中国水产科学2016，Vol.23Issue(4)：771-776,6.DOI:10.3724/SP.J.1118.2016.15488
低温胁迫诱导斑马鱼ZF4细胞ROS及MAPK相关蛋白表达的影响
Effects of cold stress on ROS production and expression of MAPK proteins in zebrafish ZF4 cells
摘要
Abstract
In our previous study, we compared the transcriptome between Antarctic fish and temperate fish and found that genes were involved in reactive oxygen species (ROS) scavenge and Mitogen-activated protein kinase (MAPK) pathway associated with the cold adaptation in fish. To further study the regulatory network and the re-active oxygen species (ROS) homeostasis under cold stress in fish, we examined the level of ROS and the expres-sion of stress-related proteins in zebrafish Danio rerio-derived ZF4 cells exposed to mildly and severely cold temperatures, 18℃ and 10℃, at various time points (1 d, 3 d, and 5 d). Results showed that (1) DCFH-DA probe method was used to determine the level of ROS in cells. We found that the level of ROS in cell positively corre-lated with the intensity of cold stress. After 3 days of cold treatment, the level of ROS in cells significantly in-creased to 1.23±0.04 (P<0.05) and 2.31±0.08 (P<0.05) times, respectively, at 18℃ and 10℃, compared with that in cells at 28℃. (2) Western blot showed that the expression of MAPKs (p-JNK p54, p-JNK p46 and p-p38) was induced under cold stress, and reached the highest at 3 d of 10℃. (3) In addition, the expression ofγH2A.X reached peak after 3 days both at 18℃ and 10℃. These findings showed that cold temperature could induce the production of ROS in ZF4 cells. The level of ROS in ZF4 cells is dependent on the stress intensity and duration. We observed the markedly induction of ROS in the cells at the first 3 days of cold treatment, while the expression of p-JNK, p-p38, andγH2A.X was markedly induced at the 3 d, indicating that 3 d of cold treatment might be a key time point to determine the protein expression in ZF4 cells.关键词
ROS/斑马鱼ZF4细胞/低温胁迫/MAPKs/γH2A.XKey words
ROS/zebrafish ZF4 cells/low temperature stress/MAPKs/γH2A.X分类
农业科技引用本文复制引用
许琼琼,韩兵社,罗军涛,李艳,侯艳雯,胡鹏,张俊芳..低温胁迫诱导斑马鱼ZF4细胞ROS及MAPK相关蛋白表达的影响[J].中国水产科学,2016,23(4):771-776,6.基金项目
国家自然科学基金项目(31372516) （31372516）
上海市自然科学基金项目(13ZR1419500) （13ZR1419500）
上海市教育委员会“东方学者”计划支持项目 （）
上海市人才发展资金项目(201457)资助 （201457）
