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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 Elsevier 2006 Journal of biotechnology Vol.126 No.3
<P><B>Abstract</B></P><P>Heat shock proteins (HSPs) play a central role in cell protection and repair upon stresses, such as that caused by heat and heavy metals. Copper sulfate inducibility of a pH<I>hsp70</I> construct expressing the enhanced green fluorescent protein (EGFP) gene under the control of the exogenous human <I>hsp70</I> promoter was tested in transfected CHSE 214 cells and transgenic zebrafish (<I>Danio rerio</I>). We developed a transient expression system, using mosaically transgenic zebrafish, which allows rapid analysis of transgenic expression. Transfected CHSE 214 cells which had been exposed to 250nM and 2.5μM copper sulfate for up to 24h showed increased EGFP expression in a dose-dependent manner. The 1.5μM copper sulfate caused stronger EGFP fluorescence than the 1.0μM copper sulfate in transgenic zebrafish. Most of the expression was spotty and was detected in the gills, dorsal and ventral retina, myotubes of the trunk, and skin epithelium. Transgenic zebrafish exposed to copper sulfate exhibited gross dysmorphogenesis, edema and trunk abnormalities, such as spinal lordosis, in vertebral development 5 days after fertilization. This transgenic zebrafish system was sensitive enough to detect copper sulfate at doses below the median lethal concentration (the LC<SUB>50</SUB> was calculated to be 1.2μM (95% confidence interval of 0.6–1.9μM)). These results indicate that zebrafish could be useful transgenic biosensor systems for the detection of xenobiotic toxicants in the environment.</P>
The Acceptance or Rejection of Syngeneic Murine Tumor Transplantability in C57BL/6 Mice
Seung Hyeok Seok,Jong Hwan Park,Min Won Baek,Hui Young Lee,Dong Jae Kim,Jae Hak Park 한국실험동물학회 2004 Laboratory Animal Research Vol.20 No.1
16.7% rejection of wild-type Lewis lung carcinoma (LLC) 1×10⁶ cells in syngeneicC57BL/6 mice by s.c, injection in the axillary region was observed, even 50.8% rejection with 1×10⁵ cells injection. To address this question, the well-characterized LLC model was chosen for this study. The growth of LLC in syngeneic C57BL/6 mice is accompanied by a weak and transient antitumor cytotoxic response that is later replaced by tumor-enhancing activity, which is correlated with the generation of suppressor cells in LLC-bearing mice. For the rejection of syngeneic tumor transplantation, ultimately "spreading" of the T cell immune response to novel antigenic epitopes needed. In summary, these in vivo transplant studies demonstrate that syngeneic tumor cell transplantation can potentiate the immune response.
In vivo alternative testing with zebrafish in ecotoxicology
Seung-Hyeok Seok,Min-Won Baek,Hui-Young Lee,김동재,나이랑,Kyoung-Jin Noh,Sung-Hoon Park,Hyun-Kyoung Lee,Byoung-Hee Lee,Jae-Hak Park 대한수의학회 2008 Journal of Veterinary Science Vol.9 No.4
Although rodents have previously been used in ecotoxicological studies, they are expensive, time-consuming, and are limited by strict legal restrictions. The present study used a zebrafish (Danio rerio) model and generated data that was useful for extrapolating toxicant effects in this system to that of humans. Here we treated embryos of the naive-type as well as a transiently transfected zebrafish liver cell line carrying a plasmid (phAhREEGFP), for comparing toxicity levels with the well-known aryl hydrocarbon receptor (AhR)-binding toxicants: 3,3’,4,4’,5-pentachlorobiphenyl (PCB126), 2,3,7,8-tetrachlorodibenzo- p-dioxin, and 3-methylcholanthrene. These toxicants induced a concentration-dependent increase in morphological disruption, indicating toxicity at early life-stages. The transient transgenic zebrafish liver cell line was sensitive enough to these toxicants to express the CYP1A1 regulated enhanced green fluorescent protein. The findings of this study demonstrated that the zebrafish in vivo model might allow for extremely rapid and reproducible toxicological profiling of early life-stage embryo development. We have also shown that the transient transgenic zebrafish liver cell line can be used for research on AhR mechanism studies.
Seok, Seung-Hyeok,Koo, Hye Cheong,Kasuga, Asako,Kim, Yeun,Lee, Eun Gae,Lee, Hyeyoung,Park, Jong-Hwan,Baek, Min-Won,Lee, Hui-Young,Kim, Dong-Jae,Lee, Byeung-Hee,Lee, Yong-Soon,Cho, Sang-Nae,Park, Jae-H Elsevier 2006 Veterinary microbiology Vol.114 No.3
<P><B>Abstract</B></P><P>Skin ulcers, scoliosis, and dropsy-like scale edema were observed in laboratory-maintained zebrafish. Affected fish had multifocal granulomas not only in internal organs such as the liver, intestine, genital organs, kidney, muscle, and spleen but also in the fin, epithelium, gills, and sclera of the eyes. Large numbers of acid-fast-rod-shaped bacteria were observed within the necrotic centers of well-demarcated, multifocal granulomas with Gram's stain and Ziehl–Neelson's stain. The size of the <I>Mycobacterium</I> spp. was 1–2μm×2–3μm with a double-layered cell wall, based upon electron-microscopical features. Definitive diagnosis of these outbreaks was obtained by culture on selective media followed by PCR-restriction fragment length polymorphism analysis (PRA) of the <I>rpoB</I> gene for species identification. The amplified 360-bp products of the <I>rpoB</I> gene of mycobacteria isolated from zebrafish were digested with <I>Msp</I>I restriction enzyme, which revealed unique band patterns matching those of <I>Mycobacterium abscessus</I> and <I>Mycobacterium chelonae</I> which are responsible for skin and soft tissue infection caused by rapidly growing mycobacteria in humans. This is the first documentation of the precise identification of zoonotic non-tuberculous mycobacteria isolated from laboratory-maintained zebrafish by the PRA of the <I>rpoB</I> gene; this study thus provides a great deal of useful epidemiological information and reduces the likelihood that epizootics will occur.</P>