VOLATILE ANESTHETIC ACTIONS--MOLECULAR GENETIC ANALYSIS

麻醉剂用于产生全身麻醉的挥发性、亲脂性化合物 在手术中对所有细胞和组织都有直接影响 仔细检查过。然而，其机制(S)和原点(S) 这些麻醉剂在任何细胞或组织中的作用尚不清楚。长的- 这项工作的射程目标是表征细胞靶标(S) 挥发性麻醉剂及对这些化合物的反应(S)。这些 将在真核生物模型中完成酿酒酵母(酵母)，在 因此有可能使用强大的分子遗传学工具 学习。我们已经证明，挥发性麻醉剂抑制酵母菌生长和 这些化合物作为酵母生长抑制剂的作用是相似的 它们作为诱导麻醉剂的作用。因此，一个前提是 这些研究的基础是对心力衰竭机制的识别(S)。 麻醉药在酵母菌中的作用应该提供对At的基本认识 这些化合物在哺乳动物细胞中的部分作用。 我们的结果表明，氨基酸转运和泛素化 在酵母的麻醉反应中起着关键作用。我们的工作假说 提示泛素化依赖于泛素化的过程 参与细胞渗透对于这种反应是至关重要的。 本申请中工作的具体目标是： 1)麻醉药对泛素代谢影响的表征。 泛素相关的几个基因中的任何一个发生突变 新陈代谢改变酵母对挥发性麻醉剂的反应。 2)麻醉药对营养物质转运影响的表征。 我们实验室的几条证据表明，氨基酸 运输在挥发性麻醉反应中起关键作用 酵母。

The volatile, lipophilic compounds used to produce general anesthesia during surgery have direct effects on all cells and tissues that have been carefully examined. However, the mechanism(s) and primary site(s) of action of these anesthetics in any cell or tissue are unknown. The long- range objective of this work is to characterize cellular target(s) of volatile anesthetics and response(s) elicited by these compounds. These will be done in the model eukaryotes Saccharomyces cerevisiae (yeast), in which it is possible to use powerful molecular genetic tools for these studies. We have shown that volatile anesthetics inhibit yeast growth and that the action of these compounds as yeast growth inhibitors parallels their actions as agents for inducing anesthesia. Thus, a premise underlying these studies is that identification of the mechanism(s) of action of anesthetics in yeast should provide fundamental insight into at least some of the actions of these compounds in mammalian cells. Our results indicate that both amino acid transport and ubiquitination play key roles in anesthetic response of yeast. Our working hypotheses suggest that ubiquitination of a ubiquitination-dependent process involving cellular permeases is critical for this response. The specific aims of the work in this application are: 1)Characterization of effects of anesthetics on ubiquitin metabolism. Mutations in any of several genes that have been implicated in ubiquitin metabolism alter the response of yeast to volatile anesthetics. 2) Characterization of effects of anesthetics on nutrient transport. Several lines of evidence from our laboratory indicate that amino acid transport plays a critical role in the volatile anesthetic response of yeast.