Department of Molecular, Cell and Cancer Biology, UMass Chan Medical School, Worcester, MA, USA.
Methods Mol Biol. 2025;2871:193-200. doi: 10.1007/978-1-0716-4217-7_18.
In Caenorhabditis elegans (C. elegans), there is a single FOXO transcription factor homolog, encoded by the gene, daf-16. As a central regulator for multiple pathways, DAF-16 integrates these signals to result in changes in longevity, development, fat storage, stress resistance, innate immunity, and reproduction. One of the main advantages of using C. elegans is the ability to study FOXO in the context of the whole animal. Therefore, manipulating the levels or the activity of daf-16 results in visible, scorable phenotypic changes. DAF-16 is the downstream target of the conserved insulin/IGF-1 signaling (IIS) pathway, a PI 3-kinase signaling cascade that ultimately controls its nuclear localization. Since the IIS pathway is a major regulator of lifespan, almost all studies of lifespan modulation examine the requirement of daf-16. More recently, lifespan analysis has been accompanied by healthspan analysis, referring to the time an animal is healthy. In this chapter, I will focus on the assays to assess lifespan and healthspan of C. elegans FOXO/daf-16, in the context of a whole animal.
在秀丽隐杆线虫(C. elegans)中,存在一个单一的 FOXO 转录因子同源物,由基因 daf-16 编码。作为多种途径的中央调节剂,DAF-16 整合这些信号,导致寿命、发育、脂肪储存、应激抗性、先天免疫和生殖的变化。使用秀丽隐杆线虫的主要优势之一是能够在整个动物体内研究 FOXO。因此,操纵 daf-16 的水平或活性会导致可见的、可评分的表型变化。DAF-16 是保守的胰岛素/IGF-1 信号(IIS)途径的下游靶标,这是一个 PI 3-激酶信号级联,最终控制其核定位。由于 IIS 途径是寿命的主要调节剂,因此几乎所有关于寿命调节的研究都检查了 daf-16 的要求。最近,寿命分析伴随着健康寿命分析,这是指动物健康的时间。在本章中,我将重点介绍评估秀丽隐杆线虫 FOXO/daf-16 整体动物寿命和健康寿命的测定法。