Molecular cloning, characterization and expression analysis of heat shock protein 90 in albino northern snakehead Channa argus

The great albino northern snakehead Channa argus is habitual to only the Sichuan Jialing Rivers System in China, making its introduction difficult to other riverine systems. Here, we characterized heat shock protein 90 (AcaHSP90) and probed its molecular responses toward the environmental stressors that C. argus can face during its introduction and breeding in the other southern latitudes of China. To serve the purpose, cDNA encoding of AcaHSP90 were cloned and characterized in albino C. argus. The cDNA was 2752bps that contained an open reading frame (ORF), encoding a 726-amino-acid polypeptide of 83.35kDa (theoretical isoelectric point [pI]: 4.89). Genomic DNA analysis showed that the AcaHSP90 gene consisted of 7 introns, five conserved amino acid blocks and other motifs or domains. The AcaHSP90 structure was highly similar with the other known HSP90s except those identified in the bacteria. The expression profiles of AcaHSP90 gene in albino C. argus were also investigated after experimentally exposed to different temperature stresses (8.5, 26 and 37°C) and infected with Edwardsiella tarda (strain NO. DL1476) at different time intervals (0, 6, 12, 24, 36, 48, 72h). In addition, the AcaHSP90 expression in different tissues of albino C. argus were also analyzed. The quantitative real-time PCR and western blot analysis revealed tissue-specific AcaHSP90 expressions in control group, and expressions were significantly stimulated in the brain, heart, kidney, liver, muscle and spleen after the heat shock (37°C), while showed no significant difference after the cold treatment (8.5°C). The mRNA levels of AcaHSP90 were also significantly upregulated in the spleen and muscle at 12h and in the kidney at 12 and 48h post pathogen injections. In a nut shell, these novel results showed tissue-specific responses of AcaHSP90 and indicated that this heat shock protein might also be sensitive to pathogen infection, but closely related to the thermal resistance in albino C. argus.