中国明对虾在胁迫条件下肝胰脏的差异蛋白质组学研究

IOCAS-IR > 海洋环流与波动重点实验室

	中国明对虾在胁迫条件下肝胰脏的差异蛋白质组学研究
其他题名	Copmarative proteomic profiles of the hepatopancreas in Fenneropenaeus chinensis response to stresses
	蒋昊
学位类型	博士
	2009-05-25
学位授予单位	中国科学院海洋研究所
学位授予地点	海洋研究所
关键词	中国明对虾蛋白质组学双向电泳免疫缺氧胁迫肝胰脏参考图谱
摘要	对虾养殖业的发展面临着病害严重和环境恶化的严重挑战。对虾细菌和病毒性疾病的频发已给对虾的养殖业造成了严重损失。养殖环境恶化造成的环境胁迫，不但影响对虾的生长性状，而且导致对虾的抵抗力下降，更容易引发病害的发生。研究对虾在外界胁迫（环境胁迫、病原感染）后蛋白表达模式的变化有助于弄清对虾的抗病抗逆机理，为抗病抗逆对虾的培育提供理论依据。肝胰脏是对虾的重要器官，参与各种重要的生理活动包括消化、吸收、储存和代谢，并且在启动对虾免疫反应起重要作用，其蛋白质表达的变化与对虾的健康状况密切相关。本研究运用蛋白质组学技术平台（双向电泳、串联质谱、数据库搜索），对中国明对虾肝胰脏蛋白进行研究，获得了一些重要进展，结果如下： 1.初步构建了覆盖范围广、分辨率高、重复性好的中国明对虾肝胰脏双向电泳参考图谱，并利用质谱分析和多种数据库搜索方法成功鉴定了122个（68种）蛋白质点，包括14个（9种）能量代谢蛋白、15个（9种）氨基酸代谢蛋白、10个（5种）碳水化合物代谢蛋白、2个（1种）多聚糖代谢蛋白、11个（5种）辅酶和维生素代谢蛋白、1个（1种）脂类代谢蛋白、5个（3种）核酸代谢蛋白、7个（3种）消化酶、3个（1种）细胞骨架蛋白、14个（8种）免疫相关蛋白、9个（6种）抗氧化蛋白、19个（7种）伴侣蛋白、4个（3种）信号转导蛋白、3个（3种）翻译功能蛋白和5个（4种）未分组蛋白。 2.在参考图谱的基础上，研究了缺氧胁迫下中国明对虾肝胰脏蛋白质表达模式的变化，成功鉴定了52个（15个上调、37个下调）差异表达的蛋白，包括11个能量产生相关蛋白、13个免疫相关蛋白、11个代谢相关蛋白、6个抗氧化蛋白、7个分子伴侣、2个细胞骨架蛋白和2个未分类蛋白，利用实时定量PCR技术，研究了缺氧胁迫下10个差异蛋白的基因在转录水平上的变化；同时研究了金属镉胁迫后中国明对虾肝胰脏蛋白质表达模式的变化，成功鉴定了29个（4个上调、25个下调）差异表达的蛋白，包括9个能量产生相关蛋白、3个免疫相关蛋白、9个代谢相关蛋白、5个抗氧化蛋白、2个分子伴侣和1个未分类蛋白。 3.分别进行了灭活鳗弧菌和WSSV感染后中国明对虾肝胰脏的差异蛋白质组学研究。灭活鳗弧菌刺激后，45个差异表达的蛋白质（4个上调、41个下调）被成功鉴定出，包括6个能量产生相关蛋白、4个免疫相关蛋白、22个代谢相关蛋白、4个抗氧化蛋白、7个分子伴侣、1个翻译相关蛋白和2个未分类蛋白；WSSV感染后， 48个差异表达的蛋白质（11个上调、37个下调）被成功鉴定出，包括7个能量产生相关蛋白、9个免疫相关蛋白、20个代谢相关蛋白、5个抗氧化蛋白和7个分子伴侣。通过对上述实验数据的分析，初步探讨了中国明对虾对虾对环境胁迫和病原感染的应答机理。为下一步筛选中国明对虾环境胁迫及免疫应答关键蛋白并研究其功能奠定了基础。
其他摘要	Shrimp diseases and the deterioration of environment are two serious challenges faced by shrimp farming in the world. The frequent shrimp diseases caused by bacteria and virus have led to serious economics loss of shrimp production. The environmental stress caused by the deterioration of aquiculture environment not only stunted normal growth of shrimp, but also decreased the immune capability leading to an increased susceptibility to infectious disease of shrimp. The studies on changes of proteins expression patterns in shrimp challenged by stresses and pathogen infection are very important to know more about the mechanisms of anti-stress and anti-disease in shrimp, and support information to anti-stress and anti-disease shrimp culture. Hepatopancreas, a main organ of decapods for digestion, absorption, storage and metabolism, play a major role in initiating the defense response in shrimp. The variations of protein expression profiles in hepatopancreas have a close relationship with the health situation of shrimp. In this study, proteomics approach (2-DE, tandem MS and database search) were used to research protein expression profiles of the hepatopancreas in Fenneropenaeus chinensis. Some important progresses are as followes: 1.A proteomic reference map with wide pH/MW range, high resolution and good reproduction for hepatopancreas of healthy adult Chinese shrimp was obtained using 2-DE. By tandem MS and multifarious database search methods, 122 spots (68 proteins) were successfully identified. Among these protein spots, 14 spots (9 proteins) participate in energy metabolism, 15 spots (9 proteins) participate in amino acid metabolism, 10 spots (5 proteins) participate in carbohydrate metabolism, 2 spots (1 proteins) participate in glycan metabolism, 11 spots (5 proteins) participate in metabolism of cofactors and vitamins, 1 spots (1 proteins) participate in lipid metabolism, 5 spots (3 proteins) participate in nucleotide metabolism, 7 spots (3 proteins) participate in digestive enzymes, 3 spots (1 proteins) participate in cytoskeleton proteins, 14 spots (8 proteins) participate in immune, 9 spots (6 proteins) are antioxidant proteins, 19 spots (7 proteins) are chaperone, 4 spots (3 proteins) participate in signal transduction, 3 spots (3 proteins) participate in translation and 5 spots (4 proteins) are ungrouped proteins. 2.Based on the reference map, the mechanisms of envionmental stress response in Chinese shrimp were investigated using proteomic approach. In hepatopancreas of Chinese shrimp, after hypoxic stress, 52 differentially expressed protein spots (15 up-regulated, 37 down-regulated) were successfully identified, including 11 protein spots related to energy production, 13 protein spots related to immune, 11 protein spots involved in the metabolism, 6 antioxidant protein spots, 7 chaperone spots, 2 cytoskeleton protein spots and 2 ungrouped protein spots; after cadmium stress, 29 differentially expressed protein spots (4 up-regulated, 25 down-regulated) were successfully identified, including 9 protein spots related to energy production, 3 protein spots related to immune, 9 protein spots involved in the metabolism, 5 antioxidant protein spots, 2 chaperone spots and 1 ungrouped protein spots. The transcription levels of 10 selected genes which encode the differentially expressed proteins after hypoxia were also analyzed by real-time PCR. 3.The mechanisms of immune response in Chinese shrimp were investigated using proteomic approach. In hepatopancreas of Chinese shrimp challenged with killed Vibrio anguillarum, 45 differentially expressed protein spots (4 up-regulated, 41 down-regulated) were successfully identified, including 6 protein spots related to energy production, 4 protein spots related to immune, 22 protein spots involved in the metabolism, 4 antioxidant protein spots, 7 chaperone spots, 1 translation related protein spot and 2 ungrouped protein spots; in hepatopancreas of Chinese shrimp challenged with white spot syndrome virus, 48 differentially expressed protein spots (11 up-regulated, 37 down-regulated) were successfully identified, including 7 protein spots related to energy production, 9 protein spots related to immune, 20 protein spots involved in the metabolism, 5 antioxidant protein spots and 7 chaperone spots. Based on these results, we discussed the mechanism of stresses response and immune response of shrimp and shed light on the relationship between environmental stresses and shrimp’s immune capability. This study set a basis for further analyses of some key genes in stresses and immune response in shrimp.
页数	133
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/869
专题	海洋环流与波动重点实验室
推荐引用方式 GB/T 7714	蒋昊. 中国明对虾在胁迫条件下肝胰脏的差异蛋白质组学研究[D]. 海洋研究所. 中国科学院海洋研究所,2009.

条目包含的文件
文件名称/大小	文献类型	版本类型	开放类型	使用许可
10001_20071800681201（6848KB）			限制开放	--	浏览