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完整幼体斑马鱼的微阵列无创神经元癫痫发作记录

Microarray Noninvasive Neuronal Seizure Recordings from Intact Larval Zebrafish.

作者信息

Meyer Michaela, Dhamne Sameer C, LaCoursiere Christopher M, Tambunan Dimira, Poduri Annapurna, Rotenberg Alexander

机构信息

Division of Epilepsy and Clinical Neurophysiology, Department of Neurology, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.

F.M. Kirby Neurobiology Center, Department of Neurology, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.

出版信息

PLoS One. 2016 Jun 9;11(6):e0156498. doi: 10.1371/journal.pone.0156498. eCollection 2016.

Abstract

Zebrafish epilepsy models are emerging tools in experimental epilepsy. Zebrafish larvae, in particular, are advantageous because they can be easily genetically altered and used for developmental and drug studies since agents applied to the bath penetrate the organism easily. Methods for electrophysiological recordings in zebrafish are new and evolving. We present a novel multi-electrode array method to non-invasively record electrical activity from up to 61 locations of an intact larval zebrafish head. This method enables transcranial noninvasive recording of extracellular field potentials (which include multi-unit activity and EEG) to identify epileptic seizures. To record from the brains of zebrafish larvae, the dorsum of the head of an intact larva was secured onto a multi-electrode array. We recorded from individual electrodes for at least three hours and quantified neuronal firing frequency, spike patterns (continuous or bursting), and synchrony of neuronal firing. Following 15 mM potassium chloride- or pentylenetetrazole-infusion into the bath, spike and burst rate increased significantly. Additionally, synchrony of neuronal firing across channels, a hallmark of epileptic seizures, also increased. Notably, the fish survived the experiment. This non-invasive method complements present invasive zebrafish neurophysiological techniques: it affords the advantages of high spatial and temporal resolution, a capacity to measure multiregional activity and neuronal synchrony in seizures, and fish survival for future experiments, such as studies of epileptogenesis and development.

摘要

斑马鱼癫痫模型正在成为实验性癫痫研究中的新兴工具。特别是斑马鱼幼体具有优势,因为它们易于进行基因改造,并且由于施加到水中的药物能够轻易穿透机体,所以可用于发育和药物研究。斑马鱼的电生理记录方法尚新且在不断发展。我们提出了一种新颖的多电极阵列方法,用于非侵入性地记录完整斑马鱼幼体头部多达61个位置的电活动。该方法能够经颅非侵入性记录细胞外场电位(包括多单位活动和脑电图)以识别癫痫发作。为了记录斑马鱼幼体的大脑活动,将完整幼体头部的背部固定在多电极阵列上。我们从各个电极记录至少三个小时,并对神经元放电频率、放电模式(连续或爆发)以及神经元放电的同步性进行量化。在向水中注入15 mM氯化钾或戊四氮后,放电和爆发率显著增加。此外,跨通道的神经元放电同步性(癫痫发作的一个标志)也增加了。值得注意的是,鱼在实验后存活下来。这种非侵入性方法补充了现有的侵入性斑马鱼神经生理学技术:它具有高空间和时间分辨率的优势,能够测量癫痫发作中的多区域活动和神经元同步性,并且鱼能存活以便用于未来的实验,如癫痫发生和发育研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ceb4/4900632/0bd650d42ae2/pone.0156498.g001.jpg

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