Genetic Screen For Oxidative Resistance in Mice

由活性氧（ROS）引起的细胞损伤被认为在许多迟发性疾病如帕金森氏症、阿尔茨海默氏症和癌症中起作用。此外，增强的生物体寿命本身与增加的ROS抗性相关，并且衰老的自由基理论假定，ROS诱导的细胞损伤的积累导致各种器官系统随年龄的下降。对ROS的关键细胞反应的分子理解由于缺乏对该过程的详细遗传解剖而受到影响。此外，很少有动物模型存在，在生物体水平上测试细胞对ROS反应的改变的影响。目前的建议使用筛选的原代成纤维细胞来源于小鼠携带随机ENU诱导的隐性突变。目标是分离导致培养的成纤维细胞中对ROS的抗性增强或降低的单基因突变。这种方法允许对ROS诱导的损伤的细胞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对ROS具有改变的敏感性的动物模型，其可立即用于衰老研究。虽然超出了这个试点赠款的范围，任何可遗传的突变体最终将被定位克隆，以确定所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.