凡纳滨对虾(Litopenaeus vannamei)造血激素的组织分布及细胞定位分析  

THE TISSUE DISTRIBUTION AND CELLULAR LOCALIZATION OF ASTAKINE IN SHRIMP LITOPENAEUS VANNAMEI

作　　者：徐萌霖 梁艳[2] 卢金凤 王晓雯[1,2] 李晨 邢婧[1] 黄倢[2] 

机构地区：[1]中国海洋大学海水养殖教育部重点实验室,青岛266003 [2]农业部海洋渔业资源可持续利用重点开放实验室中国水产科学院研究院黄海水产研究所,青岛266071

出　　处：《海洋与湖沼》2014年第1期190-196,共7页Oceanologia Et Limnologia Sinica

基　　金：国家重点基础研究发展计划(973计划)项目课题;2012CB114400号;国家自然科学基金课题;31101934号;基本科研业务费专项经费;20603022013009号;中央级科研院所基本科研业务费资助;2010-cb-06号

摘　　要：造血激素(astakine)是一种具有促进造血组织细胞分化和增殖作用的甲壳动物细胞因子,参与甲壳动物的造血活动,对于只依赖于非特异性免疫的甲壳动物抗感染能力有重要的意义。本实验室前期成功克隆凡纳滨对虾造血激素(LvAST)的基因Lvast。为了进一步了解其功能,本文通过实时荧光定量PCR技术检测了Lvast表达的组织分布,同时还用Dlight标记抗LvAST抗体进行了LvAST在对虾细胞和组织中的免疫荧光定位分析。荧光定量PCR测定结果说明,Lvast主要在肝胰腺、鳃和肌肉中大量表达,而淋巴器官和血淋巴细胞中表达量较低;免疫荧光结果显示,LvAST可同时存在于血淋巴细胞内和血淋巴细胞膜表面,不同的血淋巴细胞表面的LvAST分布呈现出明显差异,可能具有重要的血细胞分类意义。LvAST蛋白在肝胰腺、鳃、肌肉、淋巴器官等组织中均有分布,并且肝胰腺组织中分布较多,淋巴器官组织中分布较少。Astakine, a cytokine in crustaceans, involves in hematopoiesis for stimulating cell proliferation and differentiation, and plays a key role in crustacean immune responses. In our previous studies, the gene of astakine in shrimp Litopenaeus vannamei(LvAST) was cloned and expressed in vitro. To get more information about the function of LvAST, its tissue distribution was quantified by real time RCR; and additionally, its cellular localization was observed by immunofluorescence method. Results reveald that the Lvast gene distributes widely in different tissues, with relative higher expression in hepatopancreas, muscle, and gills. The cellular location recognized by Dylight 649 labeled polyclonal antibody of LvAST showed that LvAST is expressed not only in hemocytes but also on some hemocytes. In histology immunofluorescence method, we observed that the LvAST protein distributed in hepatopancreas, gills, muscle, and lymphoid organ, which is in accordance with the result of its gene expression in shrimp tissues.

关 键 词：凡纳滨对虾 造血激素 荧光定量PCR 激光共聚焦显微镜 荧光定位 

分 类 号：S917.4[农业科学—水产科学]