GENETIC SPECIFICATION AT PHYSIOLOGICAL AGING

我们的目标是：1。(a)建立培养条件

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.