生物学杂志

生物学杂志 ›› 2020, Vol. 37 ›› Issue (4): 40-.doi: 10.3969/j.issn.2095-1736.2020.04.040

• 研究报告 • 上一篇    下一篇

低温胁迫下斑马鱼胚胎成纤维细胞（ZF4）应激颗粒的探究

  

  1. 1. 上海海洋大学 水产科学国家级实验教学示范中心; 2. 水产种质资源发掘与利用教育部重点实验室;3. 海洋生物科学国际联合研究中心 中国科学技术部， 上海 201306
  • 出版日期:2020-08-18 发布日期:2020-08-10
  • 通讯作者: 陈良标，教授，博士生导师，主要从事发育、分子生物学研究，E-mail: lbchen@shou.edu.cn
  • 作者简介:程鹏丽, 硕士研究生, 主要从事分子生物学研究，E-mail:1760771678@qq.com
  • 基金资助:
    国家自然科学基金(31572611) 

Study on stress granules of zebrafish embryo fibroblasts cell (ZF4) under cold stress

  1. 1. National Demonstration Center for Experimental Fisheries Science Education (Shanghai Ocean University); 2. Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources (Shanghai Ocean University),Ministry of Education; 3. International Research Center for Marine Biosciences at Shanghai Ocean University, Ministry of Science and Technology, Shanghai 201306, China
  • Online:2020-08-18 Published:2020-08-10

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摘要： 为了探究低温胁迫下斑马鱼胚胎成纤维细胞（ZF4）应激颗粒的形成及其解聚情况，首先选取18 ℃、13 ℃和10 ℃作为低温胁迫温度，并且分别通过不同时间的低温处理；然后利用TIAL1、G3BP1（应激颗粒的标记物）分别进行细胞免疫荧光实验和RT-qPCR实验探究应激颗粒的形成和解聚；最后选取应激恢复的温度和时间，探究应激颗粒的解聚情况。结果表明，不同温度的低温胁迫，应激颗粒形成的时间不同；低温处理时间不同，应激颗粒的大小也不同。实验发现在18 ℃低温处理24 h，13 ℃低温处理4 h以及10 ℃低温处理1 h，ZF4细胞应激颗粒形成明显。说明随着胁迫温度的降低，应激颗粒形成速度也在加快。应激颗粒的形成表现为应激颗粒数量的增多和直径的变大。当ZF4细胞在13 ℃低温处理4 h，再将温度恢复到最适生长温度（28 ℃）时，发现应激颗粒会在十几分钟迅速解聚。应激颗粒的解聚表现为应激颗粒的快速溶解消散。首次发现鱼类细胞在低温下形成应激颗粒，主要成分与哺乳动物细胞类似，表明应激颗粒是动物细胞应对低温刺激的普遍反应机制。

关键词: ZF4细胞, 低温胁迫, 应激颗粒, 形成, 解聚

Abstract: To investigate the formation and depolymerization of stress granules in zebrafish embryonic fibroblasts cell (ZF4) under cold stress, 18 ℃, 13 ℃ and 10 ℃ were selected as the cold stress temperature, and the low temperature treatment was carried out at different times； and then TIAL1 and G3BP1 (marker of stress granules) were used to conduct cellular immunofluorescence experiments and RT-qPCR experiments； finally, the temperature and time of stress recovery were selected to investigate the depolymerization of stress granules (SG). The results showed that under cold stress at different temperatures, stress granules were formed at different time in ZF4 cells， and the stress granules had different sizes, with the increase of low temperature treatment time. The stress granules of ZF4 cells were formed when treated at 18 ℃ for 24 h, 13 ℃ for 4 h and 10 ℃ for 1 h. As the temperature decreased, the rate of formation of stress granules also accelerated. The formation of stress granules is manifested by a gradual increase in stress granules and an increase in the size of the diameter. When ZF4 cells were treated at 13 ℃ for 4 h and then the temperature was restored to the optimum growth temperature (28 ℃), it was found that the stress granules depolymerized rapidly in about ten minutes. Depolymerization of stress granules is manifested by the rapid dissolution of stress granules. This paper firstly found that fish cells form stress granules at low temperature, and the main components are similar to mammalian cells, indicating that stress granules are the general response mechanism of animal cells to cold stress.

Key words: ZF4 cells, cold stress, stress granules, formation, depolymerization

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