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PubMed1994

Cloning and expression of a renal Na-Pi cotransport system from flounder.

Starting with the recently published sequence of the rat renal Na-Pi cotransport system, we have cloned a corresponding cDNA from the kidney of winter flounder (Pseudopleuronectes americanus), designated flounder NaPi-II. Expression of the cognate in vitro transcribed RNA in Xenopus laevis oocytes stimulated Na-dependent Pi transport specifically and in a time- and dose-dependent manner. Apparent affinities of Na and Pi, as well as the pH dependency, were very similar to those found for the mammalian systems. The flounder NaPi-II cDNA is 2,424 base pairs long and encodes a protein of 637 amino acids. The hydropathy plot predicts eight transmembrane spanning domains. In these regions the flounder NaPi-II-deduced protein shows high homology (approximately 80%, identity, approximately 92% similarity) with the amino acid sequences reported for mammalian NaPi-II proteins. However, in the hydrophilic parts of flounder NaPi-II protein, only minimal similarity could be found between fish and mammalian systems (30% homology, 45% similarity). Northern blot analysis with flounder NaPi-II cDNA as a probe confirmed this finding: even under nonstringent washing conditions, no cross-hybridization with mRNA from rat renal cortex was observed. Interestingly, flounder intestine was found to contain high levels of mRNA corresponding to NaPi-II. Supplementary bands of 1.9 and 4.2 kb were observed on Northern blots of renal and intestinal tissue. The close functional relationship of the flounder NaPi-II protein with the previously described Na-Pi cotransport systems and the pronounced differences on the level of their primary structures provide the tools for detailed structure-function analysis of Na-Pi cotransport.

Amino Acid Sequence↗
PubMed2005

Development of non-radioisotopic immunoassay systems for measuring flounder IGF-I.

A time-resolved fluoroimmunoassay system (TR-FIA) for measuring flounder insulin-like growth factor-I (IGF-I) was developed using biotinylated flounder IGF-I, anti-fish IGF-I antiserum and europium-avidin conjugate. The detection limit per well was <5 pg/well corresponding to <0.5 ng/ml in a basic procedure for sample of 10 microl/well and to <0.08 ng/ml in a procedure modified for high volume samples (up to 70 microl/well). Specificity of the assay was validated using various IGF-Is and insulins. All IGFs except seabream IGF-I showed very low or no crossreactivity. Binding inhibition curves for flounder and seabream IGF-Is were completely identical to each other. Intra- and interassay variations ranged from coefficients of variations of 3.9% to 7.2%. Recovery tests using barfin flounder plasma varied from 82.7 to 101.6% in the added range from 20 to 160 ng/ml. This assay system was applied for measuring total plasma IGF-I in barfin flounder injected porcine growth hormone (GH). A group injected with GH at the dose of 0.05 IU/gBW showed a significant increase of total plasma IGF-I compared with those of albumin-injected (control) and initial groups. In addition, I was able to substitute time-resolved fluorometric detection in this assay system with enzymatic fluorometric detection (FIA). Binding inhibition curve for flounder IGF-I in this substituted assay system showed equal performance with that of the TR-FIA system. Correlation of IGF-I levels between TR-FIA and FIA was high (r(2)=0.957) in plasma samples from barfin flounders in various physiological conditions. Thus, the present study shows precision and efficiency of two non-radioisotopic immunoassay systems for measuring flounder IGF-I.

Animals↗
PubMed1984

Heterogeneity of hepatic aryl hydrocarbon hydroxylase activity in feral winter flounder: relevance to carcinogenicity testing.

Hepatic microsomes from winter flounder (Pseudopleuronectes americanus), treated with polycyclic aromatic hydrocarbon (PAH) inducers, had elevated activities of benzo[alpha]pyrene hydroxylase (AHH) and 7-ethoxyresorufin deethylase (7-ERD). When electrophoresed, they showed a novel or enriched polypeptide species with a monomeric molecular weight of approximately 57,000. These results are consistent with inductive responses already well-characterized in several mammalian and fish species. However, when we studied the urinary clearance of 4-chlorobiphenyl in untreated flounder, wide variations (up to twentyfold) among fish were noted. Subsequent in vitro analysis of AHH and 7-ERD activities in liver demonstrated wide variations in these monooxygenase activities in flounder caught near Mount Desert Island, Maine. In some instances, AHH activities in these feral flounder were as high as those in PAH-induced fish. Based on the response of AHH activity to 7,8-benzoflavone (ANF) added in vitro, flounder could be divided into 2 groups; one had high hepatic AHH activity which was inhibited by ANF, the other had low AHH activity which was enhanced by ANF. Examination of a large number of winter flounder (greater than 400 total) over 4 experimental seasons demonstrated this variability of hepatic AHH activity to be a recurrent characteristic of the flounder population in waters around Mount Desert Island. The hepatic AHH activities did not correlate well with any physical parameter of the fish (e.g., liver, gonad or body weight, length, or sex) or the cytochrome P-450 content of the hepatic microsomes. Our attempt to evaluate the AHH activity (high vs. low) of individual fish in vivo by urinary clearance of antipyrine was unsuccessful, due to the excretion of large amounts of unchanged antipyrine through the gills. Similar studies were performed with another marine teleost, Fundulus heteroclitus. Of the approximately 200 Fundulus examined, almost all had AHH activity inhibited by ANF; in some experiments, hepatic 7-ERD activities were further increased after treatment with the potent PAH-type inducer 3,4,5,3',4',5'-hexachlorobiphenyl but only about twofold. Collectively, these data are consistent with PAH-type induction of the hepatic cytochrome P-450-dependent monooxygenase system in some feral marine teleosts (in winter flounder and Fundulus) from Maine by environmental contaminants or food constituents. It is not known whether these chemicals are of natural or anthropogenic origin. The variation in the response appears to be related to individual exposure level (dose) or sensitivity, or both.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed2004

The gender-specific risk to liver toxicity and cancer of flounder (Platichthys flesus (L.)) at the German Wadden Sea coast.

Flatfish living in coastal areas are chronically exposed to a wide range of toxic and (pro)carcinogenic compounds derived from agriculture and industry. Flounder (Platichthys flesus (L.)) is the main target species for monitoring health effects of contamination in North Sea and Baltic Sea since the species is abundant, benthic, and inhabits shallow waters such as the Wadden Seas and estuaries along salinity gradients into fresh water. Chemical analysis in the same livers as investigated for histopathology in the present study showed positive correlation between accumulation of certain organochlorines in liver and the extend of liver injury. Toxipathic liver changes including neoplasms in female and male flounder were analysed by macroscopic and light microscopic diagnosis during a five-year survey on the basis of internationally accepted criteria agreed upon during the European BEQUALM intercalibration of liver histopathology of flatfish. Hepatocellular carcinogenesis of wild flounder principally showed sequential changes similar to experimental chemical carcinogenesis in other fish species and mammals. These ranged from early foci of altered hepatocytes (vacuolated/clear/eosinophilic, basophilic cells) and the development of adenomas. With progression to hepatocellular carcinomas, livers of wild flounder entered a multistage phase of carcinogenesis comprising of early foci, hepatocellular adenomas and carcinomas, as observed in human liver cancers. Female flounder had three-fold higher frequencies of macroscopically visible neoplasms than males of the same age classes. Histopathological diagnosis showed that hepatocellular alterations in male flounder never developed further than stages of basophilic foci and adenomas, and never into malignancies. In females, tumors of hepatocellular origin clearly dominated, occurred alone and together with cancers of bile duct epithelial cells and endothelial cells (cholangio-carcinomas, angiosarcomas). Because mutations of relevant genes could not be identified in the liver cancers that were analysed in the present study, we conclude that epigenetic events initiate carcinogenesis. Therefore, we conclude that mitogenic carcinogens found in the Elbe estuary play a dominant role for cancer development in this flounder population. Furthermore, the lower availability of reducing power (NADPH) needed for detoxification reactions in females compared to males, together with annual upregulation of the highly potent promotor 17-beta-estradiol during sexual maturation, promote progression of preneoplastic foci of initiated cells to malignant cancers in females.

Adenoma, Liver Cell↗
PubMed2006

Characterization of Japanese flounder (Paralichthys olivaceus) NK-lysin, an antimicrobial peptide.

The NK-lysin cDNA of Japanese flounder, Paralichthys olivaceus, consists of 657bp, containing an open reading frame (ORF) of 444bp, which encodes 147 amino acid residues. The amino acid sequence of Japanese flounder NK-lysin has 21% identity to porcine NK-lysin and bovine NK-lysin, 23% to equine NK-lysin, and 46% to zebrafish NK-lysin-like protein. Multiple alignments of Japanese flounder NK-lysin and other known saposin-like proteins revealed that the six cysteine residues important for structural folding are completely conserved. The Japanese flounder NK-lysin gene is approximately 2kb and consists of five exons and four introns. Japanese flounder NK-lysin mRNA constitutive expression was mainly detected in gills, heart, head kidney, intestines, peripheral blood leukocytes (PBLs), spleen and trunk kidney, and was detected at low levels in liver, muscle and ovary. However, expression was not detected in brain, skin and stomach of apparently healthy Japanese flounder. Gene expression of Japanese flounder NK-lysin was not inducible by lipopolysaccharide (LPS) treatment. A synthesized NK-lysin peptide, consisting of 27 amino acid residues, showed antimicrobial activity against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Photobacterium damselae subsp. piscicida.

Amino Acid Sequence↗
PubMed2001

Cloning and sequence analysis of the interleukin-8 gene from flounder (Paralichthys olivaceous).

A cDNA library of mRNA from flounder leukocytes stimulated with bacterial lipopolysaccharide (LPS) and hemagglutinin was constructed to clone cytokine genes of this fish. Initial screening of this library with human cytokine gene probes was not productive and clones with inserts of over 400 nucleotides (nt) were randomly sequenced, and a homologue of the vertebrate interleukin-8 (IL-8) gene was isolated. The flounder IL-8 cDNA encompassed 884 nt, including a coding region of 330 nt. Four cysteines characteristic of CXC chemokines were identified at conserved locations in the putative protein. The deduced amino acid sequence showed 36 and 35% sequence identity with counterpart genes in monkey and human, respectively, and 52% sequence similarity with these genes. However, the putative flounder IL-8 amino acid sequence showed 25% identity and 52% similarity to that of lamprey, the only other piscine IL-8 gene that has been cloned. Flounder IL-8 transcripts were detected in the head-kidney and spleen of LPS-injected flounder and leukocytes stimulated with LPS. It was not detected in the muscle or liver of LPS-injected flounder, tissues taken from non-stimulated flounder and non-stimulated leukocytes.

Amino Acid Sequence↗
PubMed1999

The detection of biomarkers of genotoxin exposure in the European flounder (Platichthys flesus) collected from the River Tyne Estuary.

The Tyne Estuary (North East England) is known to contain elevated levels of polycyclic aromatic hydrocarbons (PAH), compared with other less industrialised English waterways. Previous studies suggest that such contamination is responsible for the toxicity detected in invertebrate bioassays conducted on water and sediment samples collected from the Tyne. Here we present data from a biomonitoring study using hepatic DNA adducts (32P-postlabelling assay) and bile metabolites (synchronous fluorescence spectrometry) to investigate genotoxic exposure in flounder (Platichthys flesus) collected from three sites (Scotswood, Newcastle and Redheugh) along the Tyne Estuary. Flounder were also collected from a clean reference site, the Alde Estuary. Levels of bile metabolites (microgram kg-1 wet weight 1-OH pyrene equivalents) were elevated in flounder caught from the Tyne (Scotswood = 22,247 +/- 3408; Newcastle = 14,572 +/- 1888; Redheugh = 21,872 +/- 2935) compared with those collected from the Alde (632 +/- 56). The levels of DNA adducts (adducted nucleotides per 10(8) normal nucleotides) were also elevated in Tyne flounder (Scotswood = 24.6 +/- 3.2; Newcastle = 34.4 +/- 3.7; Redheugh = 27.6 +/- 6.3) compared with fish collected from the Alde (10.1 +/- 4.8), suggesting that a proportion of the bioavailable PAH was being converted into genotoxic metabolites. All DNA adduct profiles in flounder collected from the Tyne consisted of diagonal radioactive zones of radiolabelled adducts, which were not present in fish sampled from the Alde. The in vivo dosing of flounder with benzo[a]pyrene (BaP) to produced DNA adducts in similar chromatographic positions to the diagonal radioactive zones in the Tyne caught flounder are also described.

Animals↗
PubMed2005

Doses from the consumption of Cardiff Bay flounder containing organically bound tritium.

ICRP dose coefficients for the ingestion of organically bound tritium (OBT) by adults and children are intended for general application to unspecified forms in diet and may not be applicable to intakes of a specific form of OBT. To obtain information relating to OBT in fish from Cardiff Bay, the retention of tritium in adult rats was determined after administration as either tritiated water (HTO) or dried flounder flesh containing OBT. Two components of retention were obtained in each case. The first component, attributable to tritium equilibrating with body water, had a half-time of retention of 3 days in each case, and accounted for 97% of the intake as HTO and 70% after intake of OBT in flounder. Results were consistent with rapid catabolism of a large proportion of flounder OBT to HTO. The second component of retention, attributable to OBT in rat tissues, accounted for 3% of tritium intake as HTO and 30% after intake as flounder OBT; the half-times of retention were 10 days and 25 days, respectively. The results obtained after administration as HTO are consistent with published animal data and correlate with the ICRP assumptions for adult man of half-times of 10 days for 97% behaving as HTO in body tissues and 40 days for 3% incorporated into OBT in body tissues. The results obtained after administration of flounder OBT suggest that appropriate assumptions for retention in adult man are 70% with a 10 day half-time and 30% with a 100 day half-time. These assumptions result in an ingestion dose coefficient of 6 x 10(-11) Sv Bq(-1). This compares with the ICRP value for OBT ingestion by adults of 4.2 x 10(-11) Sv Bq(-1), based on half-times of 10 days and 40 days applied to equal proportions of retained tritium. It is proposed that a dose coefficient of 6 x 10(-11) Sv Bq(-1) should be applied to tritium in flounders from Cardiff Bay. It is further proposed that this dose coefficient should be applied to all ingestion intakes by adults relating to this source of exposure, unless specific information is available showing that a significant proportion of the intake is HTO. The same proposals apply to dose coefficients derived here for flounder OBT consumption by children.

Animals↗
PubMed1988

In vivo lactate kinetics at rest and during recovery from exhaustive exercise in coho salmon (Oncorhynchus kisutch) and starry flounder (Platichthys stellatus).

A bolus injection of [14C]lactate was used to measure lactate turnover rates at rest and during recovery from exhaustive exercise in coho salmon (Oncorhynchus kisutch) and starry flounder (Platichthys stellatus). At rest, lactate turnover rate in salmon was almost double that in flounder (1.33 versus 0.76 mumol min-1 kg-1), which reflected the higher blood lactate level in salmon (1.00 versus 0.12 mmol l-1). From 2 to 4 h after exercise, when blood lactate levels were at their peak and constant, turnover rates were elevated in both species, though to a greater extent in salmon than in flounder (11.88 versus 2.27 mumol min-1 kg-1). Lactate concentration and turnover rate were linearly correlated in both species. The higher turnover rate in salmon was solely a consequence of the higher blood lactate levels since, at similar blood lactate concentrations, turnover rates in flounder and salmon were the same. Therefore, the lower blood lactate levels in flounder after exercise were not a consequence of higher turnover. In neither species was the turnover rate adequate to account for the rate of lactate clearance from the muscle, suggesting a large portion was retained within the muscle and metabolized in situ. Furthermore, following injection of [14C]lactate, greater than 80% of the total blood activity was recovered as lactate, indicating that little label was incorporated into other products (e.g. glucose). These data suggest that the Cori cycle plays a minimal role in the metabolism of lactate in salmon and flounder. Furthermore, at least in flounder, there was no correlation between the kinetics of lactate clearance and O2 consumption, suggesting that the classical concept of 'O2 debt' is not applicable in this species.

Animals↗
PubMed2001

Feeding behavior of newly settled winter flounder (Pseudopleuronectes americanus) on calanoid copepods.

Field and laboratory investigations were conducted to examine feeding by newly settled winter flounder (Pseudopleuronectes americanus) on two co-occurring calanoid copepods, Eurytemora affinis and Acartia hudsonica. During the spring, these prey are present when winter flounder initiate their demersal lifestyle in estuaries of the northeastern United States. Epibenthic zooplankton were collected concurrently with winter flounder in the Navesink River estuary, NJ, in May 1998 and 1999. Although both calanoid species were in the estuary during the 2-year survey, E. affinis was consumed nearly to the exclusion of A. hudsonica by newly settled winter flounder. Annually, E. affinis and A. hudsonica had similar size distributions in field collections, indicating that species choice was not size selective. However, when preying on E. affinis, winter flounder preferred the larger sized organisms. In single species laboratory experiments, E. affinis and A. hudsonica were consumed equally by newly settled winter flounder (19-23 mm TL), but there were more strikes made toward E. affinis. Despite the lower catch efficiency, E. affinis was selected over A. hudsonica when the prey species were offered together in equal numbers. The selection for E. affinis over A. hudsonica by newly settled winter flounder may be the result of behavioral and/or morphological differences in the prey species.

Journal Article↗
PubMed2001

Size-related shifts in the habitat associations of young-of-the-year winter flounder (Pseudopleuronectes americanus): field observations and laboratory experiments with sediments and prey.

Field surveys and laboratory studies were used to determine the role of substrata in habitat selection by young-of-the year winter flounder. A synoptic field survey of winter flounder and sediments in the Navesink River-Sandy Hook Bay estuarine system in New Jersey demonstrated that winter flounder distribution was related to sediment grain size. Analysis using a generalized additive model indicated that the probability of capturing 10-49 mm SL winter flounder was high on sediments with a mean grain diameter of </=0.5 mm, while fish 50-95 mm were least likely to be collected on fine sediments and most commonly on sediments with a grain-size near 1.0 mm. In the laboratory, sediment preferences and the burying ability of winter flounder (15-69 mm SL) were tested by exposing fish in 10-mm size groups to a choice of azoic sediments of different sediment grain sizes. Smaller individuals (<40 mm SL) preferred fine-grained sediments while larger individuals (>/=40 mm SL) preferred coarse-grained sediments. Burying ability increased with size and all flounders avoided sediments that prevented burial. Subsequent laboratory experiments revealed that the presence of live prey (Mya arenaria) can over-ride sediment choice by winter flounder (50-68 mm SL) indicating the complexity of interrelated factors in habitat choice.

Journal Article↗
PubMed1984

Flounder intestinal absorptive cells have abundant gap junctions and may be coupled.

We noted that, unlike mammalian intestinal absorptive cells, cells of the winter flounder (Pseudopleuronectes americanus) displayed abundant gap junctions on the lateral plasma membrane. We compared the distribution of gap junctions in winter flounder to that in rabbit intestinal epithelium. We also examined for evidence of gap junction-mediated intercellular coupling by comparing the cell-to-cell variation of electrical potential difference across winter flounder intestinal cell apical membranes with that in rabbit small intestinal epithelium in which gap junctions are rare. Gap junctions were seen in 95% of flounder absorptive cells and were localized largely to the apical third of the lateral membrane. Individual gap junctions often contained several hundred uniform 9-nm intramembrane particles. Gap junction size and structure was independent of the position of individual absorptive cells on mucosal folds. These findings sharply contrasted flounder intestinal absorptive cells with rabbit small intestinal absorptive cells, in which gap junctions were rarely detected and when present consisted of few intramembrane particles. Correlating with this distribution of morphologically detectable gap junctions, rabbit small intestinal epithelial cells demonstrated marked variability in potential difference across their apical membranes, whereas those in flounder small intestine showed little variation in apical membrane potential difference. Thus, in contrast to intestinal epithelium of rabbits, flounder intestinal epithelium demonstrates morphological and functional characteristics, suggesting a substantial degree of electrical coupling.

Animals↗
PubMed2001

Spread of Neoheterobothrium hirame (Monogenea), a serious pest of olive flounder Paralichthys olivaceus, to Korea.

Neoheterobothrium hirame is a large, blood-feeding gill-worm infecting the highly prized olive flounder Paralichthys olivaceus in Japan. There is strong evidence that this worm is the primary cause of anaemia, a common and serious condition causing losses among both wild and cultured olive flounders. N. hirame was first detected in Japanese waters less than a decade ago, and its population then proliferated and spread throughout most of Japan, except Hokkaido. In neighbouring Korea, olive flounder is the most important species of cultured marine fish, and production currently exceeds that in Japan. However, until now, there have been no reports of any monogeneans or anaemia among olive flounders in Korea. Our survey conducted in 2000 of 100 cultured individuals from 4 provinces revealed 2 immature specimens of N. hirame: 1 from a land-based pond-culture system in southern Cheju Island (off the SW coast of Korea) and the other from a floating net cage near Yosu (in the mid-S part of the peninsula). The geographic range of this pathogen may have been enlarged as a result of introduction(s) of infected broodstock from Japan, but this seems unlikely. (The raising of this species in hatcheries developed in Korea in 1985, 7 years before the earliest detection of the worm in Japan.) Low numbers of flounders were also clearly anaemic. This, and the current rarity of N. hirame in Korean farms, appears to favour the hypothesis of a more recent, natural dispersal of the worm, during migrations of infected flounder across the narrow and shallow Tsushima and Korea Straits. Regardless of route of entry, we expect this pathogen will have an impact on Korean flounder fisheries equally serious to that being experienced in Japan.

Anemia↗
PubMed1986

Comparative metabolism of benzo(a)pyrene and covalent binding to hepatic DNA in English sole, starry flounder, and rat.

Metabolism of benzo(a)pyrene (BaP) in vivo and in vitro was studied using two benthic fish species, English sole (Parophrys vetulus) and starry flounder (Platichthys stellatus), and Sprague-Dawley rats. At 24 h after administration of BaP (7.9 mumol/kg of body weight) to fish either p.o. (Experiment 1) or i.p. (Experiment 2), the specific activity of binding of BaP metabolites to hepatic DNA (pmol of BaP equivalent per mg of DNA) was higher in sole [2.1 in Experiment 1; 28 +/- 5 (SE) in Experiment 2] than in flounder (0.5 in Experiment 1; 14 +/- 4 in Experiment 2). Treatment of bile with beta-glucuronidase and arylsulfatase released a significantly higher proportion of 7,8-dihydroxy-7,8-dihydro-BaP (BaP 7,8-diol) from sole bile than from flounder bile in both experiments. However, the rate of BaP metabolism and rate of formation of BaP 7,8-diol by hepatic microsomes were comparable for both fish species. Thus, the differences in both the level of DNA binding and the concentration of BaP 7,8-diol in bile of BaP-exposed sole and flounder were apparently due to differences in detoxication, rather than formation, of BaP 7,8-oxide and BaP 7,8-diol-9,10-epoxide. The rate of formation of BaP 7,8-diol by rat liver microsomes (28 +/- 1 pmol of BaP 7,8-diol formed per min per mg of protein) was comparable to that by hepatic microsomes from both fish species (50 +/- 10 for sole and 33 +/- 6 for flounder), although the rate of BaP metabolism (600 +/- 200) was approximately 3 times greater than that by the fish species (190 +/- 60 for sole and 180 +/- 40 for flounder). Thus, greater proportion of BaP was converted to BaP 7,8-diol by liver microsomes of fish species than rat. These differences in BaP metabolism in vitro help explain, in part, the substantially lower binding (0.3 +/- 0.1; Experiment 2) for hepatic DNA in BaP-exposed rat than that in either sole or flounder.

Animals↗
PubMed2005

Aromatase cytochrome P450: cloning, intron variation, and ontogeny of gene expression in southern flounder (Paralichthys lethostigma).

Aromatase cytochrome P450 (P450arom) is the enzyme complex responsible for conversion of androgens to estrogens in vertebrates. Consequently, in some fishes its activity appears critical to ovarian differentiation. Southern flounder (Paralichthys lethostigma) is a commercially important flatfish in which females grow larger than males and sex determination is temperature sensitive. Through cloning of the P450arom gene in ovary and quantitative reverse transcription-polymerase chain reaction, we developed a biomarker for early female differentiation in southern flounder. The deduced amino acid sequence for southern flounder P450arom is similar to other teleosts. Comparison of P450arom intron sequences from fish of different populations revealed substantial inter-individual variation. Adult ovary and spleen exhibited high levels of P450arom mRNA, while P450arom mRNA was only weakly detected in testes. Brain, liver, intestine, kidney, gill, muscle, and heart showed little or no P450arom mRNA expression. Gonads of wild and hatchery-produced juvenile flounder of sizes spanning the period of sex differentiation initially exhibited low levels of P450arom mRNA followed by increases in some individuals and bifurcation into two clearly segregated groups (i.e., putative males and females) beginning at approximately 65 mm in total length. Gonadal histology confirmed predictions of sex based on P450arom expression in juvenile flounder, demonstrating that the patterns of P450arom expression observed relate to sex-specific differentiation. This research represents a unique approach to assessing sex differentiation in a natural population, and a powerful technique for better understanding mechanisms of flounder sex determination and rapidly defining conditions for controlling sex for aquaculture.

Amino Acid Sequence↗
PubMed1987

Na-K-2Cl cotransport in winter flounder intestine and bovine kidney outer medulla: [3H] bumetanide binding and effects of furosemide analogues.

The effects of several sulfamoyl benzoic acid derivatives on Na-K-Cl cotransport were investigated in winter flounder intestine. The relative efficacy (IC50 values) and order of potency of these derivatives were benzmetanide, 5 X 10(-8) M greater than bumetanide 3 X 10(-7) M greater than piretanide 3 X 10(-6) M greater than furosemide 7 X 10(-6) M greater than amino piretanide 1 X 10(-5) 3-amino-4-penoxy-5-sulfamoyl benzoic acid. Binding of [3H] bumetanide was studied in microsomal membranes from winter flounder intestine and compared to that in bovine kidney outer medulla. Binding was also studied in brush-border membranes from winter flounder intestine. The estimated values for Kd and number of binding sites (n) were: bovine kidney, Kd = 1.6 X 10(-7), n = 10.5 pmol/mg protein; winter flounder intestine, Kd 1.2 X 10(-7), n = 7.3 pmol/mg protein, and brush-border membranes from winter flounder, Kd = 5.3 X 10(-7), n = 20.4 pmol/mg protein. The estimated Kd for bumetanide binding to winter flounder brush-border membranes derived from association and dissociation kinetics was 6.8 X 10(-7) M. The similarity in magnitudes of IC50 and Kd for bumetanide suggests that the brush-border cotransporter is ordinarily rate-limiting for transmural salt absorption and that bumetanide specifically binds to the cotransporter. Measurement of bumetanide binding at various concentrations of Na, K and Cl showed that optimal binding required all three ions to be present at about 5 mM concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Southern flounder hepatic and intestinal metabolism and DNA binding of benzo[a]pyrene (BaP) metabolites following dietary administration of low doses of BaP, BaP-7,8-dihydrodiol or a BaP metabolite mixture.

Certain finfish species living in chemically polluted environments exhibit a high incidence of gastrointestinal tract tumors. Carnivorous fish in such environments are likely to consume invertebrates which contain chemical procarcinogens and the invertebrate biotransformation products of these compounds. The retention in tissues, extent of DNA adduct formation in liver and intestine, and metabolite composition of bile was investigated in southern flounder following gavage with pure [3H]- or [14C]benzo[a]pyrene (BaP), pure [14C]benzo[a]pyrene-7,8-dihydrodiol (BaP-7,8D), or hepatopancreas from spiny lobsters previously dosed with [3H]- or [14C]BaP (Metab.HP). Metab.HP contained mainly polar conjugates of BaP diols, triols and tetraols. BaP-7,8D was retained in fish tissues and bile at 24 h to a greater extent (33.6% of the dose), than either BaP (19.00%) or Metab.HP (6.6%). Hepatic and intestinal DNA isolated from all dosed fish contained covalently bound radioactivity, but exposure to BaP-7,8D or BaP resulted in significantly higher binding in both tissues than exposure to Metab.HP. Hepatic DNA from BaP and BaP-7,8D-dosed flounder contained 0.24 +/- 0.07 and 0.33 +/- 0.06 pmol BaP equivalents/mg DNA respectively (mean +/- S.E.), while hepatic DNA isolated from Metab.HP-dosed flounder contained 0.006 +/- 0.002 pmol BaP equivalents/mg DNA. Binding of radioactivity to intestinal DNA was significantly higher than to hepatic DNA for flounder dosed with Metab.HP (0.026 +/- 0.003) or with BaP (0.76 +/- 0.27) but not for flounder dosed with BaP-7,8D (0.44 +/- 0.09). These studies show that dietary BaP, and metabolites likely to be present in invertebrates, can be absorbed by the southern flounder and form DNA adducts in target organs.

Animals↗
PubMed2004

The European flounder (Platichthys flesus) TP53 functions as a temperature-sensitive transcription factor which inhibits cell growth in yeast.

Numerous studies focus on biological roles of the TP53 tumor suppressor gene in mammals but little is known about the actual function of TP53 in lower vertebrates. In this study, we used an in vivo functional assay in yeast to address the transactivation capacity of the flounder TP53 protein. We showed that the flounder TP53 acts as a sequence-specific transcription factor which is able to transactivate various human promoters containing a p53-responsive element (RE). This transcriptional activity was completely abrogated in the Val147Glu TP53 mutant previously identified in two flounder hepatic hyperplasia. In addition, we showed that the wild-type (wt) flounder TP53 but not the Val147Glu mutant inhibits cell growth when expressed in yeast. We finally reported that transcription regulation and growth inhibition by the wild-type flounder TP53 is temperature-dependent. The flounder TP53 optimal temperature appeared lower than those reported for the Xenopus and human homologues.

Animals↗