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: 10776191
• 负责人: Simon C Johnson
• 金额: $ 22.88万
• 依托单位: NORTHUMBRIA UNIVERSITY
• 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 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10776191
• 关键词: Agitation; Anesthesia procedures; Anesthetics; Animals; Anxiety; Arousal; Astrocytes; Brain; Brain region; Calcium Signaling; Cell Communication; Cell Culture Techniques; Cell model; Cells; Central Nervous System; 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; Knowledge; Link; Location; LoxP-flanked allele; Maintenance; Mammals; Mediating; Medical Care Costs; Metabolic; Metabolism; Microinjections; Mitochondria; Modeling; Molecular; Mus; Neurons; Neurotransmitters; Norepinephrine; Operating Rooms; Patients; Play; Population; Prevalence; Process; Proteins; Recovery; Recycling; Reporting; Research; Risk; Role; Synapses; Viral; Volatilization; Vulnerable Populations; Wakefulness; Work; cell type; experimental study; in vivo; insight; mitochondrial dysfunction; mitochondrial metabolism; mouse model; neural; neural circuit; novel; pediatric patients; pharmacologic; pleiotropism; promoter; 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.

项目摘要 我们的总体目标是定义分子和细胞机制的作用。 挥发性麻醉药（VAS）。我们在这里提出的研究中的目标，这代表了下一步 追求这一目标是机械地定义星形胶质细胞的功能，这些功能是基本的参与 与VAS麻醉的出现过程。我们基于大量的最新和初步假设 数据，星形胶质细胞线粒体电子传输链（ETC）功能支持正常出现 维持星形胶质细胞信号传导，闪光灯闪光灯释放和对神经元的代谢支持。我们 进一步假设中枢神经系统内的特定于空间定义的星形胶质细胞种群 （CNS）通过当地支持出现的神经元突触来推动出现。 我们的实验将利用纯原代星形胶质细胞培养，这是一种细胞特异性的小鼠模型 通过CRE介导的ETC IC蛋白NDUFS4和 星形胶质细胞启动子驱动的CRE聚合酶的立体定向微注射靶向病毒递送以启用 NDUFS4在星形胶质细胞中的大脑区域特异性破坏。这些模型将使我们能够表征 由VAS影响的星形胶质细胞功能和参与出现的星形胶质细胞的种群。具体来说， 它们将允许我们：i）定义VAS对星形胶质细胞钙信号传导和线粒体功能的影响，以及 两者之间的机械联系，体外； ii）表征VA对星形胶质细胞闪光灯的影响 发布; iii）表征VA对支持神经元功能的星形胶质细胞代谢产物的影响； iv）定义 由地区定义的星形胶质细胞种群负责支持VAS麻醉的出现。 最终，这项工作将大大提高我们对VA活动机制和过程的理解 出现。 关联 尽管存在率，但VAS的确切靶标以及其多效效应的基础机制， 在很大程度上不确定。对于成千上万的患者来说，出现是每天发生的，但是出现的过程 理解很差。出现的延迟与焦虑，混乱，抵抗活动，延误有关 手术室，受伤的风险增加以及巨大的医疗费用。如果出现的机制可以 要解决，可能有可能制定积极促进意识恢复的策略。