Characterization of Metallothionein from Asian Sea Bass (Lates calcarifer, Bloch) and Application as a Biomarker for Heavy Metal Exposure in Thailand
In the present study, metallothionein (MT) proteins and cDNA were isolated from Asian sea bass (Lates calcarifer) livers. Immunochemical protocols (i.e. an enzyme-linked immunosorbent assay (ELISA) and Western blot) were developed for quantification of MT protein levels in Asian sea bass. These were applied to analyze MT protein levels in 15 different feral fish species from two different areas, one urban area Angsila and one industrial area Map Ta Phut in the Gulf of Thailand. An MT protein band was detected in Shrimp scad (Alepes djadaba) and Indian ilisha (Ilisha melastoma) from both areas. Higher MT protein expressions were seen in shrimp scad from cadmium-contaminated areas near shore as well as off shore. These results suggest that induction of hepatic MT immunoreactive proteins in the shrimp scad may be a good early warning signal for heavy metal exposure in environmental monitoring programmes in Thailand waters. Protocol for quantification of MT mRNA levels in Asian sea bass was developed using quantitative PCR. Asian sea bass were exposed in the lab to different doses of CdNO3 and sampled at different times after injection. There appears to be a bi-phasic dose-response pattern with highest MT mRNA levels in fish injected with 4 mg CdNO3 /kg. In these fish, the highest expression was seen after one day and lowest induction after three days. These results suggest that induction of MT mRNA levels in Asian sea bass liver can be used as a sensitive early warning signal for cadmium exposure in tropical waters.
Adhikari, S., Ghosh, L., Giri, B.S. and S. Ayyappan (2009). Distributions of metals in the food web of fishponds of Kolleru Lake, India. Ecotoxicology and Environmental Safety, 72: 1242-1248.
Ang, S.G. and P.S. Chong (1998). Purification of metallothionein proteins from the crab, Portunus pelagicus-selectivity of hydrophobic interaction chromatography. Talanta, 45: 693-701.
Castaño, A., Carbonell, G., Carballo, M., Fernandez, C., Boleas, S. and J.V Tarazona (1998). Sublethal effects of repeated intraperitoneal cadmium injections on rainbow trout (Oncorhynchus mykiss). Ecotoxicology and Environmental Safety, 41: 29-35.
Castro-Gonzáalez, M. and I. Méndez-Armenta (2008). Heavy metals: Implications associated to fish consumption. Environmental Toxicology and Pharmacology, 26: 263-271.
Cheung, A.P.L., Lam, T.H.J. and K.M. Chan (2004). Regulation of Tilapia metallothionein gene expression by heavy metals ions. Marine Environmental Research, 58: 389-394.
Cheung, P.L.A., Lam, V.K.L. and K.M. Chan (2005). Tilapia metallothionein genes; PCR-cloning and gene expression studies. Biochimica et Biophysica Acta, 1731: 191-201.
George, S.G., Todd, K. and J. Wright (1996). Regulation of metallothionein in Teleost: Induction of MT mRNA and protein by cadmium in hepatic and extra hepatic tissues of marine flatfish, the turbot (Scophthalmus maximus). Comparative Biochemical and Physiology, 113C(2): 109-115.
Hennies, M., Wiesmann, M., Allner, B. and H. Sauerwein (2003). Vitellogenin in carp (Cyprinus carpio) and perch (Perca fluviatilis): Purification, characterization and development of an ELISA for the detection of estrogenic effects. The Science of the Total Environment, 309: 93-103.
Honda, R.T., Araújo, R.M., Horta, B.B., Val, A.L. and M. Demasi (2005). One-step purification of metallothionein extracted from two different sources. Journal of Chromatography B, 820: 205-210.
Kachanopas-Barnette, P., Mokkongpai, P., Wassmur, B., Celander, M.C. and P. Sawangwong (2010). Molecular characterization of cytochrome P450 1A (CYP1A) in Asian sea bass (Lates calcalifer Bloch) and its application as a biomarker in the Gulf of Thailand. Asian Journal of Water, Environment and Pollution. 7(2): 43-51.
Kim, J.H., Wang, S.Y., Kim, I.C., Ki, J.S., Raisuddin, S., Lee, J.S. and K.N. Han (2008). Cloning of river pufferfish (Takifugu obscurrus) metallothionein cDNA and study of its induction profile in cadmium-exposed fish. Chemosphere, 71: 1251-1259.
Klaassen, C.D., Lui, J. and S. Choudhuri (1999). Metallothionein: An Intracellular Protein to Protect Against Cadmium Toxicity. Annual Review of Pharmacology and Toxicology, 39: 267-294.
Knapen, D., Reynders, H., Bervoets, L., Verheyen, E. and R. Blust (2007). Metallothionein gene and protein expression as a biomarker for metal pollution in natural gudgeon populations. Aquatic Toxicology, 82: 163-172.
Lacorn, M., Lahrssen, A., Rotzoll, N., Simat, T.J. and H. Steinhart (2001). Quantification of metallothionein isoforms in fish liver and its implications for biomonitoring. Environmental Toxicology and Chemistry, 20(1): 140-145.
Langston, W.J., Chesman, B.S., Burt, G.R., Pope, N.D. and J. McEvoy (2001). Metallothionein in liver of eels Anguilla anguilla from the Thames Estuary: An indicator of environmental quality? Marine Environmental Research, 53: 263-293.
Linde, A.R., Sánchez-Galán, S., Vallés-Mota, P. and E. García-Vázquez (2001). Metallothionein as bioindicator of freshwater metal pollution: European eel and brown trout. Ecotoxicology and Environmental Safety, 49: 60-63.
Mihailović, M., Petrovic, M., Grdović, N., Dinic, S., Uskoković, A., Vidaković, M., Grigorov, I., Bogojević, D., Ivanović-Matić, S., Martinović, V., Arambašić, J., Joksimović, D., Labus-Blagojević, S. and G. Poznanović (2010). CYP1A and metallothionein expression in the hepatopancreas of Merluccius merluccius and Mullus barbatus from the Adriatic sea. Journal of Serbian Chemical Society, 75(8): 1149-1159.
Montaser, M., Mahfouz, M.E., El-Shazly, S.A.M., Abdel-Rahman, G.H. and S. Bakry (2010). Toxicity of heavy metals on fish at Jeddah coast KSA: Metallothionein expression as a biomarker and histopathological study on liver and gills. Journal of Fish and Marine Science, 2(3): 174-185.
Okocha, R.C. and O.B. Adedeji (2011). Overview of Cadmium Toxicity in Fish. Journal of Applied Sciences Research, 7(7): 1195-1207.
Olsvik, P.A., Hindar, K., Zachariassen, K.E. and R.F. Anderson (2001). Brown trout (Salmo trutta) metallothionein as biomarkers for metal exposure in two Norwegian rivers. Biomarker, 6(4): 274-288.
Ruangsomboon, S. and L. Wongrat (2006). Bioaccumulation of cadmium in an experimental Aquatic food chain involving phytoplankton (Chlorella vulgaris), zooplankton (Moina macrocopa), and the predator catfish Clarias macrocephalus × C. gariepinus. Aquatic Toxicology, 78: 15-20.
Saengsupavanich, C., Coowannitwong, N., Gallardo, W.G. and C. Lertsuchatavanich (2009). Environmental performance evaluation of an industrial port and estate: ISO14001, port state control-derived indicators. Journal of Cleaner Production, 17: 154-161.
Tom, M., Chen, N., Segev, M., Herut, B. and B. Rinkevich (2004). Quantifying fish metallothionein transcript by real time PCR for its utilization as an environmental biomarker. Marine Pollution Bulletin, 48: 705-710.
Vergani, L., Grattorola, M., Donderoand, F. and A. Viarengo (2003). Expression, purification, and characterization of metallothionein-A from rainbow trout. Protein Expression and Purification, 27: 338-345.
Wu, J.P. and H.P. Chen (2005). Metallothionein induction and heavy metal accumulation in white shrimp Litopenaeus vannamei exposed to cadmium and zinc. Comparative Biochemistry and Physiology C Toxicology Pharmacology, 140(3-4): 383-394.
Wu, S.M., Shih, M. and Y.C. Ho (2007). Toxicological stress response and cadmium distribution in hybrid tilapia (Oreochromis sp.) upon cadmium exposure. Comparative Biochemistry and Physiology, Part C, 145: 218-226.
Yudkovski, Y., Rogowska-Wrzesinska, A., Yankelevich, I., Shefer, E., Herut, B. and M. Tom (2008). Quantitative immunochemical evaluation of fish metallothionein upon exposure to cadmium. Marine Environmental Research, 65: 427-436.