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转基因线虫作为微波诱导应激的生物监测器。

Transgenic nematodes as biomonitors of microwave-induced stress.

作者信息

Daniells C, Duce I, Thomas D, Sewell P, Tattersall J, de Pomerai D

机构信息

Department of Life Science, University of Nottingham, University Park, UK.

出版信息

Mutat Res. 1998 Mar 13;399(1):55-64. doi: 10.1016/s0027-5107(97)00266-2.

Abstract

Transgenic nematodes (Caenorhabditis elegans strain PC72), carrying a stress-inducible reporter gene (Escherichia coli beta-galactosidase) under the control of a C. elegans hsp16 heat-shock promoter, have been used to monitor toxicant responses both in water and soil. Because these transgenic nematodes respond both to heat and toxic chemicals by synthesising an easily detectable reporter product, they afford a useful preliminary screen for stress responses (whether thermal or non-thermal) induced by microwave radiation or other electromagnetic fields. We have used a transverse electromagnetic (TEM) cell fed from one end by a source and terminated at the other end by a matched load. Most studies were conducted using a frequency of 750 MHz, at a nominal power setting of 27 dBm. The TEM cell was held in an incubator at 25 degrees C inside a shielded room; corresponding controls were shielded and placed in the same 25 degrees C incubator; additional baseline controls were held at 15 degrees C (worm growth temperature). Stress responses were measured in terms of beta-galactosidase (reporter) induction above control levels. The time-course of response to continuous microwave radiation showed significant differences from 25 degrees C controls both at 2 and 16 h, but not at 4 or 8 h. Using a 5 x 5 multiwell plate array exposed for 2 h, the 25 microwaved samples showed highly significant responses compared with a similar control array. The wells most strongly affected were those in the rows closest to the source, whereas the most distant row did not rise above control levels, suggesting a shadow effect. These differential responses are difficult to reconcile with general heating effects, although localised power absorption affords a possible explanation. Experiments in which the frequency and/or power settings were varied suggested a greater response at 21 than at 27 dBm, both at 750 and 300 MHz, although extremely variable responses were observed at 24 dBm and 750 MHz. Thus, lower power levels tended, if anything, to induce larger responses (with the above-mentioned exception), which is opposite to the trend anticipated for any simple heating effect. These results are reproducible and data acquisition is both rapid and simple. The evidence accrued to date suggests that microwave radiation causes measurable stress to transgenic nematodes, presumably reflecting increased levels of protein damage within cells (the common signal thought to trigger hsp gene induction). The response levels observed are comparable to those observed with moderate concentrations (ppm) of metal ions such as Zn2+ and Cu2+. We conclude that this approach deserves further and more detailed investigation, but that it has already demonstrated clear biological effects of microwave radiation in terms of the activation of cellular stress responses (hsp gene induction).

摘要

携带在秀丽隐杆线虫hsp16热休克启动子控制下的应激诱导报告基因(大肠杆菌β-半乳糖苷酶)的转基因线虫(秀丽隐杆线虫PC72株)已被用于监测水和土壤中的毒物反应。由于这些转基因线虫通过合成一种易于检测的报告产物对热和有毒化学物质都有反应,它们为微波辐射或其他电磁场诱导的应激反应(无论是热应激还是非热应激)提供了一种有用的初步筛选方法。我们使用了一个横向电磁(TEM)室,一端由源馈电,另一端由匹配负载终端。大多数研究是在750 MHz频率、27 dBm的标称功率设置下进行的。TEM室置于屏蔽室内25摄氏度的培养箱中;相应的对照进行屏蔽并置于同一25摄氏度的培养箱中;额外的基线对照置于15摄氏度(线虫生长温度)。应激反应通过β-半乳糖苷酶(报告物)诱导高于对照水平来测量。对连续微波辐射的反应时间进程在2小时和16小时时与25摄氏度的对照有显著差异,但在4小时或8小时时没有。使用暴露2小时的5×5多孔板阵列,25个微波处理的样品与类似的对照阵列相比显示出高度显著的反应。受影响最强烈的孔是最靠近源的行中的孔,而最远的行没有超过对照水平,表明存在阴影效应。这些差异反应很难与一般的热效应相协调,尽管局部功率吸收提供了一种可能的解释。改变频率和/或功率设置的实验表明,在750 MHz和300 MHz时,21 dBm时的反应比27 dBm时更大,尽管在24 dBm和750 MHz时观察到极其多变的反应。因此,较低的功率水平如果有什么作用的话,往往会诱导更大的反应(有上述例外),这与任何简单热效应预期的趋势相反。这些结果是可重复的,数据采集既快速又简单。迄今为止积累的证据表明,微波辐射会给转基因线虫造成可测量的应激,大概反映了细胞内蛋白质损伤水平的增加(这是被认为触发hsp基因诱导的共同信号)。观察到的反应水平与中等浓度(ppm)的金属离子如Zn2+和Cu2+观察到的反应水平相当。我们得出结论,这种方法值得进一步和更详细的研究,但它已经在细胞应激反应的激活(hsp基因诱导)方面证明了微波辐射明显的生物学效应。

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