Name: measuring the effect of chemicals on the growth and reproduction of caenorhabditis elegans
Uploaded: 2017-10-05T15:00:00
Description: Watch this Scientific Journal Video about Measuring the Effect of Chemicals on the Growth and Reproduction of Caenorhabditis elegans at JoVE.com

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>...

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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>Caenorhabditis elegans 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>Escherichia coli 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 &#215; 10 mm Petri dish</td>
			<td>SPL Life Sciences, South Korea</td>
			<td>10035</td>
			<td></td>
		</tr>
		<tr>
			<td>90 &#215; 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 ...

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

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