摘要
目的研究不同浓度的二甲基亚砜(dimethylsulfoxide, DMSO)对斑马鱼胚胎、多巴胺能(dopaminergic,DA)神经元发育的影响。方法收集同亲代Tg(Vmat2:GFP)斑马鱼产下的胚胎,简单随机分为10组:空白对照组,0.1%、0.3%、0.5%、0.8%、1.0%、1.5%、2.0%、2.5%、3.0%DMSO(体积比)组。于浓度梯度下培养,统计存活率、畸形率、孵化率;分析行为学变化;利用激光共聚焦显微镜观察DA神经元的发育,PH3抗体检测细胞增殖及TUNEL试剂盒检测细胞凋亡。结果 DMSO浓度为2.5%时,斑马鱼存活率明显降低(P<0.01);DMSO浓度为2.0%时,斑马鱼畸形率明显升高(P<0.01)、孵化率明显降低(P<0.05);DMSO浓度为2.5%时,斑马鱼胚胎尾部摆动次数明显减少(P<0.05)。DMSO浓度为0.5%时间脑DA神经元明显减少(P<0.05);DMSO大于0.3%时,间脑细胞增殖明显减少(P<0.01);DMSO浓度≥0.8%时间脑细胞凋亡增加。结论 DMSO会引起斑马鱼胚胎发育障碍,其神经毒性造成间脑DA神经元丢失及细胞增殖的减少。
Objective To study the effect of exposures to different concentrations of dimethyl sulfoxide(DMSO)on the development of zebrafish embryos and dopaminergic(DA)neurons.Methods Tg(Vmat2:GFP)zebrafish embryos of the same generation were randomized into 10 groups at 6 h post-fertilization for exposures to 9 concentrations(0.1%,0.3%,0.5%,0.8%,1.0%,1.5%,2.0%,2.5%,and 3.0%;V/V)of DMSO for 72 h or without any exposure(blank control group).The survival rate,deformity rate,hatching rate and behavioral changes of the embryos were analyzed.The development of DA neurons was observed using laser confocal microscopy,and the cell proliferation and apoptosis were detected using PH3 antibody and an apoptosis detection kit,respectively.Results Exposure to 2.5%DMSO significantly decreased the survival rate of the zebrafish embryos(P<0.01);at the concentration of 2.0%,DMSO exposure caused an obvious increase in the malformation rate(P<0.01)and a decrease in the hatching rate of the embryos(P<0.05);DMSO at 2.5%significantly decreased the frequency of tail swing of the zebrafish embryos(P<0.05).Zebrafish embryos exposed to 0.5%DMSO showed a significantly reduced number of DA neurons(P<0.05).Exposure of the zebrafish embryos to DMSO at a concentration above 0.3%significantly reduced the proliferation of the diencephalon cells(P<0.01),and a DMSO concentration above 0.8%could obviously increase apoptosis of the dopaminergic neurons.Conclusion DMSO exposure causes developmental disorders in zebrafish embryos,and its neurotoxicity can result in loss of dopaminergic neurons in the diencephalon and suppress the cell proliferation.
作者
甘津凡
吴永梅
李淑蓉
苏炳银
GAN Jinfan;WU Yongmei;LI Shurong;SU Bingyin(Department of Anatomy,Histology and Embryology,Sichuan Provincial Key Laboratory of Development and Regeneration,Chengdu Medical College,Chengdu,Sichuan Province,610500,China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2019年第16期1538-1544,共7页
Journal of Third Military Medical University
基金
四川省教育厅重点项目(17ZA0108)
四川省科技厅应用基础研究重点项目(2017JY0138,2019YJ0366)~~
