mRNA Expression of Heat Shock protein 70 gene in the oyster(Crassostrea hongkongensis) responding to Vibiro alginolyticus challenge
CHEN Chen, ZHANG Qi-zhong, ZHANG Zhan-hui, CUI Miao
Institute of Hydrobiology, Jinan University, Engineering Research Center of Tropical and Subtropical Aquatic Ecological Engineering Ministry of Education, Key Laboratory of Aquatic Eutrophication and Control of Harmful Algal Blooms of Guangdong Higher Education Institutes, Guangzhou 510632, China
In order to investigate the role of HSP70 of oyster(Crassostrea hongkongensis) against bacterial infection,we established a real-time,quantitative fluorescent RT-PCR method to examine the expression patterns of HSP70 mRNA in mantle,digestive gland,gills,muscle,heart and haemocytes of the oyster responding to Vibiro alginolyticus challenge.The results showed that after the oysters were injected with live V.alginolyticus,HSP70 mRNA expression in gills,mantle and haemocytes was up-regulated at first,peaked at 3 hr,6 hr and 24 hr,respectively,then gradually decreased to the control level at 12 hr,24 hr sand 72 hr,respectively.The phenomenon was defined as time-dependent expression pattern of HSP70 gene.Meanwhile,the HSP70 mRNA levels in heart and digestive gland significantly increased and reached the highest values at 3 hr and 6 hr,respectively,and then dropped rapidly.But in muscle,the HSP70 mRNA expression did not increased upon the bacterial challenge during the entire experiment period.The up-regulated mRNA expression of HSP70 induced by bacterial challenge suggested an important role for HSP70 gene in immune response in oysters.
[1] Lindquist S.The heat-shock response[J].Annu Rev Genet,1986,22:631-677.
[2] Hamer B,Hamer D P,MUller W E,Batel R.Stress-70 proteins in marine mussel Mytilus galloprovincialis as biomarkers of environmental pollution:a field study[J].Envirvn Int,2004,30(7):873-882.
[3] Kiang J G,Tsokos G C.Heat shock protein 70 kDa:Molecular Biology,Biochemistry,and Physiology[J].Pharmacol Ther,1998,80(2):183-201.
[4] Steinert S A,Pickwell G V.Expression of heat shock proteins and metallothionein in mussels exposed to heat stress and metal ion challenge[J].Mar Environ Res,1988,24:211-214.
[5] 吴信忠.海洋养殖贝类病害的流行状况及其控制对策.南海资源开发研究[M].广州:广东经济出版社,1998,670-679.
[6] 吴信忠.中国海洋病害学主流研究的进展[J].太平洋学报,2005,10:49-59.
[7] 吴信忠,潘金培,江静波.贝类病害学研究进展Ⅰ.贝类微生物病学研究[J].海洋通报,1995,14(2):82-91.
[8] 李春勇.敌百虫诱导近江牡蛎(Crassostrea hongkongensis)HSC70基因表达的定量研究[D].广州:暨南大学,2007.
[9] Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△Ct method[J].Methods,2001,25(4):402-408.
[10] Sanders B M,Martin L S,Howe S R,Nelson W G,Hegre E S,Phelps D K.Tissue-specific differences in accumulation of stress proteins in Mytilus edulis exposed to a range of copper concentrations[J].Toxicol Appl Pharmacol,1994,125(2):206-213.
[11] Schlesinger M J.Heat shock proteins[J].J Biol Chan,1990,265:12111-12114.
[12] Moraga D,Meistertzheim A L,Tanguy-Royer S,Boutet I,Tanguy A,Donval A.Stress response in Cu2+ and Cd2+ exposed oysters(Crassostrea gigas):an immunehistochemical approach[J].Comp Biochem Physiol C Toxicol Pharmacol,2005,141(2):151-156.
[13] Ali K S,Dorgai L,Abraham M,Hermesz E.Tissue-and stressor-specific expression of two hsc70 genes in carp[J].Biochem Biophys Res Commun,2003,307(3):503-509.
[14] Cheng P.Liu X,Zhang G,He J.Cloning and expression analysis of a HSP70 gene from Pacific abalone(Haliotis discus hannai)[J].Fish Shellfish Immunol,2007,22:77-87.
[15] Young R A.Stress proteins and immunology[J].Annu Rev Immunol,1990,8:401-420.
[16] Forsyth R B,Candido E P M,Babich S L,Iwama G K.Stress protein expression in coho salmon with bacterial kidney disease[J].J Aquatic Animal Health,1997,9:18-25.
[17] Ackerman P A,Iwama G K.Physiological and cellular stress responses of juvenile rainbow trout to vibrio alginolyticussis[J].J Aquat Anim Health,2001,13:173-180.
[18] 周永灿,潘金培.贝类细胞和体液的防御机制研究进展[J].水产学报,1997,21(4):449-454.
[19] Song L,Wu L,Ni D,Chang Y,Xu W,Xing K.The cDNA cloning and mRNA expression of heat shock protein 70 gene in the haemocytes of bay scallop Argopecten irradians,responding to bacteria challenge and naphthalin stress[J].Fish Shellfish Immunol,2006,21:335-345.
[20] Wang Z,Wu Z,Jian J,Lu Y.Cloning and expression of heat shock protein 70 gene in the haemocytes of pearl oyster(Pinctada fucata,Gould 1850)responding to bacterial challenge[J].Fish Shellfish Immunol,2009,26:639-645.
[21] Basu N,Todgham A E,Ackerman P A,Bibeau M R,Nakano K,Schulte P M,Iwama G K.Heat shock protein genes and their functional significance in fish[J].Gene,2002,295:173-183.
[22] Sharp G J E,Secombes C J.The role of reactive oxygen species in the killing of the bacterial fish pathogen Aeromonas salmonicida by rainbow trout macrophages[J].Fish Shellfish Immunol,1993,3:119-129.
[23] Rylander M N,Feng Y S,Bass J,Dilier K.Thermally induced injury and heat-shock protein expression in cells and tissues[J].Ann N.Y Acad Sci,2005,1066:222-242.
[24] Binder R J,Vatner R,Srivastava P.The heat-shock protein receptors:some answers and more questions[J].Tissue Antigens,2004,64:442-451.
[25] Zhang J,Wang X,Guo H,Wu J,Xue Y.Effects of water soluble fractions of diesel oil on the antioxidant defenses of the goldfish[J].Carassius auratus.Ecotoxicol Environ Sqf,2004,58:110-116.
