Measuring C. elegans spatial foraging and food intake using bioluminescent bacteria

Measuring C. elegans spatial foraging and food intake using bioluminescent bacteria
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使用生物发光细菌测量线虫空间觅食和食物摄入量

DOI:
10.1101/759928
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发表时间:
2019
期刊:
--
影响因子:
--
通讯作者:
Ding S
Ding S
中科院分区:
--
文献类型:
--
作者:
Ding S

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对于大多数动物来说,进食包括两种行为:寻找食物区和发现食物区后摄入食物。线虫秀丽隐杆线虫是研究这两种行为的遗传学的有用模型。然而,大多数测量蠕虫摄食的方法都会量化觅食行为或食物摄入量,但不能同时量化两者。对荧光标记细菌的消耗进行成像可提供有关分布和消耗量的信息,但即使在斑块耗尽后,仍保留显着的背景信号,这使定量变得复杂。在这里,我们使用生物发光大肠杆菌来量化 C。线虫喂养。由于光发射与活跃的新陈代谢紧密结合,只有活细菌才能发出生物发光,因此信号在摄入后就会丢失。我们使用 N2 参考蠕虫和 eat-2 突变体对生物发光的损失进行了量化,发现与荧光损失相比,信号背景比增加了近 100 倍,并且背景更低。我们还使用聚合 npr-1 突变蠕虫来量化喂养。我们发现,npr-1突变体群体首先会相互清除细菌,然后集体寻找更多食物。同样,在高密度集群期间,只有迁移前沿的蠕虫才会与细菌接触。这些结果证明了生物发光细菌在定量摄食方面的有用性,并提出了 C 功能的卫生假设。秀丽隐杆线虫聚集和蜂拥而至。
For most animals, feeding includes two behaviours: foraging to find a food patch and food intake once a patch is found. The nematodeCaenorhabditis elegansis a useful model for studying the genetics of both behaviours. However, most methods of measuring feeding in worms quantify either foraging behaviour or food intake but not both. Imaging the depletion of fluorescently labelled bacteria provides information on both the distribution and amount of consumption, but even after patch exhaustion a prominent background signal remains, which complicates quantification. Here, we used a bioluminescentEscherichia colistrain to quantifyC. elegansfeeding. With light emission tightly coupled to active metabolism, only living bacteria are capable of bioluminescence so the signal is lost upon ingestion. We quantified the loss of bioluminescence using N2 reference worms andeat-2mutants, and found a nearly 100-fold increase in signal-to-background ratio and lower background compared to loss of fluorescence. We also quantified feeding using aggregatingnpr-1mutant worms. We found that groups ofnpr-1mutants first clear bacteria from each other before foraging collectively for more food; similarly, during high density swarming, only worms at the migrating front are in contact with bacteria. These results demonstrate the usefulness of bioluminescent bacteria for quantifying feeding and suggest a hygiene hypothesis for the function ofC. elegansaggregation and swarming.
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