Wuxi Fisheries College, Nanjing Agricultural University, Wuxi, 214081, China; Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, 214081, China.
Wuxi Fisheries College, Nanjing Agricultural University, Wuxi, 214081, China; Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, 214081, China.
Fish Shellfish Immunol. 2018 Mar;74:119-132. doi: 10.1016/j.fsi.2017.12.046. Epub 2018 Jan 4.
Heat shock proteins (HSPs) play critical roles in the process of anti-stress and immunity and are implicated in autoimmune diseases. In order to understand the comparative stress responses of HSP60 and HSP90β under intermittent thermal stress and Aeromonas hydrophila infection, we cloned their full-length cDNAs from Megalobrama amblycephala liver, predicted their secondary and tertiary structure, and examined their tissue-specific expression patterns. The full length of HSP60 and HSP90β cDNAs indicated that they included all signature sequences of corresponding protein families. They showed high homology to their counterparts in other species, and were consistent with the known classification of fishes based on phylogenetic analysis. HSP60 showed the highest expression in head-kidney, brain, and gill, while HSP90β presented higher in hindgut, liver, and brain. Significant mRNA expression differences were determined between HSP60 and HSP90β in tissues of bladder, liver, heart, and gill. During thermal stress and recovery phase, the highest expression of them were observed at the first recovery for 2 d and 1 d, respectively. The expression between them were extremely significant difference during the first recovery and second stress period. After A. hydrophila infection, their expressions were extremely significantly upregulated. The significant upregulation and rapid response indicated that they were sensitive to thermal stress and bacterial challenge. This study demonstrated that HSP60 and HSP90β might participate in innate immune and environmental responses of M. amblycephala. It indicated that they could be used as biomarkers to test the stress caused by local aquaculture environment.
热休克蛋白（HSPs）在抗应激和免疫过程中发挥着关键作用，并与自身免疫性疾病有关。为了了解 HSP60 和 HSP90β 在间歇性热应激和嗜水气单胞菌感染下的比较应激反应，我们从团头鲂肝脏中克隆了它们的全长 cDNA，预测了它们的二级和三级结构，并检测了它们的组织特异性表达模式。HSP60 和 HSP90β cDNA 的全长表明它们包含了相应蛋白家族的所有特征序列。它们与其他物种的对应物具有高度同源性，与基于系统发育分析的鱼类已知分类一致。HSP60 在头肾、脑和鳃中表达最高，而 HSP90β 在后肠、肝和脑中表达较高。在膀胱、肝、心和鳃组织中，HSP60 和 HSP90β 的 mRNA 表达存在显著差异。在热应激和恢复阶段，它们的最高表达分别出现在第一次恢复的第 2 天和第 1 天。在第一次恢复和第二次应激期间，它们之间的表达差异极显著。感染嗜水气单胞菌后，它们的表达极显著上调。这种显著的上调和快速反应表明它们对热应激和细菌挑战敏感。本研究表明，HSP60 和 HSP90β 可能参与了团头鲂的先天免疫和环境反应。这表明它们可以用作测试局部水产养殖环境引起的应激的生物标志物。
