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

项目总结 吸入麻醉药在全身麻醉中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。自首次演示乙醚诱导麻醉以来， 早在160多年前，麻醉理论就试图了解麻醉膜在麻醉中的作用。 行动。本申请寻求将质膜确立为吸入的相关靶标 麻醉剂。脂膜自发地划分成有序和不协调的脂类区域，称为 脂筏。蛋白质的棕榈酰化使蛋白质进入有序相。我们已经证明了 麻醉剂破坏活细胞膜上的这些区域，但破坏与麻醉剂无关 离子通道。 我们将验证麻醉剂破坏脂筏释放脂类修饰酶来激活的假设 钾通过化学信号进行输送。具体地说，GM1漂流隔离磷脂酶D(PLD)， RAFT的破坏释放了PLD，使酶能够找到底物并产生阴离子脂质 磷脂酸(PA)。然后，PA调节两孔结构域钾通道(K2P)Trek-1。崔克- 1也是麻醉敏感通道。我们的目的是研究麻醉药对脂筏的影响。 膜中的结构。在第二个目标中，我们将通过以下方式阐明Trek-1的激活机制 麻醉破坏膜，并在这样做的过程中建立膜作为良好的靶点 麻醉作用。在第三个目标中，我们将开发更好的成像工具和活细胞荧光分析 监测细胞中的浮筏破坏情况。

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.