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首页> 外文期刊>Genetics: A Periodical Record of Investigations Bearing on Heredity and Variation >Measuring Food Intake and Nutrient Absorption in Caenorhabditis elegans
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Measuring Food Intake and Nutrient Absorption in Caenorhabditis elegans

机译:测量秀丽隐杆线虫的食物摄入和营养吸收

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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.
机译:秀丽隐杆线虫已经成为研究饲料遗传学,食物相关行为和新陈代谢的强大模型。尽管秀丽隐杆线虫作为模型生物具有许多优势,但直接测量其细菌食物摄入量仍然具有挑战性。在这里,我们描述了两种测量线虫食物摄入量的互补方法。第一种方法是基于微量滴定板的细菌清除测定法,该方法通过量化细菌的光密度随时间变化来测量食物摄入量。第二种方法称为脉冲进料,它使用一种新的代谢脉冲标记策略通过跟踪从头蛋白质合成来测量食物的吸收。使用细菌清除率分析,我们比较了各种秀丽隐杆线虫菌株的细菌食物摄入量,结果表明长寿食用突变体的进食量大大高于先前的估计。为了证明脉冲进料测定法的适用性,我们比较了食物对两种线虫线虫血清素的同化作用。我们显示5-羟色胺增加的喂养导致以SER-7依赖性方式增加蛋白质合成,包括已知会促进衰老的蛋白质。食品中的蛋白质含量最近已成为决定食品成分如何影响衰老和健康的关键因素。通过测量从头开始的蛋白质合成,脉冲喂食测定法是一种明确确定食物成分决定蛋白质合成方式的理想方法。结合起来,这两种测定法为秀丽隐杆线虫的研究提供了强大的新工具,以研究饲喂以及食物摄入如何影响蛋白质组,进而影响生物体的生理和健康。

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