• 한국수산과학회지
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• 제43권5호
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• pp.462-473
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• 2010

Inductions of hybrids and reciprocal hybrids between Oryzias dancena and O. javanicus (ODJ and OJD) were conducted and backcross hybrids between female O. dancena and male ODJ were also produced for biological and cytogenetic analysis. Embryonic development of ODJ and OJD were compared with those of their parents. Developmental time was fastest in O. dancena and ODJ, followed by O. javanicus and OJD. Oryzias dancena hatched 11 days (d) after fertilization, ODJ at 13 d, O. javanicus at 14 d and OJD at 15 d. The abnormality of external morphology rate in ODJ was 10.6%; however, OJD showed a high degree of abnormality, over 90%. The proportion of males was 90.0% and 31.3% for ODJ and OJD, respectively. Cytogenetic analysis was conducted to obtain basic information for genetic identification of O. dancena, O. javanicus and their hybrids. The karyotypes of all experimental groups showed 2n=48 chromosomes and the fundamental number (FN) was 48. The first pair carried secondary constrictions near the centromeric regions. Erythrocyte area and volume were $9.8\;{\pm}\;0.5\;{\mu}m^2$ and $18.2\;{\pm}\;1.0\;{\mu}m^3$, respectively, for O. dancena, $8.3\;{\pm}\;0.5\;{\mu}m^2$ and $15.8\;{\pm}\;1.5\;{\mu}m^3$ in O. javanicus, and $18.3\;{\pm}\;0.5\;{\mu}m^2$ and $15.7\;{\pm}\;1.3\;{\mu}m^3$ in ODJ. Erythrocyte area and volume in ODJ were similar to those of O. javanicus. In backcross hybrids between female O. dancena and male ODJ, all embryos failed to develop and died in the late gastrula stage.

• Fisheries and Aquatic Sciences
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• 제15권4호
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• pp.317-324
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• 2012

We characterized the cytoskeletal beta-actin (${\beta}$-ACT) gene (actb) and its 5'-upstream regulatory region in the Javanese ricefish Oryzias javanicus. The gene and protein structures were deduced from amino acid sequences of the actb gene and conserved in the teleost lineage. The O. javanicus actb gene has common transcription factor binding motifs in its regulatory region found in teleostean orthologues. Following quantitative reverse transcription-PCR, actb gene transcripts were detected in all tissues examined; however, the basal expression levels were different. During early development, O. javanicus actb mRNA levels showed a gradual increase and peaked between late somitogenesis and the heartbeat stage. Microinjection of O. javanicus embryos with the actb gene promoter-driven red fluorescent protein (RFP) gene reporter vector showed a ubiquitous distribution of RFP signals, although most exhibited a mosaic pattern of transgene expression. A small number of microinjected embryos displayed a wide distribution of RFP signals over their entire body, which resembled the expression pattern of endogenous actb. Data from this study provide a basis to develop a transgenic system with ubiquitous expression of foreign genes in O. javanicus.

• Fisheries and Aquatic Sciences
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• 제15권4호
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• pp.325-335
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• 2012

The present study characterized the fast skeletal myosin light chain-2 gene (mlc2f) in the euryhaline Javanese ricefish (Oryzias javanicus: Beloniformes). Coding nucleotide and deduced amino acid sequences of Javanese ricefish mlc2f were well conserved in the vertebrate lineage. Javanese ricefish mlc2f showed a typical seven-exon structure, and its promoter exhibited transcription factor binding motifs common to most muscle-specific genes. However, Javanese ricefish mlc2f also displayed tandem duplications of intronic sequences in both intron 1 and intron 3. Based on quantitative reverse transcription-polymerase chain reaction, the mlc2f transcripts were highly predominant in skeletal muscles of adults and were differentially modulated during embryonic development. Microinjection of the mlc2f promoter-driven red fluorescent protein (RFP) reporter construct successfully exhibited heterologous expression of the fluorescent reporter, primarily in muscular areas of hatchlings, although the distribution pattern of RFP signals was not uniform due to the mosaic nature of the introduced transgene. Data from this study indicate that the Javanese ricefish mlc2f gene has undergone "intra-intronic" duplication events in a species-specific manner and that the mlc2f regulator may also be useful in heterologous expression assays of the skeletal muscles of this species.

• Molecular & Cellular Toxicology
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• 제4권4호
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• pp.293-299
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• 2008

To screen the differentially expressed genes in cadmuim-exposed marine medaka fish (Oryzias javanicus), a candidate marine test fish for ecological toxicity, the differential display polymerase chain reaction (DD-PCR) was carried out, since the genome-wide gene expression data are not available in this fish species yet. A total of 35 clones were isolated from cadmium-exposed fish and their nucleotide sequences were analyzed. The differentially expressed gene candidates were categorized to response to stimulus (3); ion binding (3); DNA binding (1); protein binding (6); carbohydrate binding (1); metabolic process (4); biological regulation (3); cellular process (2); protein synthesis (2); catalytic activity (2); sense of sight (1); immune (1); neurohormone (1); signaling activity (1); electron carrier activity (1) and others (3). For real-time quantitative RT-PCR, we selected catalase, glucose-6-phosphate dehydrogenase, heat shock protein 70, and metallothionein and confirmed that cadmium exposure enhanced induction of these four genes.

• 한국해양생명과학회지
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• 제6권1호
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• pp.31-37
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• 2021

Larvae from the marine medaka fish Oryzias javanicus were exposed with polystyrene microplastics (MPs) for 24 h. Exposure to waterborne fluorescent MPs showed clear ingestion and egestion in feces. Under constant MPs, the concentration of dissolved oxygen significantly decreased in 24 h compared to the control. Significant intracellular reactive oxygen species and malondialdehyde contents were detected in larvae, indicating oxidative stress and lipid peroxidation. Significant elevations in mRNA expressions of heat shock protein 70 and antioxidant defense system genes (glutathione reductase, glutathione peroxidase, catalase, and superoxide dismutase) were measured with increases in enzymatic activity of oxidative stress-related proteins. Taken together, the alterations to the molecular and biochemical components suggested that waterborne MPs had an oxidative stress effect on marine medaka larvae.

• 한국해양생명과학회지
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• 제9권1호
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• pp.33-40
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• 2024

To understand the detrimental effects of triclosan on Java medaka (Oryzias javanicus) embryos, fertilized embryos were exposed to different concentrations (1, 10, 50, 100, 200, 400, 600, 800, and 1,000 ㎍ l^-1) of triclosan until hatching. Then, we examined the survival rate and developmental parameters as well as alterations in antioxidant constituents and DNA damage markers. The results showed dose-dependent mortality, hatching delays, and developmental abnormalities in the embryos. Additionally, there were significant increases in oxidative stress parameters and antioxidant responses, along with elevated DNA damage. These findings suggest that sublethal concentrations of triclosan induce toxic effects through oxidative stress on Java medaka embryos, as evidenced by changes in in vivo parameters and biochemical constituents.

• Fisheries and Aquatic Sciences
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• 제14권2호
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• pp.93-104
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• 2011

Two hepcidin paralogs (ojhamp1 and ojhamp2) were isolated and characterized from a euryhaline Javanese ricefish (Oryzias javanicus: Beloniformes). The ojhamp1 cDNA encoded 90 or 91 amino acids (aa) of a typical HAMP1 preproprotein. This preproprotein is believed to cleave and yield the 66 or 67 aa-proprotein, followed by the 26 aa-mature peptide, composed of 8 conserved cysteine residues and the QSHL amino terminal motif. The ojhamp2 cDNA encoded 89 aa of HAMP2 preproprotein, cleaved to yield a 65 aa proprotein, and subsequently the 25 aa-mature peptide. The mature OJHAMP1 possessed a cationic isoelectric point (pI), whereas OJHAMP2 had an anionic charge. At the genomic level, both ojhamp1 and ojhamp2 share a conserved tripartite structure (three exons interrupted by two introns) with other vertebrate hepcidin genes. However, the ojhamp1 was shown to exist as two distinct mRNA species, encoding 90 or 91 aa, due to alternative splicing at the junction site between intron I and exon II. Both ojhamp1 and ojhamp2 transcripts were detected in a wide range of tissue types with varying levels of basal expression, although the highest expression was observed in the liver for both isoforms. Transcriptional response to bacterial challenge using Edwardsiella tarda showed that ojhamp1 was moderately upregulated in the liver but remained unchanged in the kidney. However, the ojhamp2 was significantly suppressed in both the kidney and liver, suggesting a potential diversification between the two paralogs.

• Molecular & Cellular Toxicology
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• 제5권4호
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• pp.283-290
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• 2009

Differential gene expression profiling was performed in the hepatic tissue of marine medaka fish (Oryzias javanicus) after exposure to benzo[a]pyrene (BaP), a polycyclic aromatic hydrocarbon (PAH), by heterologous hybridization using a medaka cDNA microarray. Thirty-eight differentially expressed candidate genes, of which 23 were induced and 15 repressed (P<0.01), were identified and found to be associated with cell cycle, development, endocrine/reproduction, immune, metabolism, nucleic acid/protein binding, signal transduction, or non-categorized. The presumptive physiological changes induced by BaP exposure were identified after considering the biological function of each gene candidate. The results obtained in this study will allow future studies to assess the molecular mechanisms of BaP toxicity and the development of a systems biology approach to the stress biology of organic chemicals.

• 한국어병학회지
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• 제24권2호
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• pp.131-139
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• 2011

본 연구에서는 비스페놀 A를 사용하여 바다 송사리 Oryzias dancena와 자바 송사리 O. javanicus의 잡종군(ODJ)의 성전환을 유도하였다. 부화 직후의 ODJ에 비스페놀 A를 2.5 mg/L, 5.0 mg/L와 10.0 mg/L의 농도로 56일간 처리한 결과 대조군의 경우 평균 전장이 $18.0{\pm}1.2$ mm로 나타났고 비스페놀 A 처리군에서는 2.5 mg/L로 처리한 경우 $15.0{\pm}2.0$ mm, 5.0 mg/L 처리군은 $13.7{\pm}2.5$ mm, 10.0 mg/L 처리군에서는 $12.8{\pm}2.5$ mm로 나타나 대조군과 비스페놀 A 처리군에서 성장 차이가 관찰되었다. 또한, 처리군 간에서도 비스페놀 A의 농도가 높을수록 성장이 느린 것으로 나타났다. 기형률을 조사한 결과 대조군은 13.6%였고, 비스페놀 A 처리군의 경우 2.5 mg/L는 65.4%로 대조군에 비해 매우 높게 나타났으며, 이보다 농도가 높은 5.0 mg/L와 10.0 mg/L에서는 각각 81.3%와 98.1%로 처리 농도가 높아질수록 기형률이 높게 나타났다. 성전환율을 분석하기 위하여 조직학적 분석을 한 결과 대조군에서 사용된 ODJ 50마리 중 정상 수컷이 47마리였으나, 3마리가 간성(intersex)을 나타내었으며, 비스페놀 A 농도가 2.5 mg/L인 그룹에서 76.9%인 40마리가 간성으로 나타났으며, 23.1%인 12마리가 수컷으로 나타났다. 비스페놀 A 농도 5.0 mg/L와 10.0 mg/L인 그룹에서는 52마리 중 수컷은 한 마리도 나타나지 않았으며, 모든 개체에서 간성이 나타났다.

• 한국해양생명과학회지
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• 제7권2호
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• pp.94-101
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• 2022

In this study, the marine medaka Oryzias javanicus was exposed to two concentrations of non-toxin-producing red tide dinoflagellate C. polykrikoides (1,000 and 2,000 cells ml^-1) for 96 h, and the time-course biochemical responses of antioxidant and immunity parameters were analyzed in the liver tissue. Significant ichthyotoxicity with increasing cell concentrations of C. polykrikoides and exposure period was observed for 96 h. Opercular respiratory rate was lowered in marine medaka exposed to 2,000 cells ml^-1 of C. polykrikoides. Intracellular malondialdehyde (MDA) content significantly elevated in response to both cell concentrations. In the case of glutathione (GSH) content, the levels were significantly elevated by 1,000 cells ml^-1 of C. polykrikoides, but the contents significantly depleted upon exposure to 2,000 cells ml^-1 of C. polykrikoides. Similarly, enzymatic activities of catalase (CAT) and superoxide dismutase (SOD) were increased by 1,000 cells ml^-1 of C. polykrikoides, whereas their activities were lowered by 2,000 cells ml^-1 of C. polykrikoides. Analysis of the two immunity parameters, alternative complement pathway and lysozyme, showed significantly lowered activities in 2,000 cells ml^-1 of C. polykrikoides-exposed liver tissue. These biochemical effects of C. polykrikoides on marine medaka would be helpful for understanding its acute effects in marine fish.