Van Cleef-Toedt K A, Kaplan L A, Crivello J F
Department of Physiology and Neurobiology, University of Connecticut, Storrs 06269, USA.
Cell Stress Chaperones. 2001 Oct;6(4):351-9. doi: 10.1379/1466-1268(2001)006<0351:kmmref>2.0.co;2.
Metallothionein (MT), a cysteine-rich metal binding protein, is considered to play an essential role in the regulation of intracellular metals. Induction of MT in mammalian and nonmammalian tissues following heavy metal exposure may serve as a defense mechanism and a biomarker of environmental exposure to chemical stressors such as toxic metals. In this study, MT messenger RNA (mRNA) expression was characterized in male and female nonspawning and spawning killifish (Fundulus heteroclitus) following an 8-day exposure to specific sublethal stressors, which included temperature perturbation (26 degrees C or 10 degrees C) and/or 6 ppb of waterborne cadmium chloride (CdCl2). Hepatic, gill, and intestinal MT mRNA, expressed as copy number per microgram of total RNA, was assessed by reverse transcriptase-polymerase chain reaction and electrochemiluminescence using winter flounder (Pleuronectes americanus) MT complementary DNA primers. Liver, gill, and intestine MT mRNA expression was significantly (P < 0.05) increased in nonspawning killifish exposed to 26 degrees C compared with those exposed to 19 degrees C (control). In addition, a significant (P < 0.05) increase in gill MT mRNA induction was observed in nonspawning killifish exposed to 6 ppb of waterborne CdCl2 compared with controls. The results of this study demonstrate significant MT mRNA induction in nonspawning killifish following short-term exposure to physiological and chemical stressors. Thus, further research may be necessary before the use of killifish MT mRNA induction as a biomarker of environmental chemical stress exposure alone.
金属硫蛋白(MT)是一种富含半胱氨酸的金属结合蛋白,被认为在细胞内金属调节中发挥着重要作用。重金属暴露后,哺乳动物和非哺乳动物组织中MT的诱导可能作为一种防御机制以及环境暴露于化学应激源(如有毒金属)的生物标志物。在本研究中,对雄性和雌性未产卵及已产卵的鳉鱼(Fundulus heteroclitus)暴露于特定亚致死应激源8天后的MT信使核糖核酸(mRNA)表达进行了表征,这些应激源包括温度扰动(26摄氏度或10摄氏度)和/或6 ppb的水溶性氯化镉(CdCl2)。使用冬比目鱼(Pleuronectes americanus)MT互补DNA引物,通过逆转录-聚合酶链反应和电化学发光评估肝脏、鳃和肠道中MT mRNA的表达,以每微克总RNA的拷贝数表示。与暴露于19摄氏度(对照)的未产卵鳉鱼相比,暴露于26摄氏度的未产卵鳉鱼肝脏、鳃和肠道MT mRNA表达显著增加(P < 0.05)。此外,与对照组相比,暴露于6 ppb水溶性CdCl2的未产卵鳉鱼鳃中MT mRNA诱导显著增加(P < 0.05)。本研究结果表明,未产卵鳉鱼短期暴露于生理和化学应激源后MT mRNA有显著诱导。因此,在将鳉鱼MT mRNA诱导单独用作环境化学应激暴露的生物标志物之前,可能需要进一步研究。