Xue Rui, Yang Dinglong, Han Yijing, Deng Qinyou, Wang Xin, Liu Xiangquan, Zhao Jianmin
Muping Coastal Environmental Research Station, Yantai Institute of Coastal Zone Research, Yantai Shandong, 264117, PR China; Center for Ocean Mega-science, Chinese Academy of Sciences, Qingdao Shandong, 266071, PR China; Research and Development Center for Efficient Utilization of Coastal Bioresources, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai Shandong, 264003, PR China.
Muping Coastal Environmental Research Station, Yantai Institute of Coastal Zone Research, Yantai Shandong, 264117, PR China; Center for Ocean Mega-science, Chinese Academy of Sciences, Qingdao Shandong, 266071, PR China; Research and Development Center for Efficient Utilization of Coastal Bioresources, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai Shandong, 264003, PR China.
Dev Comp Immunol. 2021 Nov;124:104176. doi: 10.1016/j.dci.2021.104176. Epub 2021 Jun 18.
The 14-3-3 proteins play important roles in various cellular processes by binding to different ligands, but little is known about these proteins in mollusks. In this study, two 14-3-3 cDNAs were identified from the Pacific abalone Haliotis discus hannai (designated 14-3-3ζ and 14-3-3ε), possessing 59.40% identity with each other. Both genes were predominantly expressed in the gills of unchallenged abalones, and their mRNA signals could also be detected in several other tissues, including the mantle, hepatopancreas and ovary. However, after Vibrio harveyi challenge, hemocytes were induced significantly (p < 0.01). Meanwhile, phagocytosis was inhibited, but apoptosis, reactive oxygen species formation, and caspase 3 expression were significantly induced (p < 0.01), and they were all suppressed with 14-3-3ζ knockdown (p < 0.01). The differences were that silencing 14-3-3ε reverted the decline in the phagocytic rate derived from bacterial infection, while ROS formation was not influenced significantly. In addition, the expression levels of several antimicrobial peptide and proinflammatory cytokine genes were also decreased with the silencing of 14-3-3 genes. However, with the knockdown of 14-3-3ζ, the expression of 14-3-3ε was further significantly increased (p < 0.01), and vice versa. Overall, our results suggested that 14-3-3ζ and 14-3-3ε should play important roles in innate immunity against V. harveyi infection.
14-3-3蛋白通过与不同配体结合在各种细胞过程中发挥重要作用，但在软体动物中对这些蛋白的了解甚少。在本研究中，从皱纹盘鲍（Haliotis discus hannai）中鉴定出两个14-3-3 cDNA（命名为14-3-3ζ和14-3-3ε），它们彼此间具有59.40%的同一性。这两个基因在未受挑战的鲍鱼的鳃中主要表达，并且在包括外套膜、肝胰腺和卵巢在内的其他几种组织中也能检测到它们的mRNA信号。然而，在哈维氏弧菌攻击后，血细胞被显著诱导（p<0.01）。同时，吞噬作用受到抑制，但凋亡、活性氧形成和半胱天冬酶3表达被显著诱导（p<0.01），并且它们在14-3-3ζ基因敲低后均受到抑制（p<0.01）。不同之处在于，沉默14-3-3ε可逆转由细菌感染引起的吞噬率下降，而活性氧形成未受到显著影响。此外，随着14-3-3基因的沉默，几种抗菌肽和促炎细胞因子基因的表达水平也降低。然而，随着14-3-3ζ基因敲低，14-3-3ε的表达进一步显著增加（p<0.01），反之亦然。总体而言，我们的结果表明14-3-3ζ和14-3-3ε在抵抗哈维氏弧菌感染的先天免疫中应发挥重要作用。
