生物学杂志

生物学杂志 ›› 2024, Vol. 41 ›› Issue (2): 67-.doi: 10.3969/j.issn.2095-1736.2024.02.067

• 研究报告 • 上一篇    下一篇

敲除kdm4ab对斑马鱼低温耐受能力的影响

张梦冉1,2， 罗军涛1,2， 张　瑶1,2， 韩兵社1,2， 张俊芳1,2   

  1. 1. 上海海洋大学 水产种质资源发掘与利用教育部重点实验室， 上海 201306；
    2.上海海洋大学 水产科学国家级实验教学示范中心， 上海 201306
  • 出版日期:2024-04-18 发布日期:2024-04-17
  • 通讯作者: 张俊芳，教授，研究方向为表观遗传学与环境适应，E-mail:jfzhang@shou.edu.cn
  • 作者简介:张梦冉，硕士研究生，研究方向为表观遗传学，E-mail:986588026@qq.com
  • 基金资助:
    上海市科技兴农重点攻关项目\[No.沪农科创字(2019)第1-2号\]；国家自然科学基金面上项目(No.81770165)；上海市教委水产一流学科建设项目

Knockout of zebrafish kdm4ab affects cold tolerance

ZHANG Mengran1,2, LUO Juntao1,2, ZHANG Yao1,2, HAN Bingshe1,2, ZHANG Junfang1,2   

  1. 1. Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean
    University, Shanghai 201306, China; 2. National Demonstration Center for Experimental Fisheries Science Education,
    Shanghai Ocean University, Shanghai 201306, China
  • Online:2024-04-18 Published:2024-04-17

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摘要： 为深入研究kdm4ab基因在鱼类低温胁迫下的作用，以斑马鱼（Danio rerio）为模式生物，利用CRISPR/Cas9基因编辑技术构建kdm4ab-/-斑马鱼品系。RT-qPCR结果显示，与WT斑马鱼相比，kdm4ab-/-斑马鱼的kdm4abmRNA表达量显著下调（P<0.0001），而kdm4c的mRNA表达量补偿性增加（P<0.05）。斑马鱼行为轨迹跟踪系统检测发现，18 ℃低温下kdm4ab-/-斑马鱼幼鱼的最大加速度显著降低（P<0.05）。8 ℃低温下kdm4ab-/-斑马鱼的半数致死时间 (LT50) 显著低于WT斑马鱼 (P<0.0001)；低温处理后kdm4ab-/-斑马鱼的γH2A.X表达量明显增加，ROS水平升高，即DNA损伤增强。研究表明，敲除kdm4ab基因降低了低温下斑马鱼的耐受能力及运动能力，这为鱼类的低温耐受机制研究提供新思路。

关键词: kdm4ab, 斑马鱼, CRISPR/Cas9, 低温

Abstract: o explore the role ofkdm4abin fish under low temperature,kdm4ab-/-zebrafish was constructed by CRISPR/Cas9. RT-qPCR showed that the mRNA expression level ofkdm4abinkdm4ab-/-zebrafish was significantly reduced (P<0.0001), while the expression of kdm4c level was compensatively increased (P<0.05). The detection of zebrafish behavior trajectory tracking system showed that the maximum acceleration ofkdm4ab-/-larval zebrafish was significantly decreased at 18 ℃ (P<0.05), compared with that of WT larval zebrafish. At 8 ℃, the LT50 ofkdm4ab-/-zebrafish was significantly lower than that of WT zebrafish (P<0.0001). After low temperature treatment, the expression of γH2A.X inkdm4ab-/-zebrafish increased significantly, and the level of ROS increased, which indicated that the DNA damage was enhanced. The results showed thatkdm4abhad a certain effect on the cold tolerance of zebrafish, which also provided a new idea for the low temperature tolerance mechanism of fish.

Key words: kdm4ab, zebrafish, CRISPR/Cas9, cold

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