Chemogenetic ablation of dopaminergic neurons leads to transient locomotor impairments in zebrafish larvae
Chemogenetic ablation of dopaminergic neurons leads to transient locomotor impairments in zebrafish larvae
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DOI：
10.1111/jnc.13214
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发表时间：
2015-10-01
影响因子：
4.7
通讯作者：
Ekker, Marc
中科院分区：
文献类型：
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作者：
Godoy, Rafael;Noble, Sandra;Ekker, Marc
为了确定多巴胺能神经元的可控性丧失对运动功能的影响，我们获得了转基因斑马鱼TG(DAT：CFP-NTR)，在多巴胺转运体(DAT)顺式调控元件的控制下表达了青色荧光蛋白-硝基还原酶融合蛋白(CFP-NTR)。胚胎和幼体斑马鱼在几组多巴胺能神经元中表达转基因，特别是在嗅球、端脑、间脑和下丘脑尾侧。注射前体药物甲硝唑(MTZ)后，受精后5天CFP阳性神经元caspase3激活，DAT阳性细胞减少。神经元的丧失与整体运动参数(如游泳距离、移动时间百分比)以及尾部弯曲参数(如最大弯曲时间和角速度)的损害相吻合。注射MTZ后，多巴胺水平一过性下降。用7DPF观察部分运动参数的恢复。然而，尽管有证据表明有新的表达dat的细胞的产生，但在7、12或14dpf时，表达dat的神经元的总数仍然减少。TG(DAT：CFP-NTR)斑马鱼提供了一种模型，可以将改变的多巴胺能神经元数量与运动功能联系起来，并研究影响多巴胺能神经元再生的因素。
To determine the impact of a controlled loss of dopaminergic neurons on locomotor function, we generated transgenic zebrafish, Tg(dat:CFP-NTR), expressing a cyan fluorescent protein-nitroreductase fusion protein (CFP-NTR) under the control of dopamine transporter (dat) cis-regulatory elements. Embryonic and larval zebrafish express the transgene in several groups of dopaminergic neurons, notably in the olfactory bulb, telencephalon, diencephalon and caudal hypothalamus. Administration of the pro-drug metronidazole (Mtz) resulted in activation of caspase 3 in CFP-positive neurons and in a reduction in dat-positive cells by 5days post-fertilization (dpf). Loss of neurons coincided with impairments in global locomotor parameters such as swimming distance, percentage of time spent moving, as well as changes in tail bend parameters such as time to maximal bend and angular velocity. Dopamine levels were transiently decreased following Mtz administration. Recovery of some of the locomotor parameters was observed by 7 dpf. However, the total numbers of dat-expressing neurons were still decreased at 7, 12, or 14 dpf, even though there was evidence for production of new dat-expressing cells. Tg(dat:CFP-NTR) zebrafish provide a model to correlate altered dopaminergic neuron numbers with locomotor function and to investigate factors influencing regeneration of dopaminergic neurons.