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使用卵滞留试验测量细菌对秀丽隐杆线虫行为的影响。

Measuring the effects of bacteria on C. elegans behavior using an egg retention assay.

作者信息

Gardner Mona, Rosell Mary, Myers Edith M

机构信息

Department of Biological and Allied Health Sciences, Fairleigh Dickinson University.

出版信息

J Vis Exp. 2013 Oct 22(80):e51203. doi: 10.3791/51203.

Abstract

C. elegans egg-laying behavior is affected by environmental cues such as osmolarity and vibration. In the total absence of food C. elegans also cease egg-laying and retain fertilized eggs in their uterus. However, the effect of different sources of food, especially pathogenic bacteria and particularly Enterococcus faecalis, on egg-laying behavior is not well characterized. The egg-in-worm (EIW) assay is a useful tool to quantify the effects of different types of bacteria, in this case E. faecalis, on egg- laying behavior. EIW assays involve counting the number of eggs retained in the uterus of C. elegans. The EIW assay involves bleaching staged, gravid adult C. elegans to remove the cuticle and separate the retained eggs from the animal. Prior to bleaching, worms are exposed to bacteria (or any type of environmental cue) for a fixed period of time. After bleaching, one is very easily able to count the number of eggs retained inside the uterus of the worms. In this assay, a quantifiable increase in egg retention after E. faecalis exposure can be easily measured. The EIW assay is a behavioral assay that may be used to screen for potentially pathogenic bacteria or the presence of environmental toxins. In addition, the EIW assay may be a tool to screen for drugs that affect neurotransmitter signaling since egg-laying behavior is modulated by neurotransmitters such as serotonin and acetylcholine.

摘要

秀丽隐杆线虫的产卵行为会受到渗透压和振动等环境线索的影响。在完全没有食物的情况下,秀丽隐杆线虫也会停止产卵,并将受精卵保留在子宫内。然而,不同食物来源,尤其是致病细菌,特别是粪肠球菌对产卵行为的影响尚未得到充分研究。虫内卵(EIW)试验是一种用于量化不同类型细菌(在这种情况下是粪肠球菌)对产卵行为影响的有用工具。EIW试验包括计算秀丽隐杆线虫子宫内保留的卵的数量。EIW试验包括对处于特定阶段的、怀孕的成年秀丽隐杆线虫进行漂白,以去除角质层,并将保留的卵与动物分离。在漂白之前,将线虫暴露于细菌(或任何类型的环境线索)中一段固定的时间。漂白后,很容易就能数出线虫子宫内保留的卵的数量。在这个试验中,可以很容易地测量出粪肠球菌暴露后卵保留量的可量化增加。EIW试验是一种行为试验,可用于筛选潜在的致病细菌或环境毒素的存在。此外,EIW试验可能是一种筛选影响神经递质信号传导的药物的工具,因为产卵行为受血清素和乙酰胆碱等神经递质的调节。

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