Measurement of Lifespan in Drosophila melanogaster
Measurement of Lifespan in Drosophila melanogaster
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DOI：
10.3791/50068
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发表时间：
2013-01-01
影响因子：
1.2
通讯作者：
Pletcher, Scott D.
中科院分区：
文献类型：
--
作者：
Linford, Nancy J.;Bilgir, Ceyda;Pletcher, Scott D.
衰老是一种现象，在几乎所有被研究的生物体中，衰老都会导致生理上的持续恶化，导致身体机能下降，患病风险增加。个体老龄化在人口水平上表现为年龄相关死亡率的增加，这通常是在实验室通过观察年龄匹配的大量个体的寿命来测量的。试图量化遗传或环境操纵对简单模式生物寿命影响程度的实验，在理解跨分类群保守的衰老方面取得了显著成功，并为延长哺乳动物寿命和预防与年龄相关的疾病提供了新的策略。醋蝇（Drosophila melanogaster）因其相对较短的寿命、饲养方便、易遗传而成为研究衰老机制的有吸引力的模式生物。然而，老龄化的人口统计测量，包括特定年龄的存活率和死亡率，极易受到实验设计和环境的微小变化的影响，因此在老龄化实验期间需要保持严格的实验室实践。这些考虑，连同对遗传背景进行仔细控制的需要，对于产生可靠的测量是必不可少的。事实上，围绕酵母、蠕虫、苍蝇和老鼠的长寿实验的推断存在许多值得注意的争议，这些实验可以追溯到环境或遗传产物（1-4）。在本协议中，我们描述了一套经过多年优化的使用实验室小瓶测量果蝇寿命的程序。我们还描述了dLife软件的使用，该软件是由我们的实验室开发的，可以下载（http://sitemaker.umich.edu/pletcherlab/software）。dLife通过整合最佳实验设计、简化苍蝇处理和数据收集以及标准化数据分析，加快了吞吐量并促进了良好实践。我们还将讨论寿命数据的设计、收集和解释中的许多潜在缺陷，并提供避免这些危险的步骤。
Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced physical performance and increased risk of disease. Individual aging is manifest at the population level as an increase in age-dependent mortality, which is often measured in the laboratory by observing lifespan in large cohorts of age-matched individuals. Experiments that seek to quantify the extent to which genetic or environmental manipulations impact lifespan in simple model organisms have been remarkably successful for understanding the aspects of aging that are conserved across taxa and for inspiring new strategies for extending lifespan and preventing age-associated disease in mammals.The vinegar fly, Drosophila melanogaster, is an attractive model organism for studying the mechanisms of aging due to its relatively short lifespan, convenient husbandry, and facile genetics. However, demographic measures of aging, including age-specific survival and mortality, are extraordinarily susceptible to even minor variations in experimental design and environment, and the maintenance of strict laboratory practices for the duration of aging experiments is required. These considerations, together with the need to practice careful control of genetic background, are essential for generating robust measurements. Indeed, there are many notable controversies surrounding inference from longevity experiments in yeast, worms, flies and mice that have been traced to environmental or genetic artifacts(1-4). In this protocol, we describe a set of procedures that have been optimized over many years of measuring longevity in Drosophila using laboratory vials. We also describe the use of the dLife software, which was developed by our laboratory and is available for download (http://sitemaker.umich.edu/pletcherlab/software). dLife accelerates throughput and promotes good practices by incorporating optimal experimental design, simplifying fly handling and data collection, and standardizing data analysis. We will also discuss the many potential pitfalls in the design, collection, and interpretation of lifespan data, and we provide steps to avoid these dangers.