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: 2.82). 12/2004; 5(2):169-77. DOI: 10.1016/j.femsyr.2004.06.015
Source: PubMed


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.

Full-text preview

Available from:
  • Source
    • "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]. "
    [Show abstract] [Hide abstract]
    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.
    Full-text · Article · May 2012 · The Scientific World Journal
  • Source
    • "To test this hypothesis, we have utilized resveratrol, a natural polyphenol found in red grapes and wine, which has previously been shown to increase AMPK activity in neurons [21]. Although several studies originally described resveratrol as an activator of sirtuin enzymes, which are NAD-dependent deacetylases [22-25] these results have been challenged based on lack of specificity in screening assays [26,27]. Moreover, several recent in vivo studies strongly suggest that resveratrol effects are independent of sirtuins. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Despite advances in our understanding of basic mechanisms driving post-surgical pain, treating incision-induced pain remains a major clinical challenge. Moreover, surgery has been implicated as a major cause of chronic pain conditions. Hence, more efficacious treatments are needed to inhibit incision-induced pain and prevent the transition to chronic pain following surgery. We reasoned that activators of AMP-activated protein kinase (AMPK) may represent a novel treatment avenue for the local treatment of incision-induced pain because AMPK activators inhibit ERK and mTOR signaling, two important pathways involved in the sensitization of peripheral nociceptors. To test this hypothesis we used a potent and efficacious activator of AMPK, resveratrol. Our results demonstrate that resveratrol profoundly inhibits ERK and mTOR signaling in sensory neurons in a time- and concentration-dependent fashion and that these effects are mediated by AMPK activation and independent of sirtuin activity. Interleukin-6 (IL-6) is thought to play an important role in incision-induced pain and resveratrol potently inhibited IL-6-mediated signaling to ERK in sensory neurons and blocked IL-6-mediated allodynia in vivo through a local mechanism of action. Using a model of incision-induced allodynia in mice, we further demonstrate that local injection of resveratrol around the surgical wound strongly attenuates incision-induced allodynia. Intraplantar IL-6 injection and plantar incision induces persistent nociceptive sensitization to PGE2 injection into the affected paw after the resolution of allodynia to the initial stimulus. We further show that resveratrol treatment at the time of IL-6 injection or plantar incision completely blocks the development of persistent nociceptive sensitization consistent with the blockade of a transition to a chronic pain state by resveratrol treatment. These results highlight the importance of signaling to translation control in peripheral sensitization of nociceptors and provide further evidence for activation of AMPK as a novel treatment avenue for acute and chronic pain states.
    Full-text · Article · Jan 2012 · Molecular Pain
  • Source
    • "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). "
    [Show abstract] [Hide abstract]
    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.
    Full-text · Article · Dec 2010 · Journal of Endocrinology
Show more