核酸染料Goodview、Gelred和Gelsafe对斑马鱼胚胎发育的毒性作用

目的研究核酸染料Goodview、Gelred和Gelsafe对斑马鱼胚胎发育的影响。方法将受精后时间（hpf）为3hpf的斑马鱼胚胎随机分组,分别加入0、0.01%、0.02%、0.04%、0.08%、0.16%和0.32%浓度的Goodview、Gelred和Gelsafe 3种核酸染料,观察3种核酸染料对斑马鱼胚胎发育的影响,记录胚胎48 hpf心率,计算72和96 hpf孵化率,96 hpf死亡率及畸形率;同时观察了4、24和48 hpf 3种染料穿透胚胎膜的情况。结果 Goodview、Gelred和Gelsafe 3种核酸染料半数致死浓度（LC50）分别为0.037%、0.092%和0.130%。3种核酸染料0.04%～0.32%浓度组的胚胎48 hpf心率、72和96 hpf孵化率、96hpf死亡率和畸形率与对照组相比都存在显著差异,3种核酸染料96 hpf死亡率、72和96 hpf孵化率还存在剂量-效应关系。Gelsafe 0.08%～0.32%浓度组部分幼鱼出现心囊水肿和脊柱弯曲,0.32%Gelred组染毒后还出现眼点发育不全,Goodview0.04～0.16%浓度组部分幼鱼除心囊水肿和脊柱弯曲外,还出现尾部发育不全等畸形表征。0.01%Goodview 4 hpf能穿透胚胎膜进入胚胎内,随着浓度和时间增加,胚胎内荧光增强。而Gelred和Gelsafe 0.32%浓度下4 hpf未发现明显荧光蓄积点,24hpf和48 hpf仍未发现。结论 3种核酸染料对斑马鱼胚胎发育有毒性,随着使用浓度的增加毒性增加。

Toxicity of nucleic acid dyes Goodview, Gelred and Gelsafe on embryonic development of zebrafish

Objective To evaluate the toxicity of nucleic acid dyes Goodview,Gelred and Gelsafe on the development of ze- brafish embryos. Methods Six kinds of concentration（O. 01% ,0. 02% ,0. 04% ,0. 08% ,0. 16% and 0. 32% ） of these nucleic acid dyes were used to evaluate the developments of zebra-fish embryos. Forty eight hours post fertilization （48 hpf） the frequency of heart- beat was recorded,72 hpf and 96 hpf hatching rate,96 hpf mortality rate and 96 hpf malformations of zebrafish embryos were calculated in the embryonic development period. Penetration of three nucleic acid dyes through the cell membrane at 4 hpf,24 hpf and 48 hpf was observed. Results The median lethal concentration （LCs0） for zebrafish embryos exposed to Goodview, Gelred and Gelsafe were 0. 037% ,0. 092% ,0. 130%. These nucleic acid dyes groups showed significant differences from control group regarding the rate of heart, mortality, hatching and malformation, and these groups exhibited dosage-response relationships in mortality rate and hatching rate. Several abnormalities and toxic symptoms caused by three nucleic acid dyes could be observed, including pericardial cyst and spinal curvature. Goodview could penetrate the embryonic membrane into embryos at 4 hpf, while Gelred and Gelsafe didn＇t. No such phene- menon was observed at 24 h and 48hpf. Conclusion Three nucleic acid dyes are toxic and concentration-depent to zebrafish embryos.