• Title/Summary/Keyword: zebrafish model

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Cloning and Characterization of Zebrafish Microsomal Epoxide Hydrolase Based on Bioinformatics (생물정보학을 이용한 Zebrafish Microsomal Epoxide Hydrolase 클로닝 및 특성연구)

  • Lee Eun-Yeol;Kim Hee-Sook
    • Microbiology and Biotechnology Letters
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    • v.34 no.2
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    • pp.129-135
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    • 2006
  • A gene encoding for a putative microsomal epoxide hydrolase (mEH) of a zebrafish, Danio rerio, was cloned and characterized. The putative mEH protein of D. rerio exhibited sequence similarity with mammalian mEH and some other bacterial EHs. A structural model for the putative mEH was constructed using homology modeling based on the crystallographic templates, 1 qo7 and 1 ehy. The catalytic triad consisting of $Asp^{233}$, $Glu^{413}$, and $His^{440}$ was identified, and the characteristic features such as two tyrosine residues and oxyanion hole were found to be highly conserved. Based on bioinformatic analysis together with EH activity assay, the putative protein was annotated as mEH of D. rerio. Enantiopure styrene oxide with enantiopurity of 99%ee and yield of 33.5% was obtained from racemic styrene oxide by the enantioselective hydrolysis activity of recombinant mEH of D. rerio for 45 min.

The Effect of Nano-scale Zn-$TiO_2$ and Pure $TiO_2$ Particles were Prepared using a Hydrothermal Method on Zebrafish Embryogenesis (수열합성법으로 제조된 Zn-$TiO_2$ 나노입자와 $TiO_2$ 나노입자가 zebrafish 배발생에 미치는 영향)

  • Yeo, Min-Kyeong;Kim, Hyo-Eun
    • Environmental Analysis Health and Toxicology
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    • v.24 no.4
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    • pp.333-339
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    • 2009
  • In this study, we investigated the biological toxicity of nano-scale Zn (0.1, 0.5, and 1 mol%)-doped $TiO_2$ and pure $TiO_2$ nanoparticles using zebrafish embryogenesis as our model organism. Zn-doped $TiO_2$ nanoparticles were prepared using a conventional hydrothermal method for the insertion of zinc into the $TiO_2$ framework. The characters of Zn-doped $TiO_2$ (0.1%, 0.5%, 1%Zn) and pure $TiO_2$ were about 7~8 nm. These sizes were smaller than 100~200 nm of $TiO_2$ was prepared using the sol-gel method. Particularly, in this study, we found no significant biological toxicity in the hatching rate and abnormal rate under expose pure $TiO_2$ and Zn-doped $TiO_2$ nanoparticles were prepared using a conventional hydrothermal method of zebrafish. It was different from the biological damage under $TiO_2$ nanoparticles were prepared using sol-gel method. We assessed that the damage was not linked to the particle's nanometer size, but rather due to the prepare method. Moreover, $TiO_2$ nanoparticles were prepared using a hydrothermal method were not shown to cause cytotoxic effects, like apoptosis and necrosis, that are the major markers of toxicity in organisms exposed to nanomaterials. Therefore, there is some relationship with biological toxicity of nanoparticles and the prepare method of nanometer size particles.

Modulation of Glial and Neuronal Migration by Lipocalin-2 in Zebrafish

  • Kim, Ho;Lee, Shin-Rye;Park, Hae-Chul;Lee, Won-Ha;Lee, Myung-Shik;Suk, Kyoung-Ho
    • IMMUNE NETWORK
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    • v.11 no.6
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    • pp.342-347
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    • 2011
  • Background: Glial cells are involved in immune and inflammatory responses in the central nervous system (CNS). Glial cells such as microglia and astrocytes also provide structural and functional support for neurons. Migration and morphological changes of CNS cells are associated with their physiological as well as pathological functions. The secreted protein lipocalin-2 (LCN2) has been previously implicated in regulation of diverse cellular processes of glia and neurons, including cell migration and morphology. Methods: Here, we employed a zebrafish model to analyze the role of LCN2 in CNS cell migration and morphology in vivo. In the first part of this study, we examined the indirect effect of LCN2 on cell migration and morphology of microglia, astrocytes, and neurons cultured in vitro. Results: Conditioned media collected from LCN2-treated astrocytes augmented migration of glia and neurons in the Boyden chamber assay. The conditioned media also increased the number of neuronal processes. Next, in order to further understand the role of LCN2 in the CNS in vivo, LCN2 was ectopically expressed in the zebrafish spinal cord. Expression of exogenous LCN2 modulated neuronal cell migration in the spinal cord of zebrafish embryos, supporting the role of LCN2 as a cell migration regulator in the CNS. Conclusion: Thus, LCN2 proteins secreted under diverse conditions may play an important role in CNS immune and inflammatory responses by controlling cell migration and morphology.

The Evaluation on the Effectiveness as a Cosmetic Material of Ascidian shell Extract Using Zebrafish model

  • Park, Sin-Ho;Kim, Bo-Ae;Yang, Jae-Chan
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.1
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    • pp.258-268
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    • 2019
  • The extracts of AS contain in alloxanthin, halocynthiaxanthin, astaxanthin and 13 kinds of carotenoids. The aim of the study was to assess the anti-oxidant activity and cell viability of AS. The anti-oxidant activity was determined by using DPPH radical inhibition activity and superoxide dismutase (SOD)-like activity. The results of cell viability assay showed that the extracts from AS were cytotoxic at concentrations above $5.0mg/m{\ell}$. This study was designed to examine inflammation induced by LPS, protection effect by UVB and the toxicity of Ascidian shell extract(ASE) as a functional cosmetic ingredient. Evaluation of embryo toxicity resulted in embryo coagulation and mortality when treated at 5.0, 10.0, $20.0mg/m{\ell}$. At the lowest concentration of $1.0mg/m{\ell}$, hatchability resulted in 100.0 % rate. The results of arrhythmia measurement in larvae showed similarity to the evaluation of embryo toxicity. This result demonstrated that toxicity is present at concentrations greater than $5.0mg/m{\ell}$. The protective effect of ASE on LPS and UVB-induced in the zebrafish was investigated. Intracellular reactive oxygen species(ROS) generated by the exposure of zebrafish to LPS, UVB-radiation were significantly decreased after treatment with ASE at $0.1mg/m{\ell}$. As a result, ASE similarly reduced UVB-induced ROS generation and cell death in live zebrafsih. Therefore, it is suggested that ASE has anti-Inflammatory effects and can possibly be used as a functional substance for skin protection in the future.

Promotion of Remyelination by Sulfasalazine in a Transgenic Zebrafish Model of Demyelination

  • Kim, Suhyun;Lee, Yun-Il;Chang, Ki-Young;Lee, Dong-Won;Cho, Sung Chun;Ha, Young Wan;Na, Ji Eun;Rhyu, Im Joo;Park, Sang Chul;Park, Hae-Chul
    • Molecules and Cells
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    • v.38 no.11
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    • pp.1013-1021
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    • 2015
  • Most of the axons in the vertebrate nervous system are surrounded by a lipid-rich membrane called myelin, which promotes rapid conduction of nerve impulses and protects the axon from being damaged. Multiple sclerosis (MS) is a chronic demyelinating disease of the CNS characterized by infiltration of immune cells and progressive damage to myelin and axons. One potential way to treat MS is to enhance the endogenous remyelination process, but at present there are no available treatments to promote remyelination in patients with demyelinating diseases. Sulfasalazine is an anti-inflammatory and immune-modulating drug that is used in rheumatology and inflammatory bowel disease. Its anti-inflammatory and immunomodulatory properties prompted us to test the ability of sulfasalazine to promote remyelination. In this study, we found that sulfasalazine promotes remyelination in the CNS of a transgenic zebrafish model of NTR/MTZ-induced demyelination. We also found that sulfasalazine treatment reduced the number of macrophages/microglia in the CNS of demyelinated zebrafish larvae, suggesting that the acceleration of remyelination is mediated by the immunomodulatory function of sulfasalazine. Our data suggest that temporal modulation of the immune response by sulfasalazine can be used to overcome MS by enhancing myelin repair and remyelination in the CNS.

Polyphenol-rich fraction from Ecklonia cava (a brown alga) processing by-product reduces LPS-induced inflammation in vitro and in vivo in a zebrafish model

  • Kim, Seo-Young;Kim, Eun-A;Kang, Min-Cheol;Lee, Ji-Hyeok;Yang, Hye-Won;Lee, Jung-Suck;Lim, Tae Il;Jeon, You-Jin
    • ALGAE
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    • v.29 no.2
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    • pp.165-174
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    • 2014
  • Ecklonia cava is a common edible brown algae that is plentiful in Jeju Island of Republic of Korea. Polyphenols from E. cava have strong anti-inflammatory activity. However, a large number of the by-products from E. cava processing are discarded. In the present study, to utilize these by-products, we assessed the anti-inflammatory activity of the polyphenol-rich fraction (PRF) from E. cava processing by-product (EPB) in lipopolysaccharide (LPS)-induced RAW264.7 macrophage cells. Four compounds, namely eckol, eckstolonol, dieckol, and phlorofucofuroeckol-A, were isolated and identified from PRF. We found that PRF suppressed the production of nitric oxide (NO), inducible nitric oxide synthase, and cyclooxygenase-2 in the LPS-induced cells. Furthermore, the protective effect of PRF was investigated in vivo in LPS-stimulated inflammation zebrafish model. PRF had a protective effect against LPS-stimulated toxicity in zebrafish embryos. In addition, PRF inhibited LPS-stimulated reactive oxygen species and NO generation. According to the results, PRF isolated from EPB could be used as a beneficial anti-inflammatory agent, instead of discard.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Streptococcus iniae shows potential as a subunit vaccine against various streptococcal species

  • Kim, Min Sun;Choi, Seung Hyuk;Kim, Ki Hong
    • Journal of fish pathology
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    • v.28 no.1
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    • pp.9-15
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    • 2015
  • The potential of Streptococcus iniae glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an antigen for a subunit vaccine was investigated using a zebrafish model. The recombinant S. iniae GAPDH was purified using His-tag column chromatography, and antisera against the recombinant GAPDH (rGAPDH) were produced by intraperitoneal immunization of rats. By immunization with S. iniae rGAPDH, the survival rates of zebrafish against an S. iniae challenge increased, suggesting that GAPDH would be an antigen capable of inducing protective immune responses in fish. Furthermore, we demonstrated using Western blotting, that the antisera against rGAPDH of S. iniae had cross-reactivity with GAPDH from Streptococcus parauberis and Lactococcus garviae, which are also culprits of streptococcosis in cultured fish in Korea. These results suggest that S. iniae GAPDH may be used as an antigen for the development of a subunit vaccine against streptococcosis caused by diverse cocci in cultured fish.

Enhanced antidiabetic efficacy and safety of compound K/β-cyclodextrin inclusion complex in zebrafish

  • Nam, Youn Hee;Le, Hoa Thi;Rodriguez, Isabel;Kim, Eun Young;Kim, Keonwoo;Jeong, Seo Yule;Woo, Sang Ho;Lee, Yeong Ro;Castaneda, Rodrigo;Hong, Jineui;Ji, Min Gun;Kim, Ung-Jin;Hong, Bin Na;Kim, Tae Woo;Kang, Tong Ho
    • Journal of Ginseng Research
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    • v.41 no.1
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    • pp.103-112
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    • 2017
  • Background: 20(S)-Protopanaxadiol 20-O-D-glucopyranoside, also called compound K (CK), exerts antidiabetic effects that are mediated by insulin secretion through adenosine triphosphate (ATP)-sensitive potassium ($K_{ATP}$) channels in pancreatic ${\beta}$-cells. However, the antidiabetic effects of CK may be limited because of its low bioavailability. Methods: In this study, we aimed to enhance the antidiabetic activity and lower the toxicity of CK by including it with ${\beta}$-cyclodextrin (CD) (CD-CK), and to determine whether the CD-CK compound enhanced pancreatic islet recovery, compared to CK alone, in an alloxan-induced diabetic zebrafish model. Furthermore, we confirmed the toxicity of CD-CK relative to CK alone by morphological changes, mitochondrial damage, and TdT-UTP nick end labeling (TUNEL) assays, and determined the ratio between the toxic and therapeutic dose for both compounds to verify the relative safety of CK and CD-CK. Results: The CD-CK conjugate ($EC_{50}=2.158{\mu}M$) enhanced the recovery of pancreatic islets, compared to CK alone ($EC_{50}=7.221{\mu}M$), as assessed in alloxan-induced diabetic zebrafish larvae. In addition, CD-CK ($LC_{50} =20.68{\mu}M$) was less toxic than CK alone ($LC_{50}=14.24{\mu}M$). The therapeutic index of CK and CD-CK was 1.98 and 9.58, respectively. Conclusion: The CD-CK inclusion complex enhanced the recovery of damaged pancreatic islets in diabetic zebrafish. The CD-CK inclusion complex has potential as an effective antidiabetic efficacy with lower toxicity.

Antioxidant and anti-inflammatory functionality of ten Sri Lankan seaweed extracts obtained by carbohydrase assisted extraction

  • Fernando, I.P. Shanura;Sanjeewa, K.K. Asanka;Samarakoon, Kalpa W.;Lee, Won Woo;Kim, Hyun-Soo;Ranasinghe, P.;Gunasekara, U.K.D.S.S.;Jeon, You-Jin
    • Food Science and Biotechnology
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    • v.27 no.6
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    • pp.1761-1769
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    • 2018
  • Enzyme-assisted extraction is a cost-effective, safe, and efficient method to obtain bioactives from plant materials. During this study, 10 different marine algae from Sri Lanka were individually extracted by using five commercial food-grade carbohydrases. The enzymatic and water extracts of the seaweeds were analyzed for their antioxidant and anti-inflammatory activities. The highest DPPH, hydrogen peroxide ($H_2O_2$) and intracellular $H_2O_2$ scavenging abilities were observed from the Celluclast extract of Sargassum polycystum (CSp). CSp exerted protective effects against oxidative stress-induced cell death in hydrogen peroxide-induced Chang cells and in model zebrafish. The Celluclast extract of Chnoospora minima (CCm) showed the strongest anti-inflammatory activity against lipopolysaccharide (LPS)-induced NO production in RAW 264.7 macrophages ($IC_{50}=44.47{\mu}g/mL$) and in model zebrafish. CCm inhibited the levels of iNOS, COX-2, $PGE_2$, and TNF-${\alpha}$ in LPS stimulated RAW 264.7 macrophages. Hence, CSp and CCm could be utilized in developing functional ingredients for foods, and cosmeceuticals.