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Primary structure of thymosin beta 12, a new member of the beta-thymosin family isolated from perch liver.

A new polypeptide termed thymosin beta 12 has been isolated from perch liver and its primary structure elucidated. This polypeptide contains 43 amino acid residues with a molecular weight of 4822 Da. The content of thymosin beta 12 from perch liver has been determined as 43 micrograms/g of tissue. The amino-terminal end of this polypeptide is blocked by an acetyl group as deciphered by fast-atom bombardment mass spectrometric analysis. Sequence analysis reveals that thymosin beta 12 is 79% homologous to thymosin beta 4, an immunomodulator which was originally isolated from calf thymus. Thymosin beta 12 also shows 84% sequence homology to thymosin beta 11, a beta 4 analog which replaces beta 4 in two species of bony fish, oscar and rainbow trout. The evolutionary implication of such results will be discussed. The isolation of a new beta 4-related peptide from perch liver which differs from beta 11 indicates that beta-thymosin peptides are widely distributed in lower vertebrate classes.

Amino Acid Sequence↗

Biomonitoring of oil spill in a boreal archipelago by xenobiotic biotransformation in perch (Perca fluviatilis).

The effect of the accidental oil spill (250 tons) in a boreal archipelago (Gulf of Bothnia, Vaasa, Finland) on xenobiotic metabolism of local perch (Perca fluviatilis) was monitored for 1.5 years. The monooxygenase (benzo[a]pyrene, 7-ethoxycoumarin O-deethylase, 7-ethoxyresorufin O-deethylase) and conjugation (UDPglucuronosyltransferase, glutathione S-transferase) activities of perch liver were determined from control areas and those areas where oil had spilled. Only a slight induction in monooxygenase activities was seen in perch caught near the oil spill 4 months after the accident. The induction of monooxygenase activities detected with the fuel oil in laboratory experiments was, however, clear. After a single dose, it rose rapidly and quickly disappeared. Conjugation enzyme activities were not affected in the laboratory.

7-Alkoxycoumarin O-Dealkylase↗

Initial purification and characterization of hepatic microsomal cytochrome P-450 from BNF-treated perch (Perca fluviatilis).

1. A procedure was developed for isolating and purifying cytochrome P-450 from hepatic microsomes of BNF-treated perch, using modified versions of the methods of Williams and Buhler (1982. Biochim. biophys. Acta 717, 398-404) and Goksøyr (1985. Biochim. biophys. Acta 850, 409-417). 2. Following chromatography on phenyl-Sepharose CL 4B and DEAE-Sepharose CL-6B, the major peaks, fractions b and c, were resolved into five fractions, possibly representing different isoenzymes, by a FPLC with a strong anion exchange column (Mono Q). 3. These fractions have been characterized on the basis of their spectral, electrophoretic and immunological properties. 4. The purified form of cytochrome P-450 in fraction V from perch liver showed a number of similarities to cytochrome P-450c, the major BNF-inducible cytochrome P-450 in cod liver. 5. Therefore we suggest that this purified form of cytochrome P-450 is a BNF-induced form in perch and that it is closely related to the gene subfamily cytochrome P-450 IA1.

Animals↗

Toxicological effects and reproductive impairments in female perch (Perca fluviatilis) exposed to leachate from Swedish refuse dumps.

We have previously found that leachate from a Swedish refuse dump caused toxicological effects, including endocrine disruption and reproductive failures, in feral female perch (Perca fluviatilis) from Molnbyggen and in brook trout (Salvelinus fontinalis) from Vadbäcken. This raised concerns that leachate-induced toxicity might affect fish in other leachate-contaminated lakes. This study looks at female perch from three different regions in Sweden, focusing on toxicological biomarkers (skin lesions, liver-somatic index (LSI), ethoxyresorufin O-deethylase (EROD) activity and DNA adducts) and reproductive biomarkers (number of mature females, gonadosomatic index (GSI), aromatase (P450arom), progesterone, 17alpha-hydroxyprogesterone (17alpha-OHP), testosterone (T) and 17beta-estradiol (E2)). Five out of seven contaminated locations had lower numbers of mature females and most of them exhibited higher frequencies of fin erosion relative to their comparable reference sites. Females from Lake Nedre Vättern exhibited these effects, as well as body sores, high EROD activity, high levels of DNA adducts in the liver and the intestinal mucosa, low GSI and low plasma levels of T and E2, similar to the anti-estrogenic effects earlier found in SM perch from Molnbyggen in 1997 and in brook trout from Vadbäcken. No effects on LSI, GSI, aromatase, or circulating steroids were found in mature females from Molnbyggen in this study. This indicated less leachate-contamination, but low numbers of mature females in Lake Siljan, at the sewage treatment plant which now processes the leachate that earlier contaminated Molnbyggen and Vadbäcken, suggested insufficient sewage treatment at this site. All females in Lake Simshyttsjön were mature, and their high GSI and LSI, low EROD activity, high level of 17alpha-OHP and low level of T, suggest an estrogenic rather than an anti-estrogenic effect. The results show that our earlier findings in Molnbyggen and Vadbäcken are not common, but neither are they unique. This study also shows that refuse dumps should be considered as potential point-sources for environmental pollutants, and that uncontrolled leachate-contamination of lakes and freshwater reservoirs could be a serious environmental hazard for both wildlife and humans.

Animals↗

Ethoxyresorufin-O-deethylase (EROD) activity and bile metabolites as contamination indicators in Baltic Sea perch: determination by HPLC.

Exposure of Baltic Sea perch (Perca fluviatilis) to organic contaminants was investigated by means of liver enzyme activity, measured as ethoxyresorufin-O-deethylase (EROD) activity, and PAH metabolites excreted to bile. Female perch used in the study were caught in coastal waters of the western Gulf of Finland. Reaction conditions were optimised for determination of EROD activity in liver. A high-performance liquid chromatography (HPLC) method with fluorescence detection was developed to separate the interfering fluorescent signal from the signal due to EROD activity. EROD activity in perch varied within a concentration range of 0.30-14 pmol min(-1)mg(-1) protein. Recent PAH exposure was indicated in enhanced levels of the bile metabolite 1-hydroxypyrene, which varied between 213 and 1149 microg kg(-1). No correlation was indicated between hepatic EROD activity and concentration of 1-hydroxypyrene.

Animals↗

The effects of salinity on naphthenic acid toxicity to yellow perch: gill and liver histopathology.

Naphthenic acids (NAs) are naturally occurring saturated linear and cyclic carboxylic acids found in petroleum, including the bitumen contained in the Athabasca Oil Sands deposit in Alberta, Canada. The processing of these oil sands leads to elevated concentrations of NAs, as well as increased salinity from produced waters as a result of ions leaching from the ores, the process aids, and the water associated with the deeper aquifers. These changes can result in waters that challenge reclamation of impacted waters associated with oil sands development. Laboratory tests examined the effects of salinity on NA toxicity using local young-of-the-year yellow perch exposed to a commercially available mixture of NAs (CNA) and an NA mixture that was extracted from oil sands process-affected water (ENA), with and without the addition of sodium sulfate (Na(2)SO(4)). Gill and liver histopathological changes were evaluated in the surviving fish after 3 weeks of exposure. At 6.8 mg/L ENA and 3.6 mg/L CNA, 100% mortality was observed, both with and without the addition of salt. Exposure of yellow perch to 25% of the NA required to give an LC100 (0.9 mg/L CNA; 1. 7 mg/L ENA) resulted in high levels of gill proliferative (epithelial, mucous, and chloride cell) changes, a response that was increased with the addition of 1g/L salt (Na2SO4) for the ENA. The significance of these changes was a reduced gill surface area, which likely caused a reduction in both the transport of NAs within the fish and the exchange of vital respiratory gases. While the gills were affected, no liver alterations were identified following NA or NA+salt exposures. Differences in the chemical composition of the NAs tested may explain the differences in the lethality and histopathology of yellow perch.

Animals↗

Basic physiological biomarkers in adult female perch (Perca fluviatilis) in a chronically polluted gradient in the Stockholm recipient (Sweden).

By measuring a battery of basic physiological biomarkers and the concentration of SigmaDDT in adult female perch (Perca fluviatilis), an assumed aquatic pollution gradient was confirmed, with the city of Stockholm (Sweden) as a point source of anthropogenic substances. The investigation included an upstream gradient, westwards through Lake Mälaren (46 km), and a downstream gradient, eastwards through the Stockholm archipelago (84 km). The results indicated a severe pollution situation in central Stockholm, with poor health status of the perch: retarded growth, increased frequency of sexually immature females, low gonadosomatic index, and disturbed visceral fat metabolism. SigmaDDT, measured as a pollution indicator, was 10-28 times higher than the background in perch from the Baltic Proper. Besides the main gradient other sources of pollution also influenced the response pattern of the measured biomarkers. In particular, there were strong indications of pollution coming from the Baltic Sea.

Analysis of Variance↗

Thyroid hormone-induced protein (TIP) gene expression by 3,5,3(')-triiodothyronine in the ovarian follicle of perch (Anabas testudineus, Bloch): modulation of 3beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4)-isomerase enzyme by TIP.

Our previous reports had shown that 3,5,3'-triiodothyronine (T(3)) induced the generation of a 52-kDa monomer protein, i.e., TIP (thyroid hormone-induced protein) in the perch ovarian follicle. TIP, in turn, increased progesterone formation by stimulating Delta(5)-3beta-HSD activity (3beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4) isomerase) [Eur. J. Endocrinol. 134 (1996) 128-135; Gen. Comp. Endocrinol. 113 (1999) 212-220]. In the present investigation, perch ovarian follicles were incubated in the absence (control) or the presence of T(3) or gonadotropin (GTH) or human chorionic gonadotropin (hCG). RNAs were isolated and allowed to hybridize with a radiolabeled TIP oligonucleotide probe prepared on the basis of the N-terminal 17-amino-acid sequence of TIP. Only RNA from T(3)-incubated follicles hybridized with the probe, while RNA from control or GTH- or hCG-incubated follicles did not hybridize with the probe. The transcript size of TIP mRNA was approximately 1.8 kb. mRNA isolated from T(3)-incubated ovarian follicles subjected to in vitro translation and Western blot analysis clearly identified a 52-kDa protein which was not found with the mRNA from the control follicles. However, both TIP and GTH stimulated progesterone secretion from perch ovarian follicles in vitro. GTH stimulation of Delta(5)-3beta-HSD was due to the stimulation of enzyme protein synthesis as a more than twofold increase in Delta(5)-3beta-HSD occurred in response to GTH. But TIP did not stimulate synthesis of Delta(5)-3beta-HSD protein. However, in vitro incubation of Delta(5)-3beta-HSD enzyme with TIP in the presence of NAD and substrate (pregnenolone) greatly stimulated enzyme activity, while incubation with GTH had no effect, indicating a modulation of Delta(5)-3beta-HSD protein from a less active to a more active state by TIP. This has been supported by another observation, in which TIP (52 kDa) and Delta(5)-3beta-HSD (45 kDa) incubation resulted in a complex of 99 kDa. This suggests a protein-protein interaction in the process of Delta(5)-3beta-HSD activation by TIP. The present work, therefore, shows some new and interesting aspects of thyroid hormone regulation of the reproductive control mechanism.

Animals↗

Polychlorinated biphenyls and some pesticides in perch (Perca fluviatilis) from inland waters of Latvia.

Polychlorinated biphenyls (PCBs), DDT-related substances, hexachlorocyclohexane (HCH) isomers and hexachlorobenzene (HCB) were analysed in perch (Perca fluviatilis) from Latvian lakes and rivers. DDE is present in the highest concentrations in all samples; 62-170 ng/g lipid weight in perch from rural areas and 460 ng/g in perch from the Riga area. Individual dominating PCB congeners were in the range 16-45 ng/g and 200-210 ng/g, respectively. The degree of contamination of rural areas in Latvia of these organohalogen substances is in the same range as in background areas in Sweden. Riga, the major urban area in Latvia, is shown to be more polluted with PCBs than other areas in Latvia.

Animals↗

Relationship between mercury accumulation in young-of-the-year yellow perch and water-level fluctuations.

A three-year (2001-2003) monitoring effort of 14 northeastern Minnesota lakes was conducted to document relationships between water-level fluctuations and mercury bioaccumulation in young-of-the-year (YOY) yellow perch (Perca flavescens) collected in the fall of each year at fixed locations. Six of those lakes are located within or adjacent to Voyageurs National Park and are influenced by dams on the outlets of Rainy and Namakan lakes. One site on Sand Point Lake coincides with a location that has nine years of previous monitoring suitable for addressing the same issue over a longer time frame. Mean mercury concentrations in YOY yellow perch at each sampling location varied significantly from year to year. For the 12-year monitoring site on Sand Point Lake, values ranged from 38 ng gww(-1) in 1998 to 200 ng gww(-1) in 2001. For the 14-lake study, annual mean concentrations ranged by nearly a factor of 2, on average, for each lake over the three years of record. One likely factor responsible for these wide variations is that annual water-level fluctuations are strongly correlated with mercury levels in YOY perch for both data sets.

Animals↗

Localization of carbonic anhydrase in swimbladder of European eel (Anguilla anguilla) and perch (Perca fluviatilis).

The distribution of carbonic anhydrase in swimbladder tissue and especially in gas gland cells of the European eel (Anguilla anguilla) and the perch (Perca fluviatilis) was analysed using histochemical staining according to Hansson (1967), with modifications proposed by Riddersträle (1991). While in the European eel, gas gland cells are distributed as a single layered epithelium over the whole secretory part of the swimbladder, the gas gland of the perch consists of a compact, richly vascularized 'multilayered' epithelium, in which gas gland cells have contact with the swimbladder lumen via small channels. In spite of these differences in organization, membranes of gas gland cells near blood vessels are richly folded in both species. A strong histochemical staining for carbonic anhydrase was observed in these membrane foldings. With prolonged incubation times a positive reaction was also observed in the cytoplasm of gas gland cells. In addition, the vascular endothelium and the erythrocytes showed a positive histochemical reaction. No staining, however, was visible in apical membranes towards the lumen of the swimbladder. In the perch, swimbladder epithelial cells outside the gas gland showed no positive staining of carbonic anhydrase. The results thus indicate that carbonic anhydrase activity is especially concentrated in membranes facing blood vessels. This suggests that a rapid equilibrium of the CO2/HCO3- reaction in the intracellular as well as in the extracellular space is essential for swimbladder function.

Air Sacs↗

Postglacial colonization shows evidence for sympatric population splitting of Eurasian perch (Perca fluviatilis L) in Lake Constance.

Previous microsatellite analysis showed that two subpopulations of perch (Perca fluviatilis L) exist in Lake Constance. This raises questions of whether (i). Lake Constance was colonized by two populations that diverged in allopatry, or (ii). the two subpopulations diverged in sympatry. Sequence analysis of a 365 bp mtDNA fragment (5'-end of the D-loop) of perch from Lake Constance and adjacent waters revealed 10 haplotypes. We suggest colonization via the Danube river, based on the frequency and dispersion of haplotypes, and knowledge of the lake's palaeohydrological development. Pairwise FST-values using mitochondrial DNA sequences showed no significant population subdivision. Our study provides strong evidence that subpopulations of perch in Lake Constance have diverged in sympatry.

Animals↗

Cloning and functional characterization of two glycine receptor alpha-subunits from the perch retina.

Glycine receptors are ligand-gated ion channel proteins mediating synaptic inhibition in the spinal cord, retina and brain of vertebrates. We have cloned and functionally characterized two glycine receptor alpha-subunits from the perch (Roccus americana) retina. Based on sequence homology with the mammalian counterparts, we termed these subunits alpha 1 and alpha 3. RT-PCR revealed the presence of both subunits in retina and brain, whereas alpha1 was predominant in spinal cord. A short splice variant of alpha1 was detected in the brain but not in the retina. Functional expression of the perch subunits in HEK-293 cells yielded robust glycine-gated currents sensitive to strychnine. The perch receptors displayed a high efficacy for taurine and GABA and thus differ from the mammalian counterparts. Because the retina is a rich source for taurine, this finding could be of physiological importance. The structural features of the ligand binding domain strongly support the notion of increased glycine/GABA discrimination in higher vertebrates.

Amino Acid Sequence↗

Dietary choline requirement of juvenile yellow perch (Perca flavescens).

We conducted an 11-wk feeding trial to determine the dietary choline requirement of juvenile yellow perch (Perca flavescens) and to investigate whether dietary phosphatidylcholine (PC) could meet this requirement. Six dietary treatments contained choline concentrations of <0.11, 0.23, 0.34, 0.75, 1.22 or 3.37 g/kg diet. Two additional diets contained 31 g of lecithin/kg diet, with or without supplemental choline chloride (4.0 g choline/kg diet). The total sulfur amino acid concentration was maintained at 1.0 g/100 g diet (methionine/cyst(e)ine, 49:51). Diets were fed to satiation twice daily to triplicate groups of yellow perch initially weighing 16.0 g/fish. Weight gain, feed intake and carcass proximate composition were significantly affected by dietary choline. Weight gains and feed intakes increased as dietary choline increased from 0 to 0.75 g/kg. Both values tended to plateau in fish fed dietary choline levels above 0.75 g/kg. Broken-line analyses of weight gain and feed intake data indicated the dietary choline requirement was 0.598 and 0.634 g/kg diet, respectively. Hepatic lipid concentrations and feed efficiency values were not significantly different. Whole-body fat concentrations increased significantly, whereas ash levels decreased significantly in fish fed increasing levels of dietary choline. Weight gain and feed intake of fish fed diets containing PC were not significantly different from fish fed 0.75 g/kg of dietary choline. However, hepatic lipid concentrations were significantly higher in fish fed the diet containing PC and no choline chloride. Thus, yellow perch require a maximum of 0.598-0.634 g of choline/kg diet for maximum growth and this requirement may potentially be met with 31 g of lecithin/kg diet.

Analysis of Variance↗

Dietary sulfur amino acid requirement of juvenile yellow perch fed the maximum cystine replacement value for methionine.

We conducted three separate experiments designed to determine the dietary methionine requirement, ability of cyst(e)ine to spare methionine, and the total sulfur amino acid requirement (TSAA) of juvenile yellow perch when fed the maximal amount of cyst(e)ine. The purified basal diet used in each experiment contained 33.6 g of crude protein/100 g diet and 12.0 g of lipid/100 g diet. In the first experiment,;>L-methionine was added to eight diets providing methionine concentrations ranging from 0.37 to 1.77 g/100 g diet in gradations of 0.2 g/100 g diet. Diets were fed for 12 wk to juvenile yellow perch initially weighing 4.7 g/fish. Broken-line analyses of weight gain and feed efficiency data indicated that the dietary methionine requirement was 1.0 g/100 g diet (3.1 g TSAA/100 g dietary protein) and 1.1 g/100 g diet (3.4 g TSAA/100 g dietary protein), respectively. In the second experiment, various ratios of L-cyst(e)ine and L-methionine were added to the basal diet and fed for 12 wk to determine the cyst(e)ine replacement value of yellow perch initially weighing 19.3 g/fish. Weight gain and feed efficiency (FE) data indicated that cyst(e)ine spared up to 51% of the methionine requirement. In the final experiment, graded levels of cyst(e)ine plus methionine in a ratio of 51:49 were added to the basal diet in gradations of 0.1 g/100 g diet (0.5 to 1.2 g TSAA/100 g diet) to determine the dietary total sulfur amino acid requirement. Diets were fed to satiation for 10 wk to fish initially weighing 8. 1 g. Broken-line analyses of weight gain, feed intake and FE data indicated that the dietary TSAA requirement was 0.85, 0.87 and 1.0 g of TSAA/100 g diet (2.5 to 3.0 g of TSAA/100 g of dietary protein), respectively. The majority of dietary TSAA requirements of fish are in the range of 2 to 4 g/100 g of dietary protein and are generally similar to those of both birds and swine, but lower than estimates for rodents.

Amino Acids, Sulfur↗

Metallo-protease activity increases prior to ovulation in brook trout (Salvelinus fontinalis) and yellow perch (Perca flavescens) follicle walls.

Proteolytic activity was measured in the follicle wall surrounding oocytes from brook trout (Salvelinus fontinalis) and yellow perch (Perca flavescens) by use of two different protease assays: sodium dodecyl sulfate-polyacrylamide gel electrophoresis (substrate-SDS-PAGE) and a chromogenic synthetic peptide for type I collagen. In brook trout follicle walls, substrate-SDS-PAGE studies demonstrated that the activity of two proteolytic enzymes (80 kDa and 66 kDa) increased significantly before ovulation. The 80 kDa enzyme decreased significantly after ovulation whereas the 66 kDa enzyme remained elevated following ovulation. In yellow perch follicle walls, substrate-SDS-PAGE studies demonstrated that the activity of the major protease (66 kDa) increased before ovulation and remained elevated after ovulation. A chromogenic synthetic peptide was used to assay collagenolytic activity in follicle walls of brook trout and yellow perch. This assay revealed that collagenolytic activity increased significantly in both species before ovulation and remained elevated after ovulation. These findings suggest that metallo-proteases are involved in digesting the follicle wall in teleosts before and after ovulation.

Animals↗

The excitatory and inhibitory amino acid receptors on horizontal cells isolated from the white perch retina.

1. The distribution and the properties of receptors to the inhibitory amino acid glycine (GLY) and the excitatory amino acid glutamate (GLU) and its analogues kainate (KA), quisqualate (QUIS), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), and N-methyl-D-aspartate (NMDA), were studied with whole-cell and outside-out patch-clamp techniques on all four types of horizontal cells isolated from the retina of white perch. 2. Glycine at concentrations above 30 microM evoked whole-cell current responses from two types of horizontal cells (H2 and H4). The other two types of horizontal cells (H1 and H3) were unresponsive to GLY (30 microM-3 mM). 3. Responses elicited by GLY from H2 and H4 cells were similar, consisting of inward currents that desensitized with a half-decay time of 0.5-2 s at glycine concentrations between 100 and 500 microM. GLY-activated currents were inhibited by the glycine receptor antagonist strychnine (STRYCH). Current responses evoked by GLY reversed at the Cl- equilibrium potential. 4. Dose-response analysis of peak currents induced by GLY revealed a Hill coefficient of 2.0 +/- 0.1 (mean +/- SD, n = 3) and an median effective concentration (EC50) of 85 +/- 2 microM (n = 3). 5. Single glycine receptor channels recorded from outside-out patches had a main-state conductance of 47 +/- 4 pS (n = 3). 6. Every type of horizontal cell from the white perch responded to GLU, KA, QUIS, and AMPA but none responded to exogenously applied NMDA (200 microM) or NMDA (200 microM) + GLY (1 microM) in a Mg+2-free bathing solution. 7. The ratio of the amplitude of responses to GLU, KA, QUIS, and AMPA remained nearly constant among all the horizontal cells tested, suggesting there might be only a single population of non-NMDA receptors on these cells. 8. QUIS and KA both elicited responses from the horizontal cells. When applied together with KA, QUIS competitively antagonized the responses of horizontal cells to KA. 9. The results demonstrated the existence of an inhomogeneous distribution of strychnine-sensitive glycine receptors and a homogeneous distribution of non-NMDA type glutamate receptors among the four types of white perch horizontal cells.

Amino Acids↗

Relationship between individual variation in morphological characters and swimming costs in brook charr (Salvelinus fontinalis) and yellow perch (Perca flavescens).

The objective of this study was to examine if individual variation in morphological characters is related to swimming costs in wild and domestic brook charr, and in wild yellow perch. Our results indicate that absolute swimming cost was higher in wild and domestic brook charr individuals having a stout body shape, and these individuals are therefore less efficient swimmers. These results are consistent with field observations that described relationships between individual variation in morphology and habitat use in salmonids. Further analyses indicated that standard metabolic rates were higher in individuals having a stout body shape, and that net swimming cost was not related to body shape. Accordingly, the higher swimming cost of stout individuals is probably an indirect consequence of an increase in standard metabolic rate. In wild yellow perch, absolute and net swimming costs were higher in individuals having a stout body shape and a low aspect caudal fin, and standard metabolic rate was not related to body shape. Therefore, in contrast to brook charr, individual variation in the swimming cost of yellow perch appears to be related to morphological characters that affect drag and thrust forces, which is consistent with previously published inter-specific observations.

Animals↗