A Toolbox for Rapid Quantitative Assessment of Chronological Lifespan and Survival in Saccharomyces cerevisiae.
Autor: | Chadwick SR; Department of Anatomy and Cell Biology, The University of Western Ontario, London, N6A 5C1, Canada., Pananos AD; Department of Applied Mathematics, University of Waterloo, Waterloo, N2L 3G1, Canada., Di Gregorio SE; Department of Pathology and Laboratory Medicine, The University of Western Ontario, London, N6A 5C1, Canada., Park AE; Department of Anatomy and Cell Biology, The University of Western Ontario, London, N6A 5C1, Canada., Etedali-Zadeh P; Department of Anatomy and Cell Biology, The University of Western Ontario, London, N6A 5C1, Canada., Duennwald ML; Department of Anatomy and Cell Biology, The University of Western Ontario, London, N6A 5C1, Canada.; Department of Pathology and Laboratory Medicine, The University of Western Ontario, London, N6A 5C1, Canada., Lajoie P; Department of Anatomy and Cell Biology, The University of Western Ontario, London, N6A 5C1, Canada. |
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Jazyk: | angličtina |
Zdroj: | Traffic (Copenhagen, Denmark) [Traffic] 2016 Jun; Vol. 17 (6), pp. 689-703. Date of Electronic Publication: 2016 Apr 01. |
DOI: | 10.1111/tra.12391 |
Abstrakt: | Saccharomyces cerevisiae is a well-established model organism to study the mechanisms of longevity. One of the two aging paradigms studied in yeast is termed chronological lifespan (CLS). CLS is defined by the amount of time non-dividing yeast cells can survive at stationary phase. Here, we propose new approaches that allow rapid and efficient quantification of survival rates in aging yeast cultures using either a fluorescent cell counter or microplate imaging. We have generated a software called analysr (Analytical Algorithm for Yeast Survival Rates) that allows automated and highly reproducible analysis of cell survival in aging yeast cultures using fluorescent data. To demonstrate the efficiency of our new experimental tools, we tested the previously characterized ability of caloric restriction to extend lifespan. Interestingly, we found that this process is independent of the expression of three central yeast heat shock proteins (Hsp26, Hsp42, Hsp104). Finally, our new assay is easily adaptable to other types of toxicity studies. Here, we assessed the toxicity of various concentrations of acetic acid, a known contributor of yeast chronological aging. These assays provide researchers with cost-effective, low- and high-content assays that can serve as an efficient complement to the time-consuming colony forming unit assay usually used in CLS studies. (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.) |
Databáze: | MEDLINE |
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