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. 2017 Sep;68:92-101. doi: 10.1016/j.fsi.2017.07.028. Epub 2017 Jul 11.
In shrimp, the biosynthesis of catecholamines, including dopamine and norepinephrine, is required for physiological and immunological responses against stress. Dopamine beta-hydroxylase (DBH), a copper-containing monooxygenase enzyme that plays an important role in catecholamine synthesis of the neuroendocrine regulatory network, was identified in Litopenaeus vannamei. In the present study, the potential role of DBH in the immunocompetence of L. vannamei was further estimated by depleting DBH by pharmaceutical inhibition of disulfiram and a gene silencing technique of L. vannamei DBH-double-stranded (ds)RNA (LvDBH-dsRNA). Immunocompetence was evaluated following the determination of the total hemocyte count, differential hemocyte count, phenoloxidase activity, respiratory bursts, superoxide dismutase activity, phagocytic activity, and the clearance efficiency as well as the susceptibility against Vibrio alginolyticus infection. At 30-120 min after shrimp had received disulfiram, they exhibited significantly reduced total hemocyte count, phenoloxidase activity of hemocytes in hemolymph, respiratory bursts of hemocytes in hemolymph and per hemocyte, phagocytic activity, clearance efficiency, and survival ratio against V. alginolyticus infection, compared to those injected with saline. In addition, the significantly lower total hemocyte count, phagocytic activity, clearance efficiency, and resistance to V. alginolyticus infection were observed in shrimp that received LvDBH-dsRNA at 3 days post injection compared to those injected with diethyl pyrocarbonate-water or non-targeting gene-dsRNA. The DBH depleted L. vannamei revealed immunosuppression and decreased the survival ratio to V. alginolyticus infection, which indicated that DBH played a crucial role in the neuroendocrine-immune regulatory network.
在虾类中,儿茶酚胺(包括多巴胺和去甲肾上腺素)的生物合成对于应对压力的生理和免疫反应至关重要。多巴胺 β-羟化酶(DBH)是一种含铜的单加氧酶,在神经内分泌调节网络中的儿茶酚胺合成中发挥重要作用,已在凡纳滨对虾(Litopenaeus vannamei)中被鉴定。在本研究中,通过使用二硫代氨基甲酸酯(disulfiram)抑制 DBH 的药物抑制和凡纳滨对虾 DBH 双链 RNA(LvDBH-dsRNA)的基因沉默技术,进一步评估了 DBH 在凡纳滨对虾免疫中的潜在作用。通过测定总血细胞计数、差异血细胞计数、酚氧化酶活性、呼吸爆发、超氧化物歧化酶活性、吞噬活性、清除效率以及对溶藻弧菌感染的敏感性来评估免疫能力。在虾类接受二硫代氨基甲酸酯后 30-120 分钟,与注射生理盐水的虾类相比,它们的总血细胞计数、血淋巴中血细胞的酚氧化酶活性、血淋巴和每个血细胞的呼吸爆发、吞噬活性、清除效率和对溶藻弧菌感染的存活率显著降低。此外,与注射二乙基焦碳酸盐-水或非靶向基因 dsRNA 的虾类相比,在注射 LvDBH-dsRNA 后 3 天的虾类中,总血细胞计数、吞噬活性、清除效率和对溶藻弧菌感染的抵抗力显著降低。DBH 耗尽的凡纳滨对虾表现出免疫抑制作用,降低了对溶藻弧菌感染的存活率,这表明 DBH 在神经内分泌-免疫调节网络中发挥着关键作用。