The role of astrocytes in emergence from volatile anesthetics

星形胶质细胞在挥发性麻醉剂苏醒中的作用

• 批准号: 10340339
• 负责人: Simon C Johnson
• 金额: $ 42.33万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10340339
• 关键词: Address; Agitation; Anesthesia procedures; Anesthetics; Animals; Anxiety; Arousal; Astrocytes; Brain; Brain region; Calcium Signaling; Cell Communication; Cell Culture Techniques; Cell model; Cells; Complex; Confusion; Conscious; Dangerousness; Data; Delirium; Development; Dose; Elderly; Electron Transport; Enterobacteria phage P1 Cre recombinase; Excision; Exposure to; Functional disorder; General Anesthesia; Glutamates; Goals; Hypersensitivity; Impairment; In Vitro; Injury; Intervention; Knockout Mice; Knowledge; Link; Location; LoxP-flanked allele; Maintenance; Mammals; Mediating; Medical Care Costs; Metabolic; Metabolism; Microinjections; Mitochondria; Modeling; Molecular; Mus; Neuraxis; Neurons; Neurotransmitters; Norepinephrine; Operating Rooms; Patients; Pharmacology; Play; Population; Prevalence; Process; Proteins; Recovery; Recycling; Reporting; Research; Risk; Role; Synapses; Viral; Vulnerable Populations; Wakefulness; Work; base; cell type; experimental study; in vivo; insight; mitochondrial dysfunction; mitochondrial metabolism; mouse model; neural circuit; novel; pediatric patients; pleiotropism; promoter; relating to nervous system; response

Project Summary Our overarching goal is to define the molecular and cellular mechanisms underlying the actions of volatile anesthetics (VAs). Our objective in the studies proposed here, which represent the next step in pursuing this goal, is to mechanistically define the functions of astrocytes that underly their involvement in the process of emergence from anesthesia with VAs. We hypothesize, based on substantial recent and preliminary data, astrocyte mitochondrial electron transport chain (ETC) function supports normal emergence through maintenance of astrocyte calcium signaling, gliotransmitter release, and metabolic support to neurons. We further hypothesize that specific, spatially defined, populations of astrocytes within the central nervous system (CNS) drive emergence through their local support of neuronal synapses involved in emergence. Our experiments will take advantage of pure primary astrocyte cell culture, a mouse model for cell-specific disruption of mitochondrial function through cre-mediated excision of the ETC complex I protein Ndufs4, and stereotactic microinjection targeted viral delivery of an astrocyte promoter driven cre-recombinase to enable brain region-specific disruption of Ndufs4 specifically in astrocytes. These models will allow us to characterize the astrocyte functions impacted by VAs and the population of astrocytes involved in emergence. Specifically, they will allow us to: i) define the effects of VAs on astrocyte calcium signaling and mitochondrial function, and the mechanistic links between the two, in vitro; ii) characterize the effects of VAs on astrocyte gliotransmitter release; iii) characterize the impact of VAs on astrocyte metabolites which support neuronal function; iv) define the astrocyte population, defined by region, responsible for supporting emergence from anesthesia with VAs. Ultimately, this work will greatly advance our understanding of the mechanisms of VA activity and the process of emergence. Relevance Despite their prevalence, the precise targets of VAs, and the mechanisms underlying their pleiotropic effects, are largely undefined. Emergence is a daily occurrence for thousands of patients, but the process of emergence is poorly understood. Delays in emergence are associated with anxiety, confusion, resistive activity, delays in the operating room, increased risks of injury, and significant medical costs. If the mechanisms of emergence can be resolved, it may be possible to develop strategies for active promotion of recovery of consciousness.

项目概要 我们的首要目标是定义潜在的分子和细胞机制 挥发性麻醉剂（VA）。我们在此提出的研究目标代表了下一步 为了实现这一目标，我们需要机械地定义星形胶质细胞的功能，这些功能是其参与 VA 麻醉苏醒的过程。我们假设，基于最近的大量和初步的 数据显示，星形胶质细胞线粒体电子传递链（ETC）功能通过以下方式支持正常出现： 维持星形胶质细胞钙信号传导、神经胶质递质释放以及对神经元的代谢支持。我们 进一步假设中枢神经系统内特定的、空间定义的星形胶质细胞群 （中枢神经系统）通过对参与涌现的神经元突触的局部支持来驱动涌现。 我们的实验将利用纯原代星形胶质细胞培养物，这是一种用于细胞特异性的小鼠模型 通过 cre 介导的 ETC 复合物 I 蛋白 Ndufs4 的切除破坏线粒体功能，以及 立体定向显微注射靶向病毒递送星形胶质细胞启动子驱动的 cre 重组酶，以实现 Ndufs4 的大脑区域特异性破坏，特别是在星形胶质细胞中。这些模型将使我们能够描述 受 VA 影响的星形胶质细胞功能以及参与出现的星形胶质细胞群。具体来说， 它们将使我们能够：i) 定义 VA 对星形胶质细胞钙信号传导和线粒体功能的影响，以及 两者之间的体外机械联系； ii) 表征 VA 对星形胶质细胞胶质递质的影响 发布; iii) 表征 VA 对支持神经元功能的星形胶质细胞代谢物的影响； iv) 定义 按区域定义的星形胶质细胞群，负责支持从 VA 麻醉中苏醒。 最终，这项工作将极大地增进我们对 VA 活性机制和过程的理解。 的出现。 关联 尽管VAs很普遍，但VAs的精确目标及其多效性作用的机制， 很大程度上是未定义的。苏醒对于成千上万的患者来说是家常便饭，但苏醒的过程 人们对此知之甚少。苏醒延迟与焦虑、混乱、抵抗活动、反应迟缓有关。 手术室、受伤风险增加以及巨额医疗费用。如果涌现机制能够 如果解决了，就有可能制定积极促进意识恢复的策略。