Zhou Aiguo, Xie Shaolin, Wang Zhenlu, Junaid Muhammad, Fan Lanfen, Wang Chao, Ye Qiao, Chen Yanfeng, Pei De-Sheng, Zou Jixing
College of Marine Sciences, South China Agricultural University, Guangzhou 510642, Guangdong, China; Qingyuan North River Fishery Science Institute, Qingyuan 511510, Guangdong, China.
College of Marine Sciences, South China Agricultural University, Guangzhou 510642, Guangdong, China.
Gene. 2017 Aug 30;626:173-181. doi: 10.1016/j.gene.2017.04.039. Epub 2017 Apr 23.
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.
白化北方黑鱼（Channa argus）仅在中国四川嘉陵江水系栖息，这使得将其引入其他水系十分困难。在此，我们对白化北方黑鱼的热休克蛋白90（AcaHSP90）进行了特性分析，并探究了其在引入和在中国其他南方纬度地区繁殖时可能面临的环境应激源下的分子反应。为此，我们克隆并分析了白化北方黑鱼中编码AcaHSP90的cDNA。该cDNA为2752bp，包含一个开放阅读框（ORF），编码一个83.35kDa（理论等电点[pI]：4.89）的726个氨基酸的多肽。基因组DNA分析表明，AcaHSP90基因由7个内含子、5个保守氨基酸块以及其他基序或结构域组成。AcaHSP90的结构与其他已知的热休克蛋白90高度相似，但与细菌中的热休克蛋白90不同。在对白化北方黑鱼进行不同温度应激（8.5、26和37°C）实验暴露以及在不同时间间隔（0、6、12、24、36、48和72小时）感染迟缓爱德华氏菌（菌株编号DL1476）后，我们还研究了AcaHSP90基因在白化北方黑鱼中的表达谱。此外，我们还分析了AcaHSP90在白化北方黑鱼不同组织中的表达。定量实时PCR和蛋白质免疫印迹分析显示，对照组中AcaHSP90存在组织特异性表达，热休克（37°C）后，脑、心、肾、肝、肌肉和脾脏中的表达显著上调，而冷处理（8.5°C）后无显著差异。病原体注射后12小时，脾脏和肌肉中AcaHSP90的mRNA水平显著上调，12小时和48小时时肾脏中AcaHSP90的mRNA水平也显著上调。总之，这些新结果显示了AcaHSP90的组织特异性反应，表明这种热休克蛋白可能也对病原体感染敏感，但与白化北方黑鱼的耐热性密切相关。
