Seasonal variations in the content of catecholamines in carp heart (Cyprinus carpio)

1. Seasonal changes in epinephrine and norepinephrine contents were demonstrated in ventricular muscle tissues of carp heart. 2. Patterns of changes in epinephrine and norepinephrine contents were similar; high during late spring-to-summer and low in winter; however, the fluctuation in epinephrine content was greater than that of norepinephrine. 3. Tyramine caused a positive inotropic effect in isolated electrically-stimulated ventricular muscles of carp heart. The inotropic effect was totally blocked by reserpine, reduced by propranolol, but not altered by atenolol. 4. These results suggest that the catecholamines released from sympathetic nerve terminals may contribute to the regulation of the seasonal change in the function of cardiac muscles in the carp.     

The complete nucleotide sequence and gene organization of carp (Cyprinus carpio) mitochondrial genome

The complete sequence of the carp mitochondrial genome of 16,575 base pairs has been determined. The carp mitochondrial genome encodes the same set of genes (13 proteins, 2 rRNAs, and 22 tRNAs) as do other vertebrate mitochondrial DNAs. Comparison of this teleostean mitochondrial genome with those of other vertebrates reveals a similar gene order and compact genomic organization. The codon usage of proteins of carp mitochondrial genome is similar to that of other vertebrates. The phylogenetic relationship for mitochondrial protein genes is more apparent than that for the mitochondrial tRNA and rRNA genes.Correspondence to: F. Huang     

The ultrastructure of leucocytes in carp (Cyprinus carpio)

Leucocytes from anterior kidney, middle kidney, spleen, thymus and peripheral blood of Common carp were examined. Lymphocytes were found to have a similar fine structure to that of mammals. Thrombocytes had a similar appearance to lymphocytes, but the former were sometimes distinguishable by vesicles in series and/or microtubules below the plasma membrane. Blast cells, monocytes and clearly identifiable plasma cells were also seen. Neutrophils had varying proportions of at least two types of granules, which often presented inclusions. Owing to their different granules, eosinophils and basophils were morphologically distinguishable, but differentiated cells with equal proportions of the two types of granules were also seen. In addition, cells of uncertain nature, possessing rod-shaped granules, were observed. The different leucocyte types showed the same morphology in the different lymphoid organs and in peripheral blood, although they were present in different proportions.     

Hormones with adrenocorticotropic and opiate-like activities from the carp (Cyprinus carpio) pituitary

Carp (Cyprinus carpio) pituitary acetone powder was extracted with a mixture of water, hydrochloric acid and acetone. An acid acetone powder was formed by adding the pituitary extract into a large volume of chilled acetone and subsequently recovering the precipitate. The powder was subjected to ion exchange chromatography on CM cellulose. Fractions adsorbed on the ion exchanger exhibited ACTH-like activity as evidenced in the ability to stimulate lipolysis in isolated hamster adipocytes and corticosterone production in isolated rat adrenal decapsular cells and also in cross-reactivity in an ACTH-specific radioimmunoassay. A portion of the ACTH-like bioactivity and immunoactivity was unadsorbed on the ion exchanger. Opiate-like activity in opiate receptor binding assay, employing [3H]D-ala2-D-leu5 enkephalin or [3H]naloxone as ligand, also resided in fractions adsorbed on CM cellulose. The data indicate a separation of ACTH-like and opiate-like activities, and the presence of opiate-like molecules with different affinities of binding to mu and delta opiate receptors.     

One-step, non-denaturing purification method of carp (Cyprinus carpio) vitellogenin

A new single-step purification procedure was developed to purify carp (Cyprinus carpio) vitellogenin (VTG), from estradiol-treated carp plasma. This method was performed by high performance liquid weak anion-exchange chromatography, using a discontinuous elution gradient of NaCl (0-0.5 M, steps of 12.5 mM/4 min). SDS and native-PAGE analysis, of treated-fish plasma and purified solution, showed the appearance of a 370 kDa phospholipoprotein, composed of two 130 kDa monomers, with all VTG characteristics. The sequencing of a 130 kDa monomer confirmed that it was carp VTG. Consequently, this procedure is a rapid method, permitting high quantities of non-denatured carp VTG to be obtained.     

Inactivation of enolase from carp (Cyprinus carpio) muscle by 2,3-butanedione

Enolase from white muscle of carp (Cyprinus carpio) is inactivated by 2,3-butanedione in borate buffer. Magnesium ions as well as substrate--2-phosphoglycerate markedly altered the rate and extent of inactivation. The partially inactivated enzyme shows unaltered Km but decreased Vmax after 10 min incubation with butanedione, however after 60 min incubation the Km value increased 2.5 fold.     

Adaptation of intestinal morphology in the temperature-acclimated carp,Cyprinus carpio L.

Summary The effects of temperature and photoperiod acclimation upon the morphology of carp intestinal mucosa have been studied using morphometric techniques. Carp intestine showed an absence of anatomical regionalisation. There was a gradual reduction in the dimensions of villi along the tract. The decrease in the dimensions of the villi was greatest in the anterior half. Temperature acclimation had no effect on intestinal-somatic indices. Acclimation to 10° C or 30° C resulted in large differences in the dimensions of villi. Cold acclimation produced significant increases in mean villus height and breadth along the entire intestine. These villus shape changes resulted in a 58% increase in total mucosal surface area and a 102% increase in total volume of villi in cold-acclimated fish relative to warm-acclimated fish. Surface area of the unmodified intestinal tube increased with cold acclimation by 28%. The total number of villi remained unchanged by thermal acclimation. Because normalisation to a nominal surface area does not take account of the possibility of differentially developed mucosal surfaces in differently acclimated animals, experiments comparing transepithelial transport rates of differently-acclimated fish, using unstripped preparations, overestimates the differences in area-specific transport capacity.     

Phospholipid content in the compact and spongious musculature of the carp heart (Cyprinus carpio)

The phospholipid content of the spongious and compact musculature of the heart of the carp (Cyprinus carpio) was compared. The content of phospholipids is higher in the spongious musculature, the greatest difference being in the content of diphosphatidylglycerol: 2.53 mumol P . g-1 wet weight in the spongious layer and 1.29 mumol P . g-1 wet weight in the compact one. In both tissues plasmalogens represent 20-27% of the tissue content of choline and ethanolamine phosphoglycerides.     

Presence of four tachykinins in an acid extract of the carp intestinal bulb (Cyprinus carpio)

1. The pharmacological and chemical properties of substance P-like peptides isolated from an acid extract of the carp intestinal bulb were examined using guinea-pig ileum longitudinal smooth muscle.2. On a Sephadex G25 column (3 × 96 cm), smooth muscle contracting material was eluted as two peaks (fraction-1 and fraction-2). The molecular weight of the fraction-1 was estimated to be 2300 and that of the fraction-2 to be 1530.3. The pharmacological properties of the contracting materials in fraction-1 and fraction-2 resembled those of substance P and neurokinin A.4. The susceptibility of the contracting activity of fraction-1 to proteolytic enzymes resembled that of physalaemin but, on the other hand, the susceptibility of that of fraction-2 resembled those of eledoisin and neurokinin A.5. Ion-exchange chromatography on sulfopropyl-Sephadex C25 indicated the presence of one contracting material in fraction-1 and three contracting materials in fraction-2. The elution positions of four materials were different from that of substance P.6. These results indicate that four tachykinins different from substance P are present in an acid extract of the carp intestinal bulb.     

Cleavage specificity of a myofibril-bound serine proteinase from carp (Cyprinus carpio) muscle

Previously, we reported the purification and characterization of a myofibril-bound serine proteinase (MBP) from carp muscle (Osatomi K, Sasai H, Cao M-J, Hara K, Ishihara T. Comp Biochem Physiol 1997;116B:159–66). In the present study, the N-terminal amino acid sequence of the enzyme was determined, which showed high identity with those of other trypsin-like serine proteases. The cleavage specificity of MBP for dibasic and monobasic residues was investigated using various fluorogenic substrates and peptides. Analyses of the cleaved peptide products showed that the enzyme hydrolyzed peptides both at monobasic and dibasic amino acid residues. Monobasic amino acid residues were hydrolyzed at the carboxyl side; dibasic residues were cleaved either at the carboxyl side of the pair or between the two basic residues and the enzyme showed a cleavage preference for the Arg-Arg pair. Unexpectedly, MBP hydrolyzed lysyl-bradykinin and methionyl–lysyl–bradykinin at the carboxyl side of Gly fairly specifically and efficiently displaying a unique cleavage. Because MBP also degraded protein substrates such as casein and myofibrillar proteins, the substrate specificity of MBP appeared not to be strictly specific.     

Fillet yield and fat content in common carp (Cyprinus carpio) produced in three Austrian carp farms with different culture methodologies

The fat content and fillet yield of common carp ( Cyprinus carpio ) from three carp farms were investigated with respect to pond management and overall climatic conditions. The mean fat content of the trimmed fillet varied from 2.7 to 6.9% and differed significantly among the three facilities (P < 0.001). It is suggested that differences in the fillet fat content may be based on the climatic conditions as well as the different management strategies (feeding, stocking biomass). The mean fillet yield was calculated as 34.0–39.0%. No significant correlation between fish weight and fillet yield (ρ = −0.16) was found. Results suggest that producing smaller fish for fillets of serving size may lead to a higher production and conclude with lower production risks and costs.     

Development and characterization of two new cell lines from common carp, Cyprinus carpio (Linn)

Two new cell lines (CCF and CCH) were established from fin and heart tissues of common carp, Cyprinus carpio. The cells were optimally maintained in Leibovitz-15 medium supplemented with 10% fetal bovine serum (FBS) and 10 ng/ml of basic fibroblastic growth factor (bFGF). The effects of temperature, concentration of FBS and bFGF on the growth of CCF and CCH cells were examined. The temperature ranged from 24 to 32°C for good growth of the cells. The growth rate of cells was higher in medium containing 10% FBS and the addition of bFGF to the medium significantly increased the growth rate. The CCF cells were found to be epithelial, while the CCH cells were fibroblastic in nature. The cytogenetic analysis of the cell lines revealed a diploid number of 100 chromosomes in C. carpio. The viability of CCF and CCH cell lines were 70 and 72%, respectively, after six months of storage in liquid nitrogen (-196° C). Molecular characterization of the cell lines using 16S rRNA and Cytochrome Oxidase Subunit I (COI) revealed the origin of the cell lines. These new cell lines will be useful for isolation of fish viruses and other in vitro biotechnological studies.     

Subcellular localization of muscle FBPase in carp (Cyprinus carpio) tissues

Subcellular localization of muscle FBPase-a regulatory enzyme of glyconeogenesis-was investigated in carp using immunohistochemistry and protein exchange method. Results of the experiments revealed that, in striated muscles, FBPase associates with alpha-actinin of the Z-line and co-localizes with aldolase. Additionally, in cardiac and smooth muscle cells FBPase is present inside the nuclei. In the light of findings on mammalian muscle FBPase, the data presented here indicates that interaction of the enzyme with specific cellular partners and nuclear presence of FBPase is a general phenomenon in contemporary vertebrates.     

Electrocardiograms recorded from the body surface of the carp, Cyprinus carpio

The electrocardiograms (ECGs) in five kinds of bipolar leads were recorded from the body surface of the carp under water. Each component constituting the ECG wave could be detected by the present method. The potentials of the ECGs recorded by the present method were relatively small, the values being 2-64 microV. The electrical axis in most carp was directed toward the right lateral side.