Functional study of kuruma shrimp immune-related genes

在这项研究中，我们利用 dsRNA 干扰阐明并表征了这些基因的体内功能。转谷氨酰胺酶（Tgase）和凝血蛋白（CP）是参与虾凝血级联的关键分子。我们利用双链RNA干扰（dsRNAi）在体内证明了转谷氨酰胺酶和凝血蛋白在虾凝血系统中的功能性参与及其对抗菌肽和酚氧化酶原系统相关基因的mRNA表达的影响。分别注射1μg和10μg Tgase-和CP-dsRNA后第1天，鳃、心脏、血细胞、肝胰腺、肠和淋巴器官中Tgase mRNA的表达受到抑制，而CP基因仅在鳃和心脏组织中受到抑制。然而，在注射后第 7 天，如 mRNA 表达、凝血和蛋白质数据所示，观察到基因和剂量的系统性基因沉默。白斑病毒和 Vibr … More io 对虾的攻击试验揭示了 Tgase 和 CP 在虾免疫系统中的关键功能。 Tgase 和 CP 沉默后，抗菌肽（crustine 和溶菌酶）和参与酚氧化酶原系统（酚氧化酶原和〓-2 巨球蛋白）的基因受到抑制，表明虾的血液凝固和体液生物防御之间存在关联。这一发现表明，螯合物和甲壳类动物中涉及酚氧化酶原系统的抗菌肽和基因的激活可能存在替代机制。总之，本研究1）提供了凝血和抗微生物肽以及参与酚氧化酶系统的基因之间的关联的证据，从而提出了激活这些分子的替代机制2）基因沉默效力结果将为dsRNA干扰在虾病和病原体的控制和管理中的应用提供预防措施。这项研究有助于更好地了解虾免疫系统的基本生物学。较少的

In this study, we elucidated and characterized the function of these genes in vivo using dsRNA interference. Transglutaminase (Tgase) and clotting protein (CP) are critical molecules involved in the blood coagulation cascade of shrimp. We demonstrate in vivo the functional involvement of transglutaminase and clotting protein in the shrimp blood coagulation system using double stranded RNA interference (dsRNAi) and their influence in the mRNA expression of antimicrobial peptides and genes involved in the prophenoloxidase system. Expression of Tgase mRNA was inhibited in gills, heart, hemocyte, hepatopancreas, intestine and lymphoid organ while the CP gene was suppressed only in gills and heart tissues on day-1 post injection with 1 μg and 10 μg of Tgase- and CP-dsRNA, respectively. However, at day-7 post injection, systemic gene silencing was observed for both genes and dosages as shown by mRNA expression, blood coagulation and protein data. Challenge test with white spot virus and Vibr … More io penaecida revealed the critical function of Tgase and CP in the immune system of shrimp. Suppression of antimicrobial peptides (crustine and lysozyme) and genes involved in the prophenoloxidase system (prophenoloxidase and 〓-2 macroglobulin) following Tgase and CP silencing demonstrate an association between blood coagulation and humoral biodefenses in shrimp. This finding suggests a possible alternative mechanism on the activation of antimicrobial peptides and genes involved in the prophenoloxidase system in chelicerates and crustaceans. In summary, this study 1)provided evidence on the association of blood coagulation and anti-microbial peptides and genes involved in the prophenoloxidase system and thus, presents an alternative mechanism on the activation of these molecules 2) gene silencing potency results will serve as a precaution on the application of dsRNA interference in the control and management of shrimp diseases and pathogens. This study contributed to the better understanding of the basic biology of the shrimp immune system. Less