• Korean Journal of Ichthyology
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• v.28 no.4
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• pp.215-222
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• 2016

Triploidy was induced in the marine medaka, Oryzias dancena by cold shock treatment ($0^{\circ}C$) of fertilized eggs for 30, 45, or 60 min, applied two minutes after fertilization. The triploid genotype was induced by each of the thermal shock regimes tested. The best result was obtained when the eggs were treated for 45 min, which induced triploidy in all the resulting fish. Triploidy was confirmed using chromosomal and flow cytometer analyses, and erythrocyte measurements. The surface areas and volumes of the erythrocytes of triploid fish were significantly larger than those of diploid fish, and their chromosome number (3N=72) was 1.5 times greater that for the diploids (2N=48). Based on a flow cytometer analysis, the triploid fish had approximately 1.5 times the cellular DNA content (2.40 pg/cell) of the diploid specimens (1.61 pg/cell). Data from this study provide the basis for the development of unique models for studying reproductive confinement in transgenic fish.

• Fisheries and Aquatic Sciences
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• v.15 no.3
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• pp.221-231
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• 2012

The marine medaka Oryzias dancena choriogenin H gene (odChgH) and its mRNA expression during estradiol-$17{\beta}$ (E2) exposure were characterized. At the amino acid level, the choriogenin H protein is predicted to possess the conserved repetitive N-terminal region, as well as zona pellucida (ZP) and Trefoil factor family (TFF) domains. At the genomic level, odChgH has an eight-exon organization with a distribution pattern of transcription factor binding sites in the 5'-upstream region, which is commonly found in other estrogen-responsive genes. The tissue distribution pattern of odChgH mRNA was found to be gender-specific, whereby females showed a higher expression level and wider tissue distribution than did males. During embryonic development, odChgH mRNA was robustly detected from the stage of visceral blood vessel formation. Experimental E2 exposure of males resulted in odChgH mRNA being induced not only in the liver, but also in other several tissues. The E2-mediated induction was fairly dose-dependent. The basal expression levels of hepatic odChgH mRNA were lower in males that were acclimated to 30 ppt salinity than in those acclimated to 0 or 15 ppt salinity. In contrast, the inducibility of odChgH mRNA during E2 exposure was greater in seawater-acclimated fish than in brackish water- or freshwater-acclimated fish.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.1
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• pp.19-30
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• 2024

Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

• Journal of the Korean Institute of Intelligent Systems
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• v.15 no.6
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• pp.682-687
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• 2005

Behavioral sequences of the medaka(Oryzias latipes) were investigated through an image system in response to medaka treated with the insecticide and medaka not treated with the insecticide, diazinon(0.1 mg/1). After much observation, behavioral patterns could be divided into 4 patterns: active smooth, active shaking, inactive smooth, and inactive shaking. These patterns were analyzed by 5 features: speed ratio, x and y axes projection, FFT to angle transition, fractal dimension, and center of mass. Each pattern was classified using decision tree. It provide a natural way to incorporate prior knowledge from human experts in fish behavior, The main focus of this study was to determine whether the decision tree could be useful in interpreting and classifying behavior patterns of the animal.

• Korean Journal of Environmental Biology
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• v.23 no.3 s.59
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• pp.293-303
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• 2005

Japanese medaka, Oryzias latipes is widely distributed in the North East Asia including Korea, Japan and east China, and commonly used for freshwater toxicity tests and cytotoxicological studies worldwide. In this study, a series of experiments were conducted to identify the potential of the fish as a standard test species for saltwater toxicity evaluation such as marine receiving waters, ocean-dumped materials and sediment pore waters etc. Hatching, growth and mortality rates of the fish were estimated with the wide ranges of salinity from freshwater to seawater (35 psu). Direct exposure of the fertilized eggs in freshwater to the wide ranges of salinity (from 0 to 35 psu) without pre- acclimation to the saltwater revealed no significant differences in hatching rates by salinities (p =0.24). On the other hand, medaka larvae hatched in freshwater and exposed to saltwater directly showed high mortality at > 25 psu treatment groups (p < 0.0001). However, there was no significant difference in mortality of medaka larvae hatched in 13.8 and 14.2 psu at the wide ranges of salinities ($0\~35$ psu). Growth rates of medaka larvae hatched in the above two salinities showed no differences in body length either from 0 to 35 psu treatment groups (p =0.64 for 13.8 psu group and p=0.32 for 14.2 psu group). The number of gill chloride cell in medaka larvae sharply increased when the larvae were exposed to high salinity. Reference tests with zinc chloride revealed 96h $LC_{50}=8.84(7.19\~10.87)mg\;L^{-1}$ using 7~10 day old medaka larvae. These were comparable or better sensitivity in comparison with the other standard test species such as North American sheepshead minnow Cyprinodon variegatus. Based on the results of these experiments, hatching rates and larvalmortality of medaka must be good toxicity parameters for seawater bioassay and the species seems to be a good standard species for both the freshwater and seawater toxicity test.

• Proceedings of the Korean Society of Toxicology Conference
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• 2001.05a
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• pp.135-135
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• 2001

Mating behavior and reproductivity of male fish were studied as an in vivo screening method of endocrine disruptors. Male medaka (Oryzias latipes) were exposed to 17$\beta$-estradiol at nominal concentrations of 2 and 20 $\mu\textrm{g}$/L for 14 days.(omitted)

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2003.05a
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• pp.158-159
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• 2003

Fluoranthene, a common polycyclicaromatic hydrocarbon (PAH), exhibits phototoxicity which may affect aquatic organisms. The eventual goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals that are toxic and persistent in the ecosystem. In this study, we investigated neural toxicity of fluoranthene in Japanese medaka (Oryzias latipes) which was correlated with its behaviors.

• Journal of Environmental Health Sciences
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• v.29 no.2
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• pp.7-15
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• 2003

Di-2-ethylhexyl phthalate (DEHP), is a widely used plasticizer known to be a suspected endocrine disrupter, but its exact effects on aquatic organisms are not yet known. When Japanese medaka (Oryzias latipes) were exposed from the time of hatching to 3 months of age to an aqueous DEHP solution at nominal concentrations of 1, 10, and 50 $\mu\textrm{g}$/l, DEHP treated female fish showed distinct reproductive effect. And the midge (Chironomus riparius.). an aquatic invertebrate, was exposed to DEHP to evaluate the effects on reproductive processes via sediment toxicity. The test endpoints included emergence, sex ratio, fecundity, and the viability of F1 offspring egg ropes. The result implied that the normal developmental and/or reproductive processes in C. riparius had been disrupted when exposed to DEHP, the effect also being displayed in the next generation. In summary, DEHP hinders the development of reproductive organs in the female Japanese medaka and C. riparius.

• Fisheries and Aquatic Sciences
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• v.19 no.9
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• pp.41.1-41.7
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• 2016

Background: The transgenic approach using estrogen-responsive regulator in fish has been given much attention as a potential means to detect and/or address estrogen-related aquatic pollutions. In order to address the development stage- and reproduction status-dependent expression patterns of the chgH-rfp transgene (red fluorescent protein transgene driven by choriogenin H promoter) in marine medaka Oryzias dancena, naturally occurring red fluorescent protein (RFP) signals under non-exposed conditions as well as the transgenically induced RFP signals under estrogen-exposed conditions were assayed. Results: Female transgenics begun to show naturally occurring RFP signals from the age of 7 weeks post hatching (WPH) without experimental estrogen exposure. Afterward, these RFP signals in female transgenics became robust with the progress of ovarian maturation. On the other hand, male transgenics did not show any naturally occurring RFP signal under non-exposed conditions irrespective of developmental stages and maturation statue. Upon exposures using estradiol-$17{\beta}$ (E2) and $17{\alpha}$-ethinylestradiol (EE2), RFP signals were significantly induced specifically in the livers of transgenic males. Conclusions: Male chgH-rfp transgenics were able to keep the "off" state of RFP expression during their entire life cycle unless exposed to exogenous estrogens. Owing to their tight regulation capability of estrogen-responsive transgene, transgenesis of chgH-rfp in male marine medaka could offer a useful model system for future ecotoxicogenomic studies regarding estrogenicity-related issues in aquatic and marine environments.

• Journal of Marine Life Science
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• v.8 no.2
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• pp.190-195
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• 2023

In this research, the marine medaka Oryzias javanicus underwent a 96 h exposure to two concentrations of the red tide dinoflagellate Karenia mikimotoi (1,000 and 5,000 cells mL^-1), and the temporal variations in biochemical responses related to antioxidant and immunity parameters were assessed in the liver tissue. The study revealed a significant increase in ichthyotoxicity with elevated cell concentrations of K. mikimotoi, especially evident at 96 h in marine medaka exposed to 5,000 cells mL^-1. At 1,000 cells mL^-1 of K. mikimotoi, the opercular respiratory rate showed a significant increase, whereas exposure to 5,000 cells mL^-1 resulted in a lowered rate. The intracellular malondialdehyde content was significantly elevated in response to both cell concentrations at 96 h. Regarding glutathione content, levels were significantly increased by exposure to both cell concentrations. Catalase and superoxide dismutase enzymatic activities experienced an increase at 1,000 cells mL^-1 of K. mikimotoi, while their activities were reduced at 5,000 cells mL^-1 at 96 h. The analysis of two immunity parameters, alternative complement pathway and lysozyme, demonstrated significantly reduced activities in the liver tissue exposed to 5,000 cells mL^-1 of K. mikimotoi. These findings aim to enhance the understanding of K. mikimotoi toxicity in marine fish by offering insights into biochemical responses associated with harmful algal blooms.