Browse > Article

Gene Structure and Altered mRNA Expression of Metallothionein in Response to Metal Exposure and Thermal Stress in Miho Spine Loach Cobitis choii (Cobitidae; Cypriniformes)  

Lee, Sang-Yoon (Department of Marine Bio-Materials & Aquaculture, Pukyong National University)
Nam, Yoon-Kwon (Department of Marine Bio-Materials & Aquaculture, Pukyong National University)
Publication Information
Korean Journal of Ichthyology / v.23, no.1, 2011 , pp. 61-69 More about this Journal
Abstract
Gene and promoter structures of metallothionein(MT) from Miho spine loach (Cobitis choii; Cypriniformes) were characterized, and the transcriptional responses to experimental exposures to heavy metals and heat stress were examined. The C. choii metallothionein displayed well-conserved features of teleostean metallothioneins at gDNA, mRNA and amino acid levels. Bioinformatic analysis predicted that the C. choii MT regulatory region potentially possessed various motifs or elements targeted by various transcription factors associated with metal-coordinating regulation (e.g., metal transcription factor-1), immune responses (e.g., nuclear factor kappa B), and thermal modulations (e.g., heat shock factor). Acute heavy-metal exposures to 0.5 or $1.0\;{\mu}M$ of cadmium (Cd), copper (Cu), manganese (Mn), nickel (Ni) or zinc (Zn) showed that MT transcription was significantly stimulated by Cd (9.6-fold relative to non-exposed control) and Cu (10.4-fold), only moderately by Mn (2.4-fold), but hardly by Ni and Zn. Elevation of water temperature from $25^{\circ}C$ to $31^{\circ}C$ caused a rapid modulation of MT mRNAs toward upregulation to 9.5-fold; however, afterward the elevated mRNA level slightly decreased during further incubation at $31^{\circ}C$ for 6 h. Results from this study suggest that MT-based expression assay could be a useful basis for better understanding the metal- and/or heat-caused stresses in this endangered fish species.
Keywords
Cobitis choii; gene structure; heat stress; heavy metals; metallothionein; mRNA expression;
Citations & Related Records
Times Cited By KSCI : 5  (Citation Analysis)
연도 인용수 순위
1 Bourdineaud, J.P., M. Baudrimont, P. Gonzalez and J.L. Moreau. 2006. Challenging the model for induction of metallothionein gene expression. Biochimie, 88: 1787-1792.   DOI   ScienceOn
2 Buckley, B.A., A.Y. Gracey and G.N. Somero. 2006. The cellular response to heat stress in the goby Gillichthys mirabilis: a cDNA microarray and protein-level analysis. J. Exp. Bio., 209: 2660-2677.   DOI   ScienceOn
3 Amiard, J.C., C. Amiard-Triquet, S. Barka, J. Pellerin and P.S. Rainbow. 2006. Metallothioneins in aquatic invertebrates: their role in metal detoxification and their use as biomarkers. Aquat. Toxicol., 76: 160-202.   DOI   ScienceOn
4 Atif, F., M. Kaur, S. Yousuf and S. Raisuddin. 2006. In vitro free radical scavenging activity of hepatic metallothionein induced in an Indian freshwater fish, Channa punctatta Bloch. Chem. Biol. Interact., 162: 172-180.   DOI   ScienceOn
5 Bang, I.C., W.J. Kim and I.R. Lee. 2009. Characterization of polymorphic microsatellite loci in the endangered Miho spine loach (Iksookimia choii) and cross-species amplification within the Cobitidae family. Mol. Ecol.Resour., 9: 281-284.   DOI   ScienceOn
6 Woo, S., S. Yum, J.H. Jung, W.J. Shim, C.H. Lee and T.K. Lee. 2006. Heavy metal-induced differential gene expression of metallothionein in Javanese medaka, Oryzias javanicus. Mar. Biotechnol., 8: 654-662.   DOI   ScienceOn
7 Yan, C.H. and K.M. Chan. 2004. Cloning of zebrafish metallothionein gene and characterization of its gene promoter region in HepG2 cell line. Biochim. Biophys. Acta., 1679: 47-58.   DOI   ScienceOn
8 Swindell, W.R. 2011. Metallothionein and the biology of aging. Ageing Res. Rev., 10: 132-145.   DOI   ScienceOn
9 Thirumoorthy, N., K.T.M. Kumar, A.S. Sundar, L. Panayappan and M. Chatterjee. 2007. Metallothionein: an overview, World J. Gastroenterol., 13: 993-996.   DOI
10 Wang, L., L. Song, D. Ni, H. Zhang and W. Liu. 2009. Alteration of metallothionein mRNA in bay scallop Argopecten irradians under cadmium exposure and bacteria challenge. Comp. Biochem. Physiol. C Pharmacol. Toxicol. Endocrinol., 149: 50-57.   DOI   ScienceOn
11 Ringwood, A.H., M.J. Hameedi, R.F. Lee, M. Brower, E.C. Peters, G.I. Scott, S.N. Luoma and R.T. Di Giulio. 1999. Bivalve biomarker workshop: overview and discussion group summaries. Biomarkers, 4: 391-399.   DOI   ScienceOn
12 Rodriguez-Ortega, M.J., A. Rodriguez-Ariza, J.L. Gomez- Ariza, A. Muñoz-Serrano and J. Lopez-Barea. 2009. Multivariate discriminant analysis distinguishes metalfrom non metal-related biomarker responses in the clam Chamaelea gallina. Mar. Pollut. Bull., 58: 64-71.   DOI   ScienceOn
13 Son, Y.-M. and H.-K. Byeon. 2005. The ichthyofauna and dynamics of the fish community in Miho stream, Korea. Kor. J. Ichthyol., 17: 271-278 (in Korean).
14 Song, H.Y., W.J. Kim, W.O. Lee and I.C. Bang. 2008. Morphological development of egg and larvae of Iksookimia choii (Cobitidae). Korean J. Limnol., 41: 104-110.
15 Lin, C.H., J.A.C. John, L.W. Ou, J.C. Chen, C.H. Lin and C.Y. Chang. 2004. Cloning and characterization of metallothionein gene in ayu Plecoglossus altivelis. Aquat. Toxicol., 66: 111-124.   DOI   ScienceOn
16 Ren, H., M. Xu, P. He, N. Muto, N. Itoh, K. Tanaka, J. Xing and M. Chu. 2006. Cloning of crucian carp (Carassius cuvieri) metallothionein-II gene and characterization of its gene promoter region. Biochem. Biophys. Res. Commun., 342: 1297-1304.   DOI   ScienceOn
17 Rhee, J.S., S. Raisuddin, D.S. Hwang, K.W. Lee, I.C. Kim and J.S. Lee. 2009. Differential expression of metallothionein (MT) gene by trace metals and endocrinedisrupting chemicals in the hermaphroditic mangrove killifish, Kryptolebias marmoratus. Ecotoxicol. Environ. Saf., 72: 206-212.   DOI   ScienceOn
18 Laity, J.H. and G.K. Andrews. 2007. Understanding the mechanisms of zinc-sensing by metal-response element binding transcription factor-1 (MTF-1). Arch. Biochem. Biophys., 463: 201-210.   DOI   ScienceOn
19 Lee, S.Y., O. Stoliar and Y.K. Nam. 2010. Transcriptional alteration of two metallothionein isoforms in mud loach (Misgurnus mizolepis) fry during acute heavy metal exposure. Fish. Aqua. Sci., 13: 112-117.
20 Lee, Y.A., Y.E. Yun, Y.K. Nam and I.C. Bang. 2008. Genetic diversity of an endangered fish Iksookimia choii (Cypriniformes), from Korea as assessed by amplified fragment length polymorphism. Kor. J. Limnol., 41: 98-103.
21 Kalman, J., I. Riba, T. Angel DelValls and J. Blasco. 2010. Comparative toxicity of cadmium in the commercial fish species Sparus aurata and Solea senegalensis. Ecotoxicol. Environ. Saf., 73: 306-311.   DOI   ScienceOn
22 Kim, K.-Y., S.Y. Lee, Y.S. Cho, I.C. Bang, D.S. Kim and Y.K. Nam. 2008. Characterization and phylogeny of two beta-cytoskeletal actins from Hemibarbus mylodon (Cyprinidae, Cypriniformes), a threatened fish species in Korea. DNA Seq., 19: 87-97.   DOI
23 Klaassen, C.D., J. Liu and B.A. Diwan. 2009. Metallothionein protection of cadmium toxicity. Toxicol. Appl. Pharmacol., 238: 215-220.   DOI   ScienceOn
24 Knapen, D., E.S. Redeker, I. Inácio, W. De Coen, E. Verheyen and R. Blust. 2005. New metallothionein mRNAs in Gobio gobio reveal at least three gene duplication events in cyprinid metallothionein evolution. Comp. Biochem. Physiol. C Pharmacol. Toxicol. Endocrinol., 140: 347-355.   DOI   ScienceOn
25 Guo, R., H. Ma, F. Gao, L. Zhong and J. Ren. 2009. Metallothionein alleviates oxidative stress-induced endoplasmic reticulum stress and myocardial dysfunction. J. Mol. Cell. Cardiol., 47: 228-237.   DOI   ScienceOn
26 He, P., M. Xu and H. Ren. 2007. Cloning and functional characterization of 5′-upstream region of metallothionein- I gene from crucian carp (Carassius cuvieri). Int. J. Biochem. Cell Biol., 39: 832-841.   DOI   ScienceOn
27 Hermesz, E., M. Abraham and J. Nemcsok. 2001. Tissuespecific expression of two metallothionein genes in common carp during cadmium exposure and temperature shock. Comp. Biochem. Physiol. C Toxicol. Pharmacol., 128: 457-465.   DOI   ScienceOn
28 Ferencz, A. and E. Hermesz. 2009. Identification of a splice variant of the metal-responsive transcription factor MTF-1 in common carp. Comp. Biochem. Physiol. C Toxicol. Pharmacol., 150: 113-117.   DOI   ScienceOn
29 Fu, Z., J. Guo, L. Jing, R. Li, T. Zhang and S. Peng. 2010. Enhanced toxicity and ROS generation by doxorubicin in primary cultures of cardiomyocytes from neonatal metallothionein-I/II null mice. Toxicol. In Vitro, 24: 1584-1591.   DOI   ScienceOn
30 Gao, D., G.T. Wang, X.T. Chen and P. Nie. 2009. Metallothionein- 2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization. Comp. Biochem. Physiol. C Toxicol. Pharmacol., 149: 18-25.   DOI   ScienceOn
31 Cho, Y.S., S.Y. Lee, K.-Y. Kim and Y.K. Nam. 2009c. Two metallothionein genes from mud loach Misgurnus mizolepis (Teleostei; Cypriniformes): Gene structure, genomic organization, and mRNA expression analysis. Comp. Biochem. Physiol. B Biochem. Mol. Biol., 153: 317-326.   DOI   ScienceOn
32 Dragun, Z., M. Podrug and B. Raspor. 2009. Combined use of bioindicators and passive samplers for the assessment of the river water contamination with metals. Arch. Environ. Contam. Toxicol., 57: 211-220.   DOI   ScienceOn
33 Falfushynska, H.I., L.L. Gnatyshyna, C.V. Priydun, O.B. Stoliar and Y.K. Nam. 2010. Variability of responses in the crucian carp Carassius carassius from two Ukrainian ponds determined by multi-marker approach. Ecotoxicol. Environ. Saf., 73: 1896-1906.   DOI   ScienceOn
34 Cho, Y.S., I.C. Bang, I.R. Lee and Y.K. Nam. 2009a. Hepatic expression of Cu/Zn-superoxide dismutase transcripts in response to acute metal exposure and heat stress in Hemibarbus mylodon (Teleostei: Cypriniformes). Fish. Aqua. Sci., 12: 179-184.
35 Cho, Y.S., S.Y. Lee, C.H. Noh, Y.K. Nam and D.S. Kim. 2006. Survey of genes responsive to long-term heat stress using a cDNA microarray analysis in mud loach (Misgurnus mizolepis) liver. Kor. J. Ichthyol., 18: 65-77.
36 Cho, Y.S., S.Y. Lee, K.H. Kim, S.K. Kim, D.S. Kim and Y.K. Nam. 2009b. Gene structure and differential modulation of multiple rockbream (Oplegnathus fasciatus) hepcidin isoforms resulting from different biological stimulations. Dev. Comp. Immunol., 33: 46-58.   DOI   ScienceOn
37 Cho, Y.S., S.Y. Lee, K.-Y. Kim, I.C. Bang, D.S. Kim and Y.K. Nam. 2008. Gene structure and expression of metallothionein during metal exposures in Hemibarbus mylodon, Ecotoxicol. Environ. Saf., 71: 125-137.   DOI   ScienceOn
38 Chen, W.Y., J.A.C. John, C.-H. Lin, H.-F. Lin, S.-C. Wu, C.-H. Lin and C.-Y. Chang. 2004. Expression of metallothionein gene during embryonic and early larval development in zebrafish. Aquat. Toxicol., 69: 215-227.   DOI   ScienceOn
39 Cheung, A.P.L., V.K.L. Lam and K.M. Chan. 2005. Tilapia metallothionein genes: PCR-cloning and gene expression studies. Biochim. Biophys. Acta., 1731: 191-201.   DOI   ScienceOn
40 Cho, Y.S., B.N. Choi, E.M. Ha, K.H. Kim, S.K. Kim, D.S. Kim and Y.K. Nam. 2005. Shark (Scyliorhinus torazame) metallothionein: cDNA cloning, genomic sequence, and expression analysis. Mar. Biotechnol., 7: 350-362.   DOI   ScienceOn
41 Bi, Y., G.X. Lin, L. Millecchia and Q. Ma. 2006. Superinduction of metallothionein I by inhibition of protein synthesis: role of a labile repressor in MTF-I mediated gene transcriptioin. J. Biochem. Mol. Toxicol., 20: 57-68.   DOI   ScienceOn