The Roles of Genetically Distinct Cortical Neuron Types in General-Anesthesia- and Sleep-Induced Slow Waves

遗传上不同的皮质神经元类型在全身麻醉和睡眠引起的慢波中的作用

• 批准号: 10601096
• 负责人: Eric D Melonakos
• 金额: $ 12.5万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-11 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10601096
• 关键词: Adrenergic Agonists; Adrenergic Receptor; Affect; Agonist; Anesthesia procedures; Anesthetics; Area; Arousal; Awareness; Bilateral; Biomimetics; Boston; Brain Stem; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium; California; Cerebral cortex; Collaborations; Community Hospitals; Conscious; Data; Dexmedetomidine; Disinhibition; Educational process of instructing; Electroencephalography; Exhibits; Faculty; Frequencies; Future; General Anesthesia; General Hospitals; General anesthetic drugs; Generations; Goals; Grant; Head; Health; Image; Individual; Interneurons; Intracellular Membranes; Ketamine; Learning; Los Angeles; Massachusetts; Measures; Medical; Membrane Potentials; Mentors; Methods; Microscope; Molecular; Molecular Target; Monitor; N-Methylaspartate; Narcolepsy; Neurons; Occupations; Parvalbumins; Pattern; Peer Review; Persons; Phase; Prefrontal Cortex; Preoptic Areas; Propofol; Protocols documentation; Rattus; Research; Rodent; Role; Sleep; Sleep Disorders; Sleeplessness; Slow-Wave Sleep; Somatostatin; Statistical Data Interpretation; Techniques; Technology; Testing; Training; Unconscious State; Universities; Vasoactive Intestinal Peptide; Writing; antagonist; calcium indicator; computational neuroscience; cortex mapping; experimental study; hippocampal pyramidal neuron; improved; information processing; inhibitory neuron; medical schools; member; neural; neural circuit; neural stimulation; non rapid eye movement; optogenetics; parabrachial nucleus; patient safety; prevent; programs; side effect

Project Summary/Abstract. Reversible loss of consciousness is a crucial part of two major medical fields: general anesthesia and sleep. General anesthetics and non-rapid-eye-movement (NREM) sleep both induce slow waves (0.1-4 Hz) in the cortical electroencephalogram (EEG). It is unknown whether slow waves generated with different anesthetic agents and during NREM sleep are generated with the same neural circuit activity. Dr. Melonakos’ preliminary data suggests that anesthetic agents with different molecular targets have distinct slow wave mechanisms (Aim 1 Hypothesis). In addition, although dexmedetomidine anesthesia shares neural circuits with NREM sleep, it may also have distinct direct cortical effects, possibly leading to different slow wave activity (Aim 2 Hypothesis). The purpose of this research is to test these hypotheses by mapping cortical neural activity with respect to the EEG slow waves of both anesthesia and NREM sleep. In order to do this, Dr. Melonakos will learn how to perform calcium imaging experiments in freely behaving rodents. He will then record calcium images from Ca2+/calmodulin-dependent protein kinase IIa-positive (CaMKIIa+), parvalbumin-positive (PV+), somatostatin-positive (SST+), and vasoactive intestinal peptide-positive (VIP+) cortical neurons during anesthesia- and sleep-induced slow waves. Propofol, ketamine, and dexmedetomidine anesthesia will be tested. Dr. Melonakos will then compare the neural activity between the anesthetics and between general anesthesia and sleep. Finally, he will identify the role of SST+ neurons in slow waves (Aim 3 Hypothesis) by (1) looking at the activity of cortical neurons following disruption of slow waves by stimulation of the parabrachial nucleus, an arousal area in the brainstem, and (2) inhibiting SST+ neurons during anesthesia- and sleep-induced slow waves. During the K99 phase of this project, Dr. Melonakos will be mentored by Drs. Christa Nehs and Emery Brown, experts in anesthesia and sleep neurocircuitry and faculty at Harvard Medical School, Massachusetts General Hospital (MGH), and Massachusetts Institute of Technology (MIT). Dr. Melonakos will also collaborate with Drs. Michael Hasselmo (Boston University), Nancy Kopell (Boston University), and Daniel Aharoni (University of California, Los Angeles). He will be trained in calcium imaging by Drs. Hasselmo and Aharoni, and statistical analysis by Dr. Brown. Dr. Kopell will guide Dr. Melonakos as he orients his findings within hypothesized slow wave mechanisms from the field of computational neuroscience. Dr. Melonakos will also learn optogenetics stimulation techniques from Dr. Nehs and in a course at MIT. The mentors, collaborators, and other members of the MGH community will also provide him with professional guidance as he nears independence, including training in grant writing, peer review, teaching, and the faculty job search. The scientific and professional training Dr. Melonakos receives will enable him to develop an independent research program to study anesthetics’ direct vs. indirect effects. The resulting understanding of slow wave mechanisms has potential to improve the protocols used to monitor general anesthesia and treat sleep disorders, thus benefiting patient safety and health.

项目摘要/摘要。可逆的意识丧失是两个主要医学领域的重要组成部分： 全身麻醉和睡眠。全身麻醉和非快速眼动 (NREM) 睡眠都会引起。 皮质脑电图（EEG）中是否有慢波（0.1-4 Hz）尚不清楚。 不同的麻醉剂和 NREM 睡眠期间的神经回路活动是相同的。 Melonakos 的初步数据表明，具有不同分子靶点的麻醉剂具有明显的缓慢作用。 波机制（目标 1 假设）此外，尽管右美托咪定麻醉共享神经回路。 对于 NREM 睡眠，它也可能具有明显的直接皮质效应，可能导致不同的慢波活动 （目标 2 假设）本研究的目的是通过绘制皮质神经活动来检验这些假设。 为了做到这一点，Melonakos 博士将研究麻醉和 NREM 睡眠的脑电图慢波。 学习如何在自由行为的啮齿动物中进行钙成像实验，然后他将记录钙图像。 来自 Ca2+/钙调蛋白依赖性蛋白激酶 IIa 阳性 (CaMKIIa+)、小白蛋白阳性 (PV+)、 生长抑素阳性 (SST+) 和血管活性肠肽阳性 (VIP+) 皮质神经元 将测试麻醉和睡眠诱导的慢波麻醉。 Melonakos 博士随后将比较麻醉剂和全身麻醉之间的神经活动 最后，他将通过 (1) 查看来确定 SST+ 神经元在慢波中的作用（目标 3 假设）。 通过刺激臂旁核破坏慢波后皮质神经元的活动， (2) 在麻醉和睡眠引起的慢波期间抑制 SST+ 神经元。 在该项目的 K99 阶段，Melonakos 博士将得到 Christa Nehs 和 Emery Brown 博士的指导， 麻醉和睡眠神经回路专家以及哈佛医学院马萨诸塞州综合医院的教员 Melonakos 博士也将与医院 (MGH) 和麻省理工学院 (MIT) 合作。 Michael Hasselmo（波士顿大学）、Nancy Kopell（波士顿大学）和 Daniel Aharoni（波士顿大学） 他将接受 Hasselmo 和 Aharoni 博士的钙成像和统计学培训。 布朗博士的分析将指导 Melonakos 博士在捕捉到的缓慢过程中确定他的发现。 Melonakos 博士还将学习计算神经科学领域的波机制。 Nehs 博士和麻省理工学院的导师、合作者和其他成员的刺激技术。 当他接近独立时，麻省总医院社区还将为他提供专业指导，包括 资助写作、同行评审、教学和教师求职方面的培训 科学和专业的培训。 Melonakos 博士获得的资助将使他能够开发一个独立的研究项目来研究麻醉剂的直接作用 对慢波机制的理解有可能改进协议。 用于监测全身麻醉和治疗睡眠障碍，从而有利于患者的安全和健康。