Genes Underlying the Response to Inhaled Anesthetics

吸入麻醉剂反应背后的基因

• 批准号: 6922900
• 负责人: JAMES Michael SONNER
• 金额: $ 38.11万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-15 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6922900
• 关键词: RNA; Xenopus; Xenopus oocyte; electrophysiology; gene environment interaction; gene expression; genetic mapping; genetic models; genetically modified animals; inhalation anesthesia; laboratory mouse; membrane channels; model design /development; polymerase chain reaction; protein structure function; single nucleotide polymorphism; spinal cord; transfection

DESCRIPTION (provided by applicant): Although volatile anesthetics have been used in surgical procedures for over 150 years, their mechanism of action remains an enigma. This research proposal aims to identify the mechanism of action of prototypic volatile anesthetics using a murine genetic model. The proposal details an approach not previously applied to investigations of mechanisms of volatile anesthetic action. Two methods identified a region on mouse chromosome 7 which influences volatile anesthetic response 1) a computational method correlating the phenotypic response to volatile anesthetics among 13 inbred mouse strains with the pattern of single nucleotide polymorphisms (SNPs) among these strains; and 2) analysis of backcross progeny derived from 2 parental strains with different anesthetic (isoflurane) responses. Eight candidate genes coding for ion channels expressed in the spinal cord, which is the neural substrate of volatile anesthetic-induced immobility, were identified within the experimentally and computationally identified region on mouse chromosome 7. This proposal has three specific aims. The first aim characterizes the 8 candidate ion channels within the identified region on chromosome 7. Ion channels isolated from two mouse strains exhibiting high and low responses to anesthetic agents will be evaluated for qualitative differences in electrophysiologic experiments on expressed channels in vitro, and for quantitative differences by evaluation of RNA expression in the spinal cord in vivo. The goal is to identify ion channels that display strain differences at a molecular level that plausibly account for the difference in anesthetic response between strains. The second aim identifies other genomic intervals that contain genes influencing anesthetic potency. Because response to volatile anesthetics is not inherited in simple Mendelian fashion, in addition to the region identified on chromosome 7, other genomic intervals must influence this response. Backcross progeny from high and low anesthetic response strains will be generated and evaluated using a SNP-based genome scan to identify these regions. The third aim produces congenic mice in which the identified chromosome 7 segment from mice with high responses to anesthetic is transferred onto the genetic background of the low responder strain, in order to quantitatively assess the effect of this region (and hence the genes in this region) on anesthetic potency.

描述（由申请人提供）： 尽管挥发性麻醉剂在外科手术中的使用已有 150 多年的历史，但其作用机制仍然是个谜。该研究计划旨在利用小鼠遗传模型确定原型挥发性麻醉剂的作用机制。该提案详细介绍了一种以前未应用于挥发性麻醉作用机制研究的方法。有两种方法确定了小鼠 7 号染色体上影响挥发性麻醉剂反应的区域：1) 一种计算方法，将 13 种近交系小鼠品系对挥发性麻醉剂的表型反应与这些品系中的单核苷酸多态性 (SNP) 模式相关联； 2) 对源自具有不同麻醉（异氟烷）反应的 2 个亲本品系的回交后代的分析。在小鼠 7 号染色体上通过实验和计算确定的区域内，鉴定出了八个编码在脊髓中表达的离子通道的候选基因，脊髓是挥发性麻醉剂引起的不动的神经基质。该提议有三个具体目标。第一个目标是表征 7 号染色体上已识别区域内的 8 个候选离子通道。从对麻醉剂表现出高反应和低反应的两种小鼠品系中分离出的离子通道将在体外电生理实验中评估表达通道的定性差异，并进行定量分析。通过评估体内脊髓中RNA表达的差异。目标是识别在分子水平上显示应变差异的离子通道，这似乎可以解释菌株之间麻醉反应的差异。第二个目标是确定包含影响麻醉效力的基因的其他基因组区间。由于对挥发性麻醉剂的反应不是以简单的孟德尔方式遗传的，因此除了 7 号染色体上确定的区域外，其他基因组区间也必定会影响这种反应。将生成高麻醉反应菌株和低麻醉反应菌株的回交后代，并使用基于 SNP 的基因组扫描进行评估，以识别这些区域。第三个目标是生产同类小鼠，其中从对麻醉剂有高反应的小鼠中鉴定出的 7 号染色体片段被转移到低反应品系的遗传背景上，以便定量评估该区域的影响（以及该区域的基因） ）关于麻醉效力。