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用一种新型长期脑电图记录方法评估kcnj10吗啡啉修饰的斑马鱼中的癫痫。

Epilepsy in kcnj10 morphant zebrafish assessed with a novel method for long-term EEG recordings.

作者信息

Zdebik Anselm A, Mahmood Fahad, Stanescu Horia C, Kleta Robert, Bockenhauer Detlef, Russell Claire

机构信息

Department of Neuroscience, Physiology and Pharmacology, University College London, London, United Kingdom ; Centre for Nephrology, University College London, London, United Kingdom.

出版信息

PLoS One. 2013 Nov 14;8(11):e79765. doi: 10.1371/journal.pone.0079765. eCollection 2013.

Abstract

We aimed to develop and validate a reliable method for stable long-term recordings of EEG activity in zebrafish, which is less prone to artifacts than current invasive techniques. EEG activity was recorded with a blunt electrolyte-filled glass pipette placed on the zebrafish head mimicking surface EEG technology in man. In addition, paralysis of agarose-embedded fish using D-tubocurarine excluded movement artifacts associated with epileptic activity. This non-invasive recording technique allowed recordings for up to one hour and produced less artifacts than impaling the zebrafish optic tectum with a patch pipette. Paralyzed fish survived, and normal heartbeat could be monitored for over 1h. Our technique allowed the demonstration of specific epileptic activity in kcnj10a morphant fish (a model for EAST syndrome) closely resembling epileptic activity induced by pentylenetetrazol. This new method documented that seizures in the zebrafish EAST model were ameliorated by pentobarbitone, but not diazepam, validating its usefulness. In conclusion, non-invasive recordings in paralyzed EAST syndrome zebrafish proved stable, reliable and robust, showing qualitatively similar frequency spectra to those obtained from pentylenetetrazol-treated fish. This technique may prove particularly useful in zebrafish epilepsy models that show infrequent or conditional seizure activity.

摘要

我们旨在开发并验证一种可靠的方法,用于对斑马鱼的脑电图(EEG)活动进行稳定的长期记录,该方法比目前的侵入性技术更不易产生伪迹。使用一根钝头的、充满电解质的玻璃微电极放置在斑马鱼头部来记录EEG活动,这模仿了人类的表面EEG技术。此外,使用筒箭毒碱使琼脂糖包埋的鱼麻痹,排除了与癫痫活动相关的运动伪迹。这种非侵入性记录技术允许记录长达一小时,并且比用膜片微电极刺入斑马鱼视顶盖产生的伪迹更少。麻痹的鱼存活下来,并且可以监测到正常心跳超过1小时。我们的技术能够在kcnj10a基因敲降鱼(一种EAST综合征模型)中展示特定的癫痫活动,该活动与戊四氮诱导的癫痫活动非常相似。这种新方法证明,戊巴比妥可改善斑马鱼EAST模型中的癫痫发作,但地西泮则不能,从而验证了其有效性。总之,对麻痹的EAST综合征斑马鱼进行的非侵入性记录被证明是稳定、可靠且强大的,其定性的频谱与戊四氮处理的鱼所获得的频谱相似。这种技术在表现出不频繁或条件性癫痫活动的斑马鱼癫痫模型中可能特别有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2704/3828195/c225c3c8d7e0/pone.0079765.g001.jpg

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