一种快速、高通量测定芽苗酵母寿命的方法

Z. Belak, T. Harkness, C. Eskiw
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引用次数: 9

摘要

出芽酵母是酿酒酵母老化研究中的一个重要模型系统。像后生动物一样,酵母的寿命通过热量限制和延长寿命的药物治疗而延长。出芽酵母衰老研究的一个主要工作是按时间顺序的寿命测定。传统上,按时间顺序的寿命测定包括定期采集老化酵母培养物的样本,在琼脂上皿出等分,并计算所得菌落。这种方法虽然高度可靠,但在消耗的材料方面是劳动密集型和昂贵的。在这里,我们报告了一种新的基于mtt的方法来评估酵母的时间顺序寿命。研究表明,该方法与菌落计数法在严格可靠地测量酵母由于热量限制而延长的寿命方面是相同的,并且能够区分已知的长寿和短寿命酵母菌株。我们已经进一步将这种方法发展成为一种高通量分析方法,可以快速筛选潜在的抗衰老化合物以及改变寿命的酵母菌株。该方法的应用允许快速鉴定酵母的抗衰老活性,并可能促进鉴定具有治疗潜力的材料,用于高等动物,最重要的是,人类。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A rapid, high-throughput method for determining chronological lifespan in budding yeast
The budding yeast Saccharomyces cerevisiae is a major model system in the study of aging. Like metazoans, yeast lifespan is extended by caloric restriction and treatment with pharmacological agents which extend lifespan. A major workhorse of aging research in budding yeast is the chronological lifespan assay. Traditionally, chronological lifespan assays consist of taking regular samples of aging yeast cultures, plating out aliquots on agar, and counting the resulting colonies. This method, while highly reliable, is labor-intensive and expensive in terms of materials consumed. Here, we report a novel MTT-based method for assessing chronological lifespan in yeast. We show that this method is equal to the colony counting method in its rigorous and reliable measurement of lifespan extension in yeast as a result of caloric restriction, and is able to distinguish known long-lived and short-lived yeast strains. We have further developed this method into a high-throughput assay that allows rapid screening of potential anti-aging compounds as well as yeast strains with altered lifespan. Application of this method permits the rapid identification of anti-aging activities in yeast and may facilitate identification of materials with therapeutic potential for higher animals and, most importantly, humans.
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