Comparative proteomic identification of the hemocyte response to cold stress in white shrimp, Litopenaeus vannamei
Comparative proteomic identification of the hemocyte response to cold stress in white shrimp, Litopenaeus vannamei
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DOI：
10.1016/j.jprot.2012.12.017
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发表时间：
2013-03-27
影响因子：
3.3
通讯作者：
Wu, Yingxia
中科院分区：
文献类型：
--
作者：
Fan, Lanfen;Wang, Anli;Wu, Yingxia
为了解甲壳动物血细胞对冷胁迫的分子响应，采用流式细胞仪和双向电泳技术研究了低温胁迫下凡纳滨对虾血细胞中蛋白质的变化。通过流式细胞仪分析，选择13 ℃冷胁迫后24 h作为冷胁迫处理的适宜温度和条件。MALDI-TOF/TOF MS分析显示，6种蛋白质的6种形式均显著上调，其中包括3种酶（胱硫醚酶、甘油醛3-磷酸脱氢酶和谷胱甘肽酶1）和一种免疫相关蛋白（癌蛋白nm 23），而3种蛋白（血蓝蛋白、血细胞转氨酶和转酮醇酶）中的24种形式在处理的虾中显著下调。结果表明，在对虾血蓝蛋白中有20个斑点被鉴定为血蓝蛋白，可能在对虾的环境调节中发挥重要作用。实时荧光定量PCR检测结果表明，冷应激后血蓝蛋白、部分血蓝蛋白mRNA、胱硫醚酶、谷胱甘肽酶和癌蛋白nm 23基因的转录水平与其翻译产物的转录水平相关，只有血细胞转氨酶的转录水平与其翻译产物的转录水平不相关。这些数据的进一步调查可能会导致更好地了解甲壳动物血细胞的分子反应冷应激。(C)2013爱思唯尔有限公司版权所有。
To understand molecular responses of crustacean hemocytes to cold stress, flow cytometer analysis and two-dimensional electrophoresis proteomic approach were used to investigate altered proteins in hemocytes of Litopenaeus vannamei during cold stress treatment. Through flow cytometer analysis, 13 degrees C for 24 h post-cold stress was selected as the suitable temperature and condition for cold stress treatment MALDI-TOF/TOF MS analysis revealed that 6 forms of 6 proteins were significantly up-regulated, including three enzymes (cystathionase, glyceraldehyde 3-phosphate dehydrogenase and glyoxalase 1) and one immune-related protein (oncoprotein nm23), whereas 24 forms of 3 proteins were significantly down-regulated in the treated shrimp (hemocyanin, hemocyte transglutaminase and transketolase). There were 20 spots identified as hemocyanin meaning that it may play important roles in environmental regulation in shrimp. Real-time fluorescence quantitative PCR confirmed that the levels of transcription of the hemocyanin, partial mRNA for hemocyanin, cystathionase, glyoxalase land oncoprotein nm23 genes were found to relate well with that of their translation products after cold stress treated, while only the levels of hemocyte transglutaminase transcripts were not corresponded with that of their translation products. Further investigation of these data may lead to better understanding of the molecular responses of crustacean hemocytes to cold stress. (C) 2013 Elsevier B.V. All rights reserved.