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: 8494703
• 负责人: RADHIKA BASHEER
• 金额: $ 15.52万
• 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 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8494703
• 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.

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。越来越多的人要么由于职业需求而被迫长时间保持清醒，要么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症、创伤后应激障碍、阿尔茨海默病和帕金森病等疾病中。介导 SD 引起注意力和认知障碍以及诱导稳态睡眠反应的中心机制对于设计治疗范例以减轻睡眠不足的有害影响至关重要，并且接近 NIMH 的使命。基底前脑 (BF) 是大脑区域之一，除了在促进觉醒方面发挥重要作用外，还因其在稳态睡眠调节以及注意力和认知方面的作用而受到认可。 BF 由多种利用乙酰胆碱、GABA 或谷氨酸的神经元组成。 BF 中神经元组成的复杂性长期以来一直阻碍我们清楚地了解每种神经元亚型对细胞外神经化学变化和皮质活动调节的因果作用，而皮质活动是 SD 期间神经元长时间激活后嗜睡增加和警觉性降低的基础。最近的研究表明，表达 GABA 能 (PARV-pos GABA) 的胆碱能神经元和小白蛋白 (PARV) 神经元在清醒期间都很活跃，并且能够调节皮质激活。然而，它们在睡眠稳态中的独特作用尚不清楚。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，但最好通过选择性操作每种神经递质特异性神经元细胞类型来研究直接因果评估。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功能作用的差异。使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图的变化，并使用体内微透析来测量细胞外神经化学变化，我们将测试以下模型：延长SD->BF胆碱能神经元激活->增加细胞外NO和腺苷->抑制唤醒活性胆碱能神经元非胆碱能神经元->嗜睡增加。这些探索性研究的成功完成将（1）验证一种新的光遗传学与体内微透析组合方法，（2）加深我们对特定 BF 神经元亚型在调节皮质脑电图和稳态睡眠反应中因果作用的理解。