Parng Chuenlei, Roy Nicole Marie, Ton Christopher, Lin Yingxin, McGrath Patricia
Phylonix Pharmaceuticals, Inc., 100 Inman St., Cambridge, MA 02139, USA.
J Pharmacol Toxicol Methods. 2007 Jan-Feb;55(1):103-12. doi: 10.1016/j.vascn.2006.04.004.
Transparency is a unique attribute of zebrafish that permits direct assessment of drug effects on the nervous system using whole mount antibody immunostaining and histochemistry.
To assess pharmacological effects of drugs on the optic nerves, motor neurons, and dopaminergic neurons, we performed whole mount immunostaining and visualized different neuronal cell types in vivo. In addition, we assessed neuronal apoptosis, proliferation, oxidation and the integrity of the myelin sheath using TUNEL staining, immunostaining and in situ hybridization. The number of dopaminergic neurons was examined and morphometric analysis was performed to quantify the staining signals for myelin basic protein and apoptosis.
We showed that compounds that induce neurotoxicity in humans caused similar neurotoxicity in zebrafish. For example, ethanol induced defects in optic nerves and motor neurons and affected neuronal proliferation; 6-hydroxydopamine caused neuronal oxidation and dopaminergic neuron loss; acrylamide induced demyelination; taxol, neomycin, TCDD and retinoic acid induced neuronal apoptosis.
Effects of drug treatment on different neurons can easily be visually assessed and quantified in intact animals. These results support the use of zebrafish as a predictive model for assessing neurotoxicity.
透明性是斑马鱼的一个独特特性，它允许使用整体抗体免疫染色和组织化学方法直接评估药物对神经系统的影响。
为了评估药物对视神经、运动神经元和多巴胺能神经元的药理作用，我们进行了整体免疫染色，并在体内观察了不同类型的神经元细胞。此外，我们使用TUNEL染色、免疫染色和原位杂交技术评估了神经元凋亡、增殖、氧化以及髓鞘的完整性。我们检查了多巴胺能神经元的数量，并进行了形态计量分析以量化髓鞘碱性蛋白和凋亡的染色信号。
我们发现，在人类中诱导神经毒性的化合物在斑马鱼中也会引起类似的神经毒性。例如，乙醇会导致视神经和运动神经元缺陷，并影响神经元增殖；6-羟基多巴胺会引起神经元氧化和多巴胺能神经元损失；丙烯酰胺会导致脱髓鞘；紫杉醇、新霉素、TCDD和视黄酸会诱导神经元凋亡。
在完整动物中，可以很容易地通过视觉评估和量化药物治疗对不同神经元的影响。这些结果支持将斑马鱼用作评估神经毒性的预测模型。
