Histone-like proteins from Atlantic cod milt: stimulatory effect on Atlantic salmon leucocytes in vivo and in vitro.
ABSTRACT This study was carried out to reveal some characteristics of cationic proteins from Atlantic cod (Gadus morhua) milt chromatin and to investigate their ability to activate Atlantic salmon (Salmo salar) macrophages. Cationic proteins extracted from cod milt chromatin were fractionated on a cation exchange chromatography column. SDS-PAGE and amino acid analyses of the resulting fractions indicated that these proteins are similar to calf thymus histones. Two cationic protein fractions were used to stimulate leucocytes from Atlantic salmon in vitro and in vivo. Increased production of superoxide, measured as reduction of nitroblue tetrazolium (NBT), was used as indication of macrophage activation. Both fractions induced elevated superoxide anion production in the macrophages after 3 and 6 days of in vitro stimulation. Intraperitoneal injection of the cationic protein fractions in Atlantic salmon (100 mg kg(-1)) four days prior to slaughtering stimulated superoxide production when assayed after one and two days of cell cultivation. In macrophages from fish slaughtered two days after injection, activation could first be seen after two days of cell cultivation.
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ABSTRACT: By the criteria of gel electrophoresis, ion-exchange chromatography, and reverse-phase HPLC, yellow perch protamine behaves as a single component. This observation was confirmed by automated Edman degradation which gave a single unambiguous amino acid sequence PRRRRHAARPVRRRRRTRRSSRVHRRRRAVRRRR. Yellow perch protamine has 34 amino acids, including 21 arginines. It has two histidines, neither of which interrupts an arginine tract. It is unusual among fish protamines in not having a serine or threonine N-terminal to the second arginine tract, and is unique in not being a mixture of components.FEBS Letters 04/1992; 299(2):166-8. · 3.58 Impact Factor
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ABSTRACT: In this work we study the presence and the composition of 24 species of bony fish, representing 18 families and 7 orders of teleosts. The species studied belonging to orders Clupeiformes and Lophiiformes display typical protamines, whereas the species belonging to orders Notacanthi-formes, Gadiformes, and Ophiidiformes contain histones instead of protamines in their sperm nuclei. In these species, one or more specific proteins are added to the pattern of canonical histones and, in the cases that have been analysed compositionally (Merluccius capensis, Cataetyx laticeps), the specific proteins seem to be related to histones. Among the order Scorpaeniformes, the species Helicolenus dactylopterus (family Scorpaenidae) presents protamines, whereas Trigla lucerna (family Triglidae) contains histones. Similarly, among the order Perciformes, the species from six families (Percichthyidae, Labridae, Trachinidae, Uranoscopidae, Gempylidae, and Scombridae) have protamines in sperm nuclei and species from two families (Sparidae and Trichiuridae) retain histones and increase the proportion of H1 histone approximately twofold. Two species of Mullus (family Mullidae, order Perciformes) display special proteins with an electrophoretic behaviour corresponding to histone H4 but an amino acid composition more basic.The results indicate that bony fishes have adopted three main alternatives to condense their nuclear sperm chromatin: namely, protamines, histones, and histone-like proteins. Moreover, these alternatives do not correspond strictly with phylogeny, favouring the hypothesis that the change “protamine → histone” (or alternatively “histone → protamine”) has occurred independently several times during the evolution of bony fishes. © 1993 Wiley-Liss, Inc.Journal of Experimental Zoology 01/1993; 265(5):575 - 586.
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ABSTRACT: The present paper describes the effect of lipopolysaccharides (LPS) fromAeromonas salmonicida and other Gram-negative bacteria on the respiratory burst, phagocytosis and bactericidal activity of head kidney macrophages from Atlantic salmon (Salmo salar L.) in vitro. Macrophages were first cultured in the presence of various concentrations of LPS from A. salmonicida for 1, 2 and 5 days and then tested for respiratory burst activity (reduction of nitroblue tetrazolium) after exposure to phorbol myristate acetate (PMA). The most marked increase in respiratory burst activity of LPS-treated macrophages was observed after 5 days of incubation with 1, 10 and 100 μg LPS ml −1. The increase appeared to be dose-dependent with a maximal response at 10 μg ml−1. At this LPS-concentration and incubation time the respiratory burst activity was 3·9 times larger in the treated macrophages than in the control macrophages. LPS from three other Gram-negative bacterial salmon pathogens and two non-fish pathogens also enhanced the respiratory burst activity of salmon macrophages. Macrophages incubated with 10 and 50 μg LPS ml −1 also showed a significant increase in PMA-stimulated H2O2-production after 5 days of incubation. LPS also stimulated the phagocytic activity of Atlantic salmon macrophages against opsonized and nonopsonized glucan particles, and glutaraldehyde-fixed sheep red blood cells. LPS-treated macrophages showed an increased ability to kill an avirulent A-layer lacking strain of A. salmonicida, but not a virulent A-layer positive strain.Fish & Shellfish Immunology. 01/1995;