Volatile Anesthetic Regulation of TASK Tandem Pore Potassium Channels

TASK 串联孔钾通道的挥发性麻醉调节

• 批准号: 7530533
• 负责人: Joseph F Cotten
• 金额: $ 13.09万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7530533
• 关键词: Acids; Agonist; Amino Acid Sequence; Anesthesia Department, Hospital; Anesthesia procedures; Anesthetics; Breathing; Bronchodilation; Cells; Chemosensitization; Clinical; Conscious; Coupled; Depressed mood; Development; Electrodes; Electrophysiology (science); Exhibits; Faculty; General Anesthesia; General Hospitals; General anesthetic drugs; Halothane; Hypoxia; Inbred F344 Rats; Ion Channel; Isoflurane; Knowledge; Lead; Mammalian Cell; Mediating; Memory; Methods; Molecular; Molecular Biology Techniques; Muscarinics; Muscle Tonus; Nervous system structure; Neuraxis; Neurosciences; Peripheral; Pharmaceutical Preparations; Pharmacology; Potassium; Potassium Channel; Principal Investigator; Program Development; Proteins; Public Health; RNA Interference; Receptor Activation; Receptor Inhibition; Regulation; Research; Research Personnel; Resistance; Role; Signaling Protein; Site; Site-Directed Mutagenesis; Testing; Thyroid Gland; Tissues; Training Programs; Vasodilation; Wakefulness; Work; Xenopus oocyte; base; cell type; clinically relevant; desflurane; medical schools; member; mutant; novel; patch clamp; programs; receptor coupling; receptor function; reconstitution; response; sevoflurane; skills; voltage clamp

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE 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.

描述（由申请人提供）：本提案描述了一项为期5年的培训计划，旨在培养一名挥发性麻醉机制方面的独立研究者。首席研究员是哈佛医学院麻醉系的一名教员，具有电生理学背景，他将获得神经科学和药理学方面的新知识和技能，足以领导他自己的独立研究项目。工作将在马萨诸塞州总医院的Mallinckrodt药理学研究单位进行，重点是挥发性麻醉机制。我们的SPECIFIC AIMS将测试挥发性麻醉剂通过抑制基础或激动剂介导的galphaq蛋白偶联受体（GqPCR）活性间接增强TASK-3功能的新假设。TASK-3串联孔钾通道是一种挥发性麻醉增强、pH和缺氧抑制的离子通道。TASK-3在中枢神经系统和外周组织中广泛表达，并在多种细胞类型的化学感应中发挥作用，包括那些参与呼吸调节的细胞。TASK-3功能也被GqPCR激活所抑制。中枢神经系统中的gqpcr调节记忆、觉醒和意识，并在周围是平滑肌张力的重要调节因子。许多gqpcr的功能被挥发性麻醉剂抑制。为了验证我们的假设，我们将使用爪蟾卵母细胞的双电极电压钳或Fischer大鼠甲状腺细胞异种表达TASK-3通道和ml或m3 gqpcr和galphaq蛋白的使用室研究。我们还将采用切除膜片钳方法，以及一些分子生物学技术，包括RNA干扰和定点诱变。长期目标：该项目将阐明挥发性麻醉剂增强TASK通道的分子机制。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物的原则，并进一步阐明这些药物诱导麻醉的机制。