ALTERED LUNG GLUTATHIONE INDUCED--INSERTION MUTAGENESIS

谷胱甘肽诱导的肺改变--插入突变

• 批准号: 3356365
• 负责人: Terrance J Kavanagh
• 金额: $ 19.05万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-30 至 1992-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3356365
• 关键词: antioxidants; embryo /fetus tissue /cell culture; endonuclease; flow cytometry; free radicals; genetic manipulation; genetic mapping; genetically modified animals; glutathione; glutathione peroxidase; glutathione reductase; glutathione transferase; hamsters; hematopoietic stem cells; laboratory mouse; ligase; lung disorder; mass tissue /cell culture; molecular cloning; mutagens; neoplastic cell culture for noncancer research; neoplastic transformation; nucleic acid probes; oxidation reduction reaction; plasmids; singlet oxygen; tissue /cell culture; transfection

Toxic oxygen species are important mediators of lung injury. An array of antioxidant defenses have evolved to protect cells against reactive oxygen species. An important component of this defensive system is glutathione. The specific aims in this study are to identify and characterize genetic factors which can influence the intracellular concentration of glutathione in mouse embryonic stem cells, and to generate transgenic mice from these cells. We hypothesize that selected mouse embryonic stems cells with alterations in glutathione content will display either increased or decreased resistance to reactive oxygen metabolites. Transgenic mice formed from such stem cells should also show altered sensitivity to free-radical damage. During the study period we will 1) isolate retrovirus-induced mutants of mouse embryonic stem cells with altered GSH content, by using a fluorescent dye specific for GSH; 2) isolate and characterize the mutated DNA that resulted in altered GSH metabolism; 3) analyse the effects of the altered GSH on cell sensitivity to free-radical damage; and 4) establish colonies of transgenic mice bearing the GSH mutations of interest. Creation of transgenic mice with genetically altered GSH content will be especially useful in future studies of free radical-induced lung injury.

有毒氧是肺损伤的重要介质。 一个 一系列抗氧化防御系统已经进化到保护细胞 对抗活性氧 其中一个重要的组成部分是 防御系统是谷胱甘肽 本研究的具体目的 是识别和描述遗传因素， 影响小鼠细胞内谷胱甘肽浓度 胚胎干细胞，并从这些细胞中产生转基因小鼠 细胞 我们假设选择的小鼠胚胎干细胞 谷胱甘肽含量的改变会显示 对活性氧代谢物的抵抗力增加或降低。 由这些干细胞形成的转基因小鼠也应该表现出 改变了对自由基损伤的敏感性 研究期间 在此期间，我们将1）分离逆转录病毒诱导的小鼠突变体 胚胎干细胞与谷胱甘肽含量改变，通过使用 GSH特异性荧光染料; 2）分离并表征 导致GSH代谢改变的突变DNA; 3）分析 GSH改变对细胞对自由基敏感性的影响 损伤;和4）建立携带 感兴趣的GSH突变。 转基因小鼠的建立 基因改变的GSH含量在未来将特别有用 自由基致肺损伤的研究。