Genetic Screen For Oxidative Resistance in Mice

活性氧簇(ROS)引起的细胞损伤被认为在许多晚发型疾病中发挥作用，如帕金森氏症、阿尔茨海默氏症和癌症。此外，随着年龄的增长，机体寿命的提高与对ROS的抵抗力增加有关，而衰老的自由基理论假设是ROS引起的细胞损伤的积累导致了各种器官系统的衰退。由于缺乏对ROS过程的详细遗传剖析，对ROS的关键细胞反应的分子理解一直受到影响。此外，几乎没有动物模型可以在生物体水平上测试细胞对ROS反应的改变的影响。目前的建议是对携带随机ENU诱导隐性突变的小鼠的原代成纤维细胞进行筛选。其目标是分离导致培养的成纤维细胞对ROS抗性增强或降低的单基因突变。这种方法允许对细胞对ROS诱导的损伤的反应进行无偏见的遗传解剖，并提供至少在一种细胞类型中改变了对ROS的敏感性的动物模型，这些模型立即可用于衰老研究。尽管超出了这项试点拨款的范围，但任何可遗传的突变最终都将被定位克隆，以确定观察到的表型的分子决定因素。

Cellular damage caused by reactive oxygen species (ROS) is thought to play a role in numerous late onset diseases such as Parkinson's, Alzheimer's and cancer. In addition, enhanced organismal longevity itself has been associated with increased resistance to ROS and the free radical theory of aging postulates that it is the accumulation of ROS induced cellular damage that leads to the decline in various organ systems with age. A molecular understanding of the critical cellular responses to ROS has suffered from lack of a detailed genetic dissection of the process. Moreover, few animal models exist in which to test the impact of alteration in the cellular response to ROS at an organismal level. The current proposal uses screening in primary fibroblasts derived from mice carrying random ENU induced recessive mutations. The goal is to isolate single gene mutations that lead to either enhance or reduced resistance to ROS in the cultured fibroblasts. This approach allows for an unbiased genetic dissection of the cellular response to ROS induced damage and provides animal models with altered sensitivity to ROS, at least in one cell type, that are immediately available for aging studies. Although beyond the scope of this pilot grant, any heritable mutants will ultimately be positionally cloned to identify the molecular determinants of the observed phenotype.