GENETIC SPECIFICATION AT PHYSIOLOGICAL AGING

我们的目标是：1.(A)建立文化条件，使 大量长寿的遗传变异可以为后续的 生理测试；(B)构建四个适当的遗传库 已经存在的长寿突变株，用于后续的生理 测试；(C)完成对这四个长期寿命的互补分析 突变株；。(D)完成这四个突变株的遗传图谱。 精神错乱。2.(A)描述所报告的变化与几年前的变化 不同的生理活动：(一)食物摄入量；(二)特定 三种溶酶体水解酶的活性，(Iii)脂褐素水平，(Iv) 呼吸率，以及(V)可见形态和发病率；(B)这些 将在(I)野生型，(Ii)衍生的长寿菌株中检测变化 通过选育线虫，以及(Iii)长寿突变株。 3.(A)确定总超氧化物歧化酶、锰超氧化物歧化酶或过氧化氢酶 年轻人的活动与老年人的平均寿命相关 长寿菌株；(B)完全互补和遗传作图 分离出三个抗百草枯(PAR)突变体，试图获得 超氧阴离子自由基代谢酶改变的突变体； (C)在3b的保证下分离更多的PAR突变体；(D)测试可能的模式 PAR突变体的抗性：(I)酶活性增加-通过检测 用超氧化物歧化酶、锰超氧化物歧化酶和过氧化氢酶的PAR突变体确定是否增加 抗药性是由于这些酶的比活性增加所致，(Ii) 减少转运-通过分析PAR突变体的放射性摄取 百草枯以确定摄取是否正常，(Iii)改变降解 百草枯-通过检测对其他产生游离的药物的抗药性 自由基，如铅黄素和(Iv)改变黄递酶专一性-通过 体外黄递酶活性测定。

Our goals are to: 1. (a) Establish culture conditions under which gram quantities of long-lived genetic variants can be maintained for subsequent physiological tests; (b) Construct appropriate genetic stocks of four already existing long-lived mutant strains for subsequent physiological testing; (c) Finish complementation analyses on these four long-lived mutant strains; (d) Complete the genetic mapping of these four mutant loci. 2. (a) Characterize the reported changes with age of several different physiological activitie: (i) rate of food intake, (ii) specific activity of three lysosomal hydrolases, (iii) lipofuscin levels, (iv) respiration rate, and (v) visible morphology and morbidity; (b) These changes will be assayed in (i) wild type, (ii) long-lived strains derived by selective breeding of C. elegans, and (iii) long-lived mutant strains. 3. (a) Determine if total superoxide dismutase (SOD), MnSOD or catalase activities of young adults are correlated with mean life-span of the long-lived strains; (b) Complete complementation and gentic mapping of three paraquat resistant (par) mutants isolated in an attempt to obtain a mutants with alterations in the superoxide radical metabolizing enzymes; (c) Isolate more par mutants if warranted by 3b; (d) Test possible modes of resistance of the par mutants: (i) increased enzyme activity - by assaying the par mutants for SOD, MnSOD, and catalase to determine if the increased resistance is due to increased specific activity of these enzymes, (ii) decreased transport - by assaying the par mutants for uptake of radioactive paraquat to determine if uptake is normal, (iii) altered degradation of paraquat - by assaying for resistance to other drugs that generate free radicals, such as plumbagin and (iv) altered diaphorase specificity - by assaying in vitro diaphorase activity.