Article

A novel assay for replicative lifespan in Saccharomyces cerevisiae

Department of Cell Biology, University of Salzburg, Hellbrunnerstrasse 34, 5020 Salzburg, Austria.
FEMS Yeast Research (Impact Factor: 2.44). 12/2004; 5(2):169-77. DOI: 10.1016/j.femsyr.2004.06.015
Source: PubMed

ABSTRACT The replicative lifespan of Saccharomyces cerevisiae is determined by both genetic and environmental factors. Many of the same factors determine the lifespan of metazoan animals. The lack of fast and reliable lifespan assays has limited the pace of yeast aging research. In this study we describe a novel strategy for assaying replicative lifespan in yeast, and apply it in a screening of mutants that are resistant to pro-oxidants. The assay reproduces the lifespan-shortening effects of deleting SIR2 and of growth in the presence of paraquat, a pro-oxidant. The lifespan-increasing activity of resveratrol is also reproduced. Compared to current assays, this new strategy promises to significantly increase the possible number of replicative-lifespan determinations.

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    • "It has been shown to be a potent antioxidant, anti-inflammatory, anticancer, and chemoprotective agent. It is reported that the possible mechanisms for its various pharmacological activities involve modulating lipid metabolism, platelet aggregation, and inflammatory response [5] [6] [7] [8] [9]. The properties of resveratrol are attributed to its ability to inhibit low-density lipoprotein oxidation, while suppressing the activity of cyclooxygenase 2 and induced nitric oxide synthase also contributes to the anti-inflammatory and antioxidant effects [10]. "
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    ABSTRACT: Microparticles of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) containing resveratrol were successfully prepared by simple emulsion/solvent evaporation. All formulations showed suitable encapsulation efficiency values higher than 80%. PHBV microparticles revealed spherical shape with rough surface and presence of pores. PCL microparticles were spherically shaped with smooth surface. Fourier-transformed infrared spectra demonstrated no chemical bond between resveratrol and polymers. X-ray powder diffraction patterns and differential scanning calorimetry analyses indicated that microencapsulation led to drug amorphization. These PHBV/PCL microparticles delayed the dissolution profile of resveratrol. Release profiles were better fitted to biexponential equation. The hypochlorous-acid-scavenging activity and 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation discoloration assay confirmed that the antioxidant activity of PHBV/PCL microparticles was kept, but was dependent on the microparticle morphology and dissolution profile. Resveratrol-loaded PHBV/PCL microparticles showed no cytotoxic effect on red blood cells.
    The Scientific World Journal 05/2012; 2012:542937. DOI:10.1100/2012/542937 · 1.73 Impact Factor
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    • "Res are mediated in part by its ability to activate SIRT1 (Lagouge et al. 2001). Studies have 13 shown that Res can extend the lifespan of S. cerevisiae (Howitz et al. 2003; Jarolim et al. 14 2004), Caenorhabditis elegans (Viswanathan et al. 2005; Wood et al. 2004), Drosophila 15 melanogaster (Bauer et al. 2004) and the vertebrate fish Nothobranchius furzeri (Valenzano 16 et al. 2006). However, in rodents, Res can delay age-related deterioration in mice without 17 extending lifespan (Pearson et al. 2008). "
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    ABSTRACT: Resveratrol (Res) has been associated with protective effects against oxidative stress. This study evaluated the effect of Res over lipid peroxidation, antioxidant defense, hepatic sirtuin 1 (SIRT1), which up-regulates antioxidant enzymes, and copper/zinc superoxide dismutase (Cu/Zn SOD) in adult offspring whose mothers were protein restricted during lactation. Lactating Wistar rats were divided into control (C) group, which were fed a normal diet (23% protein), and low-protein and high-carbohydrate (LPHC) group, which were fed a diet containing 8% protein. After weaning (21 days), C and LPHC offspring were fed a normal diet until they were 180 days old. At the 160th day, animals were separated into four groups as follows: control, control+Res, LPHC, and LPHC+Res. Resveratrol was given for 20 days (30  mg/kg per day by gavage). LPHC animals showed a higher total antioxidant capacity (TAC) without change in lipid peroxidation and SIRT1 expression. The treatment with Res increased TAC only in the control group without effect on lipid peroxidation and SIRT1. LPHC animals treated with Res had lower lipid peroxidation and higher protein and mRNA expression of SIRT1 without any further increase in TAC. No significant difference in liver Cu/Zn SOD expression was observed among the groups. In conclusion, maternal protein restriction during lactation programs the offspring for a higher antioxidant capacity, and these animals seem to respond to Res treatment with a lower lipid peroxidation and higher hepatic SIRT1 expression that we did not observe in the Res-treated controls. It is probable that the protective effect can be attributed to Res activating SIRT1, only in the LPHC-programmed group.
    Journal of Endocrinology 12/2010; 207(3):319-28. DOI:10.1677/JOE-10-0124 · 3.59 Impact Factor
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    • "Subsequently, resveratrol was shown to extend yeast, worm, and fly lifespan in a sirtuindependent manner (Howitz et al., 2003; Wood et al., 2004). Notably, the benefits of resveratrol have been disputed (Bass et al., 2007; Kaeberlein et al., 2005b), but also reproduced (Bauer et al., 2004; Greer et al., 2007; Jarolim et al., 2004; Viswanathan et al., 2005; Yang et al., 2007), in all three organisms. In addition, resveratrol extends lifespan and delays cognitive decline in N. furzeri, a short-lived species of fish (Valenzano et al., 2006), however, it has not yet been possible to assess the dependence of its effect on sirtuins in this organism. "
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    ABSTRACT: Dietary restriction (DR) delays or prevents age-related diseases and extends lifespan in species ranging from yeast to primates. Although the applicability of this regimen to humans remains uncertain, a proportional response would add more healthy years to the average life than even a cure for cancer or heart disease. Because it is unlikely that many would be willing or able to maintain a DR lifestyle, there has been intense interest in mimicking its beneficial effects on health, and potentially longevity, with drugs. To date, such efforts have been hindered primarily by our lack of mechanistic understanding of how DR works. Sirtuins, NAD(+)-dependent deacetylases and ADP-ribosyltransferases that influence lifespan in lower organisms, have been proposed to be key mediators of DR, and based on this model, the sirtuin activator resveratrol has been proposed as a candidate DR mimetic. Indeed, resveratrol extends lifespan in yeast, worms, flies, and a short-lived species of fish. In rodents, resveratrol improves health, and prevents the early mortality associated with obesity, but its precise mechanism of action remains a subject of debate, and extension of normal lifespan has not been observed. This review summarizes recent work on resveratrol, sirtuins, and their potential to mimic beneficial effects of DR.
    Mechanisms of ageing and development 02/2010; 131(4):261-9. DOI:10.1016/j.mad.2010.02.007 · 3.51 Impact Factor
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