Background

Zebrafish has emerged as a potential animal model of acute convulsion for early screening of antiepileptic agents. There is a need for alternative chronic zebrafish models of epilepsy with more correlation to the clinical condition.

New Method

Adult zebrafish were repeatedly exposed to subeffective concentrations of pentylenetetrazole (PTZ), until appearance to tonic-clonic seizures, considered as kindled. Valproic acid (VPA) exposure was given during kindling and in kindled fish in 2 different groups. The neurotransmitters level and expression of the genes associated with kindling were studied in the fish brain.

Results

There was an increase in seizure severity score at 1.25 mM concentration of PTZ, and 66.66 % of fish achieved kindling after 22 days’ exposure. A marked increase in c-fos, crebbpa and crebbpbexpression, and glutamate/GABA level was observed in the brain of kindled fish. VPA inhibited the induction of PTZ-mediated kindling and reduced seizure severity in kindled fish.

Comparison with existing method

In contrast to an existing adult zebrafish kindling method, the present protocol is of longer duration, with more similarity to clinical epilepsy. Moreover, the induction of kindling involves a simple non-invasive technique without the use of anesthesia. The protocol can be used for evaluation of both antiepileptic and antiepileptogenic agents.

Conclusion

Repeated exposure of 1.25 mM PTZ induced kindling in zebrafish, altering the brain neurotransmitter levels and gene expression. Inhibition of kindling induction and decrease in seizures in normal and kindled fish, respectively by VPA validated application of the model for preclinical testing of agents against epilepsy.

中文翻译：

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成人斑马鱼化学点燃的开发和验证：一种用于筛选抗癫痫药的简单且改良的慢性模型。

背景

斑马鱼已成为潜在的急性惊厥动物模型，可用于早期筛选抗癫痫药。需要与临床状况更多相关的替代性癫痫慢性斑马鱼模型。

新方法

成年斑马鱼反复暴露于亚有效浓度的戊四唑（PTZ）中，直到出现强直阵挛性癫痫发作（被认为是点燃的）。在点燃过程中和两组不同种类的鱼类中均给予了丙戊酸（VPA）暴露。在鱼脑中研究了与点燃相关的基因的神经递质水平和表达。

结果

在1.25 mM的PTZ浓度下，癫痫发作的严重程度评分有所提高，暴露22天后，有66.66％的鱼实现了点燃。在点燃的鱼的大脑中观察到c-fos，crebbpa和crebbpb表达以及谷氨酸/ GABA水平显着增加。VPA抑制了点燃的鱼中PTZ介导的点燃的诱导并降低了癫痫发作的严重程度。

与现有方法的比较

与现有的成年斑马鱼点燃方法相反，本协议的持续时间更长，与临床癫痫病的相似性更高。此外，引燃涉及不使用麻醉的简单非侵入性技术。该协议可用于评估抗癫痫药和抗癫痫药。

结论

1.25 mM PTZ的反复暴露会引起斑马鱼的点燃，从而改变大脑神经递质的水平和基因表达。通过VPA分别抑制正常鱼和被点燃鱼类的点燃诱导和减少癫痫发作，验证了该模型在临床前测试抗癫痫药物的应用。