Temperature-dependent changes of growth rate and protein components were investigated for primary cultured cells derived from goldfish caudal fin. When the culture temperature was shifted from 20ЊC to 35ЊC and 40ЊC, the growth rate was increased at 35ЊC as compared with that at 20ЊC, but no cell growth was observed at 40ЊC. The differential scanning calorimetry demonstrated the onset of the endothermic reaction for goldfish cellular components at 40ЊC. Therefore, the temperature shift to 40ЊC was found to be of severe heat shock for goldfish cultured cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that, although expression of 70-kDa components was slightly induced at 35ЊC, the temperature shift to 40ЊC markedly induced the expression of the 30-kDa component in addition to that of 70-kDa component. The N-terminal amino acid sequencing identified the 30-and 70-kDa components to be heat shock protein (Hsp)-30 and Hsp70, respectively. Northern blot analysis revealed that the enhanced Hsp30 messenger ribonucleic acid (mRNA) levels were only observed at 40ЊC, whereas Hsp70 mRNA was slightly accumulated at 35ЊC. These results indicated that Hsp30 might have important functions under severe heat stress condition.
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