草鱼hsp70和hsp90对温度急性变化的响应
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中国海洋大学水产学院海水养殖教育部重点实验室,厦门大学 海洋与地球科学学院,中国海洋大学水产学院海水养殖教育部重点实验室,中国海洋大学水产学院海水养殖教育部重点实验室

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国家“九七三”重点基础研究发展计划(2009CB118706)


Effect of rapid temperature change on expression of hsp70 andhsp90 in grass carp(Ctenopharyngodon idella)
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The Key Laboratory of Mariculture, Ministry of Education, Fisheries College, Ocean University of China, Qingdao,College of Oceanography and Earth Science, Xiamen University,The Key Laboratory of Mariculture, Ministry of Education, Fisheries College, Ocean University of China, Qingdao,The Key Laboratory of Mariculture, Ministry of Education, Fisheries College, Ocean University of China, Qingdao

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    摘要:

    为研究不同温度条件下草鱼热休克蛋白(hsp70和hsp90)的表达模式,了解草鱼对温度的耐受和适应机理,实验将在20 ℃下驯化的草鱼在7个实验温度(22,24,26,28,30,32和34 ℃)中热激3 h,然后20 ℃恢复2 h,取肝脏,肌肉和鳃测定hsp70和hsp90表达。结果表明,hsp70和hsp90表达量随着温度升高而上升,当温度达到34 ℃时,肌肉与鳃热休克蛋白基因表达量显著下降,草鱼耗氧率、热休克蛋白表达和死亡率之间的关系符合氧限制热耐受理论(OCLTT)。基于OCLTT理论,草鱼生理临界温度为28 ℃,当温度超过28 ℃,热休克蛋白表达所需能量主要由无氧代谢提供,进而导致体内氧自由基和变性蛋白的增加,影响草鱼的生长和存活。

    Abstract:

    Grass carp(Ctenopharyngodon idella)is one of the most important cultured fishes in China.During transportation in summer,massive mortality often occurs.As molecular chaperones,Hsps assist cells in their recovery from stress and promote cytoprotection.Limited reports described the expression of Hsps in grass carp at high temperatures,and the relationship between oxygen consumption,expression of heat shock protein and mortality needs to be investigated based on the oxygen- and capability- limited thermal tolerance theory(OCLTT)to further elucidate the thermal adaptation of this species to high temperature in view of integrated biology.In the present study,the expressions of hsp70 and hsp90 after rapid temperature increase were studied.Grass carps were acclimated at 20 ℃ and then exposed to designated temperatures(20,22,24,26,28,30,32,34 ℃)for 3 h heat shock.After 2 h- recovery at 20 ℃,liver,muscle and gill of tested fish were sampled and hsp70 and hsp90 were determined using semi-quantitive real time PCR.Expression of hsps increased with temperature increase until 32 ℃ in muscle and gill,and then decreased at 34 ℃.In liver,expression of hsps kept increasing in all temperature treatments.These results indicated that hsp70 and hsp90 were sensitive to temperature increase in grass carp.Analyzed heat shock proteins data with previous results of oxygen consumption and lethal temperatures of grass carp,the physiological adaptations of grass carp could be explained by oxygen and capability- limited thermal tolerance theory(OCLTT),which pointed out that the imbalance between oxygen demand and tissue oxygen supply ability was the primary factor which limited organisms’ thermal tolerance and the ability of aerobic metabolism greatly limited the survival of organism beyond critical temperatures.With temperature increase,the maximum value of oxygen consumption in grass carp occurred at 28 ℃,and then the oxygen consumption decreased when temperature was beyond 28 ℃,indicating a transformation from aerobic metabolism to anaerobic metabolism at this critical temperature.The expressions of heat shock proteins kept increasing until 32 ℃ in muscle and gill in grass carp,and provided defense against high temperature.However,this protection of heat shock proteins was limited and then death occurred when temperature was beyond 32 ℃.The change of metabolism,increase of oxygen free radicals and the consumption of energy due to stress proteins synthesis of grass carp were closely related to the decrease of capability of immune response when water temperature was beyond 28 ℃.In general,high water temperature could alter the metabolic pathway,induce heat shock protein expression,reduce immune response,and then cause large scale mortality.Therefore,appropriate ways should be applied to maintain water temperature under 28 ℃ to avoid large-scale mortality during transportation.

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周鑫,董云伟,王芳,董双林.草鱼hsp70和hsp90对温度急性变化的响应[J].水产学报,2013,37(2):216~221

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  • 收稿日期:2012-02-05
  • 最后修改日期:2012-12-01
  • 录用日期:2013-01-17
  • 在线发布日期: 2013-02-22
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