Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Psychiatr. Jul 19, 2020; 10(7): 150-161
Published online Jul 19, 2020. doi: 10.5498/wjp.v10.i7.150
Comparison of three administration modes for establishing a zebrafish seizure model induced by N-Methyl-D-aspartic acid
Xin-Yi Long, Shuang Wang, Zhi-Wen Luo, Xu Zhang, Hong Xu
Xin-Yi Long, Shuang Wang, Queen Mary School of Nanchang University, Nanchang 330006, Jiangxi Province, China
Zhi-Wen Luo, Department of Sports Medicine, Huashan Hospital, Fudan University, Shanghai 200433, China
Xu Zhang, Affiliated Eye Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China
Hong Xu, Institute of Life Science, Nanchang University, Nanchang 330031, Jiangxi Province, China
Author contributions: Long XY, Wang S, Luo ZW and Zhang X conceived the study; Wang S, Long XY and Luo ZW performed the experiments; Long XY, Wang S and Luo ZW analyzed the data; Luo ZW, Wang S and Long XY drafted the manuscript; All authors read and approved the final version of the manuscript; Xu H provided the idea and funding; Long XY and Wang S contributed equally.
Supported by the National Natural Science Foundation of China, No. 81760216, No. 81160144 and No. 31171044.
Institutional animal care and use committee statement: The study was approved by the Ethical Review Committee of Nanchang University.
Conflict-of-interest statement: All other authors have nothing to disclose.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Corresponding author: Hong Xu, PhD, Professor, Institute of Life Science, Nanchang University, Xuefu Avenue 1299, Nanchang 330031, Jiangxi Province, China.
Received: December 31, 2019
Peer-review started: December 31, 2019
First decision: April 3, 2020
Revised: April 25, 2020
Accepted: May 26, 2020
Article in press: May 26, 2020
Published online: July 19, 2020

Epilepsy is a complex neurological disorder characterized by recurrent, unprovoked seizures resulting from the sudden abnormal discharge of brain neurons. It leads to transient brain dysfunction, manifested by abnormal physical movements and consciousness. It can occur at any age, affecting approximately 65 million worldwide, one third of which are still estimated to suffer from refractory seizures. There is an urgent need for further establishment of seizure models in animals, which provides an approach to model epilepsy and could be used to identify novel anti-epileptic therapeutics in the future.


To compare three administration modes for establishing a seizure model caused by N-Methyl-D-aspartic acid (NMDA) in zebrafish.


Three administration routes of NMDA, including immersion, intravitreal injection and intraperitoneal injection, were compared with regard to their effects on inducing seizure-like behaviors in adult zebrafish. We evaluated neurotoxicity by observing behavioral changes in zebrafish and graded those behaviors with a seizure score. In addition, the protective effects of MK-801 (Dizocilpine) and natural active constituent resveratrol against NMDA-induced alterations were studied.


The three NMDA-administration methods triggered different patterns of the epileptic process in adult zebrafish. Seizure scores were increased after increasing NMDA concentration regardless of the mode of administration. However, the curve of immersion continuously rose to a high plateau (after 50 min), while the curves of intravitreal injection and intraperitoneal injection showed a spike in the early stage (10-20 min) followed by a steady decrease in seizure scores. Furthermore, pretreatment with resveratrol and MK-801 significantly delayed seizure onset time and lowered seizure scores.


By comparing the three methods of administration, intravitreal injection of NMDA was the most suitable for establishing an acute epileptic model in zebrafish. Thus, intraperitoneal injection in zebrafish can be applied to simulate diseases such as epilepsy. In addition, NMDA immersion may be an appropriate method to induce persistent seizures. Moreover, MK-801 and resveratrol showed strong anti-epileptic effects; thus, both of them may be clinically valuable treatments for epilepsy.

Keywords: Seizure, Zebrafish, N-Methyl-D-aspartic acid, Administration modes, Resveratrol, MK-801

Core tip: This is the first study to systematically compare the three main administration modes to establish a seizure model in zebrafish. A newly developed zebrafish model with acute and sustained experimental epileptic behavior enables us to study and identify potential mechanisms and screen anti-epileptic drugs. Direct administration of N-Methyl-D-aspartic acid stimulates abnormal excitations of brain nerve cells to simulate epileptic seizures. This study demonstrated that intravitreal injection can be used to establish an acute epilepsy model and immersion can be used as a persistent epilepsy model. The protective effects of resveratrol and MK-801 on the epileptic process were also confirmed, which may have clinical application value.