Volatile Anesthetic Regulation of TASK Tandem Pore Potassium Channels

该提案描述了一项为期5年的培训方案，以培养一名独立的调查员。 易挥发的麻醉机制。首席调查员是该系的一名教员 麻醉，哈佛医学院，具有电生理学背景，谁将获得新的 神经科学和药理学方面的知识和技能足以领导他自己的独立研究 程序。工作将在马萨诸塞州的Mallinckrodt药理学研究单位进行 重点研究挥发性麻醉机制的综合医院。； 我们的特定目标将检验挥发性麻醉剂增强TASK-3功能的新假设 间接通过抑制基础或激动剂介导的Galpha-Q蛋白偶联受体(GqPCR)活性。这个 TASK-3串联孔钾通道是一种增强pH和抑制缺氧的挥发性麻醉剂 离子通道。TASK-3在中枢神经系统和外周组织中广泛表达，并具有一种 在各种类型的细胞中，包括那些参与调节呼吸的细胞中，发挥化学感觉的作用。任务-3 功能也被GqPCR激活所抑制。中枢神经系统中的GqPCRs调节记忆、觉醒和 意识和外周是调节肌肉张力的重要因素。许多人的功能 GqPCRs被挥发性麻醉药抑制。为了验证我们的假设，我们将使用双电极电压 非洲爪哇卵母细胞钳或Ussing小室Fischer大鼠甲状腺细胞的异种研究 表达TASK-3通道和m1或m3 GqPCRs和Galpha-Q蛋白。我们还将雇用被切除的 膜片钳方法以及RNA干扰和定点定位等几种分子生物学技术 诱变。长期目标：该项目将阐明挥发性物质 麻醉剂增强了任务通道。与公共卫生的相关性：这项工作将有助于建立 指导开发不影响呼吸的更安全麻醉药的原则并进一步阐明 这些药物诱导麻醉的机制。我

This proposal describes a 5-year training program for the development of an independent investigator in volatile anesthetic mechanisms. The principal investigator is a faculty member in the Department of Anesthesia, Harvard Medical School, with a background in electrophysiology,who will acquire new knowledge and skills in neuroscience and pharmacology sufficient to lead his own independent research program. Work will be conducted in the Mallinckrodt Pharmacology Research Unit at Massachussets General Hospital which focuses on volatile anesthetic mechanisms. ; Our SPECIFIC AIMS wil test the novel hypothesis that volatile anesthetics potentiate TASK-3 function indirectly by inhibition of basal or agonst-mediated Galpha-q protein coupled receptor (GqPCR) activity. The TASK-3 tandem pore potassium channel is a volatile anesthetic-potentiated and pH- and hypoxia-inhibited ion channel. TASK-3 is widely expressed in the central nervous system and in peripheral tissue and has a role in chemosensing in a variety of cell types, including those involved in regulation of breathing. TASK-3 function is also inhibited by GqPCR activation. GqPCRs in the CNS regulate memory, wakefulness, and consciousness and peripherally are important regulators of smooth muscle tone. The function of many GqPCRs are inhibited by volatile anesthetics. To test our hypothesis we will use two-electrode voltage clamp of Xenopus oocytes or Ussing Chamber studies of Fischer Rat Thyroid cells heterologously expressing TASK-3 channels and ml or m3 GqPCRs and Galpha-q protein. We will also employ the excised patch-clamp method, and several molecular biology techniques including RNA interference and site-directed mutagenesis. LONG TERM GOALS: This project will clairfy the molecular mechanism by which volatile anesthetics potentiate TASK channels. RELEVANCE TO PUBLIC HEALTH: This work will help establish principles to guide development of safer anesthetic drugs that do not effect breathing and further elucidate the mechanism by which these drugs induce anesthesia. i