title: A Toolbox for Rapid Quantitative Assessment of Chronological Lifespan and Survival in Saccharomyces cerevisiae.
abstract: 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.
authors.affiliation: Department of Anatomy and Cell Biology, The University of Western Ontario, London, N6A 5C1, Canada.
authors.forename: Sarah R
authors.initials: SR
authors.lastname: Chadwick
authors.affiliation: Department of Applied Mathematics, University of Waterloo, Waterloo, N2L 3G1, Canada.
authors.forename: Athanasios D
authors.initials: AD
authors.lastname: Pananos
authors.affiliation: Department of Pathology and Laboratory Medicine, The University of Western Ontario, London, N6A 5C1, Canada.
authors.forename: Sonja E
authors.initials: SE
authors.lastname: Di Gregorio
authors.affiliation: Department of Anatomy and Cell Biology, The University of Western Ontario, London, N6A 5C1, Canada.
authors.forename: Anna E
authors.initials: AE
authors.lastname: Park
authors.affiliation: Department of Anatomy and Cell Biology, The University of Western Ontario, London, N6A 5C1, Canada.
authors.forename: Parnian
authors.initials: P
authors.lastname: Etedali-Zadeh
authors.affiliation: 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.
authors.forename: Martin L
authors.initials: ML
authors.lastname: Duennwald
authors.affiliation: Department of Anatomy and Cell Biology, The University of Western Ontario, London, N6A 5C1, Canada.
authors.forename: Patrick
authors.initials: P
authors.lastname: Lajoie
mesh_terms: D019342:Acetic Acid / Q000633:toxicity; D049109:Cell Proliferation; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D012984:Software
chemical_list: D006360:Heat-Shock Proteins; D029701:Saccharomyces cerevisiae Proteins; D019342:Acetic Acid
keywords: caloric restriction; cell viability; chronological lifespan; computer-assisted analysis; fluorescence; heat shock proteins; yeast