GENETIC SPECIFICATION AT PHYSIOLOGICAL AGING

生理衰老的遗传规范

• 批准号: 3116462
• 负责人: Thomas E. Johnson
• 金额: $ 14万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-08-01 至 1988-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3116462
• 关键词: aging; catalase; cell age; cellular respiration; cytogenetics; drug resistance; flow cytometry; gene complementation; genetic manipulation; genetic mapping; genetic strain; life cycle; lipofuscin; longevity; lysosomes; morphology; nutrient intake activity; paraquat; physiology; radiotracer; statistics /biometry; superoxide dismutase; thin layer chromatography; tissue /cell culture

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.

我们的目标是：1. (a)建立培养条件， 可以维持大量的长寿遗传变异， （B）构建适当的四个遗传原种， 已经存在的长寿命突变株用于随后的生理 （c）完成对这四种长寿植物的互补分析 （d）完成这四种突变株的遗传图谱绘制 的位点 2. (a)描述所报告的随年龄变化的特征， 不同的生理活动：（i）食物摄入率，（ii）特定 三种溶酶体水解酶的活性，（iii）脂褐素水平，（iv） 呼吸速率，和（v）可见的形态和发病率;（B）这些 将在（i）野生型，（ii）长寿命菌株中测定变化 通过对C.线虫，和（iii）长寿突变株。 3. (a)确定是否存在总超氧化物歧化酶（SOD）、MnSOD或过氧化氢酶 年轻人的活动与他们的平均寿命相关。 （B）完全互补和遗传作图 分离出三种百草枯抗性突变体， 超氧自由基代谢酶发生改变的突变体; (c)如果3b有必要，分离更多的par突变体;（d） PAR突变体的抗性：（i）增加的酶活性-通过测定 对SOD、MnSOD和过氧化氢酶的PAR突变体进行测定，以确定是否增加了 抗性是由于这些酶的比活性增加，（ii） 运输减少-通过测定PAR突变体对放射性的摄取 百草枯，以确定摄取是否正常，（三）百草枯降解的改变， 百草枯-通过测定对其他药物的耐药性， 自由基，如白花丹素和（iv）改变心肌黄酶的特异性-通过 测定体外心肌黄酶活性。