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克氏原螯虾(Procambarus clarkii) CypA基因应答WSSV侵染的表达研究
朱俊杰1,2, 林锋3, 王岩1, 张海琪3
1.浙江大学 杭州 310058;2.湖州师范学院 湖州 313000;3.浙江省淡水水产研究所 湖州 313001
摘要:
克氏原螯虾亲环素蛋白A(Procambarus clarkii Cyclopllilin A,简称PcCypA)是一种存在于克氏原螯虾(P. clarkii)中高度保守的重要蛋白,具有多种生物学功能。为了初步阐明CypA在病毒侵染克氏原螯虾过程中的免疫应答机制,本研究利用qRT-PCR等技术分析了克氏原螯虾在WSSV病毒感染后,PcCypA在不同组织中的转录表达情况。研究结果表明克氏原螯虾在感染WSSV后,心脏和肌肉组织中都出现了肌纤维断裂溶解的情况,在肝胰腺组织中出现了脂肪颗粒脱落、细胞空泡化和细胞膜溶解现象。同时,不同组织中的PcCypA基因表达存在明显的时序差异,以24hpi为界,0—24hpi为注射应激期,在此期间,肝胰腺、心脏等组织中的PcCypA转录表达量均有上调;24—36hpi为应激恢复期,绝大多数的组织中PcCypA转录表达均出现了回落,但在心脏、肌肉、血淋巴组织中出现了上调,说明这三个组织对WSSV感染的响应可能早于其他组织;48hpi之后为病毒感染响应阶段,在此期间鳃和肌肉组织中PcCypA表达量显著提升,而在肝胰腺和精巢组织中却出现表达下调;96—120hpi为感染响应后期阶段,肝胰腺和肠组织的PcCypA转录水平再次上调。因而,可以初步推断PcCypA在病毒WSSV侵染克氏原螯虾过程中参与了机体的免疫应答,且在不同的组织中PcCypA的转录表达水平存在显著差异。
关键词:  克氏原螯虾  CypA  qRT-PCR  WSSV
DOI:10.11693/hyhz20180400091
分类号:Q789
基金项目:浙江省重大科技攻关项目,2014C02011号。
EXPRESSION OF CypA AFTER WSSV CHALLENGE IN RED SWAMP CRAYFISH, PROCAMBARUS CLARKII
ZHU Jun-Jie1,2, LIN Feng3, WANG Yan1, ZHANG Hai-Qi3
1.College of Animal Sciences, Zhejiang University, Hangzhou 310058, China;2.College of Life Sciences, Huzhou University, Huzhou 313000, China;3.Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China
Abstract:
Cyclophilins A is a family of highly conserved and multi-functional proteins that exist extensively from microorganisms to mammals. In order to explore the immune function of CypA in Procambarus clarkii after WSSV challenge, real-time quantitative PCR was used to detect the expression characteristics after WSSV challenge. Histologic observation under a light microscope showed that the heart, the hepatopancreas and the striate muscle of the abdomen were severely affected. PcCypA transcriptional levels were significantly up-regulated in the heart and the hepatopancreas tissues at 0-24hpi. After 48hpi, CypA transcripts in the gill and the muscle were significantly increased, while transcripts in the hepatopancreas and the testicle decreased. During 96-120hpi, CypA transcripts in the hepatopancreas and the intestine were significantly increased again. The results suggested that CypA might be involved in the immune response of P. clarkii.
Key words:  Procambarus clarkii  CypA  qRT-PCR  WSSV
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