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At least 145 records · Page 8Linked to original sources

In vitro responses of rainbow trout (Oncorhynchus mykiss) somatotrophs to carp growth hormone-releasing factor (GRF) and somatostatin.

To study the hypothalamic control of growth hormone (GH) release in lower vertebrates, we employed an in vitro technique using a monolayer cell culture system of rainbow trout pituitary glands. Two newly purified carp brain growth hormone-releasing factors, carp GRF(1-45) and carp GRF(1-29), and synthetic somatostatin-14 (SST-14) were applied to the cultured pituitary cells. The results indicate that: (1) The carp GRFs had a dose-related potency in stimulating growth hormone release. The dose of half maximum effect (ED50) for carp GRF(1-45) was 0.107 nM, and an equal potency for carp GRF(1-29) was 0.388 nM. (2) SST-14 inhibited GH release having a dose-dependent potency with an ED50 of 0.186 nM. (3) Osmotic pressure did not influence SST-14 inhibited GH secretion but did affect spontaneous GH release. (4) The response of cultured cells was not affected by length of incubation period with SST-14 or carp GRF but was affected by cell density.

Animals↗

Investigations into the ubiquitous nature of high or low immune responsiveness after divergent selection for antibody production in common carp (Cyprinus carpio L.).

This paper reports on the selection of individual carp with a high or low antibody response, in combination with reproduction by gynogenesis, in order to develop well-characterised inbred carp lines consisting of practically unlimited numbers of carp with the same genotype. Two homozygous progenies, previously characterised as having a high or low immune response to dinitrophenyl keyhole limpet haemocyanin (DNP-KLH), were immunised with either a T-dependent (DNP-human serum albumin (DNP-HSA)) or T-independent (trinitrophenyl lipopolysaccharide (TNP-LPS)) hapten-carrier complex. In comparison with the antibody response after DNP-KLH immunisation, the response to DNP-HSA was observed to be highly variable and did not differ between the divergently selected progenies. This suggests that the divergent selection for antibody production to DNP-KLH has been carrier-specific. Immunisation with T-independent TNP-LPS induced a very rapid response which differed between the high and low responders, and likely measured changes in the DNP-specific precursor pool of B cells caused by the selection. A number of selected individuals with a high immune response to DNP-KLH were infected with Trypanoplasma borreli, a haemoflagellate parasite of carp, to examine a possible relationship between the increase in immune responsiveness and disease resistance, but no change could be detected. However, individual homozygous carp were able to escape inbreeding depression and survive the infection. Such carp would be likely candidates for gynogenetic reproduction to obtain viable inbred carp lines.

Animals↗

Plasma electrolyte changes after ingestion of bile extract of the grass carp (Ctenopharyngodon idellus) in rats.

Some Chinese people believe that eating the gallbladder of the grass carp (Ctenopharyngodon idellus) may improve their visual acuity. However, ingestion of grass-carp bile, in severe cases, may kill human beings and experimental animals. The cause of death after eating grass-carp bile was studied in rats. The bile acid fraction was prepared from the n-hexane-insoluble and alcohol-soluble parts of grass-carp and hog bile. The bile extract (6 ml/kg, 15%) was administrated by gastric intubation to conscious female Long-Evans rats. Ingestion of grass-carp bile extract killed all the experimental rats in 2-8 h and was associated with a decrease in blood pressure and an increase in plasma potassium, hydrogen ions, blood urea nitrogen and hematocrit. However, rats that ingested hog bile or saline showed no significant changes in the measured parameters. If the rats were fed cholestyramine-treated grass-carp bile extract, they survived without significant change in the measured parameters. This report demonstrates that a drastic increase in plasma potassium and hydrogen ions probably caused the death of the rats, and also shows that bile acid (alcohol) in grass-carp bile is related to the toxicity.

Animals↗

Carp liver DNase--isolation, further characterization and interaction with endogenous actin.

Deoxyribonuclease I (DNase I)-like enzyme from the liver of the carp (Cyprinus carpio) was purified to homogeneity and further characterized. Ion exchange chromatography on DEAE-cellulose, molecular filtration on Sephacryl S-300 and Con A-Sepharose affinity chromatography were applied for enzyme isolation. Carp liver DNase, similarly to DNase I from bovine pancreas, was found to be an endonuclease that hydrolyses linear DNA from salmon sperm as well as circular DNA forms--plasmid and cosmid. The purified enzyme is a glycoprotein and shows microheterogeneity, as observed in DNase zymograms prepared after native and two-dimensional electrophoresis (2D-PAGE). The composition of sugar component of the enzyme was characterized. Special attention was focused on the ability of carp liver DNase to interact with carp liver actin. The carp liver enzyme was inhibited by endogenous actin. The estimated binding constant of carp liver DNase to carp liver actin was calculated to be 1.1 x 10(6) M(-1).

Actins↗

Evolutionary conservation of alternative activation of macrophages: structural and functional characterization of arginase 1 and 2 in carp (Cyprinus carpio L.).

Classically activated macrophages (caMF) play an important role in type-I immune responses and alternatively activated macrophages (aaMF) function in type-II immune responses. While the classical activation of fish macrophages has been well described, the existence of aaMF has not yet been described for teleosts. Arginase is the characteristic enzyme in aaMF and two isoforms have been described for mammals. To study the presence of aaMF in a primitive vertebrate species we cloned arginase 1 and 2 cDNA of common carp. Carp arginase 1 is a 340 aa protein with 63% aa sequence identity to human arginase 1. Carp arginase 2 is a 347 aa protein with 63% aa sequence identity to human arginase 2. Three highly homologous arginase 2 genes were found, each showing only single non-synonymous substitutions. Basal arginase 1 expression is mainly found in carp mid kidney. In contrast, arginase 2 was expressed in all organs examined with the highest basal gene expression in liver. Cultured carp head kidney-derived macrophages were used to study aaMF in vitro. Carp macrophages showed significant arginase activity which could be induced by dibutyryl cyclic adenosine mono phosphate (cAMP) and specifically inhibited by NG-hydroxy-L-arginine (NOHA). At the gene level, arginase 2 gene expression was upregulated by cAMP stimulation, while arginase 1 gene expression was not influenced. LPS stimulation did not alter the arginase 1 or 2 expression, inducible nitric oxide synthase (iNOS) expression was, however, upregulated. This expression of iNOS was used as a measure of classical activation of carp macrophages. Thus, in contrast to mammals, fish arginase 2 and not arginase 1 is differentially regulated and likely involved in the alternative activation of fish macrophages. Our data suggest there may be an evolutionary conservation of the presence of aaMF down to teleost fish.

Amino Acid Sequence↗

High glucuronidation activity of environmental estrogens in the carp (Cyprinus carpino) intestine.

Many adverse effects on carp reproductive organs have been reported to be caused by exposure to environmental estrogens, such as nonylphenol and bisphenol A, which contaminate the aquatic environment. The glucuronidation activities of xenoestrogens (bisphenol A and diethylstilbestrol) and phytoestrogens (coumestrol, genistein and biochanin A), but not nonylphenol and octylphenol, were observed in microsomes prepared from carp organs. The highest levels of glucuronidation of environmental estrogens, for which the optimum temperature was 25-30 degrees C, were observed in the intestinal microsomes of 2-year-old carp. These activities in carp intestine increased developmentally, and the maximum levels corresponded to 5-10 % of that in rat liver microsomes. However, the glucuronidation of phytoestrogen by carp intestinal microsomes corresponded to that of rat liver microsomes. Only bisphenol A-glucuronide was excreted from the everted intestine, indicating that bisphenol A is metabolized in the carp intestine mainly as glucuronide. These results suggest that glucuronidation by carp intestine plays an important role for the detoxification of xenoestrogens and phytoestrogens, except for nonylphenol and octylphenol.

Aging↗

Cytochrome p450 induction and gonadal status alteration in common carp (Cyprinus carpio) associated with the discharge of dioxin contaminated effluent to the Hikiji River, Kanagawa Prefecture, Japan.

Accumulations of polychlorinated dibenzo-p-dioxins, dibenzofurans and coplanar polychlorinated biphenyls were analyzed in common carp (Cyprinus carpio) collected in the Hikiji River, Kanagawa Prefecture, Japan in which dioxin contaminated effluent was released during the period starting from November 1992 to March 2000. Higher levels of 2,3,7,8-tetrachlorinated dibenzo-p-dioxin toxic equivalents were observed in carps collected downstream to the dioxin release site (contaminated site) than the reference site. Modulations of cytochrome p450 (CYP) enzyme in liver, serum estrogen concentration and gonadal somatic index (GSI) were also measured as biomarkers for the contaminants. Total CYP content in livers was markedly higher in male and female carps from the contaminated site relative to the reference site fish. The expression level of the cytochrome p450 1A and Ethoxyresorufin O-deethylase activity were significantly higher in female carps from the contaminated site than from the reference site. A lower level of plasma estrogen was observed in carps from the contaminated site. The GSI in female carps from the contaminated site was smaller than that recorded at the reference site. The present study indicates that dioxins released to the Hikiji River might induce the CYP enzyme and inhibit the reproductive functions in common carps dwelling downstream from the release site.

Animals↗

Terminal components of carp complement constituting a membrane attack complex.

The membrane attack complex (MAC) of carp complement was extracted with deoxycholate from rabbit erythrocytes lysed by carp serum and purified by a two-step chromatographic procedure. On two-dimensional SDS-PAGE of carp MAC, eight bands were detected. The band of M(r) 91,000 was identified as carp C9 by western blotting using anti-carp C9, and two bands of M(r) 62,000 and one band of M(r) 22,000 were confirmed as those of carp C8 alpha, C8 beta and C8 gamma, respectively, by their N-terminal amino acid sequences. The bands of M(r) 102,000 and 73,000, which generated from a 180,000 band under reducing conditions, were those corresponding to human C5b alpha and C5b beta, respectively. The remaining bands of M(r) 115,000 and 106,000 were identified as those corresponding to human C6 and C7, as determined by their molecular size, single-chain structures and similarities in N-terminal amino acid sequences to their mammalian counterparts. Densitometric scan of the gels showed the molar ratio of C5b, C6, C7, C8 and C9 in carp MAC to be 1:1:1:1:4. Based on these results, it appears that, as with mammals, the cytolytic pathway of bony fish complement is composed of five terminal components from C5 to C9.

Amino Acid Sequence↗

Molecular cloning of a carp Jak3 from activated leukocytes.

Janus kinases (Jaks) are involved in a signalling pathway leading to tyrosine phosphorylation, which is the key step in transducing cytokine signals from the external environment to the nucleus. We report here the molecular cloning of carp Jak3. A partial cDNA of the Jak homologue was initially identified from a cDNA pool obtained by subtracting the cDNAs from Con A-induced carp head kidney leukocytes with those from uninduced carp head kidney leukocytes. The entire coding sequence was assembled by sequencing both strands of cDNA clones amplified by using anchored PCR reaction. The complete cDNA was cloned by specific primers designed against the partial cDNAs and then sequenced to assure accuracy of the complete sequence. Sequence analysis reveals an ORF encoding a protein of 1026 amino acids, with the unique presence of two carboxy-terminal kinase and pseudokinase domains. The cDNA clone shows approximately 52-50%, 50-48.2%, 42-39% and 35-39% amino acid identity with the known Jak3s, Jak2s, Jak1s and Tyk2s, respectively. The sequence of the amino acid residues within the N-terminal JH domains of the carp Jak homologue shows higher similarities with the regions of human Jak3 than with human Jak2. The carp Jak is expressed as a single 4. 5Kb transcript at low levels in the head kidney and the spleen, with the RNA level increasing when leukocytes are activated with con A. The trace transcript was also detected by RT-PCR in the carp liver, muscle and skin, but not in the egg sample. Sequence analysis and pattern of expression suggest that this cDNA represents the carp Jak3 (accession number in GenBank: AF148993).

Amino Acid Sequence↗

Bioavailability of Cd to the common carp, Cyprinus carpio, in the presence of humic acid.

The uptake of cadmium by the common carp, Cyprinus carpio, was studied in chemically defined freshwater in the absence and presence of commercial humic acid. This was done to evaluate whether the cadmium uptake by carp in the presence of humic acid was related to the ambient Cd2+ -ion activity or whether the complexed metal also contributed to the uptake. Uptake of Cd during a 3-h period of exposure was used as a measure of the biological availability of the metal. The uptake rate data for Cd in total fish and gills obtained in the absence (control) and presence (treatment) of humic acid were analyzed using a Michaelis-Menten model for mediated transport. The Michaelis-Menten parameters KM and Vmax obtained in the control and the treatment experiment were compared for each of the two investigated carp compartments (total fish and gills). The model parameter estimates for Cd uptake by total carp in the treatment experiment (KM = 0.41 +/- 0.11 micromol l(-1); Vmax = 0.66 +/- 0.13 micromol kg(-1) h(-1)) were not significantly different from the model parameters in the control experiment (KM = 0.34+/-0.06 micromol l(-1); Vmax = 0.58+/-0.07 micromol kg(-1) h(-1)) on the basis of Welch's approximate t-test. Similarly, the Michaelis-Menten model parameter estimates for Cd uptake by carp gills in the treatment experiment (KM = 0.15 +/- 0.06 micromol l(-1); Vmax = 5.14 +/- 1.07 micromol kg(-1) h(-1)) were not significantly different from the model parameters derived from the control experiment (KM =0.27 +/- 0.09 micromol l(-1); Vmax = 7.63+/-1.38 micromol kg(-1) h(-1)). This indicated that the Cd uptake rate by total carp and in carp gills in the presence of commercial humic acid followed the measured variations in Cd2+ -ion activity as predicted by the control experiment.

Animals↗

Isolation and characterisation of tilapia beta-actin promoter and comparison of its activity with carp beta-actin promoter.

The regulatory sequence including proximal promoter, untranslated exon 1 and intron 1 of the beta-actin gene from tilapia (Oreochromis niloticus) has been isolated and spliced to a beta-galactosidase reporter gene to test its activity. Comparisons of promoter activity have been carried out with three different constructs: (1) 1.6 kb tilapia beta-actin regulatory sequence, (2) 1.5 kb carp beta-actin regulatory sequence, and (3) 4.7 kb carp beta-actin regulatory sequence. Although the 1.6 kb tilapia beta-actin regulatory sequence gave slightly different expression patterns in tilapia embryos assayed by in situ X-gal staining, no difference was observed in expression level when the tilapia sequence was compared with the 4.7 kb carp beta-actin regulatory sequence by quantitative assay. In comparison with the 1.5 kb carp beta-actin regulatory sequence, the 1.6 kb tilapia beta-actin regulatory sequence gave higher expression levels in tilapia embryos, while a reverse result was observed in zebrafish embryos. In cell transfection experiments, the 1.6 kb tilapia beta-actin regulatory sequence showed three to four times better activity in blue gill cells than either the 4.7 kb carp beta-actin or the 1.5 kb carp beta-actin regulatory sequences. The 1.6 kb tilapia beta-actin regulatory sequence also drove higher reporter gene activity in somatic cells of tilapia than did the 4.7 kb carp beta-actin regulatory sequence following direct injection of constructs into muscle. Therefore, taken together, the data demonstrate that the tilapia beta-actin promoter can be used as an efficient regulatory sequence to produce autotransgenic tilapia.

Actins↗

Cloning of black carp beta-actin gene and primarily detecting the function of its promoter region.

A 3 338 bp DNA fragment including the open reading frame and 5'-flanking region of beta-actin gene for black carp genome was obtained through PCR amplification. Analysis of the sequencing results indicated the ORF of black carp beta-actin gene encoding a 375 amino acid protein that shares a high degree of conservation to other known actins. The black carp beta-actin sequence showed 100% identity to common carp, grass carp, and zebrafish, 99.2% identity to human and Norway rat beta-actin gene, 98.9% and 98.1% identity to chicken and Kenyan clawed frog beta-actin gene, respectively. The promoter region of black carp beta-actin gene was inserted into the promoterless pEGFP1 vector. The recombinant plasmid was microinjected into the fertilized eggs of mud loach before two-cell stage as well as transfected into HeLa cell line. GFP expression was found in 50% of mud loach embryos and 2/3 HeLa cells. The GFP expression could be observed in every part of the mud loach embryos, and in some embryos, the GFP was expressed in the whole body. Thus, the usefulness of black carp beta-actin promoter as a ubiquitous expression promoter was confirmed using the EGFP as a reporter gene.

5' Flanking Region↗

Carp (Cyprinus carpio) vitellogenin: purification and development of a simultaneous chemiluminescent immunoassay.

Vitellogenin (Vg) was purified from the serum of vitellogenic female carp (Cyprinus carpio) by hydroxylapatite column chromatography and gel filtration. Vg had an apparent molecular mass of 490 kDa and appeared as two bands corresponding to 190 and 156 kDa after SDS-PAGE under reducing conditions. These bands were immunoreacted in Western blotting using antiserum against carp lipovitellin (anti-Lv) which is an egg yolk protein derived from Vg. The amino acid composition of carp Vg was similar to previous reports of cyprinids. The chemiluminescent immunoassay (CLIA) for carp Vg was developed to quantify serum Vg using purified carp Vg and anti-Lv. Its measurable range was from 1.95 to 1000 ng/ml. The dilution curve in the CLIA of vitellogenic female serum was parallel to the standard curve of purified Vg. The coefficient variations of intra- and inter-assay were less than 5%, respectively. Furthermore, the assay had cross-reactivity with the sera of other female cyprinids (crucian carp and Japanese dace). In fish diets-experiments, Vg was detected in all fish in the fish meal containing soybean (20%) group, but was not detected in almost all of the fish in the fish meal-group. This suggests that a soybean based-diet may induce Vg production in the serum of cultivated carp.

Animals↗

Parasite assemblages of crucian carp (Carassius carassius)--is depauperate composition explained by lack of parasite exchange, extreme environmental conditions or host unsuitability?

Recent studies on parasite communities have considered the factors which contribute to variability in parasite infections among populations of one host species. This paper examined parasite assemblages of crucian carp (Carassius carassius) in a natural set-up of two distinctive environments inhabited by the same host species and differing in respect to composition of other fish species: ponds, where extreme conditions prevent other fish species from occurring and lakes, where crucian carp coexist with other fish species. Our aim was to evaluate the significance of parasite exchange, environmental conditions and host suitability for the depauperate parasite assemblages observed in pond crucian carp. As predicted, crucian carp had more diverse parasite species composition in lakes, which supports the hypotheses of increased opportunities for parasite exchange with other fish species and better environmental conditions for the parasites. However, for instance, diplostomids were found only from few fish individuals in very low numbers. Experimental exposure trials with the eye fluke Diplostomum spathaceum in the laboratory indicated that crucian carp were totally resistant to infection, which is exceptional and suggests that the low number of these parasites in crucian carp was due to physiological unsuitability of the host. To obtain a better understanding on the mechanisms underlying the formation of parasite assemblages in these fish, further studies on the relationships between the unique physiology of crucian carp and infection success by other parasite species are needed.

Acanthocephala↗

Differential scanning calorimetry and CD spectrometry of acclimation temperature-associated types of carp light meromyosin.

Differential scanning calorimetry and CD spectrometry were employed to study the thermal unfolding of light meromyosin (LMM) prepared from carp acclimated to different temperatures. The transition temperatures given by the major peaks at pH 8.0 in 0.6 M KCl for LMM from carp acclimated to 10 degrees C were 32.5 and 39.5 degrees C with the calorimetric enthalpies (DeltaHcal) of 269 and 52 kcal/mol, respectively. LMM from carp acclimated to 20 degrees C exhibited three peaks of transition temperatures at 34.5, 40.2, and 46.9 with DeltaHcal of 152, 20, and 10 kcal/mol, respectively. On the other hand, LMM from carp acclimated to 30 degrees C showed two different patterns. The first experiment gave two transition temperatures at 39.2 and 47.3 degrees C with DeltaHcal of 231 and 39 kcal/mol, respectively. The second series of experiments resulted in showing three peaks of 34.4, 39.5, and 47.5 degrees C with DeltaHcal of 117, 123, and 28 kcal/mol, respectively. N-terminal amino acid sequence analysis revealed that LMM at the second series of experiments with the 30 degrees C-acclimated carp contained component(s) predominant in the 20 degrees C-acclimated carp. Thermal unfolding responsible for these transition temperatures was well explained by melting of alpha-helices which could be determined by far-ultraviolet CD spectroscopy. These results clearly demonstrate that the 30 degrees C-acclimated carp contained the most thermostable LMM.

Acclimatization↗

Homology model and molecular dynamics simulation of carp ovum cystatin.

In this study, a homology model of carp ovum cystatin was constructed based on the crystal structure of chicken egg white cystatin. The results of amino acid sequence alignment indicate that these two proteins exhibit 36.11% of sequence identity. The resultant homology model reveals that carp ovum cystatin shares similar folds as chicken egg white cystatin, particularly in the conserved regions of Q48-V49-G52 and P98-W99 and the locations of two disulfide bonds, C67-C76 and C90-C110. However, the results of 1 ns molecular dynamics simulations show that carp ovum cystatin exhibits less structural integrity than chicken egg white cystatin in explicit water at 300 K. The relatively hydrophilic Met62 of carp ovum cystatin, corresponding to the hydrophobic Leu68 of human cystatin C and Ile66 of chicken egg white cystatin, may destabilize the hydrophobic core and form a dimeric structure more easily through domain swapping. A total of 16 positively charged residues are equally distributed on the surface of carp ovum cystatin, resulting in agglutination with the negatively charged spermatozoa via electrostatic interaction. Thus, carp ovum cystatin is considered to be important in preventing carp eggs from polyspermy.

Agglutination↗

Lipid deposition in intestine as a possible cause of malabsorption of nutrients in zinc-deficient common carp (Cyprinus carpio).

An experiment was performed to examine the interaction between Zn deficiency and lipid intake in carp. The carp were given a high-lipid diet that was either Zn-deficient (ZD) or Zn-supplemented (ZS), or were pair-fed (PF) the ZS diet to the intake of the ZD group. After 8 weeks the carp were killed and measurements were made of intestinal glucose uptake, levels of DNA, RNA and triacylglycerol, and alkaline phosphatase (EC 3.1.3.1) activity in liver and intestine samples. A further group of similar carp were given the same diets but at week 8 were transferred to low-lipid diets, with the exception of half the ZD group. After a further 8 weeks of treatment, carps were killed for biochemical studies. Intestinal [14C]glucose uptake, levels of DNA, RNA and alkaline phosphatase activity in intestine and liver were significantly (P < 0.05) lower in the high-lipid ZD group than in the high-lipid ZS and PF diet groups. The triacylglycerol concentration in the intestine was higher in the high-lipid ZD group than in the other two groups. When the carp were given the corresponding low-lipid diets, the variables measured in intestine and liver of the ZD group were close to those of the other groups. The results of this study demonstrate that lipid, when present in excess in the diet, accumulates in the intestine under Zn-deficient conditions and may reduce the absorption of glucose in carp. The reduced RNA and DNA levels and alkaline phosphatase activity in liver and intestine of ZD fish compared with those of ZS fish given high-lipid diets is proposed to be due to the malabsorption of nutrients linked with lipid deposition in the intestine, rather than their dependence on the level of Zn in the diet.

Alkaline Phosphatase↗

Effects of adding various oils to the diet on growth, feed conversion and chemical composition of carp (Cyprinus carpio).

The suitability of various vegetable and fish oils was tested as feed components for one-summer-old carp fingerlings. Five diets were used: a basal diet, which served as control feed, was supplemented with 10% corn-germ oil, 10% sunflower oil, 10% fish oil or 10% rapeseed oil, respectively. By means of the lipid supplementation the crude protein content decreased from 35.5% in the basal diet to 31.9% in the test diets, whereas the gross energy level increased from 16.3 to 18.9 MJ/kg. The diets were fed over a period of 84 feeding days and water temperature was 23 degrees C. Weight gain, feed conversion ratio and protein utilization were superior in all groups of carp which received the fat-enriched high energy diets. Considering the gain in % of the initial weight, the control fish reached less than 400% whereas all carp in the test groups, which consumed the oil-enriched diets, reached 455 to 485%. A significant influence of the kind of lipid on growth and feed conversion could not be recognized. The lipid-enriched diets with higher energy content resulted in increased fat levels of the carp. Fatty acid composition of the carp reflected that of the diets. In the control group and the fish oil group the levels of palmitic, palmitoleic, eicosapentaenoic and docosahexaenoic acid were especially high corresponding to the fatty acid profile of the diets. In carp fed the diets supplied with corn-germ oil and sunflower oil linoleic acid was dominant whereas the carp fed the rapeseed diet were characterized by a high level of oleic acid.

Aging↗