Liu Zhaoqun, Wang Lingling, Zhou Zhi, Liu Yu, Dong Miren, Wang Weilin, Song Xiaorui, Wang Mengqiang, Gao Qiang, Song Linsheng
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; Functional Laboratory of Marine Fisheries Science and Food Production Process, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200, China; University of Chinese Academy of Sciences, Beijing 100049, China.
Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian 116023, China.
Dev Comp Immunol. 2017 Aug;73:52-60. doi: 10.1016/j.dci.2017.03.005. Epub 2017 Mar 7.
Bacterial infection and heat stress, as two major environmental threats of marine molluscs, could affect larval development and dramatically promote mortality of oysters. In the present study, next-generation sequencing, together with determinations of mRNA expression and measurements of enzyme activities, were employed to understand the response patterns of oyster larvae under acute heat and bacterial stress. After RNA-seq, a total of 9472 differentially expressed genes including 4895 significantly up-regulated ones and 4577 significantly down-regulated ones were obtained from 12 transcriptome libraries. GO overrepresentation analysis of the up-regulated genes revealed that the neuroendocrine immunomodulation pathway was activated after acute heat and bacterial stimulation, in which the catecholaminergic regulation played an important role. GO overrepresentation analysis of the down-regulated genes suggested that the immune capacity of Crassostrea gigas larvae was suppressed under stress, which was further validated since superoxide dismutase (SOD) and phenoloxidase (PO) activities in the total protein extract of larvae decreased dramatically after stress. Moreover, the shell formation of trochophore was inhibited and severe mortality was caused after acute heat and bacterial stress. These results collectively indicated that acute heat and bacterial stress could significantly inhibit larval development and suppress immune response of oyster C. gigas larvae. And the neuroendocrine immunomodulation, especially the catecholaminergic regulation, played an indispensable role in the stress response of molluscan larvae.
细菌感染和热应激是海洋软体动物面临的两大主要环境威胁,它们会影响幼体发育,并显著提高牡蛎的死亡率。在本研究中,采用新一代测序技术,结合mRNA表达测定和酶活性测量,来了解牡蛎幼体在急性热应激和细菌应激下的反应模式。RNA测序后,从12个转录组文库中总共获得了9472个差异表达基因,其中包括4895个显著上调基因和4577个显著下调基因。对上调基因的GO富集分析表明,急性热应激和细菌刺激后神经内分泌免疫调节途径被激活,其中儿茶酚胺能调节发挥了重要作用。对下调基因的GO富集分析表明,应激状态下太平洋牡蛎幼体的免疫能力受到抑制,这一点也通过幼体总蛋白提取物中的超氧化物歧化酶(SOD)和酚氧化酶(PO)活性在应激后显著降低得到了进一步验证。此外,急性热应激和细菌应激后担轮幼虫的贝壳形成受到抑制,并导致严重的死亡率。这些结果共同表明,急性热应激和细菌应激可显著抑制太平洋牡蛎幼体的发育并抑制其免疫反应。神经内分泌免疫调节,尤其是儿茶酚胺能调节,在软体动物幼体的应激反应中发挥了不可或缺的作用。