Gomez-Amaro Rafael L, Valentine Elizabeth R, Carretero Maria, LeBoeuf Sarah E, Rangaraju Sunitha, Broaddus Caroline D, Solis Gregory M, Williamson James R, Petrascheck Michael
Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037 Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California 92037 Department of Molecular and Cellular Neuroscience, The Scripps Research Institute, La Jolla, California 92037.
Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, California 92037.
Genetics. 2015 Jun;200(2):443-54. doi: 10.1534/genetics.115.175851. Epub 2015 Apr 21.
Caenorhabditis elegans has emerged as a powerful model to study the genetics of feeding, food-related behaviors, and metabolism. Despite the many advantages of C. elegans as a model organism, direct measurement of its bacterial food intake remains challenging. Here, we describe two complementary methods that measure the food intake of C. elegans. The first method is a microtiter plate-based bacterial clearing assay that measures food intake by quantifying the change in the optical density of bacteria over time. The second method, termed pulse feeding, measures the absorption of food by tracking de novo protein synthesis using a novel metabolic pulse-labeling strategy. Using the bacterial clearance assay, we compare the bacterial food intake of various C. elegans strains and show that long-lived eat mutants eat substantially more than previous estimates. To demonstrate the applicability of the pulse-feeding assay, we compare the assimilation of food for two C. elegans strains in response to serotonin. We show that serotonin-increased feeding leads to increased protein synthesis in a SER-7-dependent manner, including proteins known to promote aging. Protein content in the food has recently emerged as critical factor in determining how food composition affects aging and health. The pulse-feeding assay, by measuring de novo protein synthesis, represents an ideal method to unequivocally establish how the composition of food dictates protein synthesis. In combination, these two assays provide new and powerful tools for C. elegans research to investigate feeding and how food intake affects the proteome and thus the physiology and health of an organism.
秀丽隐杆线虫已成为研究进食、与食物相关行为及新陈代谢遗传学的强大模型。尽管秀丽隐杆线虫作为模式生物有诸多优势,但直接测量其细菌食物摄入量仍具有挑战性。在此,我们描述了两种互补的方法来测量秀丽隐杆线虫的食物摄入量。第一种方法是基于微量滴定板的细菌清除试验,通过量化细菌光密度随时间的变化来测量食物摄入量。第二种方法称为脉冲喂食,采用一种新颖的代谢脉冲标记策略,通过追踪从头合成蛋白质来测量食物的吸收情况。利用细菌清除试验,我们比较了各种秀丽隐杆线虫菌株的细菌食物摄入量,结果表明长寿的进食突变体的食量比之前估计的要多得多。为了证明脉冲喂食试验的适用性,我们比较了两种秀丽隐杆线虫菌株对血清素的食物同化情况。我们发现血清素增加进食会以依赖SER-7的方式导致蛋白质合成增加,包括已知会促进衰老的蛋白质。食物中的蛋白质含量最近已成为决定食物组成如何影响衰老和健康的关键因素。脉冲喂食试验通过测量从头合成蛋白质,是明确确定食物组成如何决定蛋白质合成的理想方法。这两种试验相结合,为秀丽隐杆线虫研究提供了新的强大工具,用于研究进食以及食物摄入量如何影响蛋白质组,进而影响生物体的生理和健康。