| 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的蛋白在肝胰腺、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。 |
| 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 |
| DOI：10.11693/hyhz20121206001 |
| 分类号: |
| 基金项目:国家重点基础研究发展计划（973计划）,国家自然科学基金项目（面上项目，重点项目，重大项目），中央级科研院所基本科研业务费 |
附件 |
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| The tissue distribution and cellular localization of astakine in shrimp Litopenaeus vannamei |
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xumenglin
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Fisheries college，Ocean University of China
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| Abstract: |
| Astakine, a kind of cytokine in crustaceans, involves in hematopoiesis and can stimulate the proliferation, differentiation, and survival of hematopoietic tissue cells in freshwater crayfish, plays a key role in crustacean immune responses. In our previous studies, the gene of Litopenaeus vannamei astakine (LvAST) was cloned and expressed in vitro. The recombined LvAST was demonstrated to be able to inhibit the infection of white spot syndrome virus (WSSV). In order to get more information about the function of LvAST, this paper reports its tissue distribution that quantified by real time RCR in five different tissues of shrimp L. vannamei, as well its cellular localization which observed by immunofluorescence location method. The results revealed that expression of the gene Lvast had a wide tissue distribution, but its expression was enriched in hepatopancreas, muscle and gills. The cellular location results by immunofluorescence with LvAST polyclonal antibody showed that LvAST was expressed not only in almost all of the hemocytes but also on some of the cell surface.The distribution of LvAST protein was also observed in hepatopancreas, gills, muscle, and lymphoid organ by the immunofluorescence. And the major distribution was measured in hepatopancreas, while the minor was in lymphoid organ. |
| Key words: Litopenaeus vannamei, Astakine, Immunofluorescence, Location, laser-scanning confocal microscope |
