高浓度葡萄糖对斑马鱼胚胎及多巴胺神经元发育的影响

Impact of high concentration of glucose on development of zebrafish embryos and dopaminergic neurons

目的研究高浓度葡萄糖在斑马鱼胚胎早期发育中的作用以及对斑马鱼多巴胺神经元发育的影响。方法实验分为3组:空白对照组,葡萄糖实验(25 mmol/L葡萄糖)组,甘露醇对照(25 mmol/L甘露醇)组。25 mmol/L葡萄糖溶液处理Vmat-GFP转基因斑马鱼胚胎,当胚胎发育到第3天时,激光共聚焦显微镜观察多巴胺神经元的发育,荧光定量PCR检测PD相关基因的表达。结果与空白对照组、甘露醇组相比,25 mmol/L葡萄糖溶液处理胚胎后,胚胎发育迟缓,心脏畸形,多巴胺神经元荧光增强,端脑部位多巴胺神经元数量增加,PD(帕金森病)相关基因表达水平升高,其中parkin基因表达差异具有统计学意义(P<0.05)。结论 25 mmol/L葡萄糖溶液对胚胎的发育具有毒性作用,但在胚胎发育的早期对多巴胺神经元具有营养作用。

Objective To study the effect of high concentration of glucose on the early development of embryos and dopaminergic neurons in zebrafish. Methods Vmat-GFP transgenic zebrafish was used. After fertilization, the embryos were divided into control, mannitol group （25 mmol/L ） and glucose group （25 retool/L）. In 3 d after fertilization, the morphology of embryonic dopaminergic neurons was visualized under laser scanning confocal microscope （LSCM）. The expression of PD （Parkinson＇ s disease）-related genes was measured using fluorescent quantitative polymerase chain reaction （PCR）. Results Compared with control and mannitol group, the embryonic development in glucose group was retarded with cardiac abnormalities. Enhanced fluorescent intensity of Vmat-GFP and increased number of dopaminergic neurons in the teleneephalon were also observed. In glucose group the expression of PD-related genes was upregulated, and particularly, upregulation of PARKIN was significant （ P 〈 0.05 ）. Conclusion High concentration of glucose （25 mmol/L） is toxic to embryonic development, but neurotrophic for dopaminergie neurons at the early stage of embryos in zebrafish.