长期高氨氮暴露对海水青鳉毒理效应的蛋白质组学研究

doi:10.11978/2016126

热带海洋学报 ›› 2017, Vol. 36 ›› Issue (5): 40-48.doi: 10.11978/2016126CSTR: 32234.14.2016126

长期高氨氮暴露对海水青鳉毒理效应的蛋白质组学研究

高娜^1,^2,³(), 朱丽梅^1,², 于德良^3,⁴, 张黎^1,²()

1. 中国科学院南海海洋研究所, 中国科学院热带海洋生物资源与生态重点实验室, 广东广州 510301
2. 中国科学院南海海洋研究所, 广东省应用海洋生物学重点实验室, 广东广州 510301
3. 中国科学院大学, 北京 100049
4. 中国科学院烟台海岸带研究所, 中国科学院海岸带环境过程与生态修复重点实验室, 山东省海岸带环境过程重点实验室, 山东烟台 264003

收稿日期:2016-12-02 修回日期:2017-04-21 出版日期:2017-09-20 发布日期:2017-09-22
作者简介:
作者简介：高娜(1988—), 女, 山东省新泰市人, 博士研究生, 主要从事海洋生物学研究。E-mail: gaona12@mails.ucas.ac.cn
基金资助:
国家重点基础研究发展规划项目(2015CB452904);国家自然科学基金项目(41376161、31501862);中国科学院百人计划项目;广东省科技计划项目(2014B030301064)

Proteomic responses of marine medaka (Oryzias melastigma) to chronic high environment ammonia exposure

Na GAO^1,^2,³(), Limei ZHU^1,², Deliang YU^3,⁴, Li ZHANG^1,²()

1. Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China
2. Guangdong Provincial Key Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China
3. University of Chinese Academy Sciences, Beijing 100049, China
4. Key Laboratory of Coastal Zone Environmental Processes and Ecological Restoration, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Shandong Provincial Key Laboratory of Coastal Zone Environmental Processes, Yantai 264003, China

Received:2016-12-02 Revised:2017-04-21 Online:2017-09-20 Published:2017-09-22
About author:
Author:QIU Chunhua.E-mail: qiuchh3@mail. sysu.edu.cn
Supported by:
State Key Development Program for Basic Research of China (2015CB452904);National Natural Science Foundation of China (41376161, 31501862);100 Talents Program of the Chinese Academy of Sciences to ZHANG Li;Science and Technology Planning Project of Guangdong Province, China (2014B030301064).

摘要/Abstract

摘要：

氨氮是大多数硬骨鱼的氮代谢产物, 但是环境中氨氮浓度过高也会对鱼类产生毒性作用。该研究结果显示, 海水青鳉暴露于0.6和1.1mmol·L^-1 NH₄Cl 4周后的死亡率分别为21.25%和40%。这说明高浓度氨氮暴露对海水青鳉具有明显的毒性作用, 而且暴露浓度越高, 毒性越大。利用蛋白质组学的研究技术研究海水青鳉在长期氨氮暴露后的蛋白质响应变化, 发现在0.6mmol·L^-1 NH₄Cl暴露条件下有16种蛋白的表达发生明显的变化, 而在1.1mmol·L^-1 NH₄Cl暴露条件下有7种蛋白的表达发生明显的变化。这些蛋白主要参与氧化应激的产生, 影响海水青鳉细胞结构组成、肌肉收缩、视觉通路、代谢调节和免疫调节, 引起神经毒性。该研究利用蛋白质组学的研究方法研究了长期氨氮暴露对海水青鳉的毒理效应, 为利用蛋白质组学探讨环境污染物的毒性效应提供了更深刻的认识。

关键词: 长期氨氮暴露, 蛋白质组学, 海水青鳉, 毒理效应

Abstract:

Abstract: Most teleost fish excretes nitrogen waste as ammonia, but high environment ammonia is toxic to fish. In this study, we demonstrated that the death rate of marine medaka Oryzias melastigma exposed to 0.6 and 1.1 mmol·L^-1 NH₄Cl for four weeks were 21.25% and 40.00%, respectively, suggesting high environment ammonia is toxic to the fish. Besides, the higher concentration the ambient ammonia, the more toxic it is to the fish. We investigated the responses of chronic ammonia exposure in marine medaka using proteomics. Sixteen proteins were found to be remarkably altered in the fish exposed to 0.6 mmol·L^-1 NH₄Cl and seven proteins, in the fish exposed to 1.1 mmol·L^-1 NH₄Cl. The function of these proteins included inducing oxidative stress, neurotoxicity, disturbance in cell structure, muscle contraction, visual pathway, metabolic and immunological regulation. This is the first report of studying the toxicological effect of ammonia on marine medaka using proteomics. It provides important insights into toxicological effects of environmental contaminant using proteomics.

Key words: chronic ammonia exposure, proteomic, marine medaka, toxicological effect

高娜, 朱丽梅, 于德良, 张黎. 长期高氨氮暴露对海水青鳉毒理效应的蛋白质组学研究[J]. 热带海洋学报, 2017, 36(5): 40-48.

Na GAO, Limei ZHU, Deliang YU, Li ZHANG. Proteomic responses of marine medaka (Oryzias melastigma) to chronic high environment ammonia exposure[J]. Journal of Tropical Oceanography, 2017, 36(5): 40-48.

图/表 4

图 1

图2

表1

长期氨氮暴露后海水青鳉体内差异表达蛋白质谱鉴定结果"

	蛋白点数	组别	蛋白名称		序列号	分子量/u	等电点	蛋白得分	序列覆盖率	匹配肽数	倍数变化
氧化应激	94	A	低分子量蛋白: 类线粒体的过氧化物还原酶5	青鳉	gi\|432899770	20491	8.68	114	0.18	2	4.26
	140	B	类蛋白酶体β亚基4	青鳉	gi\|432881057	28758	6.32	138	0.20	3	2.58
	178	B	胰蛋白酶原	青鳉	gi\|145966014	26708	5.16	255	0.12	1	1.63
细胞结构	285	A	类透明质酸蛋白聚糖连接蛋白1	青鳉	gi\|432874420	37805	8.18	63	0.03	1	-1.54
细胞结构	288	B	类透明质酸蛋白聚糖连接蛋白1	青鳉	gi\|432874420	37805	8.18	76	0.03	1	-1.71
肌肉收缩	92	A	类丝切蛋白-2	青鳉	gi\|432936587	18889	6.62	426	0.37	4	1.60
	343	A	类原肌球蛋白α1链亚型4	青鳉	gi\|432861692	31829	4.77	268	0.19	3	-1.70
	333	A	肌钙蛋白T, 类快骨骼肌型亚型2	青鳉	gi\|432860165	27699	9.41	96	0.10	2	-1.55
	403	B	类钙腔蛋白B	青鳉	gi\|432863493	29610	4.42	152	0.18	3	-1.92
视觉通路	176	A	中间丝蛋白ON3亚型2	青鳉	gi\|432864505	57350	5.06	69	0.02	1	1.63
	90	A	α-晶状体A链	青鳉	gi\|432958610	19919	5.95	184	0.44	6	1.70
	135	A	类β-晶状体B1链	青鳉	gi\|432878256	28885	8.36	70	0.23	4	4.88
	161	A	类β-晶状体A2链亚型1	青鳉	gi\|432964694	23855	6.09	377	0.51	5	2.07
代谢调节	17	A	14ku载脂蛋白部分片段	海水青鳉	gi\|328963182	11405	6.23	134	0.32	1	-1.53
	375	A	类线粒体ATP合成酶β亚基	青鳉	gi\|432849647	55238	5.10	562	0.24	6	2.15
	386	A	类α-烯醇酶	青鳉	gi\|432866138	45323	6.36	431	0.30	5	1.99
	207	B	类异戊烯基二磷酸Δ异构酶1	青鳉	gi\|432927313	32545	6.06	110	0.23	3	-3.27
免疫调节	153	A	类组织蛋白酶B	青鳉	gi\|432852559	37145	5.93	90	0.05	1	2.07
	42	A	类半乳凝素9	青鳉	gi\|432852360	23618	8.81	90	0.16	2	2.81
	559	B	类胎球蛋白B	大黄鱼	gi\|734621215	57367	5.99	63	0.01	1	2.75
神经毒性	18	A	类S100蛋白A	青鳉	gi\|432908621	11360	5.62	60	0.08	1	1.90
	391	A	突触囊泡膜蛋白VAT-1同系物	青鳉	gi\|432922308	46708	5.40	150	0.10	3	1.53
	847	B	类S100B亚型1	青鳉	gi\|432848584	11052	4.43	67	0.17	1	-2.70

表1

图3

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长期高氨氮暴露对海水青鳉毒理效应的蛋白质组学研究

Proteomic responses of marine medaka (Oryzias melastigma) to chronic high environment ammonia exposure

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