Essential function of transglutaminase and clotting protein in shrimp immunity
Essential function of transglutaminase and clotting protein in shrimp immunity
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DOI：
10.1016/j.molimm.2007.09.016
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发表时间：
2008-03-01
影响因子：
3.6
通讯作者：
Aoki, Takashi
中科院分区：
文献类型：
--
作者：
Maningas, Mary Beth Bacano;Kondo, Hidehiro;Aoki, Takashi
凝血是无脊椎动物的一种保守的防御机制，在马蹄蟹和淡水小龙虾中已经得到了很好的研究，但在虾中的定义不够明确。转谷氨酰胺酶(TGase)和凝血蛋白(CP)是参与对虾凝血系统的分子。在这里，我们利用RNA干扰在体内证明了TGase和CP在对虾凝血系统中的功能参与。注射1mgTGase-dsRNA和10mgCP-dsRNA后1d，TGase基因在鳃、心脏、血细胞、肝胰腺、肠和淋巴器官中的表达受到抑制，而CP基因仅在鳃和心脏组织中表达受到抑制。然而，在注射后第7天，基因和剂量都观察到系统性的基因沉默，从mRNA的表达来看。我们还展示了dsRNA沉默蛋白表达以及抑制凝血的有效性。这种在转录、翻译和表型水平上的沉默在虾系统中是第一次被记录下来。对虾白斑病毒和对虾弧菌的攻击试验表明，TGase和CP是对虾抵抗细菌和病毒感染的免疫功能的关键分子。(C)2007爱思唯尔有限公司。保留所有权利。
Blood coagulation is a conserved defense mechanism among invertebrates and it has been well studied in horseshoe crab and freshwater crayfish but is ill defined in shrimp. Transglutaminase (TGase) and clotting protein (CP) are molecules involved in the blood clotting system of shrimp. Here, we demonstrate in vivo the functional involvement of TGase and CP in the shrimp blood coagulation system using RNA interference. 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 mu g and 10 mu g 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. We also show the efficiency of dsRNA silencing the protein expression as well as inhibited blood coagulation. Such silencing at the transcription, translation and phenotypic level is the first to be documented in the shrimp system. Challenge test with white spot virus and Vibrio penaecida revealed that TGase and CP are critical molecules for the immune function of shrimp against bacterial and viral infection. (c) 2007 Elsevier Ltd. All rights reserved.