Regulation of Tilapia metallothionein gene expression by heavy metal ions

Tilapia is a common fish species inhabiting inland waters and estuarine regions in Hong Kong and Southeast Asia, and useful for bio-monitoring of metal pollution. Metallothionein (MT) gene expression in fish tissues has been useful to sub-lethal risk assessment as biomarker of exposure to metal ions in fishes inhabiting metal contaminated area. To investigate metal inductions of Tilapia MT gene expression in vivo, Tilapias were injected with different concentrations of heavy metals and tissues were then removed for quantitative PCR assay using mimic PCR methods. All of the metal ions tested (Cu²⁺, Cd²⁺, Hg²⁺, Ni²⁺, Pb²⁺ and Zn²⁺) were able to induce hepatic MT mRNA levels. Renal MT mRNA levels of Cd²⁺ and Zn²⁺ treated fish was not induced with significant fold induction, however MT mRNA levels in gills were sensitive to the administrations of these metal ions. These data indicated that Tilapia MT mRNA levels in gills and liver are sensitive biomarker of exposure to various metal ions.     

重金属对扁额细首纽虫金属硫蛋白的影响

生物体内金属硫蛋白含量变化,可作为环境中重金属污染的生物标志物.通过暴露实验研究了扁额细首纽虫Cephalothrix simula暴露在不同浓度的铜,锌和镉溶液96 h,其体内金属硫蛋白含量的变化规律.结果表明3种重金属离子的胁迫作用均导致金属硫蛋白含量增加,在实验初期(6～24 h),金属硫蛋白含量达到高峰;金属硫蛋白含量与暴露溶液金属浓度具有明显的剂量～效应关系,金属离子浓度越高,金属硫蛋白含量越高;锌离子诱导金属硫蛋白合成的作用大于铜、镉离子.     

Regulation of Tilapia metallothionein gene expression by heavy metal ions

Tilapia is a common fish species inhabiting inland waters and estuarine regions in Hong Kong and Southeast Asia, and useful for bio-monitoring of metal pollution. Metallothionein (MT) gene expression in fish tissues has been useful to sub-lethal risk assessment as biomarker of exposure to metal ions in fishes inhabiting metal contaminated area. To investigate metal inductions of Tilapia MT gene expression in vivo, Tilapias were injected with different concentrations of heavy metals and tissues were then removed for quantitative PCR assay using mimic PCR methods. All of the metal ions tested (Cu(2+), Cd(2+), Hg(2+), Ni(2+), Pb(2+) and Zn(2+)) were able to induce hepatic MT mRNA levels. Renal MT mRNA levels of Cd(2+) and Zn(2+) treated fish was not induced with significant fold induction, however MT mRNA levels in gills were sensitive to the administrations of these metal ions. These data indicated that Tilapia MT mRNA levels in gills and liver are sensitive biomarker of exposure to various metal ions.     

Study on the Adsorption of Metal Ions by Immobilized Marine Algae with the Existence of Clay

The process of adsorption of metal ions by immobilized marine algae with the existence of clay was investigated. It can be noted from the results that, after mixing with clay, the adsorption rate increases rapidly with the increasing amount of the marine algae. When pH=5, the best ratio between the clay and the marine algae is 1：4 for Pb^2＋. The result of in situ handling of the waste water containing heavy metals shows that the average adsorption rates of heavy metal irons Cu^2＋, Cd^2＋, Pb^2＋ and Ni^2＋ are all over 70 %.     

亚甲基蓝在水体系中的光化学降解研究

研究水体系中亚甲基蓝的光化学降解.结果表明,在高压汞灯照射下,亚甲基蓝在人工海水中降解得最快,蒸馏水次之,而在天然海水中降解得最慢.通过对比研究发现.重金属离子(Cu2+,Zn2+,Cd2+,Hg2+)和腐殖酸能够在一定程度上抑制亚甲基蓝的光降解;而丙酮能促进亚甲基蓝的光降解.由此可见,重金属离子和腐殖酸可能是造成亚甲基蓝在天然海水中降解缓慢的主要因素之一.     

Metal uptake in zebrafish embryo-larvae exposed to metal-contaminated sediments

Heavy metal contamination in coastal and estuarine regions of inner Tolo Harbour, as well as some river and nullah areas, is one of the major water-pollution issues in Hong Kong. Zebrafish (Danio rerio) embryo-larvae was used to study metal uptake from coastal sediments collected from Fo Tan River (industrial area, Sha Tin), Tai Po River (residential area, Tai Po) and Wu Kai Sa (rural area, Tolo Harbour). Exposure experiments (7-days) were carried out using different concentrations (0.1%, 0.5%, 1% and 0% as control) of sediments that were added to aquaria containing fertilized zebrafish eggs until they hatched to become larvae. Uptake of heavy metals (Cd, Cu, Pb and Zn) was determined in whole embryo-larvae following exposure. Significant levels of Cd, Cu, Pb and Zn were detected in the embryo-larvae exposed to sediments from Tai Po River. However, significant levels of only Cd and Cu were found in embryo-larvae exposed to sediments from Fo Tan River.     

四种金属离子对中国对虾无节幼体生长的毒性影响

对虾工厂化育苗在我国沿海已经普及,对虾人工养殖也在沿海地区蓬勃发展,到1992年养殖面积已达200多万亩,人工育苗场全国约有上千个。养殖对虾已成为我国水产业中规模最大、经济效益最高的水产品。但近几年来沿海地区的工业飞速发展,沿岸海水均受到了不同程度的污染,各对虾育苗场卵孵化率普遍下降。今年我国养殖对虾又出现了大面积暴发性死亡,原因是复杂的,但作者认为,治虾病之本应是治水质。众所周知,一切生物的幼体阶段都是最敏感的时期,海水中的金属离子对对虾幼体生长的影响已有报道(吴彰宽等,1988; Lawrence et al.,1981),但其实验环境、方法及条件各有不同。作者于1992年5月至1993年4月在海阳育苗场作了现场实验,本文根据实验结果分析和研究了Zn、Cd、Pb、Cu金属离子对中国对虾受精卵及无节幼体生物毒性的影响;不同pH值海水加入不同浓度的混合金属离子,对无节幼体生长的毒性影响;不同盐度的海水加入不同浓度的混合金属离子对无节幼体生长毒性的影响。     

Thermodynamic Analysis of Heavy Metals Biosorption by Two Macroalgae

The thermodynamic process of two macroalgae, Sargassum fusiforme and Lamin aria japonica, absorbing heavy, metal ions Cu^2 , Pb^2 , Cd^2 and Ni^2 . has been studied. The result indicates that the absorption isotherms of these two macroalgae clearly accord with the Landmuir adsorption model. The absorptive processes of S. fusiforme and L. japonica for Cu^2 are endothermal, and at 35℃, the adsorption heat of these two algae is 59.5kJ/mol and 76.8 kJ/mol respectively. Temperature could affect the algae‘s adsorption capacity. Their adsorption capacity increases with temperature (25℃ and 35℃).     

Mycobacteria, but not mercury, induces metallothionein (MT) protein in striped bass, Morone saxitilis, phagocytes, while both stimuli induce MT in channel catfish, Ictalurus punctatus, phagocytes

Recent advances in molecular immunology indicate that the expression of inducible pro-inflammatory proteins is increased in vertebrates in response to both infectious disease agents and various xenobiotics. For example, iNOS, COX-2, and CYP1A are induced by both inflammation and AhR ligands. Moreover, the expression of these proteins in response to stimuli varies among individuals within populations. Little is known of the differences among fish in the inducibility of proinflammatory proteins in response to both infectious agents and xenobiotics. Through random screening of a striped bass, Morone saxitilis, peritoneal macrophage cDNA library, a full length metallothionein (MT) gene was cloned and sequenced. MT is a low-molecular weight (6-8 kDa), cysteine-rich metal binding protein. Metals are required by pathogenic bacteria for growth, and by the host defense system by serving as a catalyst for the generation of reactive oxygen intermediates (ROIs) by phagocytes. A recombinant striped bass MT (rMT) was expressed and purified, then used to generate a specific mAb (MT-16). MT protein expression was followed in freshly isolated striped bass and channel catfish, Ictalurus punctatus, phagocytes after in vitro exposure to the naturally occurring intracellular pathogen Mycobacteria fortuitum or to 0.1 and 1 microM mercury (Hg), as HgCl(2). MT expression was increased by 24 h in both channel catfish and striped bass phagocytes as a result of exposure to M. fortuitum cells. On the other hand, MT was induced by Hg in channel catfish cells, but not those of striped bass. These results indicate that metal homeostasis in phagocytes is different between catfish and striped bass. In addition, these data suggest that care should be taken to distinguish between inflammation-induced vs. metal-induced MT when using MT expression as a biomarker of metal exposure.     

Metallothionein and mercury tolerance in the killifish, fundulus heteroclitus

Synthesis of metallothionein (MT) as a possible mechanism by which Fundulus heteroclitus adapts to heavy metal pollution has been investigated in relation to responses to methylmercuric chloride (meHg) and HgCl₂(Hg²⁺) at different life stages.Clutches of embryos from unpolluted sites exhibit teratogenic responses to a standard dose of meHg or of Hg²⁺ which range from little or no effect (tolerant) to severely affected or almost complete inhibition of development (susceptible). At the time of hatching, untreated embryos from tolerant clutches had twice as much of a protein which co-migrates in SDS-PAGE gels with Fundulus MT as do those of susceptible clutches. This difference could be a mechanism for conveying tolerance to Hg²⁺ and also to meHg if demethylation occurs. However, eggs at the time of deposition had virtually no MT, suggesting that the protein is synthesized too late to provide tolerance in the early stages of development which are critical in the etiology of malformations. Treatment of embryos with non-teratogenic doses of either meHg or Hg²⁺ did not increase the amount of MT-like protein. This may indicate that the thionein gene is already fully active, and would also explain observations that Hg²⁺ pretreatment of embryos does not enhance tolerance to Hg²⁺ in the larval stage.MT, isolated from livers of Hg²⁺- or Cu²⁺-dosed or control adults, is associated with Cu and Zn but not with Hg, although it can bind Hg²⁺in vitro. Fundulus, like other fish, methylates much (ca. 75 %) of its Hg burden, and meHg is considered by some not to react with MT. Fundulus liver has its Hg burden distributed between homogenate sediment (25 %), the cytosol (60 %) and the remainder in lipid. The liver responds to Hg²⁺ treatment by increasing lipid content. This may allow sequestration of some lipid-soluble meHg. Of the cytosolic Hg, most is associated with larger proteins and some (ca. 25 %) is dialyzable.These data suggest that metallothionein is not a significant factor in Hg tolerance in Fundulus adults, and has a questionable role in embryonic and larval tolerance to Hg²⁺ and none to the more toxic meHg.     

The effect of estrogen on cadmium distribution in rainbow trout (Oncorhynchus mykiss)

The effects of injections of 17β-estradiol (E2) and Cd, on the distribution of Cd and the induction of metallothionein (MT) mRNA and vitellogenin mRNA was investigated. Bone and liver were the main organs accumulating Cd. However, E2 redirected the metal accumulation from the bone and liver to the gill, gut and muscle upon exposure to E2. Cd did not induce the hepatic MT mRNA levels in animals treated with E2. The VTG mRNA levels were also reduced following co-injection of E2 and Cd. However, the kidney responded to Cd exposure by upregulating MT mRNA even in the presence of E2 treatment. In the liver the reduced MT mRNA induction led to a redistribution of CD from MT to non-MT proteins. The toxicological significance of these alterations in Cd handling remains to be determined.     

Mycobacteria, but not mercury, induces metallothionein (MT) protein in striped bass, Morone saxitilis, phagocytes, while both stimuli induce MT in channel catfish, Ictalurus punctatus, phagocytes

Recent advances in molecular immunology indicate that the expression of inducible pro-inflammatory proteins is increased in vertebrates in response to both infectious disease agents and various xenobiotics. For example, iNOS, COX-2, and CYP1A are induced by both inflammation and AhR ligands. Moreover, the expression of these proteins in response to stimuli varies among individuals within populations. Little is known of the differences among fish in the inducibility of proinflammatory proteins in response to both infectious agents and xenobiotics. Through random screening of a striped bass, Morone saxitilis, peritoneal macrophage cDNA library, a full length metallothionein (MT) gene was cloned and sequenced. MT is a low-molecular weight (6–8 kDa), cysteine-rich metal binding protein. Metals are required by pathogenic bacteria for growth, and by the host defense system by serving as a catalyst for the generation of reactive oxygen intermediates (ROIs) by phagocytes. A recombinant striped bass MT (rMT) was expressed and purified, then used to generate a specific mAb (MT-16). MT protein expression was followed in freshly isolated striped bass and channel catfish, Ictalurus punctatus, phagocytes after in vitro exposure to the naturally occurring intracellular pathogen Mycobacteria fortuitum or to 0.1 and 1 μM mercury (Hg), as HgCl₂. MT expression was increased by 24 h in both channel catfish and striped bass phagocytes as a result of exposure to M. fortuitum cells. On the other hand, MT was induced by Hg in channel catfish cells, but not those of striped bass. These results indicate that metal homeostasis in phagocytes is different between catfish and striped bass. In addition, these data suggest that care should be taken to distinguish between inflammation-induced vs. metal-induced MT when using MT expression as a biomarker of metal exposure.     

Metallothionein in liver of eels Anguilla anguilla from the Thames Estuary: an indicator of environmental quality?

The purpose of this study was to examine the role of the cysteine-rich protein metallothionein (MT) in the detoxification and metabolism of metals in livers of eels Anguilla anguilla from the Thames Estuary, and to assess the value of MT measurements in environmental monitoring. Hepatic MT levels and associated metal concentrations were determined in A. anguilla collected on three occasions during 1998 at sites along the tidal Thames (from freshwater to the sea) and also from a reference site (Tamar Estuary) in southwest England. MT was present at basal levels of 2 mg g(-1) dry weight (dw), predominantly in association with the essential metals Cu and Zn. MT concentrations were variable within each eel 'population' but were generally highest (up to 11 mg g(-1) in individual eels) at the more contaminated upper- and mid-estuarine sites in the Thames (e.g. Brentford and Kew). Lowest MT levels were those in eels from Blythe Sands at the mouth of the estuary. Season, sex, reproductive status and salinity may have influenced MT levels to some extent, although metals were the most significant factor. Hepatic MT concentrations were highly correlated with associated metal burdens (Zn, Cu, Ag and Cd),--notably reflecting Cu and Ag enrichment in the upper- and mid-estuary. With the exception of Zn, metals in the supernatant fraction of eel livers were primarily associated with MT. Proportions of Cu, Ag and Cd bound to MT increased as a function of accumulated metal burdens, with no indication of saturation. Thus, despite causing induction of MT, excess bioavailable Cu, Ag and Cd appear to be successfully detoxified in eels over the range of environmental contamination encountered along the Thames Estuary. Paradoxically, it may be argued from conventional doctrine that the presence of raised levels of MT, at upstream sites, constitutes a response to contamination and, by definition, signals that the fish are affected by metals; however, obvious signs of deleterious effects were not detected. Superficially, from liver MT data, it would seem that eel populations have become biochemically adapted to metal contamination. The approach tested here has proved viable in helping to interpret trends in bioavailability and the toxicological significance of accumulated metal burdens in Thames eels. A. anguilla is a common inhabitant of estuarine and fresh waters throughout Europe, and related species occur world wide. Therefore, determination of hepatic MT (and associated metals) in eels could be useful for monitoring responses to metal exposure and environmental quality on a much broader basis. Ideally these determinations should be performed in tandem with other sublethal-effects measurements to maximise information.     

用显微注射方法分析牙鲆肌肉发育调节基因MyoD, Myf5的启动子序列

克隆了牙鲆(Paralichthys olivaceus)肌肉发育调节基因MyoD和Myf5的启动子序列,分别为608 bp和1 073 bp。对序列进行分析发现MyoD和Myf5的启动子含有参与肌肉发育调控的基因的结合位点。将MyoD和Myf5的启动子连接到GFP载体上,构建了重组质粒MyoDP-GFP和Myf5P-GFP,并注射到一细胞或两细胞的斑马鱼胚胎中,对这两个基因的启动子进行了瞬时表达研究。结果表明,牙鲆MyoD和Myf5的启动子可以驱动绿色荧光蛋白特异地在斑马鱼肌肉纤维中表达,因此牙鲆608 bpMyoD和1073 bpMyf5的启动子包含着可以驱动MyoD和Myf5正常表达的必需元件,它们是肌肉特异的,并且可以跨物种行使其功能。     

重金属Cd2+、Cu2+对文蛤的急性毒性

用静态急性毒性试验的方法,研究24h、48h和96h内重金属离子Cd2+、Cr+对文蛤的毒性影响.结果表明:Cd2+对文蛤的24h、48h、96h LC50和安全浓度分别为197.35mg/L、40.47mg/L、13.18mg/L和0.13mg/L;Cu2+的24h、48h、96h LC50和安全浓度分别为1.66m...