Measuring Food Intake and Nutrient Absorption in Caenorhabditis elegans.
Measuring Food Intake and Nutrient Absorption in Caenorhabditis elegans.
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DOI：
10.1534/genetics.115.175851
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发表时间：
2015-06
期刊：
影响因子：
3.3
通讯作者：
Petrascheck M
中科院分区：
文献类型：
--
作者：
Gomez-Amaro RL;Valentine ER;Carretero M;LeBoeuf SE;Rangaraju S;Broaddus CD;Solis GM;Williamson JR;Petrascheck M
秀丽隐杆线虫已成为研究摄食、食物相关行为和代谢遗传学的有力模型。尽管C.作为模式生物，直接测量其细菌食物摄入量仍然具有挑战性。在这里，我们描述了两种互补的方法，测量食物摄入量的C。优雅的第一种方法是基于微量滴定板的细菌清除测定，其通过定量细菌随时间的光密度变化来测量食物摄入。第二种方法，称为脉冲喂养，通过使用新的代谢脉冲标记策略跟踪从头蛋白质合成来测量食物的吸收。采用细菌清除试验，比较了不同条件下的C。线虫菌株，并表明长寿吃突变吃大大超过以前的估计。为了证明脉冲喂养试验的适用性，我们比较了两种C。elegans应变in response响应to serotonin血清素.我们发现，增加摄食可导致SER-7依赖性蛋白质合成增加，包括已知促进衰老的蛋白质。食物中的蛋白质含量最近成为决定食物成分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蛋白质合成，代表了明确确定食物组成如何决定蛋白质合成的理想方法。结合起来，这两种检测方法提供了新的和强大的工具，C。线虫的研究，以调查喂养和食物摄入如何影响蛋白质组，从而影响生物体的生理和健康。
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.