• Environmental Analysis Health and Toxicology
• /
• v.20 no.1
• /
• pp.13-21
• /
• 2005

The purpose of this study was to determine the effects of bisphenol A (BPA), which is known to have estrogenic activity, on the early development of medaka fish (Oryzias latipes). The fertilized eggs of medaka were treated with BPA at different concentrations for 3 weeks. Embryonic growth, deformation, hatching success, and gonadal differentiation were determined to observe the effects of this chemical. Also we tried to measure the estrogenic activity of bisphenol A using ELISA and RT-PCR methods. By using this techniques, we evaluated the induction of vitellogenin, an estrogen-regulated gene from the whole body-homogenates of larvae. At results, a reduced blood circulation was seen in embryos and peritoneal edema and hindrance of yolk-sac absorption were observed in larvae of treated group. However, BPA at the concentrations tested (2～200 ㎍/L) did not have severe adverse effects on the early life-stages. According to the observation of gonadal histology, inter-sex or sex -reversal was not found in all test fish. After the exposure was ended, vitellogenin mRNA and protein levels were measured in larvae and then their levels were found to be increased in treated group with 200㎍/L. These results indicate that BPA can induce the expression of vitellogenin in early life-stages as well as in adult male fish.

• Journal of Animal Reproduction and Biotechnology
• /
• v.35 no.1
• /
• pp.65-72
• /
• 2020

Fish ovarian germline stem cells (OGSCs) that have the abilities to self-renew and differentiate into functional gametes can be used in various researches and applications. A main issue to be solved for effective utilization of fish OGSCs is the development of their stable in vitro culture condition, but only few researches about fish OGSC culture have been reported so far. In this study, in order to find the clues to develop the culture condition for OGSCs from Japanese medaka (Oryzias latipes), we tried to establish somatic cell lines as a candidate for the feeder cells and evaluated its supporting effects on the culture of ovarian cell populations from O. latipes. As the results, the somatic cell lines could be established only from the embryonic tissues among three tissues derived from embryos, fins and ovaries. Three embryonic cell lines were tested as a feeder cell for the culture of ovarian cell population and all three cell lines induced cell aggregation formation of the cultured ovarian cells whereas the feeder-free condition did not. Furthermore, a significant cellular proliferation was observed in the ovarian cells cultured on two of three cell lines. As a trial to increase the capacity of the cell lines as a feeder cell that supports the proliferation of the cultured ovarian cells, we subsequently established a stable line that expresses the foreign O. latipes fibroblast growth factor 2 (FGF2) from an embryonic cell line and evaluated its effectiveness as a feeder cell. The ovarian cells cultured on FGF2 expressing feeder cells still formed cell aggregates but did not show a significant increase in cellular proliferation compared to those cultured on non-transformed feeder cells. The results from this study will provide the fundamental information for in vitro culture of medaka OGSCs.

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

• Development and Reproduction
• /
• v.22 no.3
• /
• pp.275-281
• /
• 2018

The purpose of this study is to examine fluctuating asymmetry of eye diameter, maxilla length, operculum length, and the number of pectoral fin ray and pelvic fin ray between ploidy and sex in diploid and triploid marine medaka, Oryzias dancena. In all experimental groups, eye diameter and maxilla length showed no significant difference between left side and right side (p>0.05). Results of operculum length in triploid male group and pectoral fin ray's number in diploid male group showed similarity ones with results of operculum length in triploid female group and pectoral fin ray's number in diploid female group (p<0.05). However, the operculum length in diploid male group and pectoral fin ray's number in triploid male group showed consinderable difference with those of operculum length in diploid female group and pectoral fin in triploid female group. Findings of pelvic fin ray's number in all groups were similar to those of pectoral fin ray's number in all groups (p<0.01).

• Environmental Analysis Health and Toxicology
• /
• v.18 no.1
• /
• pp.21-25
• /
• 2003

카바메이트계 농약인 카보푸란은 어류에 대한 독성이 매우 높으며, 낮은 농도에서 어류의 척추기형이나 행동이상을 유발한다. 이러한 카보푸란의 독성기전을 밝히기 위한 일환으로 신경물질대사와 관련이 깊은 acetylcholine esterase(AChE)와 monoamine oxidase(MAO)에 미치는 농약의 효과를 송사리(Oryzias latipes; Medaka fish)를 이용하여 평가하였다. Medaka fish에 대한 카보푸란의 반수치사농도(LC$_{50}$)는 2.5 ppm이었으며, 1 ppb 카보푸란에 24시간 노출된 경우, AChE 효소활성이 머리와 몸통부위에서 각각 30, 20%씩 감소되었다. 한편, MAO 효소활성은 카보푸란의 농도가 증가함에 따라 머리부위에서는 감소한 반면, 몸통부위에서는 증가하는 경향을 보였다. 특히 카보푸란의 농도가 1 ppb에서도 송사리의 MAO 효소활성이 영향을 받는 것으로 나타나, 카보푸란에 의한 송사리의 행동이상은 AChE 활성 뿐 아니라 MAO활성의 변화에 의한 복합적인 효과일 가능성이 높다.

• Journal of the Korean Institute of Intelligent Systems
• /
• v.15 no.6
• /
• pp.682-687
• /
• 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
• /
• v.26 no.3
• /
• pp.220-225
• /
• 2008

In this study, alterations in whole proteome expression patterns in the head of Japanese medaka (Oryzias latipes) was investigated following acute or chronic exposure to benzo(a)pyrene (BaP) (25 ${\mu}g$ L$^{-1}$) for 48 hrs and 15 days, respectively. The results showed that 9 and 6 protein spots were statistically different, relative to controls, in response to acute and chronic BaP exposure, respectively. In the acute exposure group, 5 spots were up regulated and 4 spots were downregulated, while in the chronic exposure group, 4 spots were upregulated and 2 spots were downregulated. Three of these spots were common to both the acute and chronic BaP exposure groups and were identified using LC-MS/MS followed by database searching. These 3 spots were found to be associated with structural proteins belonging to the actin and keratin families. These data suggest that acute and chronic exposures to BaP may affect tissue morphology in the head of Japanese medaka.

• Proceedings of the Korea Society of Environmental Toocicology Conference
• /
• 2003.10a
• /
• pp.23-24
• /
• 2003

The 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. Japanese medaka was treated by copper and fluoranthene of appropriate sublethal concentrations after starvation for 48 hr. In this study we investigated neural toxicity of copper and fluoranthene in Japanese medaka (Oryzias latipes) along with comparative analysis of corresponding behavioral responses. The untreated individuals showed common behavioral characteristics (i.e., smooth and linear movements). Locomotive activity of the fish was monitored using an image processing and automatic data acquisition system. When treated with copper (100 ppb), the fish showed shaking patterns more frequently. As the concentration of copper increased to 1,000 ppb, activity decreased, and the fish showed an erratic movement. The treated with fluoranthene, however, showed stopping and abrupt change of orientation (100 ppb), and severely reduced locomotive activity and enhanced surfacing activity (1,000 ppb).

• Fisheries and Aquatic Sciences
• /
• v.16 no.3
• /
• pp.177-185
• /
• 2013

The development of species-specific fish cell lines has become a valuable tool for biological research. In recent years, marine medaka Oryzias dancena has been recognized as a good experimental model fish but there are no reports of establishment of cell lines from this fish. In this study, two cell lines from O. dancena blastula embryos were established from 41 total trials (4.9%). The two cell lines displayed typical in vitro morphology and have been cultured for >121 passages, which corresponds to 293 days. The doubling times of the cell lines were 29.84 and 28.59 h, respectively, and both possessed the potential to expand in a clonal manner, albeit with significant differences between the two cell lines. The absence of any of the four main medium supplements; i.e., fish serum, fetal bovine serum, basic fibroblast growth factor, and medaka embryo extract, significantly inhibited growth. The proportion of cells possessing normal chromosome number was 45% and 46.7% of the cell lines, respectively. Taken together, two cell lines that proliferate continuously were established from marine medaka and these cell lines may provide a basic tool for characterizing the unique features of this fish species.

• Environmental Analysis Health and Toxicology
• /
• v.23 no.3
• /
• pp.221-230
• /
• 2008

Chlorpyrifos (CPF) is a widely used organophosphate insecticide. Japanese medaka, Oryzias latipes was selected to investigate the effects of chlorpyrifos on survival rate, hatching rate, reproductivity and histopathological changes. Adult fish were exposed to CPF at concentrations of 20, 40, 80 and $160{\mu}g/L$, and the test duration was 4 weeks. An experiment with eggs was also performed under CPF exposure continued for 7 days covering from the fecundation to end-hatching stages. Survival rate of the adult medaka decreased in CPF $80{\mu}g/L$ and $160{\mu}g/L$ treatment groups compared with control, solvent control, $20{\mu}g/L$ or $40{\mu}g/L$L treatment group. Spontaneous start of feeding significantly decreased compared with the control (p < 0.01) when exposed to CPF at above 40 $\mu g/L$. Total spawning decreased by the 20 or $40{\mu}g/L$ CPF treatment compared with the control or solvent control in the test with eggs obtained from untreated adult medaka, and abnormal eggs increased in those CPF-treated groups. There were certain adverse effects at above $40{\mu}g/L$ determined from liver and gonad histological examinations. The results indicate that CPF has insignificant toxic effects in Japanese medaka at less than $20{\mu}g/L$ in long term exposure.