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610500 成都, 成都医学院人体解剖与组织胚胎学教研室,发育与再生四川省重点实验室
[ 苏炳银,E-mail:subingyinn@163.com SU Bingyin, E-mail: subingyinn@163.com
收稿:2019-03-15,
修回:2019-05-07,
纸质出版:2019-08-30
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甘津凡, 吴永梅, 李淑蓉, 等. 不同浓度的二甲基亚砜对斑马鱼胚胎及其多巴胺能神经元发育的影响[J]. 第三军医大学学报, 2019,41(16):1538-1544.
Jinfan GAN, Yongmei WU, Shurong LI, et al. Effects of different concentrations of dimethylsulfoxide on development of zebrafish embryos and dopamine neurons[J]. Journal of Third Military Medical University, 2019, 41(16): 1538-1544.
甘津凡, 吴永梅, 李淑蓉, 等. 不同浓度的二甲基亚砜对斑马鱼胚胎及其多巴胺能神经元发育的影响[J]. 第三军医大学学报, 2019,41(16):1538-1544. DOI: 10.16016/j.1000-5404.201903087.
Jinfan GAN, Yongmei WU, Shurong LI, et al. Effects of different concentrations of dimethylsulfoxide on development of zebrafish embryos and dopamine neurons[J]. Journal of Third Military Medical University, 2019, 41(16): 1538-1544. DOI: 10.16016/j.1000-5404.201903087.
目的
2
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
2
收集同亲代
Tg
(
Vmat
2:
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
2
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
2
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
2
To study the effect of exposures to different concentrations of dimethyl sulfoxide (DMSO) on the development of zebrafish embryos and dopaminergic (DA) neurons.
Methods
2
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 (bl
ank 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
2
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
2
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.
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