• 한국어병학회지
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• 제22권1호
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• pp.45-52
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• 2009

Zebrafish was firstly applied to an experimental model for scuticociliatosis caused by Philasterides dicentrarchi, a facultative parasitic ciliate in cultured marine fish. The susceptibility of zebrafish to infection of P. dicentrarchi was assessed by intraperitoneal injection of the ciliates, which produced typical symptoms of scuticociliatosis and significant mortality. The potential use of zebrafish as a model to evaluate the vaccine efficacy against scuticociliatosis was analyzed by immunization of zebrafish with the ciliates lysate. Furthermore, the effect of different adjuvants, such as Quillaja saponin (QS), Montanide, and Freund’s incomplete adjuvant (FIA) on the protective efficacy of the vaccine was investigated. Groups of zebrafish injected with QS or Montanide alone showed higher survival of fish against challenge test compared to control fish. The results suggest that adjuvant-mediated enhancement of innate immune responses play important roles in protection of fish against scuticociliatosis. The considerably high survival in the fish immunized with the antigen alone indicates that the ciliate lysate itself is highly immunogenic to zebrafish, which can elicit protective immune responses. The protective potential of the antigen, ciliate lysate, was enforced through combined administration with adjuvants including QS, Montinide and FIA. No or low mortalities in the groups of fish immunized with the antigen plus adjuvants suggests that the adaptive immune responses of zebrafish might be accelerated by the adjuvants or the protective potential of the antigen and adjuvants might synergistically interact. In spite of several shortcomings such as difficulties in sampling of serum and leucocytes enough to routine immunological analyses, zebrafsih might be the most convenient experimental animal for scuticociliatosis.

• Animal cells and systems
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• 제16권3호
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• pp.198-206
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• 2012

Anxiety in zebrafish can be determined by examining their bottom-dwelling and light-avoidance behavior. This study determines the effects of physostigmine and scopolamine on anxiety in zebrafish by measuring swimming frequency for three horizontal layers and three vertical columns of a water test tank illuminated by a light source located above the central surface of the tank. In the 1 h session, zebrafish in the control group preferred the bottom layer the most and the center column the least. Zebrafish treated with 2-20 ${\mu}M$ physostigmine were more likely to prefer the to layer than controls, and there were significant pairwise differences between physostigmine-treated zebrafish and controls, indicating the anxiolytic effect of physostigmine. Further, 10 and $20{\mu}M$ physostigmine-treated zebrafish no longer avoided the center column. Scopolamine had no anxiolytic effect on bottom-dwelling and light-avoidance behaviors but suppressed the anxiolytic effect of physostigmine. In terms of their preference for various zones formed by layers and columns, zebrafish in the control group preferred the bottom left and right zones the most. Physostigmine had a positive effect on the preference for the top center zone, which was suppressed by scopolamine pretreatment. The results suggest that the level of anxiety in zebrafish can be reduced by activating acetylcholinergic neurotransmitter systems, which is mediated in part by muscarinic receptors.

• Journal of Applied Biological Chemistry
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• 제58권1호
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• pp.51-54
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• 2015

오디에서 분리한 flavonoid가 zebrafish에 alloxan을 처리하여 손상된 췌장섬을 어느 정도 회복 시키는지 검토하였다. Zebrafish의 배아에 alloxan을 처리하면 확실하게 췌장섬의 크기가 감소하였다(p <0.001). Rutin, isoquercetin 및 quercetin의 처리에 의해 췌장섬의 크기가 유의적으로 증가하였으며(p <0.001), 형광세포의 fluorescence intensity도 뚜렷하게 증가하였다.

• Fisheries and Aquatic Sciences
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• 제20권12호
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• pp.32.1-32.7
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• 2017

Recent in vitro studies have demonstrated that extract of soft coral Dendronephthya gigantea (SCDE) had strong anti-inflammatory activities. However, the direct effects of SCDE on anti-inflammatory activities in vivo model remained to be determined. Therefore, the present study was designed to assess in vivo anti-inflammatory effect of SCDE using lipopolysaccharide (LPS)-stimulated zebrafish model. We also investigated whether SCDE has toxic effects in zebrafish model. The survival, heart beat rate, and developmental abnormalities were no significant change in the zebrafish embryos exposed to at a concentration below $100{\mu}g/ml$ of SCDE. However, lethal toxicity was caused after exposure to 200 and $400{\mu}g/ml$ of SCDE. Treating zebrafish model with LPS treatment significantly increased the reactive oxygen species (ROS) and nitric oxide (NO) generation. However, SCDE inhibited this LPS-stimulated ROS and NO generation in a dose-dependent manner. These results show that SCDE alleviated inflammation by inhibiting the ROS and NO generation induced by LPS treatment. In addition, SCDE has a protective effect against the cell damage induced by LPS exposure in zebrafish embryos. This outcome could explain the profound anti-inflammatory effect of SCDE both in vitro as well as in vivo, suggesting that the SCDE might be a strong anti-inflammatory agent.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.457-461
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• 2011

JAZF1 (Juxtaposed with Another Zinc Finger gene 1) transcription factor are Zn-finger proteins that bind to the nuclear orphan receptor TAK/TR4 (Nakajima et al., 2004). The nuclear orphan receptor TAK1/TR4 functions as a positive as well as a negative regulator of transcription. It was recently reported that congenital cardiovascular malformations are significantly more frequent in Neurofibromatosis 1 (NF1) patients with microdeletion syndrome than in those with classical NF1. JAZF1 was expressed in adult heart of patients with microdeletion syndrome. JAZF1 is highly conserved among various species include zebrafish. We hypothesized that the expression of zebrafish Jazf1 may lead to severe forms of congenital heart disease that allow the survival of newborns and adults. In this study, we created Jazf1 transgenic zebrafish which over-express zebrafish Jazf1 cDNA under control of the CMV promoter. Our results suggested that Jazf1 expression may play an important role in zebrafish cardiac development.

• 생약학회지
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• 제50권1호
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• pp.25-31
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• 2019

The zebrafish (Danio rerio) is an established model organism for several pathophysiological conditions which are related to human diseases. In this study, we tested the preventive effect of eight herbal extracts, which show the inhibitory effect of advanced glycation end products (AGEs) or aldose reductase (AR) in our previous study, on high glucose (HG)-induced retinal vessel dilation in larval zebrafish and analyzed the change of hyaloid vasculature. HG-induced zebrafish hyaloid vasculatures were significantly increased in the thickness compared to untreated zebrafish (P<0.001, n=6~10). Eight herbal extracts were found to have significant retinal vessel dilation on the inhibitory activity. Particularly, Brandisia hancei (twigs and fruits), Castanopsis orthacantha (leaves and twigs), Litsea japonica (leaves and twigs), Spenceria ramalana (whole plant), and Synedrella nodiflora (leaves and stems) showed potent inhibitory activity against retinal vessel dilation in HG-induced larval zebrafish.

• 한국동물생명공학회지
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• 제36권1호
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• pp.17-24
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• 2021

In this study, the potential toxicity of isoprocarb was demonstrated using zebrafish embryos. We treated isoprocarb (0, 29, and 58 mg/L) to the zebrafish embryos for 72 h then, we estimated morphological changes and apoptotic cell numbers. The increasing extent of apoptosis from the anterior to posterior region of developing zebrafish larvae was correlated with toxicity in the overall development process, including growth and normal organ formation. The appearance of abnormalities in the isoprocarb-treated groups in comparison to normal developing zebrafish larvae was verified using quantitative image analysis based on ImageJ software program. The vascular system comprising a complex interconnection of blood vessels was visualized in vessel-fluorescent transgenic zebrafish (fli1:eGFP). The main vasculature was malformed on isoprocarb treatment, and this was also related to cardiac defects. Taken together, normal embryonic development in zebrafish was interrupted owing to the acute toxicity of isoprocarb.

• 생명과학회지
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• 제16권4호
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• pp.571-578
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• 2006

Mitochondrial serine protease로 알려진 human HtrA2 (hHtrA2)는 apoptosis 유도 과정에서 중요한 역할을 담당하고 있을 뿐만 아니라 hHtrA2가 motor neuron degeneration과 관련이 있다는 최근 연구 결과가 있으나, hHtrA2의 생리적 기능은 아직 명확하게 밝혀져 있지 않다. 이와 같이 생체내에서 필수적인 업무를 담당하는 hHtrA2의 기능을 심도 있게 연구하기 위해서는 적절한 동물모델 시스템이 필요하나 이에 대한 연구도 미흡한 실정이다. 따라서 본 연구에서는 hHtrA2의 기능 분석을 위한 기본적인 실험으로 zebrafish라는 동물모델을 선택하여 hHtrA2의 발현 시스템을 정립하였다. 먼저 zebrafish에 hHtrA2를 발현시키기 위하여 zebrafish에서 일반적으로 사용되는 발현 시스템인 pCS2+ vector에 hHtrA2와 GFP를 cloning하고 plasmid를 HEK293 cell에 transfection한 후, hHtrA2-GFP fusion 단백질의 발현을 immunoblot과 immunofluorescence staining assay로 확인한 바 약 64 kDa의 hHtrA2 단백질의 발현을 확인할 수 있었다. Zebrafish에서 hHtrA2-GFP fusion 단백질의 발현양상은 immunofluorescence microscope으로 확인하였다. hHtrA2-GFP DNA와 mRNA를 zebrafish embyro에 microinjection하여 두 가지 component의 발현을 비교 분석한 결과, DNA는 dot 형태로 mRNA는 몸 전체에 퍼져보이는 형태로 발현 양상의 차이는 있었으나 둘 다 zebrafish embryo에서 잘 발현되는 것을 알 수 있다. 다음 DNA를 주 component로 microinjection하여 zebrafish embryo에서 발현을 확인한 결과 hHtrA2는 72 hpf 까지 발현이 지속되는 것을 확인하였다. 본 연구에서 정립한 hHtrA2의 zebrafish 발현 조건은 앞으로 zebrafish에서 hHtrA2의 생리적 기능을 심도있고 정확하게 연구하는 데 있어 기본적인 자료로 활용 할 수 있을 것이다.

• 생명과학회지
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• 제25권2호
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• pp.151-157
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• 2015

Zebrafish는 약리학 연구에 널리 사용되는 실험 동물 중 하나이다. Oxytetracycline은 넓은 항균 범위를 가지는 항생제로써 단백질 합성을 억제하여 정균 작용을 가진 항생제이다. 본 연구의 목적은 OTC에 노출이 되었을 때 zebrafish에게서 나타나는 행동변화와 내분비계의 변화를 관찰하였다. OTC (50, 100 or 200 mg/l)에 노출된 zebrafish의 행동변화를 확인하기 위해 novel tank test와 open field test를 진행하였다. 또한 내분비계의 변화를 확인하기 위하여 whole-body cortisol의 수치를 cortisol kit를 이용하여 측정하였다. Novel tank test 결과 상층부에서 머문 시간과 부동 시간에서 OTC를 약욕하였을 때 통계적으로 유의성이 있는 증가현상이 확인되었다. 반면에, 총 이동거리, 과행동 시간, 유영 속도 그리고 상하 변위에서 OTC를 약욕하였을 때 통계적으로 유의성 있는 감소 현상이 확인되었다. Open field test 결과 부동시간에서 OTC를 약욕 하였을 때 통계적으로 유의성 있는 증가 현상이 확인 되었다. 그러나 OTC를 약욕 하였을 때 총 이동거리, 움직인 시간, 유영 속도에서 통계적으로 유의성이 있는 감소 현상이 확인 되었다. Whole-body cortisol에서는 zebrafish에게 약욕을 하였을 때 증가하는 경향이 관찰되었다. 이러한 결과를 통하여 OTC가 zebrafish에게 화학적 스트레스로 작용한 것으로 사료된다.

• 생명과학회지
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• 제26권2호
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• pp.181-189
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• 2016

본 연구에서는 발아전처리 제초제인 Dithiopyr (DTP)를 시험물질로 사용하여 급성 독성시험을 평가하고, zebrafish의 행동에 미치는 영향을 연구하였다. 급성 독성시험 결과 zebrafish에서의 DTP의 LC₅₀값은 14.49 μM이었다. 또한 DTP 노출이 유영 패턴에 미치는 영향을 관찰하기 위하여 아치사농도(2.5-20 μM)에서 novel tank test (NTT)와 open field test (OFT)의 두 가지 행동실험을 실시하였다. NTT를 실시한 결과, 하층부의 머무는 시간 및 최초로 상층부에 들어간 시간, 회전각, 부동 시간의 경우 control 그룹과 비교하여 통계적으로 유의성 있게 농도 의존적으로 증가하였다(p<0.05). 또한 상층부에 진입한 횟수 및 평균 유영속도에서 통계적인 유의성 있는 감소현상이 확인되었다(p<0.05). OFT를 실시한 결과, 총 이동 거리 및 평균 유영속도에서 유의성이 있는 감소현상이 확인되었다(p<0.05). 부동 시간 및 회전각, 굴곡유영에서는 유의성 있는 증가 현상이 확인되었다(p<0.05). 또한 Whole-body cortisol level 측정 결과, 저농도 노출 그룹에서만 유의성 있는 증가가 관찰되었으나(p<0.05), 5와 10 μM에서는 유의성 있는 차이가 관찰되지 않아 농도의존적으로 감소하는 것을 확인하였다.