigms for alleviate such deleterious effects of sleep loss, and is close to the NIMH mission. One of the brain regions, the basal forebrain (BF), in addition to its important role in promoting wakefulness, is also recognized for its role i homeostatic sleep regulation as well as in attention and cognition. The BF consists of a variety of neurons that utilize acetylcholine or GABA or glutamate. The complexity of the neuronal composition in BF has long prevented a clear understanding of the causal role of each neuronal subtype on extracellular neurochemical alterations and modulation of cortical activity that underlies increased sleepiness and reduced alertness following prolonged neuronal activation during SD. Recent studies demonstrated that both cholinergic and parvalbumin (PARV) expressing GABAergic (PARV-pos GABA) neurons are active during wakefulness and are capable of modulating cortical activation. However, their distinct roles in sleep homeostasis are not clear. While neurotoxic lesions of cholinergic and GABAergic neurons have underlined the importance of these neurons in wakefulness and homeostatic sleep response, a direct cause and effect evaluation is best studied by selective manipulation of each neurotransmitter-specific neuronal cell types. The overall goals of this application is to discern the differences in the functional role of cholinergic and PARV-pos GABAergic neuronal activation in homeostatic sleep regulation. Using the state-of-the-art optogenetic technology to selectively manipulate the activities of cholinergic and PARV-GABA neurons combined with simultaneous polysomnographic recordings to monitor changes in cortical EEG and in vivo microdialysis for measuring extracellular neurochemical changes we will test the following model: Prolonged SD->BF cholinergic neuronal activation->increase extracellular NO and adenosine->inhibition of wake active cholinergic and non-cholinergic neurons->increased sleepiness. Successful completion of these exploratory studies will (1) validate a novel combinatorial method of performing optogenetics with in vivo microdialysis, (2) extend our understanding of the causal role of specific BF neuronal subtypes in modulating cortical EEG and homeostatic sleep response. PUBLIC HEALTH RELEVANCE: Sleep deprivation is a wide spread problem with negative impacts on performance efficiency, health and cognition. The current application proposes to use in vivo microdialysis with the state-of-the-art method of optogenetics to selectively manipulate two major wake-active neuronal cell types to discern their role in modulating the homeostatic sleep factors that increase the propensity to sleep and decrease attention and cognition. Understanding the neuronal components involved in mediating neurochemical changes is important towards designing targeted treatment plans to counteract the effects of sleep deprivation.

描述(由申请人提供)：鉴于睡眠剥夺(SD)与人类健康、福祉和认知表现的相关性，稳态睡眠调节的生物学基础是睡眠研究的一个重要方面。越来越多的人要么出于职业需求，要么被迫长时间保持清醒，或者在一天中生理上非最佳的时间保持清醒。此外，与睡眠不足相关的认知障碍经常出现在抑郁障碍、创伤后应激障碍、阿尔茨海默氏症和帕金森氏症等疾病中。调节SD引起注意力和认知障碍以及诱导稳态睡眠反应的中枢机制对于设计治疗范例以减轻睡眠缺失的有害影响至关重要，并且接近NIMH的任务。作为大脑区域之一的基底前脑(BF)，除了在促进觉醒方面发挥重要作用外，还被认为在自我平衡睡眠调节以及注意力和认知方面发挥着重要作用。BF由利用乙酰胆碱、GABA或谷氨酸的各种神经元组成。BF中神经元成分的复杂性长期以来阻碍了对每种神经元亚型在细胞外神经化学变化和皮质活动调节中的因果作用的清楚了解，这是SD期间神经元长期激活后嗜睡增加和警觉性降低的基础。最近的研究表明，胆碱能和小白蛋白(PARV)表达GABA能神经元(PARV-pos GABA)在觉醒时都是活跃的，并能够调节皮质的激活。然而，它们在睡眠稳态中的不同作用尚不清楚。虽然胆碱能和GABA能神经元的神经毒性损伤强调了这些神经元在觉醒和稳态睡眠反应中的重要性，但直接的因果评估最好通过选择性地处理每种神经递质特定的神经细胞类型来研究。这项应用的总体目标是辨别胆碱能和PARV-pos GABA能神经元激活在稳态睡眠调节中的功能作用的差异。利用最先进的光遗传学技术选择性地操纵胆碱能和PARV-GABA神经元的活动，结合同步多导睡眠图记录来监测皮质脑电的变化和体内微透析测量细胞外神经化学变化的变化，我们将测试下列模型：延长SD-&GT；BF胆碱能神经元激活-&GT；增加细胞外NO和腺苷-&GT；抑制觉醒活跃的胆碱能和非胆碱能神经元-&GT；增加嗜睡。这些探索性研究的成功完成将(1)验证一种利用体内微透析进行光遗传学的新组合方法，(2)扩大我们对特定BF神经元亚型在调节皮质脑电和稳态睡眠反应中的因果作用的理解。 公共健康相关性：睡眠剥夺是一个广泛存在的问题，对工作效率、健康和认知都有负面影响。目前的应用建议使用体内微透析和最先进的光遗传学方法来选择性地操纵两种主要的觉醒活性神经细胞类型，以识别它们在调节动态平衡睡眠因素中的作用，这些因素增加睡眠倾向，降低注意力和认知。了解参与调节神经化学变化的神经元成分对于设计有针对性的治疗计划以抵消睡眠剥夺的影响非常重要。