Charles Perkins Centre, The University of Sydney, Camperdown, NSW 2006, Australia.
School of Life & Environmental Sciences, The University of Sydney, Camperdown, NSW 2006, Australia.
Biotechniques. 2019 Sep;67(3):123-125. doi: 10.2144/btn-2019-0062. Epub 2019 Aug 6.
Metabolism is often studied in animal models, with the fruit fly model offering ease of genetic manipulation and high-throughput studies. Fly metabolism is typically studied using end-point assays that are simple but destructive, and do not provide information on the utilization of specific nutrients. To address these limitations, we adapted existing gas-trapping protocols to measure the oxidation of radiolabeled substrates (such as glucose) in multi-well plates. This protocol is cost effective, simple, and offers precise control over experimental diet and measurement time, thus being amenable to high-throughput studies. Furthermore, it is nondestructive, enabling time-course experiments and multiplexing with other parameters. Overall, this protocol is useful for merging fly genetics with metabolic studies to understand whole organism responses to different macronutrients.
代谢通常在动物模型中进行研究,而果蝇模型具有易于进行遗传操作和高通量研究的特点。通常使用终点测定法来研究果蝇的代谢,这些方法虽然简单但具有破坏性,并且无法提供有关特定营养素利用情况的信息。为了解决这些限制,我们对现有的气体捕获方案进行了改编,以测量多孔板中放射性标记底物(如葡萄糖)的氧化。该方案具有成本效益、简单、可以精确控制实验饮食和测量时间,因此适用于高通量研究。此外,它是非破坏性的,能够进行时程实验并与其他参数进行多重分析。总的来说,该方案可用于将果蝇遗传学与代谢研究相结合,以了解生物体对不同宏量营养素的整体反应。