Multiparameter Behavioral Analyses Provide Insights to Mechanisms of Cyanide Resistance in Caenorhabditis elegans
Multiparameter Behavioral Analyses Provide Insights to Mechanisms of Cyanide Resistance in Caenorhabditis elegans
复制标题
多参数行为分析为秀丽隐杆线虫的氰化物抗性机制提供了见解
DOI：
10.1093/toxsci/kft138
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发表时间：
2013
影响因子：
3.8
通讯作者：
Powell-Coffman, Jo Anne
中科院分区：
文献类型：
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作者：
Saldanha, Jenifer N.;Parashar, Archana;Pandey, Santosh;Powell-Coffman, Jo Anne
环境毒物影响发育，行为，并最终生存。线虫秀丽隐杆线虫已被证明是一个非常强大的毒理学研究模型。在这里，我们开发新的技术来描述高时空分辨率的氰化物毒性的影响。重要的是，我们使用这些方法来研究抗氰化物的遗传基础。缺乏EGL-9氧传感酶的秀丽隐杆线虫已被证明对病原体铜绿假单胞菌PAO 1产生的氰化氢（HCN）气体具有抗性。我们证明yegl-9突变体的抗氰性完全依赖于HIF-1缺氧诱导因子，并由thecysl-2半胱氨酸合酶介导，该酶可能在清除氰的代谢途径中起作用。此外，cysl-2的表达与在每个遗传背景中表现出的抗氰化物的程度相关。我们发现，每个突变体表现出类似的相对电阻HCN气体板或水溶液氰化钾在微流控室。微流体装置的设计与实时成像相结合，解决了突变表型和毒物化学性质所带来的一系列挑战。微流控分析产生了一组具有增加的分辨率的行为参数，其描述了C中的氰化物毒性和抗性。这在分析微妙的表型时特别有用。对C. elegansbehavior具有巨大的潜力，可作为真实的监测毒物或化学干预的影响以及研究支撑毒物抗性的生物网络的一种手段。
Environmental toxicants influence development, behavior, and ultimately survival. The nematodeCaenorhabditis eleganshas proven to be an exceptionally powerful model for toxicological studies. Here, we develop novel technologies to describe the effects of cyanide toxicity with high spatiotemporal resolution. Importantly, we use these methods to examine the genetic underpinnings of cyanide resistance.Caenorhabditis elegansthat lack the EGL-9 oxygen sensing enzyme have been shown to be resistant to hydrogen cyanide (HCN) gas produced by the pathogenPseudomonas aeruginosaPAO1. We demonstrate that the cyanide resistance exhibited byegl-9mutants is completely dependent on the HIF-1 hypoxia-inducible factor and is mediated by thecysl-2cysteine synthase, which likely functions in metabolic pathways that inactivate cyanide. Further, the expression ofcysl-2correlates with the degree of cyanide resistance exhibited in each genetic background. We find that each mutant exhibits similar relative resistance to HCN gas on plates or to aqueous potassium cyanide in microfluidic chambers. The design of the microfluidic devices, in combination with real-time imaging, addresses a series of challenges presented by mutant phenotypes and by the chemical nature of the toxicant. The microfluidic assay produces a set of behavioral parameters with increased resolution that describe cyanide toxicity and resistance inC. elegans, and this is particularly useful in analyzing subtle phenotypes. These multiparameter analyses ofC. elegansbehavior hold great potential as a means to monitor the effects of toxicants or chemical interventions in real time and to study the biological networks that underpin toxicant resistance.