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用于快速定量评估酿酒酵母按时间顺序排列的寿命和存活率的工具箱。

A Toolbox for Rapid Quantitative Assessment of Chronological Lifespan and Survival in Saccharomyces cerevisiae.

作者信息

Chadwick Sarah R, Pananos Athanasios D, Di Gregorio Sonja E, Park Anna E, Etedali-Zadeh Parnian, Duennwald Martin L, Lajoie Patrick

机构信息

Department of Anatomy and Cell Biology, The University of Western Ontario, London, N6A 5C1, Canada.

Department of Applied Mathematics, University of Waterloo, Waterloo, N2L 3G1, Canada.

出版信息

Traffic. 2016 Jun;17(6):689-703. doi: 10.1111/tra.12391. Epub 2016 Apr 1.

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.

摘要

酿酒酵母是一种用于研究长寿机制的成熟模式生物。酵母中研究的两种衰老模式之一被称为时序寿命(CLS)。CLS由非分裂酵母细胞在稳定期存活的时间量来定义。在此,我们提出了新方法,可使用荧光细胞计数器或微孔板成像快速有效地量化衰老酵母培养物中的存活率。我们开发了一个名为analysr(酵母存活率分析算法)的软件,它能利用荧光数据对衰老酵母培养物中的细胞存活进行自动化且高度可重复的分析。为了证明我们新实验工具的效率,我们测试了热量限制延长寿命的先前已表征的能力。有趣的是,我们发现这个过程与三种核心酵母热休克蛋白(Hsp26、Hsp42、Hsp104)的表达无关。最后,我们的新检测方法很容易适用于其他类型的毒性研究。在此,我们评估了不同浓度乙酸的毒性,乙酸是酵母时序衰老的已知促成因素。这些检测为研究人员提供了具有成本效益的低含量和高含量检测方法,可作为CLS研究中通常使用的耗时的菌落形成单位检测的有效补充。

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