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Analysis of life span by DNA transfection of antioxidant defense genes in oxygen-dependent short-lived mutants.

通过 DNA 转染氧依赖性短寿命突变体抗氧化防御基因来分析寿命。

基本信息

批准号：
05834015
负责人：
ISHII Naoaki
金额：
$ 1.47万
依托单位：
Tokai University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

To investigate the possible role of oxygen free radicals in aging, antioxidant defense genes and mutants of the nematode C.elegans which are hypersensitive to oxygen were isolated. Then we tried to inject these genes into these oxygen-sensitive mutants to know their roles.Results(1) We cloned a Mn-superoxide dismutase (SOD) gene(2) An oxygen-sensitive mutant mev-1 has a short life span even under atmospheric condition and the life span changes depending on concentration of oxygen. This mutant has about half the wild-type level of Cu, Zn-SOD activity. The mev-1 mutant accumulated more fluorescent material at a greater rate than does wild type. Furthermore, the accumulation rates depended on concentration of oxygen. Since this phenotype has been widely used as an aging marker, these results suggested which mev-1 may be a mutant of premature aging.(3) It has already been found a Cu, Zu-SOD (sod-1) on the chromosome II.Recently, another Cu, Zu-SOD (sod-4) has been found on the chromosome III.The mev-1 gene may code the sod-4 itself because these genes are located at the same region on the chromosome III.By RT-PCR method, there were no difference in the mRNA level between the wild type and mev-1 mutant.Microinjection of the sod-4 gene in the mev-1 mutants to know the role of the gene is in progress.(4) Another oxygen-sensitive mutnat mev-3 of which development, fecundity and mean life span was retarded, reduced and shortened, respectively, when animals were incubated under high concentration of oxygen. Molecular cloning of the gene is in progress.

为了研究氧自由基在衰老过程中的作用，分离了对氧敏感的秀丽隐杆线虫的抗氧化防御基因和突变体。然后我们试图将这些基因注入这些氧敏感突变体中，以了解它们的作用。结果(1)克隆了一个锰超氧化物歧化酶（SOD）基因(2)氧敏感突变体mev-1即使在大气条件下也具有较短的寿命，且寿命随氧浓度的变化而变化。该突变体的Cu、Zn-SOD活性约为野生型的一半。mev-1突变体比野生型积累更多荧光物质的速度更快。此外，积累速率与氧浓度有关。由于该表型已被广泛用作衰老标记，这些结果表明mev-1可能是早衰的突变体。(3)在II号染色体上已经发现了Cu， Zu-SOD （sod-1）。最近，在第三染色体上发现了另一个Cu， Zu-SOD （sod-4）。mev-1基因可能编码sod-4基因本身，因为这些基因位于染色体III上的同一区域。通过RT-PCR方法，野生型和mev-1突变体的mRNA水平没有差异。在mev-1突变体中微量注射sod-4基因以了解该基因的作用正在进行中。(4)另一种氧敏感突变体mev-3，在高浓度氧气条件下孵育，其发育迟缓，繁殖力降低，平均寿命缩短。基因的分子克隆正在进行中。