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  • 1
    ISSN: 1573-5168
    Keywords: temperature ; acclimation ; cytokine ; enhancer ; goldfish ; hemopexin ; hepatopancreas ; immune system ; plasma protein ; lipopolysaccharide ; transcription ; Wap65
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of the Wap65 mRNA was compared between hepatopancreas, intestine, ovary, heart, gill, eye, skin, hemocytes, muscle, and brain from 30 °C-acclimated goldfish (Carassius auratus) by reverse transcription PCR, demonstrating the highest levels of expression of the gene in the hepatopancreas. Other tissues expressing the gene were ovary, gill, eye, and muscle, whereas Wap65 mRNA could not be detected in intestine, heart, skin, and brain. In Northern blot analysis, levels of Wap65 mRNA in the hepatopancreas increased after raising water temperature from 10 to 30 °C. The transcript levels reached a maximum of 40-fold on day 3 following temperature shift. On day 21 after raising water temperature, levels were 10-fold compared with those of the 10 °C-acclimated fish. Subsequently, in order to elucidate the transcriptional mechanisms of Wap65, a part of the Wap65 gene was cloned from a genomic library. The 5′-flanking region and introns of this gene contained enhancer motifs including cytokine responsive elements previously identified in mammals. The suggesting that some of these motifs are involved in the transcriptional regulation of Wap65, was tested by administering of lipopolysaccharide (LPS) to the 10 °C-acclimated goldfish. This resulted in the accumulation of Wap65 mRNA in the hepatopancreas, suggesting the involvement of immune associated factors in the regulation of transcription of Wap65.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 160 (1990), S. 233-239 
    ISSN: 1432-136X
    Keywords: Temperature ; Acclimation ; Myosin ; Myosin heavy chain ; ATPase activity ; Carp
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Myosins were isolated from dorsal ordinary muscles of carp acclimated to 10°C and 30°C for a minimum of 5 weeks and examined for their ATPase activities. Ca2+-ATPase activity was different between myosins from cold-and warm-acclimated carp, especially at KCl concentrations ranging from 0.1 to 0.2 M, when measured at pH 7.0. The highest activity was 0.32 μmol Pi·min-1·mg-1 at 0.2 M KCl for cold-acclimated carp and 0.47 μmol Pi·min-1·mg-1 at 0.1 M KCl for warm-acclimated fish. The pH-dependency of Ca2+-ATPase activity at 0.5 M KCl for both carp was, however, similar exhibiting two maxima around 0.3 μmol Pi·min-1·mg-1 at pH 6 and 0.4 μmol Pi·min-1·mg-1 at pH 9. K+(EDTA)-ATPase activity at pH 7.0 neither exhibited differences between both myosins. It increased with increasing KCl concentration showing the highest value of about 0.4 μmol Pi·min-1·mg-1 at 0.6–0.7 M KCl. Actin-activated myosin Mg2+-ATPase activity was markedly different between cold-and warm-acclimated carp. The maximum initial velocity was 0.53 μmol Pi·min-1·mg-1 myosin at pH 7.0 and 0.05 M KCl for cold-acclimated carp, which was 1.6 times as high as that for warm-acclimated carp. These differences were in good agreement with those obtained with myofibrillar Mg2+-ATPase activity between both carp. No differences were, however, observed in myosin affinity to actin. Differences in myosin properties between cold- and warm-acclimated carp were further evidenced by its thermal stability. The inactivation rate constant of myosin Ca2+-ATPase was 25·10-4·s-1 at 30°C and pH 7.0 for cold-acclimated carp, which was about 4 times as high as that for warm-acclimated carp. Light chain composition did not differ between both carp myosins. The differences in a primary structure of the heavy chain subunit was, however, clearly demonstrated between both myosins by peptide mapping.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 163 (1993), S. 349-354 
    ISSN: 1432-136X
    Keywords: Temperature ; Acclimation ; Heat-shock protein ; Expression ; Goldfish, Carassius auratus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Goldfish Carassius auratus were acclimated to either 10 or 30°C for a minimum of 5 weeks. A 65-kDa protein specific to warm-temperature-acclimated fish was extracted from the gel with 70% formic acid after two-dimensional electrophoresis of the muscle cytoplasmic protein fraction. The 65-kDa protein thus prepared to homogeneity was used to raise specific antibodies in rabbit by conventional methods. The antibody produced exhibited specific reaction with a protein having the same molecular weight from brain and liver tissue, suggesting that the 65-kDa protein is a ubiquitous cytosolic component in warm-acclimated goldfish. When water temperature was increased from 20 to 30°C over a 20-h period, a prominent amount of the 65-kDa protein was observed in muscle tissue extracts within 5 days of additional rearing; this was demonstrated by immunoblotting with the specific antibody. The N-terminal amino acid sequence of the 65-kDa protein was determined as Asp-Glu-Pro-Gln-Gly-His-Gln-His (or Asp)-Glu-Leu, differing from that of a family of known heat-shock proteins having about 70 kDa in molecular mass (hsp 70). No interaction between ATP and the 65-kDa protein revealed by ATP-agarose affinity chromatography further confirmed the different properties of the 65-kDa protein from those of hsp 70.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 163 (1993), S. 445-451 
    ISSN: 1432-136X
    Keywords: Multiple hemoglobins ; Subunit structure ; α-chain ; β-chain ; Carp, Cyprinus carpio
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Three hemoglobin components in carp designated CI, CII, and CIII, were isolated by DEAE-Toyopearl ion-exchange chromatography. Constituent globin chains, α1, α2, β1 and β2, were analyzed by urea-Triton acid polyacrylamide gel electrophoresis and isolated by high performance liquid chromatography with a reversed-phase column. Tryptic peptide mapping indicated that the α-globin chains of the three hemoglobin components have slightly different structures. In addition, N-terminal amino acid sequence analysis indicated that the β1-globin chain has a primary structure different from that of the β2-chain. A series of hybridization experiments between isolated hemoglobins, together with such structural properties of globin chains, suggested that the three hemoglobins have the following compositions: CI (α1 α2 β 2 1 ), CII (α1 α2 β1 β2), and CIII (α1 α2 β 2 2 ). Hemoglobin CII was a hybrid between the two types each of α- and β-chain and could be constructed in vitro from two hemoglobin components CI and CIII.
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