The role of lipid raft disruption in the activation of TREK-1 channels by anesthetics

脂筏破坏在麻醉剂激活 TREK-1 通道中的作用

• 批准号: 10158518
• 负责人: Scott B Hansen
• 金额: $ 43.9万
• 依托单位: SCRIPPS FLORIDA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2022-04-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10158518
• 关键词: Absence of pain sensation; Anesthesia procedures; Anesthetics; Binding; Binding Sites; Biological Assay; Biological Process; Catalysis; Cell membrane; Cells; Chemicals; Cholesterol; Dose; Electrophysiology (science); Encapsulated; Enzymes; Ethers; Exhibits; Exposure to; General Anesthesia; General anesthetic drugs; Health; Homologous Gene; Human; Hydrophobicity; Image; Imaging Device; Inhalation Anesthetics; Ion Channel; Link; Lipids; Luciferases; Membrane; Membrane Lipids; Membrane Microdomains; Modeling; Modification; Molecular; Monitor; Mutagenesis; Nerve; Neurosciences; Oranges; Pain; Pain management; Palmitic Acylation Site; Pathway interactions; Phase; Phosphatidic Acid; Phosphatidylinositol 4,5-Diphosphate; Phospholipase D; Potassium Channel; Production; Proteins; Reporter; Resolution; Role; Signal Transduction; Structure; Testing; Therapeutic; United States; Work; chronic pain; effective therapy; enantiomer; lipid disorder; novel; palmitoylation; phospholipase D2; potassium channel protein TREK-1; protein biomarkers; response; sensor; theories

PROJECT SUMMARY The molecular mechanism of inhaled anesthetics in general anesthesia remains an important unanswered question in neuroscience and human health. Since the first demonstration of ether-induced anesthesia more than 160 years ago, theories of anesthesia have sought to understand the role of the membrane in anesthetic action. This application seeks to establishes the plasma membrane as a relevant target for inhaled anesthetics. Lipid membranes spontaneously partition into regions of ordered and discorded lipids known as lipid rafts. Palmitoylation of proteins drives the proteins into the ordered phase. We have shown that anesthetics disrupt these domains in live cell membranes, but disruption has not been linked to anesthetic ion channels. We will test the hypothesis that anesthetics disruption of lipid rafts releases lipid modifying enzymes to activate potassium channels through a chemical signal. Specifically, GM1 rafts sequester phospholipase D (PLD), disruption of the raft releases PLD allowing the enzyme to find its substrate and generate anionic lipid phosphatidic acid (PA). The PA then regulates the two-pore domain potassium channel (K2P) TREK-1. TREK- 1 is also an anesthetic sensitive channel. We aim to characterize the effects of anesthetics on lipid raft structure in the membrane. In a second aim, we will elucidate the mechanism of TREK-1 activation through anesthetic disruption of the membrane and in so doing establish the membrane as a bon a fide target of anesthetic action. In a third aim we will develop better imaging tools and a live cell fluorescent assay for monitoring raft disruption in cells.

项目概要 全身麻醉中吸入麻醉药的分子机制仍然是一个重要的未解之谜 神经科学和人类健康的问题。自从首次演示乙醚诱导麻醉以来 160 多年前，麻醉理论就试图了解膜在麻醉中的作用 行动。该应用旨在将质膜确立为吸入性污染物的相关目标。 麻醉剂。脂质膜自发地分成有序和不协调的脂质区域，称为 脂筏。蛋白质的棕榈酰化驱动蛋白质进入有序相。我们已经证明 麻醉剂会破坏活细胞膜中的这些区域，但破坏与麻醉无关 离子通道。 我们将检验以下假设：麻醉剂破坏脂筏会释放脂质修饰酶以激活 钾通道通过化学信号。具体来说，GM1 筏隔离磷脂酶 D (PLD)， 筏的破坏释放 PLD，使酶能够找到其底物并产生阴离子脂质 磷脂酸（PA）。 PA 然后调节双孔域钾通道 (K2P) TREK-1。跋涉- 1也是麻醉敏感通道。我们的目标是表征麻醉剂对脂筏的影响 膜中的结构。第二个目标是，我们将通过以下方式阐明 TREK-1 激活机制： 对膜进行麻醉性破坏，从而使膜成为真正的目标 麻醉作用。第三个目标是，我们将开发更好的成像工具和活细胞荧光测定法 监测细胞中筏的破坏。