梨状皮层注射红藻氨酸建立癫痫小鼠模型
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徐州医科大学

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江苏省自然科学基金(编号:BK20170251),江苏省高等学校自然科学基金(编号:21KJB180011),徐州市科技局社会发展项目(编号:ZYSB20210410),江苏省实验动物协会科研基金(编号:DWXH202206),徐州市科技创新重点研发计划-面上项目(编号:KC22078)


Kainic acid was injected into the piriform cortex to establish a mouse model of epilepsy
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Xuzhou Medical University

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    摘要:

    目的:建立和评价KA(Kainic acid,红藻氨酸)在梨状皮层微量注射激发癫痫小鼠模型。方法:利用c-fos(即刻早期基因)免疫荧光染色技术、TUNEL(TdT-mediated dUTP Nick-End Labeling,原位末端转移酶标记)染色技术在急性癫痫小鼠模型中找到癫痫相关脑区梨状皮层。在梨状皮层脑立体定位注射KA后,观察小鼠癫痫发作程度(Racine评分)和癫痫发作潜伏期,行为学实验包括水迷宫检测小鼠学习记忆能力和新物体识别实验检测小鼠探索能力,癫痫发作结果和行为学实验结果均与海马立体定位注射KA的癫痫模型作对比。利用电生理技术检测新模型小鼠神经元电活动。结果:c-fos免疫荧光和TUNEL染色结果显示,在急性癫痫小鼠模型中除海马外,梨状皮层脑区神经元也被大量激活,且发生凋亡;在梨状皮层注射KA后成功激发癫痫,与海马注射KA的癫痫模型相比,癫痫发作评分没有显著性差异且发作潜伏期更短;行为学结果显示,梨状皮层注射KA的癫痫模型小鼠空间记忆能力降低,新物体探索能力降低,该结果与海马注射KA的癫痫模型相比无显著性差异;膜片钳实验显示梨状皮层注射KA的癫痫模型神经元放电频率增加、幅值降低、静息膜电位上升。结论:在梨状皮层微量注射KA后成功激发癫痫,这为癫痫发病机制的研究提供了一种新的动物模型。

    Abstract:

    This study aims to establish and evaluate epileptic kindled by kainic acid (KA) microinjected into piriform cortex. Methods: Using c-fos immunofluorescence staining and TdT-mediated dUTP Nick-End Labeling (TUNEL) staining techniques to find the piriform cortex associated with epilepsy in a mouse model of acute epilepsy. Racine score and seizure latency were observed after injection of kainic acid in piriform cortex. Behavioral experiments included testing the learning and memory abilities of mice in a water maze and testing the exploration ability of mice in a new object recognition experiment. The results of seizures and behavioral experiments were compared with the epileptic model of stereotactic injection of kainic acid into the hippocampus. The electrical activities of new model mouse neurons were measured by electrophysiological techniques. Results: The results of c-fos immunofluorescence and TUNEL staining showed that in addition to the hippocampus, neurons in the piriform cortex and hippocampus were also significantly activated and apoptosis occurred in the acute epilepsy mouse model; After injecting KA into the piriform cortex, the epilepsy was successfully ignited. Compared with the epilepsy model injected KA into the hippocampus, there was no significant difference in the epileptic seizure score and the seizure latency was shorter; The behavioral results showed that the epileptic model mice injected KA into the piriform cortex had a reduced spatial memory ability and a reduced ability to explore new objects. These results showed no significant difference compared to the epileptic model injected KA into the hippocampus. Patch clamp recording test showed an increase of firing frequency and the resting membrane potential, and a decrease of amplitude of neuronal discharge. Conclusion: Microinjection of KA into the piriform cortex has successfully induced epilepsy, which provides a new animal model for studying the pathogenesis of epilepsy.

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  • 收稿日期:2022-12-14
  • 最后修改日期:2023-04-17
  • 录用日期:2023-08-09
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