1-苯基-2-硫脲对斑马鱼胚胎发育与黑色素生成的影响  

Developmental and Melanogenesis Effects of 1-phenyl 2-thiourea(PTU) in Zebrafish Embryos

作　　者：张利军[1] 史慧勤[1] 苑晓燕[1] 余寿忠[1] 赵君[1] 彭双清[1] 

机构地区：[1]军事医学科学院疾病预防控制所毒理学评价研究中心,北京100071

出　　处：《中国比较医学杂志》2012年第9期21-25,共5页Chinese Journal of Comparative Medicine

基　　金：国家"重大新药创制"科技重大专项(2009ZX09501-034);国家973计划(2011CB503803)

摘　　要：目的 1-苯基-2-硫脲(PTU)可抑制斑马鱼胚胎黑色素的产生,保持斑马鱼透明,便于形态观察和信号检测。本文研究了PTU对斑马鱼胚胎发育的影响和抑制斑马鱼胚胎黑色素生成,保持斑马鱼透明性的最佳浓度。方法用不同浓度PTU处理23 hpf(受精后,hours post fertilization,hpf)斑马鱼胚胎,作用57 h后观察80 hpf斑马鱼的形态学、生理学改变,计算死亡率和孵化率,测量心率和静脉窦-动脉球之间的距离。结果浓度为0.197mmol/L、0.296 mmol/L PTU可以有效抑制黑色素生成,保持斑马鱼整体透明,对斑马鱼心血管系统结构和生理功能无影响,且不影响斑马鱼正常孵化过程。随着PTU浓度的增加,斑马鱼死亡率增加,孵化率下降,出现心包水肿,心脏畸形等改变,心率下降,静脉窦-动脉球之间的距离增大。结论浓度不高于0.296 mmol/L的PTU溶液能有效抑制斑马鱼黑色素生成,对斑马鱼心血管毒性研究无影响。Objective To generate transparent zebrafish, embryos may be treated with 1 - phenyl 2 - thiourea （PTU） to inhibit melanogenesis during embryogenesis. When working with techniques designed to study gene expression during embryogenesis, especially in teleosts, signal detection may be aggravated by pigmentation. Here we focus on the effects of PTU on embryo, and describes an optimized protocol for generating transparent zebrafish using PTU. Methods zebrafish embryos at the stage of 23 hpf （ hours post fertilization） were treated with serial concentrations of PTU. Morphological and physiological endpoints were selected and quantified for unexposed and exposed embryos. Mortality, heart rate, hatching rate and SV-BA distance were detected until 80 hpf. Results Exposure to 0. 197mmol/L and 0. 296 mmol/L PTU completely inhibited pigmentation while avoiding problems with embryo mortality, teratogenesis, reduced hatching frequency, and heart rate. The highest PTU concentration resulted in reduced hatching frequency and embryo mortality. We observed pericardial and yolk sac edema. Conclusion Optimized conditions can achieve complete inhibition of pigmentation for generating transparent zebrafish while avoiding toxic effects of PTU treatment.

关 键 词：1-苯基-2-硫脲 斑马鱼 心率 孵化率 

分 类 号：R965.1[医药卫生—药理学] R332[医药卫生—药学]