Belak Zachery R, Harkness Troy, Eskiw Christopher H
Department of Food and Bioproduct Sciences, College of Agriculture and Bioresources, University of Saskatchewan, Saskatoon, SaskatchewanS7N 5A8, Canada.
Department of Biochemistry, Microbiology and Immunology (BMI), College of Medicine, University of Saskatchewan, Saskatoon, SaskatchewanS7N 5A8, Canada.
J Biol Methods. 2018 Dec 18;5(4):e106. doi: 10.14440/jbm.2018.272. eCollection 2018.
The budding yeast 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.
芽殖酵母是衰老研究中的一个主要模型系统。与后生动物一样,酵母的寿命可通过热量限制和使用延长寿命的药物来延长。芽殖酵母衰老研究的一个主要方法是时序寿命测定。传统上,时序寿命测定包括定期采集衰老酵母培养物的样本,将等分试样接种在琼脂上,并对产生的菌落进行计数。这种方法虽然高度可靠,但劳动强度大,且在材料消耗方面成本高昂。在此,我们报告一种基于MTT的新型方法,用于评估酵母的时序寿命。我们表明,该方法在严格可靠地测量热量限制导致的酵母寿命延长方面与菌落计数法相当,并且能够区分已知的长寿和短寿酵母菌株。我们进一步将该方法发展成一种高通量测定法,可快速筛选潜在的抗衰老化合物以及寿命改变的酵母菌株。应用该方法可快速鉴定酵母中的抗衰老活性,并可能有助于鉴定对高等动物尤其是人类具有治疗潜力的物质。