Name: forebrain electrophysiological recording in larval zebrafish
Uploaded: 2013-01-24T12:00:00
Description: Watch this Scientific Journal Video about Forebrain Electrophysiological Recording in Larval Zebrafish at JoVE.com

作者想，感谢他们及早建立斑马鱼在实验室的彼得·卡斯特罗和马修Dinday。这项工作是由美国国立卫生EUREKA（＃R01NS079214-01）资助。

1。鸡蛋的生产和收集<ol><li>斑马鱼的饲养标准程序，前面描述的8。简单地说，成年斑马鱼在养殖箱分频器的地方。第二天早上，在房间里的灯都分频器从坦克和鱼类育种允许不受干扰的交配时间约20至60分钟。 </li><li>从繁殖罐的鸡蛋被收集在过滤器并和用鸡蛋水漂洗。卵，然后转移到陪替氏培养皿用鸡蛋水。用移液管未受精的卵子和碎片被删除。 </li><li>将培养皿包含收集鸡蛋的培养箱?...

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	<li>Clark, K. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stressing+zebrafish+for+behavioral+genetics.">Stressing zebrafish for behavioral genetics.</a> Reviews in Neuroscience. 22 (1), 49 (2011).</li><li>Rinkwitz, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zebrafish:+an+integrative+system+for+neurogenomics+and+neurosciences.">Zebrafish: an integrative system for neurogenomics and neurosciences.</a> Progress in Neurobiology. 93 (2), 231 (2011).</li><li>Penberthy, W. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+zebrafish+as+a+model+for+human+disease.">The zebrafish as a model for human disease.</a> Frontiers in Bioscience. 7, d1439 (2002).</li><li>Letamendia, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+validation+of+an+automated+high-throughput+system+for+zebrafish+in+vivo+screenings.">Development and validation of an automated high-throughput system for zebrafish in vivo screenings.</a> PLoS One. 7, e36690 (2012).</li><li>Nasevicius, A., Ekker, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effective+targeted+gene+'knockdown'+in+zebrafish.">Effective targeted gene 'knockdown' in zebrafish.</a> Nature Genetics. 26 (2), 216 (2000).</li><li>Haffter, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mutations+affecting+development+of+the+zebrafish+inner+ear+and+lateral+line.">Mutations affecting development of the zebrafish inner ear and lateral line.</a> Development. 123, 1 (1996).</li><li>Suster, M. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenesis+in+zebrafish+with+the+tol2+transposon+system.">Transgenesis in zebrafish with the tol2 transposon system.</a> Methods Molecular Biology. 561, 41 (2009).</li><li>Rosen, J. N., Sweeney, M. F., Mably, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microinjection+of+Zebrafish+Embryos+to+Analyze+Gene+Function.">Microinjection of Zebrafish Embryos to Analyze Gene Function.</a> J. Vis. Exp. (25), e1115 (2009).</li><li>Baraban, S. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pentylenetetrazole+induced+changes+in+zebrafish+behavior,+neural+activity+and+c-fos+expression.">Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression.</a> Neuroscience. 131 (3), 759 (2005).</li><li>Baraban, S. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+large-scale+mutagenesis+screen+to+identify+seizure-resistant+zebrafish.">A large-scale mutagenesis screen to identify seizure-resistant zebrafish.</a> Epilepsia. 48 (6), 1151 (2007).</li><li>Williams, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+radiotelemetry+to+evaluate+electrographic+seizures+in+rats+with+kainate-induced+epilepsy.">The use of radiotelemetry to evaluate electrographic seizures in rats with kainate-induced epilepsy.</a> Journal of Neuroscience Methods. 155 (1), 39 (2006).</li><li>Marsh, E. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interictal+EEG+spikes+identify+the+region+of+electrographic+seizure+onset+in+some,+but+not+all,+pediatric+epilepsy+patients.">Interictal EEG spikes identify the region of electrographic seizure onset in some, but not all, pediatric epilepsy patients.</a> Epilepsia. 51 (4), 592 (2010).</li><li>Zhu, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+and+application+of+modularly+assembled+zinc-finger+nucleases+in+zebrafish.">Evaluation and application of modularly assembled zinc-finger nucleases in zebrafish.</a> Development. 138 (20), 4555 (2011).</li></ol>

这里介绍的方法使细胞外记录大脑活动的一个非常敏感和快速的分析。这些录音是类似的脑电图（EEG）监测常用的评估存在的异常放电（ 即癫痫发作）在啮齿类动物模型癫痫11和患者12。细胞外记录可以结合药理的操作，如下所示。也可用于这些类型的记录，以评估潜在的癫痫表型的转基因斑马鱼。确定，在ENU的突变屏幕（见斑马鱼资源中心， <a href="http://zebrafish.org/zirc/home...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td>琼脂糖低熔点</td><td>费舍尔的科学</td><td> BP1360-100 </td><td>溶解于1.2％的胚胎媒体</td></tr><tr><td>记录媒体</td><td>费舍尔的科学</td><td> BP3581，P330-3，BP410，BP214-500，D16-1，C77-500 </td><td> 1毫摩尔NaCl，2.9 mM KCl中，10mM的HEPES，1.2毫摩尔MgCl 2，10mM的葡萄糖，2.1 mM的CaCl 2的 pH值至约7.3，用1N NaOH </td></tr><tr><td>三卡因</td><td>银色实验室</td><td> MS-222 </td><td> 0.02％ </td></tr><tr><td> α-银环蛇毒素</td><td> TOCRIS生命科学</td><td> 2133 </td><td> 1毫克/毫升</td></tr><tr><td>毛细玻璃管</td><td>华纳仪器</td><td> G120TF -3 </td><td>拉出到2 -7MΩ的电阻</td></tr><tr><td>膜片钳放大器</td><td>华纳仪器</td><td> PC-505B </td><td>使用华纳放大器的电流钳模式;在2 mV / PA和贝塞尔滤波器在2K的增益设置。同类机型可根据制造商的说明。 </td></tr><tr><td>过滤器/放大器</td><td>天鹅座技术</td><td> FLA-01 </td><td>我们使用一个新天鹅前置放大器增益设置为10〜20;截止频率设置为1-2K;陷波滤波器。同类机型可根据制造商的说明。 </td></tr><tr><td>轴突A / D板和Axoscope的软件</td><td> Molecular Devices公司</td><td>轴突Digidata 1320A 8.2 Axoscope </td><td>数据被收集在Axoscope采用无间隙采集模式，在10 kHz采样。比较模型和程序可以根据制造商的说明使用。 </td></tr><tr><td>蛋水</td><td&gt;即时海洋</td><td></td><td> 3克即溶海洋海盐，2毫升10毫升去离子水，0.1％的亚甲基蓝</td></tr></tbody></table>

forebrain electrophysiological recording in larval zebrafish

癫痫会影响近3万人在美国和世界各地多达50万人。定义为发生自发的无端发作，癫痫，可以获取到大脑或基因突变的结果的一种侮辱。在动物模型癫痫发作的努力，主要用于收购侮辱（惊厥药，刺激或脑损伤）和遗传操作（反义击倒，同源重组或转基因）在啮齿类动物。斑马鱼是脊椎动物模型系统1-3，可以提供一个有价值的替代老鼠癫痫研究。斑马鱼被广泛应用于脊椎动物的遗传学研究或发展，对哺乳动物的遗传相似性，表现出高度表达的同系物〜85％的已知的人类单基因遗传性癫痫突变。由于它们的小尺寸（长4-6毫米），斑马鱼幼虫可以保持低的流体体积为100微升，在早期的发展和阿拉进行期间，在多井板。试剂可直接加入到该溶液中，在胚胎发育，简化给药，从而实现快速的在体内筛选试验化合物4。可用于合成寡核苷酸（吗啉代），致突变，锌指核酸酶和转基因方法，以迅速产生基因敲除或突变的斑马鱼5-7。这些属性提供了前所未有的斑马鱼研究的统计动力分析优势的研究啮齿动物的神经系统疾病，如癫痫。 ，因为癫痫研究的“金标准”是描述监测和分析的异常放电起源于一个中央的大脑结构（ 即癫痫发作），斑马鱼幼虫的大脑活动的方法，有效地记录在这里。该方法是一种适应传统的细胞外记录技术，并允许稳定的长期监测在完整的斑马鱼幼虫的大脑活动。小号充足的记录上所显示的急性发作引起的惊厥药和自发发作在转基因鱼浴应用。

在图1中所示的实施例的电子照相记录的前脑琼脂嵌入的斑马鱼幼虫的癫痫样放电。在这些样本中的大振幅的多穗型脉冲放电引起的惊厥药浴应用，40毫米毛果芸香碱（A; 6 DPF）或1毫米印防己毒素（B; 8 DPF）。在这些记录中，固定，和琼脂嵌入式斑马鱼的连续监测90分钟。鱼这些记录的条件下继续生存24小时。药物添加到泳季介质和通常扩散到琼脂在30至45分钟之内引出在幼虫斑马鱼前脑?...

Watch this Scientific Journal Video about Forebrain Electrophysiological Recording in Larval Zebrafish at JoVE.com

Forebrain Electrophysiological Recording in Larval Zebrafish

外场电位记录在幼虫斑马鱼的前脑的一个简单的方法来描述。该方法提供了一个强大的<em在体内</em&gt;读出癫痫样活动。这种技术可以使用转基因斑马鱼幼虫携带癫痫相关基因或癫痫发作诱发的惊厥药给药。

斑马鱼幼虫的前脑的电生理记录

Epilepsy affects nearly 3 million people in the United States and up to 50 million people worldwide. Defined as the occurrence of spontaneous unprovoked ...

Research

JoVE Journal

Neuroscience

Time-lapse Live Imaging of Clonally Related Neural Progenitor Cells in the Developing Zebrafish Forebrain

在发展中国家斑马鱼的前脑的神经祖细胞克隆相关的定时实时成像

Precise patterns of division, migration and differentiation of neural progenitor cells are crucial for proper brain development and function1,2. To ...

科研

神经科学

Simultaneous Electrophysiological Recording and Calcium Imaging of Suprachiasmatic Nucleus Neurons

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in ...

Electrophysiological Recording in the Brain of Intact Adult Zebrafish

在完整的成年斑马鱼大脑电生理记录

Previously, electrophysiological studies in adult zebrafish have been limited to slice preparations or to eye cup preparations and electrorentinogram ...

A Method for Systematic Electrochemical and Electrophysiological Evaluation of Neural Recording Electrodes

对于系统电化学神经记录电极和电生理评价方法

New materials and designs for neural implants are typically tested separately, with a demonstration of performance but without reference to other implant ...

Electrophysiological Recording From Drosophila Labellar Taste Sensilla

Electrophysiological Recording From Drosophila Labellar Taste Sensilla

电生理记录从<em&gt;果蝇</em&gt; Labellar味感器

The peripheral taste response of insects can be powerfully investigated with electrophysiological techniques. The method described here allows the ...

Spinal Cord Transection in the Larval Zebrafish

在斑马鱼幼虫脊髓横断

Mammals fail in sensory and motor recovery following spinal cord injury due to lack of axonal regrowth below the level of injury as well as an inability ...

Simultaneous Electrophysiological Recording and Micro-injections of Inhibitory Agents in the Rodent Brain

同时电生理记录和抑制剂在啮齿动物大脑微注射

Here we describe a method for the construction of a single-use &#8220;injectrode&#8221; using commercially accessible and affordable parts. A probing ...

Electrophysiological Recording from Drosophila Trichoid Sensilla in Response to Odorants of Low Volatility

Electrophysiological Recording from Drosophila Trichoid Sensilla in Response to Odorants of Low Volatility

电生理记录<em&gt;果蝇</em&gt; Trichoid Sensilla响应低挥发性气味

Insects rely on their sense of smell to guide a wide range of behaviors that are critical for their survival, such as food-seeking, predator avoidance, ...

In Vivo Calcium Imaging of Lateral-line Hair Cells in Larval Zebrafish

幼虫斑马鱼外系头发细胞的活体钙成像

Sensory hair cells are mechanoreceptors found in the inner ear that are required for hearing and balance. Hair cells are activated in response to sensory ...

Investigating Long-term Synaptic Plasticity in Interlamellar Hippocampus CA1 by Electrophysiological Field Recording

通过电生理场记录研究跨体海马CA1的长期突触可塑性

The study of synaptic plasticity in the hippocampus has focused on the use of the CA3-CA1 lamellar network. Less attention has been given to the ...