Isoflurane: Identification of Key New Targets

异氟烷：关键新靶点的识别

基本信息

批准号：
7816990
负责人：
AARON P. FOX
金额：
$ 41.05万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7816990
关键词：
Affinity Anesthesia procedures Anesthetics Animals Base Pairing Biological Assay Catecholamines Cell Line Cells Chromaffin Cells Data Data Set Depressed mood Dose Drug effect disorder Electric Capacitance Etomidate Exhibits Future General anesthetic drugs Generations Genes Glutamates Goals Hippocampus (Brain)Human Isoflurane Knock-in Mouse Membrane Lipids Membrane Potentials Membrane Proteins Modeling Molecular Monitor Mus Mutant Strains Mice Mutate Mutation Nervous System Physiology Neuraxis Neurons Operative Surgical Procedures PC12 Cells Pain Pharmaceutical Preparations Physiological Play Property Propofol Proteins RNA RNA Interference Rattus Reflex action Research Resistance Role S-nitro-N-acetylpenicillamine Site Structure of superior cervical ganglion Synapses Techniques Testing behavior test clinically relevant clinically significant design experience fluidity insight knock-down neurotransmitter release receptor research study response stable cell line success synaptotagmin I syntaxin 1A vesicle-associated membrane protein

项目摘要

DESCRIPTION (provided by applicant): Despite a great deal of research, a complete understanding of the actions of general anesthetics is still not available. The objective of this research is to advance our understanding of isoflurane, with the hope of gaining insights into all anesthetics. Isoflurane, a halogenated volatile anesthetic, is thought to produce anesthesia by depressing central nervous system function. Many anesthetics, including isoflurane, are thought to modulate and/ or directly activate GABAA receptors. Isoflurane is also known to have effects on other channels and receptors. In a set of preliminary studies we observed that isoflurane, at clinically relevant concentrations, dramatically inhibited the neurotransmitter release machinery in PC12 cells. Etomidate and propofol also inhibited the release machinery in PC12 cells, suggesting that inhibition of neurotransmitter release may be an important general action of anesthetics. Because neurotransmitter release mechanisms are strongly conserved between PC12 and neurons we hypothesize that anesthetics interfere with the neuronal neurotransmitter release machinery and this may represent an important site of action for anesthesia in intact animals. The experiments outlined in this application will be carried out in cultured hippocampal neurons, PC12 cells and mutant mice. This proposal has three goals. First, we will determine whether isoflurane interferes with neurotransmitter release in hippocampal neurons at clinically relevant concentrations. Next, the anesthetic site of action will be identified. We start by examining syntaxin 1A and unc-13, but other sites including SNAP- 25, synaptobrevin and synaptotagmin 1, will be investigated as well. Details about why these sites were selected are provided in this application. Once neurotransmitter release machinery targets are identified they will be mutated in order to suppress the interaction between isoflurane and the target. This information will be used to design knock-in mice with the same mutations, which will permit us to determine, using a battery of physiological and behavioral tests, whether these animals still respond to isoflurane (or other anesthetics) or whether their responses to the anesthetics are altered.Project Narrative: Isoflurane is a widely used volatile anesthetic. Our goal is to find out how isoflurane produces anesthesia. In particular, we will explore the effect on the neurotransmitter release machinery, as our preliminary data suggests this may be a critical site of action for isoflurane and other anesthetics. This may allow for the design of new more beneficial drugs in the future.

描述（由申请人提供）：尽管进行了大量研究，但仍然无法完全了解全身麻醉药的作用。这项研究的目的是增进我们对异氟烷的了解，希望深入了解所有麻醉剂。异氟醚是一种卤化挥发性麻醉剂，被认为通过抑制中枢神经系统功能产生麻醉作用。许多麻醉剂，包括异氟烷，被认为可以调节和/或直接激活 GABAA 受体。众所周知，异氟醚对其他通道和受体也有影响。在一组初步研究中，我们观察到临床相关浓度的异氟烷可显着抑制 PC12 细胞中的神经递质释放机制。依托咪酯和丙泊酚还抑制 PC12 细胞中的释放机制，表明抑制神经递质释放可能是麻醉药的重要一般作用。由于 PC12 和神经元之间的神经递质释放机制高度保守，我们推测麻醉剂会干扰神经元的神经递质释放机制，这可能代表完整动物麻醉的重要作用位点。本申请中概述的实验将在培养的海马神经元、PC12 细胞和突变小鼠中进行。该提案有三个目标。首先，我们将确定临床相关浓度的异氟烷是否会干扰海马神经元的神经递质释放。接下来，将确定麻醉作用部位。我们首先检查 Syntaxin 1A 和 unc-13，但也将研究其他位点，包括 SNAP-25、synaptobrevin 和 synaptotagmin 1。本申请中提供了有关选择这些站点的详细信息。一旦确定了神经递质释放机制目标，它们就会发生突变，以抑制异氟烷和目标之间的相互作用。这些信息将用于设计具有相同突变的敲入小鼠，这将使我们能够使用一系列生理和行为测试来确定这些动物是否仍然对异氟烷（或其他麻醉剂）有反应，或者它们是否对异氟烷有反应。麻醉剂被改变。项目叙述：异氟烷是一种广泛使用的挥发性麻醉剂。我们的目标是找出异氟烷如何产生麻醉作用。特别是，我们将探讨对神经递质释放机制的影响，因为我们的初步数据表明这可能是异氟烷和其他麻醉剂的关键作用部位。这可能有助于将来设计出更有益的新药物。