Li Zhuo, Ai Fangting, Zhang Jing, Yu Zhenyang, Yin Daqiang
College of Environmental Sciences and Engineering, Tongji University; Shanghai Institute of Pollution Control and Ecological Security.
Jiaxing Tongji Institute of Environment.
J Vis Exp. 2019 Jul 29(149). doi: 10.3791/59367.
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.
化学品毒性信息在其应用和废物管理中至关重要。对于低浓度化学品，长期影响对于判断其对环境和人类健康的后果非常重要。在证明长期影响方面，近期研究中化学品对多代的影响提供了新的见解。在此，我们描述了使用自由生活的线虫秀丽隐杆线虫研究化学品对多代影响的实验方案。介绍了两个方面：（1）跨代（TG）和（2）多代效应研究，后者又分为多代暴露（MGE）和多代残留（MGR）效应研究。TG效应研究稳健，其简单目的是确定亲代接触化学品是否会对后代产生任何残留影响。在对亲代测量影响后，使用次氯酸钠溶液杀死亲代并保留后代，以便于对后代进行影响测量。TG效应研究用于确定当亲代接触污染物时后代是否受到影响。MGE和MGR效应研究系统地用于确定连续几代暴露是否会导致后代产生适应性反应。通过仔细挑选和转移来区分世代，以便于对每一代进行影响测量。我们还结合了测量运动行为、繁殖、寿命、生化和基因表达变化的实验方案。还给出了一些示例实验来说明跨代和多代效应研究。
