复制性寿命和时序性寿命测定

Replicative and chronological life-span assays.

作者信息

Postnikoff Spike D L, Harkness Troy A A

机构信息

Department of Anatomy and Cell Biology, University of Saskatchewan, 107 Wiggins Road, Saskatoon, SK, Canada, S7N 5E5.

出版信息

Methods Mol Biol. 2014;1163:223-7. doi: 10.1007/978-1-4939-0799-1_17.

DOI:10.1007/978-1-4939-0799-1_17

PMID:24841311

Abstract

Life-span assays in yeast are invaluable in characterizing the functions of gene products on cellular aging. Replicative life-span (RLS) is a measure of the number of divisions an individual cell can undergo. In this assay daughter cells are removed using a tetrad dissection microscope with a micromanipulator and scored. Chronological life-span (CLS) measures the length of time nondividing cells survive. A culture is grown to stationary phase with samples removed over time to assess the survival within the population. The strength of the yeast system lies in the ease of genetically manipulating genes of interest and the evolutionarily conserved nature of the genes found to influence longevity. Here, we describe methods used to measure yeast RLS and CLS.

摘要

酵母中的寿命测定对于表征基因产物在细胞衰老中的功能非常重要。复制寿命（RLS）是衡量单个细胞能够经历的分裂次数的指标。在这个测定中，使用带有显微操作器的四分体解剖显微镜去除子细胞并进行计数。时序寿命（CLS）衡量不分裂细胞存活的时间长度。将培养物培养至稳定期，随着时间的推移取出样品以评估群体中的存活率。酵母系统的优势在于易于对感兴趣的基因进行遗传操作，以及发现影响寿命的基因具有进化保守性。在这里，我们描述了用于测量酵母RLS和CLS的方法。