DETECTING DNA DAMAGE--INDIVIDUAL GERM CELL

最近开发的一种微凝胶碱性电泳技术提供了 一种非常灵敏的评估DNA损伤存在的方法 (单链断裂和/或碱不稳定部位)。 因为不需要切除修复过程来演示 DNA损伤的存在，因为样本量可能非常小(1 单细胞凝胶(SCG)技术应优于其他技术 目前用于评估生殖细胞DNA损伤的方法。此外， 由于数据是在单个单元的级别上收集的，因此对 细胞间的损伤差异是可以传导的，使其 比基于混合细胞样本的生化技术更有信息量。这个 拟议项目的目的是评估该项目的适宜性 染毒雄性小鼠细胞DNA损伤的检测技术 可能是诱变剂。该项目的重点将是各种方法 用于从睾丸中分离生殖细胞，并研究了 经选择的诱变剂(丙烯酰胺、 顺铂、环磷酰胺和甲磺酸甲酯)。

A recently developed microgel alkaline electrophoresis technique provides an extremely sensitive method for evaluating the presence of DNA damage (single strand breaks and/or alkali-labile sites) in individual cells. Because excision repair processes are not required to demonstrate the presence of DNA lesions and because sample sizes can be extremely small (1 to <105), the single cell gel (SCG) technique should be superior to other methods currently used to evaluate DNA damage in germ cells. Furthermore, since data are collected at the level of the single cell, an assessment of intercellular differences in damage can be conducted, making it more informative than biochemical techniques based on pooled cell samples. The purpose of the proposed project is to evaluate the suitability of this technique for detecting DNA damage in the cells of male mice exposed to possible mutagens. The focus of the project will be on the various methods used to isolate germ cells from the testis, and the kinetics of the induction of DNA damage induced by selected mutagenic agents (acrylamide, cis-platinum, cyclophosphamide, and methylmethansulphonate).