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基础是睡眠研究的重要方面。越来越多的人由于职业需求而被迫长期保持清醒，或者在一天中的生物学上不最佳时期。此外，在抑郁症，创伤后应激障碍，阿尔茨海默氏症和帕金森氏病等疾病中，经常观察到与睡眠失眠相关的认知障碍。介导SD影响的中心机制引起注意力和认知障碍以及稳态睡眠反应的诱导对于减轻睡眠损失的这种有害影响的设计治疗范式至关重要，并且接近NIMH任务。大脑区域之一，基础前脑（BF）除了其在促进清醒中的重要作用外，它的作用I I i稳态睡眠调节以及注意力和认知的作用也得到了认可。 BF由使用乙酰胆碱或GABA或谷氨酸的多种神经元组成。 BF中神经元组成的复杂性长期以来一直无法清楚地了解每个神经元亚型在细胞外神经化学改变和皮质活性的调节中的因果作用，这是SD期间延长嗜睡和延长的神经元激活后的警觉性降低的基础。最近的研究表明，表达GABA能（PARV-POS GABA）神经元的胆碱能和白蛋白（PARV）在清醒期间都活跃，并且能够调节皮质激活。但是，它们在睡眠体内平衡中的独特作用尚不清楚。尽管胆碱能和GABA能神经元的神经毒性病变强调了这些神经元在觉醒和稳态睡眠反应中的重要性，但通过选择性操纵每种神经递质特异性神经元细胞类型，可以最好地研究直接原因和效果评估。该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差异。 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腺苷 - >抑制活性胆碱能和非胆碱能神经元 - >增加嗜睡。这些探索性研究的成功完成将（1）验证一种使用体内微透析进行光遗传学的新型组合方法，（2）（2）扩展了我们对调节皮质EEG和稳态睡眠反应的特定BF神经元亚型的因果作用的理解。 公共卫生相关性：睡眠剥夺是一个广泛的传播问题，对绩效效率，健康和认知的负面影响。当前的应用建议使用最先进的光遗传学方法在体内微透析中使用，以选择性地操纵两种主要的唤醒活性神经元细胞类型，以识别其在调节体内稳态睡眠因素中的作用，从而增加睡眠的倾向，从而增加睡眠的倾向并减少注意力和认知。了解介导神经化学变化涉及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很重要。