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PubMed2003

ras oncogene mutations in diethylnitrosamine-induced hepatic tumors in medaka (Oryzias latipes), a teleost fish.

Medaka fish are an established non-mammalian research model for the study of liver carcinogenesis and exposure to environmental pollutants. Studies have emphasized the development of hepatic neoplasms in medaka following exposure to model carcinogens. To date however, little information is known regarding the mechanisms underlying initiation of hepatic tumors in this species. The aim of this study was to relate our understanding of diethylnitrosamine (DEN)-induced tumor formation to ras gene activation in hepatic neoplasms of exposed medaka. Initial studies were conducted to identify medaka ras exons 1 and 2 by reverse transcriptase polymerase chain reaction (RT-PCR). Amplification of ras exons 1 and 2 from untreated medaka liver resulted in the identification of three polymorphic ras sequence variants exhibiting a high degree of homology to other teleost and mammalian ras genes. Exposure of medaka to 159 ppm of DEN resulted in a wide range of hepatic neoplasms including: hepatocellular adenomas, hepatocellular carcinomas, cholangiomas, and mixed hepatocholangiocellular carcinomas. Individual liver tumors were examined for oncogenically activating ras mutations by probing genomic DNA with probes specific for activating point mutations or by direct cloning and sequencing of ras transcripts using RT-PCR. Using allele-specific oligonucleotide (ASO) analysis, a single point mutation was detected in codon 12 position two in 8/25 (32%) tumors examined. Mutated ras alleles were additionally detected in 12 of 39 (30%) medaka liver tumors by sequence analysis. Ten of the 12 mutations identified contained a single point mutation at codon 12 resulting in a Gly to Asp amino acid substitution. Two unique mutations were identified at codon 16 resulting in either Lys to Asn or Lys to Thr amino acid substitutions. Our results show that ras mutations are induced by DEN and are present in over 30% of the fish that developed tumors. A ras mutation incidence of 30% is similar to that reported in mammalian species exposed to DEN. While mutations at codon 12 have previously been reported, the present study is the first in vivo report of ras point mutations at codon 16.

Amino Acid Sequence↗
PubMed2002

Development and application of a monoclonal antibody-based sandwich ELISA for quantification of Japanese medaka (Oryzias latipes) vitellogenin.

Vitellogenin (Vtg) was purified from ascitic fluid of a 17beta-estradiol (E2)-treated female Japanese medaka by anion-exchange chromatography. The molecular mass of medaka Vtg by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), corresponding to the Vtg monomer, was 200 kDa. BALB/c mice were immunized with purified-Vtg and two hybridoma clones producing specific antibodies against medaka Vtg were selected. The specificity of these monoclonal antibodies (mAbs) was evaluated by Western blot analysis of the plasma proteins separated on SDS-PAGE, and no cross-reactivity was observed with plasma proteins from control males. A sandwich enzyme-linked immunosorbent assay (ELISA) for quantification of medaka Vtg was developed using these mAbs. The assay range was between 1 and 100 ng/ml, and the intra- and inter-assay variations determined from plasma samples were within 7.7 and 8.5%, respectively. Recovery of medaka Vtg added to plasma was 92-111%. In a plasma dilution test, plots of Vtg concentration gave a straight line. After exposure of male medaka to E2 (10 ng/l), Vtg appeared in liver and plasma on the first day and reached a maximum on the 3rd to 5th day. The sandwich ELISA could be useful for the detection of estrogenic properties, and the medaka Vtg bioassay could be a very sensitive and good tool for screening of endocrine disrupting compounds.

Animals↗
PubMed2005

Purification and gene cloning of Fundulus heteroclitus hatching enzyme. A hatching enzyme system composed of high choriolytic enzyme and low choriolytic enzyme is conserved between two different teleosts, Fundulus heteroclitus and medaka Oryzias latipes.

Two cDNA homologues of medaka hatching enzyme -- high choriolytic enzyme (HCE) and low choriolytic enzyme (LCE) -- were cloned from Fundulus heteroclitus embryos. Amino acid sequences of the mature forms of Fundulus HCE (FHCE) and LCE (FLCE) were 77.9% and 63.3% identical to those of medaka HCE and LCE, respectively. In addition, phylogenetic analysis clearly showed that FHCE and FLCE belonged to the clades of HCE and LCE, respectively. Exon-intron structures of FHCE and FLCE genes were similar to those of medaka HCE (intronless) and LCE (8-exon-7-intron) genes, respectively. Northern blotting and whole-mount in situ hybridization showed that both genes were concurrently expressed in hatching gland cells. Their spatio-temporal expression pattern was basically similar to that of medaka hatching enzyme genes. We separately purified two isoforms of FHCE, FHCE1 and FHCE2, from hatching liquid through gel filtration and cation exchange column chromatography in the HPLC system. The two isoforms, slightly different in molecular weight and in MCA-peptide-cleaving activity, swelled the inner layer of chorion by their limited proteolysis, like the medaka HCE isoforms. In addition, we identified FLCE by TOF-MS. Similar to the medaka LCE, FLCE hardly digested intact chorion. FHCE and FLCE together, when incubated with chorion, rapidly and completely digested the chorion, suggesting their synergistic effect in chorion digestion. Such a cooperative digestion was confirmed by electron microscopic observation. The results suggest that a hatching enzyme system composed of HCE and LCE is conserved between two different teleosts Fundulus and medaka.

Amino Acid Sequence↗
PubMed2004

Quantifying the ontogeny of optokinetic and vestibuloocular behaviors in zebrafish, medaka, and goldfish.

We quantitatively studied the ontogeny of oculomotor behavior in larval fish as a foundation for studies linking oculomotor structure and function with genetics. Horizontal optokinetic and vestibuloocular reflexes (OKR and VOR, respectively) were measured in three different species (goldfish, zebrafish, and medaka) during the first month after hatching. For all sizes of medaka, and most zebrafish, Bode plots of OKR (0.065-3.0 Hz, +/-10 degrees/s) revealed that eye velocity closely followed stimulus velocity (gain > 0.8) at low frequency but dropped sharply above 1 Hz (gain < 0.3 at 3 Hz). Goldfish showed increased gain proportional to size across frequencies. Linearity testing with steps and sinusoids showed excellent visual performance (gain > 0.8) in medaka almost from hatching; but zebrafish and goldfish exhibited progressive improvement, with only the largest equaling medaka performance. Monocular visual stimulation in zebrafish and goldfish produced gains of 0.5 versus <0.1 for the eye viewing a moving versus stationary stimulus pattern but 0.25 versus <0.1 in medaka. Angular VOR appeared much later than OKR, initially at only high accelerations (>200 degrees /s at 0.5 Hz), first in medaka followed by larger (8.11 mm) zebrafish; but it was virtually nonexistent in goldfish. Velocity storage was not observed except for an eye velocity build-up in the largest medaka. In summary, a robust OKR was achieved shortly after hatching in all three species. In contrast, larval fish seem to be unique among vertebrates tested in their lack of significant angular VOR at stages where active movement is required for feeding and survival.

Animals↗
PubMed2004

Increased kidney, liver, and testicular cell death after chronic exposure to 17alpha-ethinylestradiol in medaka (Oryzias latipes).

Sublethal effects observed in fish exposed to environmental estrogens may be mediated via stimulation of cell death. To investigate whether cell death is induced in fish after chronic exposure to estrogenic chemicals, Japanese medaka (Oryzias latipes) were exposed from hatch until sexual maturity to 10 ng/L 17alpha-ethinylestradiol (EE2) or acetone solvent (control). Cell death was evaluated in blinded histological sections of whole medaka using terminal dideoxynucleotidyl-mediated dUTP nick end-labeling (TUNEL), which labels nuclei of cells containing apoptotic or necrotic (fragmented) DNA. The major impact of EE2 exposure in both male and female medaka was to significantly increase the number of TUNEL-positive hepatocytes and kidney tubule cells compared to control. Cell morphology was consistent with apoptosis in the liver and cloudy swelling or necrosis in the tubule cells. The number of TUNEL-positive interstitial (hematopoietic) and glomerular cells was significantly greater in the kidneys of EE2-exposed male, but not female, medaka. The EE2 exposure also significantly increased the number of TUNEL-positive testicular cells in medaka compared to corresponding controls, namely Leydig cells, Sertoli cells, spermatocytes, and spermatids. In medaka with gonadal intersex, areas of fibrosis and areas containing female gonadal cells were relatively unstained with TUNEL. No effect of EE2 exposure on the number of TUNEL-positive ovarian somatic cells or on the rate of female ovarian follicle atresia was found. These results suggest that chronic exposure to EE2 in medaka is hepatotoxic and nephrotoxic in both sexes, whereas gonadal toxicity is specific to males.

Animals↗
PubMed2005

Effects of polychlorinated biphenyls on the schooling behavior of Japanese medaka (Oryzias latipes).

The effects of polychlorinated biphenyls (PCBs; Kanechlor 400) on the schooling behavior of Japanese medaka (Oryzias latipes) were tested after feeding with various concentrations of PCBs (0, 1, 5, 25, and 125 microg/g). To test schooling, three PCB-exposed medaka and three untreated fish were placed in one chamber, and their swimming trajectories were recorded. Swimming velocity, turning angle, and nearest-neighbor distance (NND) were analyzed individually, and the polarization and expanse of each schooling group were analyzed. Fractal dimension analysis was performed for trajectory, swimming velocity, and turning angle. Six behaviors (school formed by six swimming medaka, school formed by six stationary medaka, school formed by three to five swimming medaka, school formed by three to five stationary medaka, swimming without schooling, and stationary without schooling) were quantified with time. Behavioral parameters were the same for PCB-exposed and unexposed medaka in the same schooling group. Swimming velocity decreased in a dose-dependent manner, and NND in the group exposed to 25 microg/g of PCBs was significantly longer (126%) than that in the controls. The fractal dimensions of trajectory and turning angle increased significantly in the highest PCB-exposure group, indicating that schools containing PCB-exposed individuals frequently changed direction. The PCBs shortened the time of school formed by six swimming fish, which was 23.7% of the control value in the group exposed to 25 microg/g of PCBs. The frequency of collisions between individuals was significantly greater in the highest-exposure group (31.3) than in the controls (13.3). Thus, PCB exposure influences the behavior of unexposed fish in the same school, consequently affecting schooling behavior.

Animals↗
PubMed2001

Estrogenic influences of estradiol-17 beta, p-nonylphenol and bis-phenol-A on Japanese medaka (Oryzias latipes) at detected environmental concentrations.

Mature male medaka were continuously exposed to 0.005, 0.0-5 or 1.0 ppb of estradiol-17 beta (E2 or 0.1, 10 or 100 ppb of p-nonylphenol (NP) or bis-phenol-A (BPA). Female-specific proteins (Fsp) were induced in medaka exposed to 0.005 ppb of E2, 0.1 ppb of NP, or 10 ppb of BPA. Concentrations of 0.005 pbb of E2 and 0.1 ppb of NP corresponded to concentrations of these chemicals detected in river water in Japan. The abilities of the 3 chemicals to induce Fsp were E2 > NP > BPA. Embryonic medaka were exposed to E2, NP and BPA under conditions of static-renewal for 200-230 days until pre-maturity. Survival ratios of medaka exposed to E2 and NP declined in concentrations more than 25 ppb and 50 ppb, respectively. The groups of medaka exposed to E2 had individuals with testis-ova or abnormal gonad. There was no male in exposure to 1.0 ppb E2. When exposed to 100 ppb of NP or BPA, abnormal gonad was also detected. Abnormal anal fin (female-like) was observed in male exposed to 100 ppb of NP. The LC50 values for each of the 3 chemicals were much higher than the concentrations detected in water in the environment--the 3 chemicals were considered to have no lethal effect on medaka in aquatic environments. However, exposures to E2 or NP at environmental concentrations induced Fsp. BPA also had the ability to affect medaka as an environmental estrogen, although its extrogenic activity was weaker than that of E2 or NP.

Animals↗
PubMed1994

An efficient expression vector for transgenic medaka construction.

The transparency and external fertilization of the eggs of medaka (Oryzias latipes) make them ideally suitable for investigating molecular interactions that occur during vertebrate development. Genetically engineered medaka is a potential tool for such studies. It requires several types of suitable expression vectors. To obtain abundant and ubiquitous expression of foreign genes in medaka embryos, we have designed an expression vector that contains the proximal promoter and enhancer elements and polyadenylation signal of the medaka beta-actin gene. The utility of this "all-medaka" expression vector was examined using the Escherichia coli lacZ gene as a reporter gene. Most of the injected embryo showed high gene expression, and several embryos showed ubiquitous expression even at six days after injection. Of nine individuals derived from the injected embryos and grown until adult stage, one produced expression-positive F1 fish. The transgene was identified in these F1 using polymerase chain reaction (PCR). These data revealed that the expression vector based on the expression cassette from the medaka beta-actin gene should be useful for making transgenic medaka. The cloned gene in this cassette vector is stably transmittable and efficiently expressible.

Actins↗
PubMed1996

Isolation, characterization, and expression of cDNAs encoding the medaka (Oryzias latipes) ovarian follicle cytochrome P-450 aromatase.

Two cDNA clones of cytochrome P-450 aromatase (P-450arom) were isolated from a medaka (Oryzias latipes, a daily spawner) ovarian follicle cDNA library using a medaka P-450arom genomic DNA fragment as a probe. The first, 1,809-bp insert (S81f) contains an 1,554-bp open reading frame encoding a 518-amino-acid polypeptide. The second, 1,852-bp (S52f) insert possesses an open reading frame identical to that of the S81f insert, except for the absence of the heme-binding region as the result of an additional A residue at the position of nucleotide 1,827. Expression of the S81f cDNA, but not of the S52f cDNA, in nonsteroidogenic COS-1 cells leads to production of a steroidogenic enzyme which is capable of converting exogenous testosterone to estrogen. The P-450arom genomic DNA fragment hybridizes to a single mRNA in medaka ovarian follicle RNA. Changes in level of P-450arom transcripts and P-450arom enzyme activity were determined in medaka ovarian follicles collected at 16 stages of development. A close correlation was found between these two profiles, both being high in midvitellogenic follicles and low in postvitellogenic follicles collected during oocyte maturation. These findings, together with those of our previous study showing that actinomycin D prevented gonadotropin-induced aromatase activation by medaka ovarian follicles, suggest that aromatase activity is regulated at the transcriptional level in medaka vitellogenic follicles.

Animals↗
PubMed2001

Use of Japanese medaka (Oryzias latipes) and Tilapia (Oreochromis mossambicus) in toxicity tests on different industrial effluents in Taiwan.

In Taiwan, aquatic toxicity tests for industrial effluents are not required for discharge permits. However, relying on traditional chemical and physical characteristics of an effluent to monitor and regulate such discharges to manage water quality of a receiving water is insufficient. In this study, we used two fish species, Japanese medaka (Oryzias latipes) and tilapia (Oreochromis mossambicus), and three toxic endpoints, including acute and subacute toxicity, to determine toxicity of seven different types of industrial effluents. Prior to the study, two reference toxicants were tested on two fish species. The LC50s of CdCl2 for tilapia and medaka juveniles were 29.6 +/- 15.3 mg/L and 2.2 +/- 1.2 mg/L, respectively. The sensitivity of medaka embryo mortality and hatching inhibition to CdCl2 were about the same, with the LC50 and EC50 of 0.3 +/- 0.1 mg/L and 0.1 +/- 0.1 mg/L, respectively. The LC50s for tilapia and medaka juveniles to sodium dodecyl sulfate (SDS) were 19.7 +/- 10.6 mg/L and 12.5 +/- 5.9 mg/L. The medaka embryo was less sensitive to SDS than to CdCl2. The embryo's LC50 for SDS was 5.8 +/- 2.8 mg/L and the hatching inhibition EC50 was 1.3 +/- 1.1 mg/L. Results of toxicity tests on different effluents showed that the electroplating effluent was the most toxic, followed by acrylonitrile manufacturing and pulp/paper mill discharges. The LC50s of the electroplating effluent to different assays were in the range of several percents of the whole effluent. The pulp/paper effluent was toxic only to the medaka embryo. The rest of the industrial effluents tested showed either moderate or no toxicity to the animals.

Animals↗
PubMed2006

Mycobacterium marinum produces long-term chronic infections in medaka: a new animal model for studying human tuberculosis.

Human infection by Mycobacterium tuberculosis is endemic, with approximately 2 billion infected and is the most common cause of adult death due to an infectious agent. Because of the slow growth rate of M. tuberculosis and risk to researchers, other species of Mycobacterium have been employed as alternative model systems to study human tuberculosis (TB). Mycobacterium marinum may be a good surrogate pathogen, conferring TB-like chronic infections in some fish. Medaka (Oryzias latipes) has been established for over five decades as a laboratory fish model for toxicology, genotoxicity, teratogenesis, carcinogenesis, classical genetics and embryology. We are investigating if medaka might also serve as a host for M. marinum in order to model human TB. We show that both acute and chronic infections are inducible in a dose dependent manner. Colonization of target organs and systemic granuloma formation has been demonstrated through the use of histology. M. marinum expressing green fluorescent protein (Gfp) was used to monitor bacterial colonization of these organs in fresh tissues as well as in intact animals. Moreover, we have employed the See-Through fish line, a variety of medaka devoid of major pigments, to monitor real-time disease progression, in living animals. We have also compared the susceptibility of another prominent fish model, zebrafish (Danio rerio), to our medaka-M. marinum model. We determined the course of infections in zebrafish is significantly more severe than in medaka. Together, these results indicate that the medaka-M. marinum model provides unique advantages for studying chronic mycobacteriosis.

Animals↗
PubMed2006

Effects of elevated temperature and nickel pollution on the immune status of Japanese medaka.

Changes in a host's environment (i.e. physical or chemical) can alter normal immune function. In aquatic organisms, exposure to stress can result in significant changes in innate immunity. In the natural environment, fish are exposed to multiple stressors simultaneously. Temperature change and/or chemical exposure as individual environmental stressors have been shown in various fish species to alter all aspects of the immune response. These same stressors have also been shown to alter plasma steroid levels in exposed fish. For this study, the effects of elevated temperature and nickel pollution on specific immune parameters of Japanese medaka (Oryzias latipes) were determined. Fish were exposed for 1, 7 or 14d to either: waterborne nickel (Ni) at the nominal concentration of 125ppb; a 5 degrees C (+/-0.5 degrees C) rapid increase in water temperature; or, both potential stressors in combination. Medaka maintained at room temperature (25 degrees C+/-1 degrees C) served as the controls. Altered function of the innate and adaptive arms of the immune response was evaluated by assessing kidney macrophage-mediated superoxide (O(2)(-)) production and splenic T-cell proliferation, respectively. Plasma cortisol levels were analysed in the same fish as a marker of the physiological stress response. While kidney cell number was unaffected by exposure of fish to either stressor alone or both factors in combination, spleen cellularity was decreased (compared to control fish) in medaka exposed for 1d to thermal stress in combination with Ni, and to a lesser extent to thermal stress alone. T-lymphocyte proliferation by medaka splenocytes was not affected by any exposure paradigm. Unstimulated intracellular O(2)(-) production by kidney phagocytes was significantly elevated (compared to control) in medaka exposed for 1d to either thermal stress alone or temperature change in combination with Ni; by 7d, only the stressor combination significantly increased baseline O(2)(-) production. Resting levels of extracellular O(2)(-) production was significantly reduced in fish maintained for 1d at the elevated temperature. Effects on phorbol 12-myristate 13 acetate (PMA)-stimulated intracellular and extracellular O(2)(-) production were less dramatic than those observed for resting phagocytes. Exposure of medaka to elevated temperature for 14d tended (p<0.06) to reduce PMA-stimulated intracellular O(2)(-) production (compared to the time-matched control). Although exposure of fish for 14d to elevated temperature only slightly reduced stimulated extracellular O(2)(-) production, exposure for the same duration to Ni alone significantly depressed oxyradical production by kidney phagocytes (compared to the time-matched controls). Decreased plasma cortisol levels were observed in fish exposed for 7d to either an elevated water temperature or Ni (compared to the time-matched control); by 14d of exposure, no significant treatment-induced effects on cortisol levels were observed. These findings add to the growing body of literature seeking to determine what effects, if any, exposure to multiple aquatic pollution-induced effects have upon fish health and the health of impacted ecosystems.

Animals↗
PubMed2005

Genetic linkage map of medaka with polymerase chain reaction length polymorphisms.

With recent improvements in genetic and genomic infrastructures, great interest has been taken in genetic dissection of multi-factorial traits. A genetic map consisting of markers that are highly polymorphic and rapidly genotyped is essential for the genetic mapping of such a complex trait. Medaka, Oryzias latipes, is an excellent model system for genetic studies. To promote genetic mapping of complex traits in medaka we developed the first high-throughput and genome-wide marker set in the organism by using its genomic information and the bioinformatic techniques. We tested 545 primer pairs and obtained 265 co-dominant markers between two inbred strains, HNI and Hd-rR. Our map, consisting of 231 uniquely mapped markers, covers 1257.3 centimorgan (cM) of the medaka genome with an average interval distance of 5.4 cM. Furthermore, the newly designed markers were examined for polymorphisms among six medaka inbred strains: HNI, Hd-rR and four additional strains. Most of our markers are simple sequence length polymorphisms (SSLPs) and can be rapidly genotyped by an automated system under a single polymerase chain reaction (PCR) condition. Together with the genotyping data of six medaka inbred strains, our new marker set provides a powerful tool for genome-wide analysis of complex biological phenomena found widely in medaka populations.

Animals↗
PubMed2006

Anti-androgen flutamide affects gonadal development and reproduction in medaka (Oryzias latipes).

The effects of an anti-androgenic chemical on the reproduction of medaka (Oryzias latipes) were examined. Six mating pairs of medaka were exposed to flutamide (FLU) at measured concentrations of 0.101, 0.202, 0.397, 0.787, or 1.56 mg/L for 3 weeks. During the exposure period, one female died in the 1.56-mg/L treatment group, but no lesions or pathological abnormalities were observed. The fecundity and fertility of paired medaka were significantly decreased at 1.56 mg/L compared with those of the controls. Histological examination showed intersex gonads (testis-ova) in males exposed to FLU at 0.202, 0.397, and 0.787 mg/L. However, concentrations of hepatic vitellogenin (Vtg) in both sexes were not statistically different from those in the controls. These results demonstrate that FLU affected gonadal development in male medaka and reproductive capacity in paired medaka. This is the first report of testis-ova in male medaka exposed to FLU.

Androgen Antagonists↗
PubMed2004

Twist functions in vertebral column formation in medaka, Oryzias latipes.

Medaka twist, a basic helix-loop-helix (bHLH) transcription factor, is expressed in the sclerotome during embryogenesis. We previously established a line of twist-EGFP transgenic medaka, whose EGFP expression is regulated by the twist promoter; therefore, we could observe the behavior of sclerotomal cells in vivo. In the transgenic medaka embryos, EGFP-positive sclerotomal cells migrated dorsally around the notochord and the neural tube, where at a later stage the vertebral column would be formed. This finding strongly suggests that twist-expressing sclerotomal cells participate in vertebral column formation in medaka. To clarify the function of twist gene in the sclerotome, we performed knockdown analysis of twist by using two kinds of morpholino antisense oligonucleotides targeted against twist (MO1 and MO2). Both the MO1 and MO2 morphants exhibited absence of neural arches, which are bilaterally paired, dorsomedially oriented bones on the dorsal aspect of the centrum. In addition, MO2, which blocks translation of only endogenous twist mRNA in the twist-EGFP transgenic medaka, did not affect the migration pattern of EGFP-positive cells, revealing that the migration of sclerotome-derived cells were normal in the absence of twist gene function. These results demonstrate that medaka twist functions in vertebral column formation by regulating the sclerotomal cell differentiation.

Amino Acid Sequence↗
PubMed2006

Protection against nonylphenol-induced cell death by DJ-1 in cultured Japanese medaka (Oryzias latipes) cells.

The Japanese medaka (Oryzias latipes) has been used to investigate diverse aspects of toxicology, genetics and developmental biology and to monitor biological changes caused by endocrine disruptors. In this study, we analyzed a medaka homolog of human DJ-1 (meDJ-1) in cultured medaka cells into which nonylphenol (NP) was added. Like human DJ-1, meDJ-1 was found to be oxidized by treatment with H(2)O(2) and its pI was shifted to acidic points. NP was found to induce cell death with kinetics similar to that of H(2)O(2) in cultured medaka OLHE-13 cells. After OLHE-13 cells had been treated with sub-lethal concentrations of H(2)O(2) and NP, production of reactive oxygen species and oxidation of meDJ-1 were observed. meDJ-1 knockdown by short interfering RNA rendered OLHE-13 cells susceptible to H(2)O(2) and NP-induced cell death, suggesting a protective role of DJ-1 against oxidative stress-induced cell death in medaka cells. These results suggest that meDJ-1 is a suitable biomarker for oxidative stress reactions in medaka.

Amino Acid Sequence↗
PubMed1998

Carcinogenic effects of 1,2-dibromoethane (ethylene dibromide; EDB) in Japanese medaka (Oryzias latipes).

The carcinogenicity of 1,2-dibromoethane (ethylene dibromide; EDB) was investigated in the Japanese medaka (Oryzias latipes), a small fish species. EDB was administered in water continuously for 97 days to a low concentration group, for 73 days to an intermediate concentration group, and intermittently for 24 h once each week over 97 days to a high concentration group. Medaka were 7 days old at the beginning of the tests. Mean measured EDB concentrations in the ambient water were 0.13 mg l-1, 6.20 mg l-1, and 18.58 mg l-1 in the low, intermediate, and high concentration groups, respectively. Two control groups, one inside and one outside the exposure apparatus, were used. Samples were examined histologically at 24, 36, and 58 weeks from the beginning of the tests. EDB was clearly carcinogenic to medaka in the intermediate and high concentration groups causing (1) hepatocellular adenomas and carcinomas, (2) cholangiomas, (3) chloangiocarcinomas, and (4) gall bladder papillary adenomas and adenocarcinomas. In separate studies, medaka exposed to 1.0 mg l-1 EDB for 2 to 5 weeks had elevated hepatic glutathione S-transferase activities, possibly indicating induction of a pathway that forms the reactive metabolite of EDB in mammals. SDS-PAGE of hepatic cytosolic fractions of EDB-exposed medaka showed a pronounced increase in a band at 26,000 Da, the expected position for GSH-S-transferase. Although little is known about EDB's mechanisms of action, medaka appear exceptionally sensitive to the carcinogenic effects of EDB and could serve as a model test species for studying similar compounds.

Adenoma↗
PubMed1999

Medaka (Oryzias latipes) FTZ-F1 potentially regulates the transcription of P-450 aromatase in ovarian follicles: cDNA cloning and functional characterization.

Our previous findings suggest the activity of cytochrome P-450 aromatase (P-450arom), the enzyme which converts testosterone to estradiol-17beta, in the ovarian follicle of medaka (Oryzias latipes) is regulated at the transcriptional level. In this study, we cloned a cDNA encoding a FTZ-F1-like protein (mdFtz-F1) from ovarian follicles of medaka. In vitro translated mdFTZ-F1, and nuclear extract from medaka ovarian follicles, formed complexes with oligonucleotide probes containing putative orphan nuclear receptor binding motifs, which are present in the promoter region of the medaka P-450arom gene. The expression pattern of mdFtz-F1 transcripts during oogenesis coincides with that of P-450arom transcripts. Transfection assays further suggest a potential transcriptional regulatory activity of mdFTZ-F1 upon the medaka P-450arom promoter. Taken together, these results suggest a potential role of mdFTZ-F1 in the transcriptional regulation of P-450arom in the ovarian follicle of medaka.

Amino Acid Sequence↗