Andalusian Center for Developmental Biology, Consejo Superior de Investigaciones Científicas/Junta de Andalucía/Universidad Pablo de Olavide, Departament of Molecular Biology and Biochemical Engineering, Carretera de Utrera, km 1, 41013, Seville, Spain.
Institut de Biochimie et Génétique Cellulaires - C.N.R.S. UMR 5095 and Université de Bordeaux, 1, rue Camille Saint-Saëns, 33077, Bordeaux Cedex, France.
Sci Rep. 2018 Feb 26;8(1):3633. doi: 10.1038/s41598-018-21964-z.
The study of mechanisms that govern feeding behaviour and its related disorders is a matter of global health interest. The roundworm Caenorhabditis elegans is becoming a model organism of choice to study these conserved pathways. C. elegans feeding depends on the contraction of the pharynx (pumping). Thanks to the worm transparency, pumping can be directly observed under a stereoscope. Therefore, C. elegans feeding has been historically investigated by counting pharyngeal pumping or by other indirect approaches. However, those methods are short-term, time-consuming and unsuitable for independent measurements of sizable numbers of individuals. Although some particular devices and long-term methods have been lately reported, they fail in the automated, scalable and/or continuous aspects. Here we present an automated bioluminescence-based method for the analysis and continuous monitoring of worm feeding in a multi-well format. We validate the method using genetic, environmental and pharmacological modulators of pharyngeal pumping. This flexible methodology allows studying food intake at specific time-points or during longer periods of time, in single worms or in populations at any developmental stage. Additionally, changes in feeding rates in response to differential metabolic status or external environmental cues can be monitored in real time, allowing accurate kinetic measurements.
研究控制进食行为及其相关障碍的机制是全球健康关注的问题。秀丽隐杆线虫（Caenorhabditis elegans）作为一种模式生物，正在成为研究这些保守途径的首选。线虫的进食依赖于咽部的收缩（抽吸）。由于线虫的透明性，可以在立体显微镜下直接观察到抽吸。因此，线虫的进食行为在历史上一直通过计数咽部抽吸或其他间接方法来研究。然而，这些方法是短期的、耗时的，并且不适合对大量个体进行独立测量。尽管最近有一些特定的设备和长期的方法被报道，但它们在自动化、可扩展和/或连续方面存在不足。在这里，我们提出了一种基于生物发光的自动化方法，用于在多孔板格式中分析和连续监测线虫的进食。我们使用咽部抽吸的遗传、环境和药理学调节剂来验证该方法。这种灵活的方法允许在特定时间点或在更长的时间内、在单个线虫或任何发育阶段的群体中研究食物摄入。此外，还可以实时监测对不同代谢状态或外部环境线索的进食率变化，从而进行精确的动力学测量。
