Specific activation of the human HSP70 promoter by copper sulfate in mosaic transgenic zebrafish

Seok, Seung Hyeok; Park, Jong Hwan; Baek, Min Won; Lee, Hui Young; Kim, Dong Jae; Uhm, Hyung Min; Hong, Jung Joo; Na, Yi Rang; Jin, Bo Hwan; Ryu, Doug-Young; Park, Jae Hak

doi:10.1016/j.jbiotec.2006.04.029

Cited by 24 publications

(15 citation statements)

References 37 publications

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“…Prolonged incubation induced the expression of EGFP in liver (where MT is not expressed in developing fi sh, Chen et al, 2004). Our results also differ from those previously reported, in which mosaic transgenic fish carrying EGFP under the control of the exogenous human hsp70 promoter were exposed to copper (Seok et al, 2006). These authors show mosaic EGFP expression in gills, skin epithelium, auditory epithelium and myotubes.…”

Section: Discussioncontrasting

confidence: 99%

“…This is in contrast to the stable transgenic Tg(hsp70:egfp) line used in our experiments that expresses EGFP in the entire embryo. In addition, these authors reported a calculated LC50 of 1.2 μM of copper in contrast to the value of 13.82 μM calculated in our experiments, a difference of an order of magnitude (Seok et al, 2006). There are several possible explanations for these differences such as the strain of fi sh used (AB strain in our study versus a non-typifi ed strain), ion composition of the media (E3 in our experiments versus Ringer's solution) and water hardness (Long et al, 2004;Van Genderen et al, 2007).…”

Section: Discussioncontrasting

confidence: 99%

“…This protein has been reported to be induced by a variety of heavy metals including Zn, Cd, Hg, Ag and Cu (Murata et al, 1999;Williams et al, 1990) and this activation is mediated by the heat shock transcription factor HSF-1 (Mosser et al, 1988;Murata et al, 1999). Recent work has shown that copper can activate the human hsp70 promoter in mosaic transgenic fi sh (Seok et al, 2006), and previous reports show that heat shock and cadmium induce the expression of EGFP in Tg(hsp70:egfp) fi sh expressing EGFP driven by the hsp70 gene promoter (Blechinger et al, 2002a;Blechinger et al, 2002b;Blechinger et al, 2007;Matz and Krone, 2007). Using these transgenic fish, we found that waterborne copper induces EGFP expression in a concentration-dependent manner.…”

Section: Discussionmentioning

confidence: 99%

“…However, there is little information on the effects of copper on early fish development and its potential toxicity at sub-lethal concentrations (around 5 μM of Cu) (Hernandez et al, 2006;Sandahl et al, 2007;Seok et al, 2006). In adult fi sh, copper uptake is largely regulated by the gills (Grosell et al, 2004;Kamunde et al, 2003;Matsuo et al, 2004;Taylor et al, 2003), but in developing embryos, prior to gill development; other mechanisms could account for this process.…”

Section: Introductionmentioning

confidence: 99%

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Sublethal concentrations of waterborne copper induce cellular stress and cell death in zebrafish embryos and larvae

et al. 2011

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Copper is an essential ion that forms part of the active sites of many proteins. At the same time, an excess of this metal produces free radicals that are toxic for cells and organisms. Fish have been used extensively to study the effects of metals, including copper, present in food or the environment. It has been shown that different metals induce different adaptive responses in adult fi sh. However, until now, scant information has been available about the responses that are induced by waterborne copper during early life stages of fi sh. Here, acute toxicity tests and LC50 curves have been generated for zebrafi sh larvae exposed to dissolved copper sulphate at different concentrations and for different treatment times. We determined that the larvae incorporate and accumulate copper present in the medium in a concentrationdependent manner, resulting in changes in gene expression. Using a transgenic fi sh line that expresses enhanced green fl uorescent protein (EGFP) under the hsp70 promoter, we monitored tissue-specifi c stress responses to waterborne copper by following expression of the reporter. Furthermore, TUNEL assays revealed which tissues are more susceptible to cell death after exposure to copper. Our results establish a framework for the analysis of whole-organism management of excess external copper in developing aquatic animals.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Sublethal concentrations of waterborne copper induce cellular stress and cell death in zebrafish embryos and larvae

et al. 2011

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“…These results suggest that the regulatory components of the PtHSP70 promoter may be conserved among these species and that the PtHSP70 promoter may be applied as a universal promoter to other Porphyra species. Previous studies have reported that HSP70 proteins are highly conserved and that its promoter can be active in heterologous hosts (Seok et al 2006;Spena et al 1985;Whitley et al 1999). Thus, these previous studies support our results showing the ability of the PtHSP70 promoter to drive expression similarly in both P. tenera and P. yezoensis.…”

Section: Discussionmentioning

confidence: 99%

Development of an expression system using the heat shock protein 70 promoter in the red macroalga, Porphyra tenera

et al. 2011

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Porphyra is a commercially valuable source of food and drugs, and represents an important model organism for algal research. However, genetic research on P. tenera has been limited due to lack of a heterologous gene expression system. In the present study, we isolated a native promoter, the PtHSP70 promoter, for efficient expression of foreign genes in this organism. This promoter lies approximately 1 kb upstream of the coding sequence for Heat Shock Protein 70 (HSP70) and was isolated using adapter-mediated genomic PCR. Promoter activity was evaluated using the synthetic GUS gene (PyGUS) with optimized codons for Porphyra yezoensis. Interestingly, the PtHSP70 promoter allowed equivalent expression of PyGUS in both P. tenera and P. yezoensis, whereas the GAPDH promoter from P. yezoensis was not fully functional in P. tenera. These data suggest that the PtHSP70 promoter has a more conserved regulatory mechanism than the PyGAPDH promoter between these species.We also established an efficient transient transformation system for P. tenera by evaluating various transformation parameters such as quantity of gold particles, pressure of helium and vacuum, developmental stages of leafy gametophytes, and target distance. Under the optimal conditions of transient transformation, the frequency of GUS expression was determined by histochemical staining to be 30-50 cells per bombardment. Therefore, the new transient transformation system using the PtHSP70 promoter can be used for foreign gene expression in P. tenera, which may advance the development of P. tenera as a model organism.

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Potential Applications of Transgenic Fish to Environmental Monitoring and Toxicology

Lam

Sukardi

et al. 2011

Aquaculture Biotechnology

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Specific activation of the human HSP70 promoter by copper sulfate in mosaic transgenic zebrafish

Cited by 24 publications

References 37 publications

Sublethal concentrations of waterborne copper induce cellular stress and cell death in zebrafish embryos and larvae

Sublethal concentrations of waterborne copper induce cellular stress and cell death in zebrafish embryos and larvae

Development of an expression system using the heat shock protein 70 promoter in the red macroalga, Porphyra tenera

Potential Applications of Transgenic Fish to Environmental Monitoring and Toxicology

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