Department of Aquaculture, National Pingtung University of Science and Technology, Pingtung, 91201, Taiwan, ROC.
Department of Aquaculture, National Pingtung University of Science and Technology, Pingtung, 91201, Taiwan, ROC.
Fish Shellfish Immunol. 2019 Apr;87:590-599. doi: 10.1016/j.fsi.2019.02.005. Epub 2019 Feb 8.
Complementary (c)DNA encoding novel protein kinase C (PKC) messenger (m)RNA of the white shrimp Litopenaeus vannamei, consisted of 2454-bp cDNA containing an open reading frame (ORF) of 2232 bp, belonging to the novel (n)PKC family of proteins characterized by their containing two phorbol ester/diacylglycerol-binding domains (C1 domain), a C2 domain, and a catalytic domain of the serine/threonine kinase, designated LvnPKC. A comparison of amino acid sequences showed that LvnPKC was closely related to arthropod nPKC. LvnPKC cDNA was detected in all tested tissues with a real-time PCR including the hepatopancreas, gills, muscles, subcuticular epithelium, abdominal nerve, thoracic nerve, brain, the stomach, heart, and especially in hemocytes and the intestines. Moreover, significantly upregulated LvnPKC expression was only observed in the eyestalk, brain, and hepatopancreas of shrimp transferred from 28 °C to 18 °C for 30 min. Induction of LvnPKC expression in hemocytes of L. vannamei injected with Vibrio alginolyticus at 10 cfu shrimp was detected earlier than in those injected with 10 cfu shrimp. Shrimp received LvnPKC-dsRNA for 1 days specifically depleted the expression of LvnPKC mRNA in hemocytes compared those of diethylpyrocarbonate water treatment. After that, significantly decreased expressions of lipopolysaccharide - and β-1,3-glucan-binding protein, prophenoloxidase-activating enzyme, peroxinectin, prophenoloxidase I, and prophenoloxidase II in the prophenoloxidase-activating system; lysozyme and cytosolic manganese superoxide dismutase and mitochondrial manganese superoxide dismutase in the antioxidant system were observed. We therefore concluded that LvnPKC is involved in immune defense of L. vannamei exposed to hypothermal stress or infected with V. alginolyticus.
编码凡纳滨对虾新型蛋白激酶 C（PKC）信使（m）RNA 的互补（c）DNA，由 2454 个碱基对的 cDNA 组成，包含 2232 个碱基对的开放阅读框（ORF），属于新型（n）PKC 蛋白家族，其特征是含有两个佛波酯/二酰基甘油结合域（C1 结构域）、一个 C2 结构域和丝氨酸/苏氨酸激酶的催化结构域，命名为 LvnPKC。氨基酸序列比较表明，LvnPKC 与节肢动物 nPKC 密切相关。实时 PCR 检测到 LvnPKC cDNA 存在于所有测试组织中，包括肝胰腺、鳃、肌肉、皮下上皮、腹部神经、胸部神经、脑、胃、心脏，特别是在血细胞和肠道中。此外，仅在从 28°C 转移到 18°C 30 分钟的对虾眼柄、脑和肝胰腺中观察到 LvnPKC 表达显著上调。在注射 10 个 cfus 对虾的 Vibrio alginolyticus 后，L. vannamei 血细胞中 LvnPKC 的诱导表达比注射 10 个 cfus 对虾时更早。用 LvnPKC-dsRNA 处理 1 天的对虾，血细胞中 LvnPKC mRNA 的表达明显低于二乙基焦碳酸盐水处理的对虾。之后，在酚氧化酶激活系统中，脂多糖和β-1,3-葡聚糖结合蛋白、原酚氧化酶激活酶、过氧化物酶、原酚氧化酶 I 和原酚氧化酶 II 的表达显著降低；在抗氧化系统中，溶菌酶和细胞质锰超氧化物歧化酶和线粒体锰超氧化物歧化酶的表达也观察到。因此，我们得出结论，LvnPKC 参与了凡纳滨对虾在低温胁迫或感染 V. alginolyticus 时的免疫防御。
