Roh Ji-Yeon, Park Yeon-Jung, Choi Jinhee
Faculty of Environmental Engineering, College of Urban Science, University of Seoul, 90 Jeonnong-dong, Dongdaemun-gu, Seoul 130-743, Republic of Korea.
Environ Toxicol Pharmacol. 2009 Nov;28(3):409-13. doi: 10.1016/j.etap.2009.07.006. Epub 2009 Jul 17.
To test the applicability of Caenorhabditis elegans mutant for toxicity screening, the sensitivity of cadmium (Cd) in C. elegans was investigated on 14 mutant strains using median lethal concentration (LC50) tests, with further analysis on growth and reproduction conducted on five selected strains. The 24h LC50 of Cd observed on the wildtype and mutant strains of C. elegans was in the order of age-1(hx546)>mtl-2(gk125)>sod-3(gk235)>daf-21(p673)>cyp35a2(gk317)>skn-1(or13)>daf-12(rh62rh157)>hsp-16.2(gk249)>daf-18(e1375)>ctl-2(ok1137)>wildtype(N2)>sod-1(or13)>daf-16(mu86)>cep-1(gk138)>cdr-2(ok1996). Compared to the wildtype response, a decreased reproduction potential was observed in mtl-2(gk125), sod-3(gk235), cdr-2(ok1996) and cep-1(gk138) strains. To gain a mechanistic understanding of different sensitivities of the mutant strains, a time-course gene expression analysis was also performed on the five genes. A dramatic increase in the expression of the mtl-2 gene due to Cd exposure confirmed the importance of this gene in C. elegans Cd toxicity. An increased expression of the sod-3 gene at the longer exposure time period (48h) suggests that oxidative stress may not be a direct toxic mechanism, but may rather be a consequence of Cd toxicity. Even though, LC50 values for the age-1(hx546) mutant strain were the highest among the tested strains, the response on the reproduction potential in age-1(hx546) mutant was unchanged compared to the wildtype, and the age-1 gene expression remained unaltered on exposure to Cd, which may be interpreted as the maintenance of age-1 expression level is needed for the exertion of Cd toxicity; however, the role of the age-1 gene in Cd toxicity may not be via a reproduction-related pathway. The overall results suggest that the C. elegans mutant assay seems to be a promising tool for the study of toxic mechanisms, as well as for toxicity screening in ecotoxicological research.
为了测试秀丽隐杆线虫突变体在毒性筛选中的适用性,利用半数致死浓度(LC50)试验研究了14种突变体菌株对镉(Cd)的敏感性,并对5种选定菌株进行了生长和繁殖的进一步分析。在秀丽隐杆线虫的野生型和突变体菌株上观察到的Cd的24小时LC50顺序为:age-1(hx546)>mtl-2(gk125)>sod-3(gk235)>daf-21(p673)>cyp35a2(gk317)>skn-1(or13)>daf-12(rh62rh157)>hsp-16.2(gk249)>daf-18(e1375)>ctl-2(ok1137)>野生型(N2)>sod-1(or13)>daf-16(mu86)>cep-1(gk138)>cdr-2(ok1996)。与野生型反应相比,在mtl-2(gk125)、sod-3(gk235)、cdr-2(ok1996)和cep-1(gk138)菌株中观察到繁殖潜力下降。为了从机制上理解突变体菌株的不同敏感性,还对这五个基因进行了时间进程基因表达分析。由于Cd暴露导致mtl-2基因表达急剧增加,证实了该基因在秀丽隐杆线虫Cd毒性中的重要性。在较长暴露时间段(48小时)sod-3基因表达增加表明氧化应激可能不是直接的毒性机制,而可能是Cd毒性的结果。尽管age-1(hx546)突变体菌株的LC50值在测试菌株中最高,但age-1(hx546)突变体对繁殖潜力的反应与野生型相比没有变化,并且age-1基因表达在暴露于Cd时保持不变,这可以解释为发挥Cd毒性需要维持age-1表达水平;然而,age-1基因在Cd毒性中的作用可能不是通过与繁殖相关的途径。总体结果表明,秀丽隐杆线虫突变体试验似乎是研究毒性机制以及生态毒理学研究中进行毒性筛选的一种有前途的工具。