ALTERED LUNG GLUTATHIONE INDUCED--INSERTION MUTAGENESIS

有毒氧是肺损伤的重要介质。一个 一系列的抗氧化剂防御已经进化到保护细胞 对抗活性氧物种。这其中的一个重要组成部分 防御系统是谷胱甘肽。这项研究的具体目的 就是识别和表征可以 对小鼠细胞内谷胱甘肽浓度的影响 胚胎干细胞，并用这些细胞产生转基因小鼠 细胞。我们假设选择了小鼠胚胎干细胞 随着谷胱甘肽含量的变化，将显示 增加或减少对活性氧代谢物的抵抗力。 由这种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。在研究期间 期间我们将1)分离逆转录病毒诱导的小鼠突变体 GSH含量改变的胚胎干细胞，通过使用 GSH特异性荧光染料；2)分离鉴定 导致谷胱甘肽代谢改变的突变DNA；3)分析 谷胱甘肽变化对细胞对自由基敏感性的影响 以及4)建立携带该基因的转基因小鼠群体 感兴趣的GSH突变。转基因小鼠的建立 转基因的谷胱甘肽含量在未来将特别有用 自由基致肺损伤的研究进展。

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.