首页> 中文期刊> 《中国水产科学》 >HSP90抑制剂根赤壳菌素对斑马鱼胚胎发育的影响

HSP90抑制剂根赤壳菌素对斑马鱼胚胎发育的影响

         

摘要

To study the role of heat shock protein 90 (HSP90) in the development of zebrafish embryos,we examined the mRNA levels of two HSP90 activity marker genes,namely bag3 (BCL2-associated athanogene 3) and hspb1 (heat shock protein beta-l),at different developmental stages.Furthermore,we observed the marker gene expression and development of zebrafish embryos at different stages after treating with 2 μmol/L,5 μmol/L,and 10 μmol/L of the HSP90 inhibitor radicicol.Analysis by quantitative polymerase chain reaction showed a strong increase in the mRNA levels of bag3 and hspb1 when zebrafish embryos were treated with radicicol at 12 or 24 hours post-fertilization (hpf).Western blot results showed that the protein level of HSP70 was upregulated when zebrafish embryos were treated with 5 μmol/L radicicol at 24 hpf,indicating that the activity of HSP90 was inhibited.Observations of embryonic development showed that the radicicol-treated embryos had a delayed development and survival rates of 95%,77%,and 35% after treatment with 2 μmol/L,5 μtmol/L,and 10 μmol/L radicicol,respectively.The embryos manifested some abnormalities including reduced pigmentation,an enlarged pericardial membrane,and muscular dystrophy after treatment with 5 μmol/L radicicol at 72 hpf.The results showed that HSP90 plays an important role on the development of zebrafish embryos.The radicicol concentration of 5 μtmol/L appeared optimal,because it strongly inhibited HSP90 without decreasing the survival rate of zebrafish embryos below 75%.Various morphological changes appeared in the radicicol-treated embryos,laying a foundation for the further study of HSP90's functions in zebrafish embryos.%为研究热休克蛋白90(heat shock protein 90,HSP90)在斑马鱼(Danio rerio)胚胎发育中的作用,本实验采用2 μmol/L、5μmol/L、10 μmol/L的HSP90抑制剂根赤壳菌素(radicicol)对斑马鱼发育期胚胎进行处理,监测斑马鱼胚胎不同发育时期两个HSP90功能抑制标志基因BAG3 (BCL2-associated athanogene3)和HSPB1(heat shock proteinbeta-1)的mRNA的表达水平,并观察不同发育时期的胚胎发育状况.结果如下:(1)实时荧光定量PCR结果显示,BAG3和HSPB1 mRNA水平在根赤壳菌素处理胚胎12 hpf (hours post-fertilization,hpf)或24 hpf后显著增高,Western blot检测到5 μmol/L根赤壳菌素处理胚胎24 hpf后HSP70表达上调,证明该实验条件下HSP90功能受到抑制;(2)根赤壳菌素处理后斑马鱼胚胎发育变缓,胚胎成活率统计显示:2 μmol/L、5 μmol/L、10 μmol/L根赤壳菌素处理24 hpf胚胎成活率分别是95%、77%、35%,成活率随根赤壳菌素浓度的增高而降低;(3)5 μmol/L根赤壳菌素处理72 hpf可见部分个体色素沉积减少、心包膜增大、肌肉萎缩等发育畸形.研究结果证明HSP90在斑马鱼胚胎发育过程中发挥重要的作用,根赤壳菌素在5 μmol/L时已达到较佳抑制效果且成活率较高,而且胚胎发育出现了各种形态学变化,为后期研究HSP90调节斑马鱼胚胎发育奠定了基础.

著录项

  • 来源
    《中国水产科学》 |2017年第5期|988-994|共7页
  • 作者单位

    省部共建水产种质资源发掘与利用教育部重点实验室,上海海洋大学水产与生命学院,上海201306;

    省部共建水产种质资源发掘与利用教育部重点实验室,上海海洋大学水产与生命学院,上海201306;

    省部共建水产种质资源发掘与利用教育部重点实验室,上海海洋大学水产与生命学院,上海201306;

    省部共建水产种质资源发掘与利用教育部重点实验室,上海海洋大学水产与生命学院,上海201306;

    省部共建水产种质资源发掘与利用教育部重点实验室,上海海洋大学水产与生命学院,上海201306;

    省部共建水产种质资源发掘与利用教育部重点实验室,上海海洋大学水产与生命学院,上海201306;

    省部共建水产种质资源发掘与利用教育部重点实验室,上海海洋大学水产与生命学院,上海201306;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 水产生物学;
  • 关键词

    斑马鱼; 热休克蛋白90; 根赤壳菌素; 色素沉积; 发育;

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