Name: measuring the effect of chemicals on the growth and reproduction of caenorhabditis elegans
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这项研究得到了韩国科学和技术研究所校内研究补助金 (2E27513) 和农业、粮食和农村事务部 (315067-03) 资助的高增值食品技术发展方案 (IPET) 的支持。

注意: 整个实验必须在20和 #176 保持的干净的隔离实验室中进行; 在蠕虫和细菌处理过程中, 低粉尘和最小污染的 C。为此, 必须在酒精灯的火焰或使用干净的长凳上进行实验. 
 1. 维护 c. 线虫 和鸡蛋准备化学测试 <ol> <li> 维护 线虫 N2 (var) 在 NGM 琼脂板上喂养的活 大肠杆菌 OP50 在20和 #176; C 12 , 13 . </li>
	<li> 使用漂白剂溶液处理的龄期同步卵 5 或时间蛋铺设方法 6 14 , 15 
	注: 在准备鸡蛋之前, 用热灭活的 大肠杆菌 OP50 涂依托含 NGM 板, 如下所述。热灭活的 大肠杆菌 OP50 用于将 大肠杆菌 6 、 16 的新陈代谢活动降至最低。
</ol> 2。制备热灭活的 大肠杆菌 OP50 在毒性试验中, 为饲料中的 线虫 <ol> <li> 准备500毫升的双酵母胰 (戴阳天) 培养基, 如表1所述, 并接种单一菌落的 大肠杆菌 OP50 培养Luria-Bertani (LB) 琼脂平板 5 . </li>
	<li> 孵育 大肠杆菌 OP50 在14小时的震动孵化器中, 37 和 #176; C 和 150 rpm. </li>
	<li> 离心30分钟后在 3220 x g 和 20 #176; C、取出所有上清戴阳天培养基, 并添加混25毫升的 s-缓冲器、250和 #181; 1 米 MgSO 4 , 25 和 #181; 5 毫克/毫升胆固醇 (溶于乙醇)。所有这些解决方案都是无菌的. </li>
	<li> 重细菌并将其转移到50毫升的一次性塑料管中. </li>
	<li> 热失活, 孵育悬浮 大肠杆菌 OP50 在水浴30分钟, 在65和 #176; C. </li>
	<li> 冷却到室温并存储在4和 #176; C 直到使用。这可以存储长达一个月. </li>
</ol> 3。制备含有1% 二甲基亚砜或750和 #181 的 NGM 板; M 依托 <ol> <li> 准备两个干净的200毫升玻璃瓶。加入0.6 克氯化钠, 0.5 克蛋白胨, 3.4 克琼脂, 195 毫升蒸馏水, 一瓶磁力搅拌器。在另一个空瓶子里加一个磁力搅拌器. </li>
	<li> 将这两个瓶子在121和 #176 上压釜15分钟, 然后在水浴中冷却30分钟, 在55和 #176; c. </li>
	<li> 添加 0.2 mL 1 米 CaCl 2 , 0.2 毫升的5毫克/毫升胆固醇, 0.2 毫升 1 m MgSO 4 , 5 毫升 1 m KPO 4 (所有这些解决方案是无菌), 然后混合他们的磁搅拌在热板上, 在55和 #176; C. </li>
	<li> 将混合 NGM 介质分成两个相同体积的瓶子 (每瓶100毫升)。然后, 将1毫升的亚甲基亚砜添加到一个瓶子中, 再用磁力搅拌再混合。将另一个瓶子放在55和 #176 的水浴里, 直到下一步。分3毫升的含亚砜的培养基, 每 35 x 10 mm 2 培养皿。从这一步可以制成大约33含亚砜的 NGM 板. </li>
	<li> 混合其他瓶子使用磁性搅拌, 添加1毫升的75毫米依托 (股票溶解在二甲基亚砜), 再次混合, 并重复分程序 (步骤 3.4)。从这一步中可以得到大约33依托 (750 和 #181; M)-包含 NGM 板. 
	注意: 在相关的前一篇文章 6 中, 我们测试了0、250、500和1000和 #181; 依托。根据这一数据, 本文选取了 750 #181; 依托剂量. </li>
	<li> 将含有化学成分的 NGM 板冷却至室温, 将其置于室温下约3小时, 并将其存储在4和 #176; C 直到使用. 
	注意: 在光敏化学品的情况下, 阻挡光, 以尽量减少光分解. 
	注: 可选, 做一个正常的 NGM 板没有化学品的鸡蛋铺设化验, 可以执行使用两种不同的化学处理条件如下所述. </li>
	<li> 添加100和 #181; 热灭活 大肠杆菌 OP50 (如2节所述) 并在每个 NGM 板上传播。允许在20和 #176 的 c 线虫 孵化器中过夜, 用于化学测试. </li>
</ol> 4。测量 C. 线虫 增长 <ol> <li> 将年龄同步的 线虫 的卵转移到 NGM 板上, 再配以1% 亚砜或750和 #181; 我依托. </li>
	<li> 孵育 c. 线虫 在20和 #176 的孵化器中; c 为4天。用立体显微镜观察蠕虫, 每天拍摄显微图像。通常20X 放大 (1X 物镜, 2X 变焦放大, 和10X 目镜镜头) 是适合的增长观察的 C. 线虫 。还采取显微镜阶段千分尺的显微图像的蠕虫大小校准. 
	注意: 饥饿影响毒性试验。因此, 饲料有足够的细菌或调整和 #160; 在开始时转移了 C. 线虫 卵数。观察到3天, 排除饥饿的可能性. </li>
	<li> 使用 ImageJ 软件在每个时间点测量用亚砜或依托处理的 C. 线虫 的体长. 
	注: 对于每个样本测量, 50 蠕虫是足够的统计分析。
	<ol> <li> 在 ImageJ 中, 打开显微镜阶段千分尺图像 (#39; 文件和 #39; #8594; #39; 打开和 #39;). 
		注: 千分尺图像和蠕虫图像 (步骤 4.2) 必须是相同的放大倍数. </li>
		<li> 通过使用和 #39、直线和 #39; 工具, 在千分尺校准图像上拖动1000和 #181;
		<li> 单击和 #39; 分析和 #39; #8594; #39; 设置规模和 #39; 输入1000和 #181; 我进入和 #39; 已知的距离和 #39; 框和 #39; 长度和 #39 的单位; 框, 分别。检查和 #39; 全局和 #39; 单击并 #39; 确定 #39;. </li>
		<li> 打开蠕虫图像 (#39; 文件和 #39; #8594; #39; 打开和 #39;)。使用和 #39; 手绘线和 #39; 工具, 沿每个蠕虫的特征绘制一条线。通过单击和 #39 获取正文长度数据; 分析和 #39; #8594; #39; 测量和 #39;. </li>
		<li> 对50蠕虫重复这些步骤. </li>
	</ol> </li> </ol> 5。 C. 线虫 产卵试验 <ol> <li> 在包含亚甲基亚砜或依托的 NGM 板上孵育年龄同步的卵, 在64小时 (第一次出生之前), 直到20和 #176 的年轻成人阶段; c. 
	注意: 在生长测量实验中使用蠕虫是可选的。这是方便做的生长测量和鸡蛋铺设化验一起. </li>
	<li> 将5只成年蠕虫转移到新的无化学品的 NGM 板 (条件 1, 在一定时间内进行化学处理, 从卵到幼成体阶段) 或新的 NGM 板含有相同的化学预处理 (条件 2, 连续曝光化学品通过整个实验期), 然后孵化为24小时. 
	注意: 建议每种治疗方法使用 3-5 NGM 板进行复制;这些复制可用于统计分析. </li>
	<li> 转移到新的 NGM 板如上所述, 每天计数的鸡蛋数。计数蠕虫爬行, 死亡, 并在内部孵化ed. </li>
	<li> 重复这些步骤, 直到蠕虫不再下蛋, 通常在5天内. </li>
	<li> 计算每个盘中每条蠕虫的产卵数, 并在5天内每天对这些值求和, 以确定产卵的总数量. 
	注意: 排除从计算中抓取和内部孵化的蠕虫的数量. </li>
</ol>

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作者没有什么可透露的。

在这篇文章中, 我们描述的毒性评价的化学品在线虫, 土壤线虫, 使用依托作为毒物的例子。为此, 我们使用了两个实验条件。在第一个集合中, C. 线虫生长在含有从卵到幼成虫阶段的依托板上, 然后允许蠕虫在正常的 NGM 板上产卵, 而不使用化学物质。在第二个实验组中, 在整个实验期间, 用依托连续处理线虫。通过比较两个实验条件下的卵数, 我们可以确定被测化合物的生殖毒性...

毒理学评价对制药、营养和妆的发展以及各种环境毒素的风险评估都是必不可少的。啮齿类动物模型是其中一个最普遍的在体内实验系统为这毒理学研究;另外, 哺乳有机体, 如C. 线虫也被广泛使用。非哺乳动物毒理学评价模型是有益的, 因为不仅动物伦理问题, 而且它们的方便性和实用性考虑 cost-effectiveness, 可维护性, 速度和重现性1,2 ,3,4。
线虫是一种土壤圆形蠕虫, 在各种基础和应用生物学和化学研究中被作为动物模型来开发。它是一个1毫米长, 透明的线虫, 这是简单地维持在固体或液体线虫生长培养基 (NGM) 喂养的细菌菌株大肠杆菌OP50。c. 线虫的生命周期较短, 野生类型的 N2 c. 线虫约有300只卵。因此, 它很容易被传播到用作实验材料3,4,5。线虫在许多药物和环境污染物的毒理学研究中也得到了广泛的应用6,7,8,9。
由于许多抗癌药物的目标是迅速分裂的癌细胞, 他们也可以损害迅速分裂正常细胞, 如骨髓, 肠上皮, 毛囊细胞。例如, 拓扑抑制抗癌药物靶向癌细胞的 DNA 复制过程;因此, 它们也抑制了快速分裂正常细胞。每一个生物体都有 topoisomerases, 而这些拓扑抑制剂最有可能影响环境生态系统6,10,11。因此, 使用模型动物的药物毒理学评价平台对药品的开发和环境风险评估都是有价值的。
在本文中, 我们描述的详细的协议, 以测试毒性的依托, 这是一个临床抗癌剂的目标拓扑 II, 作为一个模型有毒化学品在线虫。为此, 我们将描述的测量方法的身体大小和总数量的鸡蛋奠定了在线虫处理的依托。

<table><tbody><tr><td>Agar</td><td>Affymetrix, USA</td><td>10906</td><td></td></tr><tr><td>&lt;em&gt;Caenorhabditis elegans&lt;/em&gt; N2</td><td>Caenorhabditis Genetics Center (CGC)</td><td></td><td>Wild type</td></tr><tr><td>Cholesterol</td><td>Sigma, USA</td><td>C3045</td><td></td></tr><tr><td>Dimethyl sulfoxide</td><td>Sigma, USA</td><td>D2650</td><td></td></tr><tr><td>&lt;em&gt;Escherichia coli &lt;/em&gt;OP50</td><td>Caenorhabditis Genetics Center (CGC)</td><td></td><td></td></tr><tr><td>Etoposide</td><td>Sigma, USA</td><td>E1383</td><td></td></tr><tr><td>Image J software (ver 1.4)</td><td>Natinoal Institute of Health, USA</td><td></td><td>https://imagej.nih.gov/ij/</td></tr><tr><td>Microscope camera</td><td>Jenopitk, Progress Gryphax, Germany</td><td></td><td></td></tr><tr><td>Peptone</td><td>Merck, USA</td><td>107213</td><td></td></tr><tr><td>35 &amp;times; 10 mm Petri dish</td><td>SPL Life Sciences, South Korea</td><td>10035</td><td></td></tr><tr><td>90 &amp;times; 15 mm Petri dish</td><td>SPL Life Sciences, South Korea</td><td>10090</td><td></td></tr><tr><td>Stereo microscope</td><td>Nikon, Japan</td><td>SMZ800N</td><td></td></tr><tr><td>Yeast extract</td><td>Becton Dickinson, USA</td><td>212750</td><td></td></tr></tbody></table>

measuring the effect of chemicals on the growth and reproduction of caenorhabditis elegans

毒理学评价对于了解化学物质对生物科学在基础和应用生物学领域的影响至关重要。哺乳土壤圆形蠕虫,线虫线虫, 是一个有价值的模型生物毒理学研究, 因为它的方便和缺乏动物伦理问题与哺乳动物的动物系统相比。在本议定书中, 详细介绍了在线虫中对化学品进行毒理学评价的过程。一种临床抗癌药物, 依托, 它的目标人类拓扑 II 和抑制 DNA 复制的人癌细胞, 被选为一个模型试验化学。年龄同步的线虫卵被暴露在二甲基亚砜 (亚砜) 或依托, 然后通过立体显微镜观察, 每天4天监测c. 线虫的生长情况。用依托和亚甲基亚砜处理的从线虫中得到的卵总数也用立体显微镜计算。依托治疗显著影响了线虫的生长和繁殖。通过对不同药剂处理期的蚯蚓卵总数的比较, 可以断定, 化学物质对线虫繁殖的生殖毒性是可逆的或不可逆转的。这些协议可能有助于发展各种药物和风险评估的环境毒物。

依托 (24-96 h) 的治疗明显延缓了线虫的生长。经过 96 h 的孵化, 依托处理的蠕虫生长到0.86 毫米的体长, 而车辆处理的蠕虫增长到1.04 毫米 (图 1)。在立体显微镜观察下, 生长迟缓也有明显的观察 (图 2)。我们开始在72小时的孵化中看到由汽车处理过的蠕虫的卵。另一方面, 在96小时的依托治疗后观察卵子。从这些数据, 我们推?...

Watch this Scientific Journal Video about Measuring the Effect of Chemicals on the Growth and Reproduction of Caenorhabditis elegans at JoVE.com

Measuring the Effect of Chemicals on the Growth and Reproduction of Caenorhabditis elegans

一个基本的协议, 以评估化学品的毒性在模型动物,线虫线虫, 描述。该方法对药品的开发和各种环境污染物的风险评估都具有很好的实用价值。

测定化学物质对线虫线虫生长和繁殖的影响

Toxicological evaluation is crucial for understanding the effects of chemicals on living organisms in basic and applied biological science fields. A ...

measuring-effect-chemicals-on-growth-reproduction-caenorhabditis

Research

JoVE Journal

Medicine

Measuring the Effect of Chemicals on the Growth and Reproduction of Caenorhabditis elegans

16.9K 次观看.  Korea Institute of Science and Technology. 一个基本的协议, 以评估化学品的毒性在模型动物,线虫线虫, 描述。该方法对药品的开发和各种环境污染物的风险评估都具有很好的实用价值。

视频: Measuring the Effect of Chemicals on the Growth and Reproduction of Caenorhabditis elegans

Comprehensive Assessment of Germline Chemical Toxicity Using the Nematode Caenorhabditis elegans

Identifying the reproductive toxicity of the thousands of chemicals present in our environment has been one of the most tantalizing challenges in the field of environmental health. This is due in part to the paucity of model systems that can (1) accurately recapitulate keys features of reproductive processes and (2) do so in a medium- to high-throughput fashion, without the need for a high number of vertebrate animals.
We describe here an assay in the nematode C. elegans that allows the rapid identification of germline toxicants by monitoring the induction of aneuploid embryos. By making use of a GFP reporter line, errors in chromosome segregation resulting from germline disruption are easily visualized and quantified by automated fluorescence microscopy. Thus the screening of a particular set of compounds for its toxicity can be performed in a 96- to 384-well plate format in a matter of days. Secondary analysis of positive hits can be performed to determine whether the chromosome abnormalities originated from meiotic disruption or from early embryonic chromosome segregation errors. Altogether, this assay represents a fast first-pass strategy for the rapid assessment of germline dysfunction following chemical exposure.

Comprehensive Assessment of Germline Chemical Toxicity Using the Nematode Caenorhabditis elegans

We describe the detailed steps of a high-throughput chemical assay in the nematode Caenorhabditis elegans used to assess germline toxicity. In this assay, disruption of germline function following chemical exposure is monitored using a fluorescent reporter specific to aneuploid embryos.

生殖系化学毒性的利用线虫综合评估<em&gt;秀丽隐杆线虫</em

Identifying the reproductive toxicity of the thousands of chemicals present in our environment has been one of the most tantalizing challenges in the ...

科研

发育生物学

A Simple Method for High Throughput Chemical Screening in Caenorhabditis Elegans

Caenorhabditis elegans is a useful organism for testing chemical effects on physiology. Whole organism small molecule screens offer significant advantages for identifying biologically active chemical structures that can modify complex phenotypes such as lifespan. Described here is a simple protocol for producing hundreds of 96-well culture plates with fairly consistent numbers of C. elegans in each well. Next, we specified how to use these cultures to screen thousands of chemicals for effects on the lifespan of the nematode C. elegans. This protocol makes use of temperature sensitive sterile strains, agar plate conditions, and simple animal handling to facilitate the rapid and high throughput production of synchronized animal cultures for screening.

A Simple Method for High Throughput Chemical Screening in Caenorhabditis Elegans

Here we describe a simple protocol for rapidly producing hundreds of nematode growth media agar, 96-well culture plates with consistent numbers of Caenorhhabditis elegans per well. These cultures are useful for the phenotypic screening of whole organisms. We focus here on using these cultures to screen chemicals for pro-longevity effects.

一种简单的高通量化学筛选方法在秀丽线虫中的应用

Caenorhabditis elegans is a useful organism for testing chemical effects on physiology. Whole organism small molecule screens offer significant advantages ...

生物化学

A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans

Applying toxicity testing of chemicals in higher order organisms, such as mice or rats, is time-consuming and expensive, due to their long lifespan and maintenance issues. On the contrary, the nematode Caenorhabditis elegans (C. elegans) has advantages to make it an ideal choice for toxicity testing: a short lifespan, easy cultivation, and efficient reproduction. Here, we describe a protocol for the automatic phenotypic profiling of C. elegans in a 384-well plate. The nematode worms are cultured in a 384-well plate with liquid medium and chemical treatment, and videos are taken of each well to quantify the chemical influence on 33 worm features. Experimental results demonstrate that the quantified phenotype features can classify and predict the acute toxicity for different chemical compounds and establish a priority list for further traditional chemical toxicity assessment tests in a rodent model.

A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans

A quantitative method has been developed to identify and predict the acute toxicity of chemicals by automatically analyzing the phenotypic profiling of Caenorhabditis elegans. This protocol describes how to treat worms with chemicals in a 384-well plate, capture videos, and quantify toxicological related phenotypes.

一种用自动表型分析预测线虫化学毒性的高通量测定

Applying toxicity testing of chemicals in higher order organisms, such as mice or rats, is time-consuming and expensive, due to their long lifespan and ...

环境科学

Using Caenorhabditis elegans for Studying Trans- and Multi-Generational Effects of Toxicants

Information about toxicities of chemicals are essential in their application and waste management. For chemicals at low concentrations, the long-term effects are very important in judging their consequences in the environment and on human health. In demonstrating long-term influences, effects of chemicals over generations in recent studies provide new insight. Here, we describe protocols for studying effects of chemicals over multiple generations using free-living nematode Caenorhabditis elegans. Two aspects are presented: (1) trans-generational (TG) and (2) multi-generational effect studies, the latter of which is separated to multi-generational exposure (MGE) and multi-generational residual (MGR) effect studies. The TG effect study is robust with a simple purpose to determine whether chemical exposure to parents can result in any residual consequences on offspring. After the effects are measured on parents, sodium hypochlorite solutions are used to kill the parents and keep the offspring so as to facilitate effect measurement on the offspring. The TG effect study is used to determine whether the offspring are affected when their parent is exposed to the pollutants. The MGE and MGR effect study is systematical used to determine whether continuous generational exposure can result in adaptive responses in offspring over generations. Careful pick-up and transfer are used to distinguish generations to facilitate effect measurement on each generation. We also combined protocols to measure locomotion behavior, reproduction, lifespan, biochemical and gene expression changes. Some example experiments are also presented to illustrate the trans- and multi-generational effect studies.

Using Caenorhabditis elegans for Studying Trans- and Multi-Generational Effects of Toxicants

Trans- and multi-generational effects of persistent chemicals are essential in judging their long-term consequences in the environment and on the human health. We provide novel detailed methods for studying trans- and multi-generational effects using free-living nematode Caenorhabditis elegans.

使用卡内哈布迪炎酶研究有毒物质的跨代和多代效应

Information about toxicities of chemicals are essential in their application and waste management. For chemicals at low concentrations, the long-term ...

测量 秀丽隐杆 线虫食物摄入量和寿命的高通量测定

Quantifying Food Intake in Caenorhabditis elegans by Measuring Bacterial Clearance

Feeding is an essential biological process for an organism&#39;s growth, reproduction, and survival. This assay aims to measure the food intake of Caenorhabditis elegans&#160;(C. elegans), an important parameter when studying the genetics of aging or metabolism. In most species, feeding is determined by measuring the difference between the amount of food provided and the amount left after a given time interval. The method presented here uses the same strategy to determine the feeding of C. elegans. It measures the amount of bacteria, the food source of C. elegans, cleared within 72 h. This method uses 96-well microtiter plates and has allowed the screening of hundreds of drugs for their ability to modulate food intake at a speed and depth not possible in other animal models. The strength of this assay is that it allows to measure feeding and lifespan simultaneously and directly measures the disappearance of food and, thus, is based on the same principles used for other organisms, facilitating species-to-species comparison.

作者聚焦：探索脂毒性与 HFpEF 之间的关系 

This protocol describes an assay to quantify Caenorhabditis elegans feeding rate based on measuring the clearance of bacteria in liquid culture.

通过测量细菌清除率来量化 秀丽隐杆线虫 的食物摄入量

Feeding is an essential biological process for an organism&#39;s growth, reproduction, and survival. This assay aims to measure the food intake of ...

行为学

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the development of more effective treatments is the relative lack of understanding of the molecular underpinnings of the effects of ethanol on behavior. The nematode, Caenorhabditis elegans (C.&#160;elegans), provides a useful model in which to generate and test hypotheses about the molecular effects of ethanol. Here, we describe an assay that has been developed and used to examine the roles of particular genes and environmental factors in behavioral responses to ethanol, in which locomotion is the behavioral output. Ethanol dose-dependently causes an acute depression of crawling on an agar surface. The effects are dynamic; animals exposed to a high concentration demonstrate an initial strong depression of crawling, referred to here as initial sensitivity, and then partially recover locomotion speed despite the continued presence of the drug. This ethanol-induced behavioral plasticity is referred to here as the development of acute functional tolerance. This assay has been used to demonstrate that these two phenotypes are distinct and genetically separable. The straightforward locomotion assay described here is suitable for examining the effects of both genetic and environmental manipulations on these acute behavioral responses to ethanol in C.&#160;elegans.

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

C. elegans is a useful model for studying the effects of ethanol on behavior. We present a behavioral assay that quantifies the effects of ethanol on the locomotion speed of crawling worms; both initial sensitivity and the development of acute functional tolerance to ethanol can be measured with this assay.

测量乙醇的影响上的步态速度的测定<em&gt;秀丽隐杆线虫</em

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the ...

神经科学

Measuring Caenorhabditis elegans Life Span in 96 Well Microtiter Plates

Lifespan is a biological process regulated by several genetic pathways. One strategy to investigate the biology of aging is to study animals that harbor mutations in components of age-regulatory pathways. If these mutations perturb the function of the age-regulatory pathway and therefore alter the lifespan of the entire organism, they provide important mechanistic insights1-3.
Another strategy to investigate the regulation of lifespan is to use small molecules to perturb age-regulatory pathways. To date, a number of molecules are known to extend lifespan in various model organisms and are used as tools to study the biology of aging4-16. The number of molecules identified thus far is small compared to the genetic "toolset" that is available to study the biology of aging.
Caenorhabditis elegans is one of the principle models used to study aging because of its excellent genetics and short lifespan of three weeks. More recently, C.elegans has emerged as a model organism for phenotype based drug screens5,7,16-20 because of its small size and its ability to grow in microtiter plates.
Here we present an assay to measure C.elegans lifespan in 96 well microtiter plates. The assay was developed and successfully used to screen large libraries for molecules that extend C.elegans lifespan7. The reliability of the assay was evaluated in multiple tests: first, by measuring the lifespan of wild type animals grown at different temperatures; second, by measuring the lifespan of mutants with altered lifespans; third, by measuring changes in lifespan in response to different concentrations of the antidepressant Mirtazepine. Mirtazepine has previously been shown to extend lifespan in C.elegans7. The results of these tests show that the assay is able to replicate previous findings from other assays and is quantitative. The microtiter format also makes this lifespan assay compatible with automated liquid handling systems and allows integration into automated platforms.

Measuring Caenorhabditis elegans Life Span in 96 Well Microtiter Plates

In this protocol we present a method to measure Caenorhabditis elegans lifespan in 96 well microtiter plates.

测量线虫寿命在96孔板

Lifespan is a biological process regulated by several genetic pathways. One strategy to investigate the biology of aging is to study animals that harbor ...

生物学

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

C. elegans egg-laying behavior is affected by environmental cues such as osmolarity1 and vibration2. In the total absence of food C. elegans also cease egg-laying and retain fertilized eggs in their uterus3. 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. elegans4. 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 acetylcholine5-9.

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

An egg-in-worm (EIW) assay is a useful method to quantify egg-laying behavior. Alterations in egg laying can be a behavioral response of the model organism Caenorhabditis elegans to potentially harmful environmental substances such as those produced by pathogenic bacteria.

测量细菌的影响<em&gt; C。线虫</em&gt;行为使用一个鸡蛋保留含量

C. elegans egg-laying behavior is  affected by environmental cues such as osmolarity1 and  vibration2. In the total absence of food C. elegans also cease ...

测定化学物质对线虫线虫生长和繁殖的影响

摘要

探索更多视频

此视频中的章节

生殖系化学毒性的利用线虫综合评估

一种简单的高通量化学筛选方法在秀丽线虫中的应用

一种用自动表型分析预测线虫化学毒性的高通量测定

使用卡内哈布迪炎酶研究有毒物质的跨代和多代效应

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

测量乙醇的影响上的步态速度的测定

测量线虫寿命在96孔板

测量细菌的影响

测定化学物质对线虫线虫生长和繁殖的影响

摘要

探索更多视频

此视频中的章节

生殖系化学毒性的利用线虫综合评估

一种简单的高通量化学筛选方法在秀丽线虫中的应用

一种用自动表型分析预测线虫化学毒性的高通量测定

使用卡内哈布迪炎酶研究有毒物质的跨代和多代效应

通过测量细菌清除率来量化 秀丽隐杆线虫 的食物摄入量

通过测量细菌清除率来量化 秀丽隐杆线虫 的食物摄入量

通过测量细菌清除率来量化 秀丽隐杆线虫 的食物摄入量

测量乙醇的影响上的步态速度的测定

测量线虫寿命在96孔板

测量细菌的影响

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量