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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;
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