Name: a behavioral assay to measure responsiveness of zebrafish to changes in light intensities
Uploaded: 2008-10-03T00:00:00
Description: Watch this Scientific Journal Video about A Behavioral Assay to Measure Responsiveness of Zebrafish to Changes in Light Intensities at JoVE.com

这项工作是由全国健康教育资助EY0081和5T3​​2UY07145研究所和圣殿骑士眼科基金会的支持。贾森Rihel是一个生命科学研究基金会的布里斯托尔 - 施贵宝研究员。

该协议列出的步骤来执行在自己的实验室光增量和递减的斑马鱼幼虫的视觉运动反应。斑马鱼是一个伟大的行为研究模型系统。他们易于维护，他们有很大的离合器大小，并迅速发展。例如，斑马鱼幼虫的眼睛是顺应轻发展的第3天，在这时候，他们表现出了惊吓反应。 第1部分：电镀成96孔板个别鱼<ol><li> WT幼虫生长在一个黑暗/光?...

<ol>
	<li>Allwardt, A. B., Lall, B. A., Brockerhoff, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synapse+formation+is+arrested+in+retinal+photoreceptors+of+the+zebrafish+nrc+mutant.">Synapse formation is arrested in retinal photoreceptors of the zebrafish nrc mutant.</a> J Neurosci. 21, 2330-2330 (2001).</li><li>Emran, F., Rihel, J., Adolph, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=OFF+ganglion+cells+cannot+drive+the+optokinetic+reflex+in+zebrafish.">OFF ganglion cells cannot drive the optokinetic reflex in zebrafish.</a> Proceedings of the National Academy of Sciences of the United States of America. 104, 19126-19126 (2007).</li><li>Prober, D. A., Rihel, J., Onah, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypocretin/orexin+overexpression+induces+an+insomnia-like+phenotype+in+zebrafish.">Hypocretin/orexin overexpression induces an insomnia-like phenotype in zebrafish.</a> J Neurosci. 26, 13400-13400 (2006).</li><li>Westerfield, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The zebrafish book: a guide for the laboratory use of zebrafish. , (2000).</li></ol>

我们在影片中展示的实验程序是代表所有的WT鱼。然而，这些实验可以做类似于突变鱼（见代表结果部分）。我们建议板聚鱼和在棋盘大纲为最优控制的目的相同板上的突变鱼。 当使用相同的板聚鱼突变的鱼，确保您写下什么类型的鱼，每孔镀。这里有一个如何保持鱼镀在每口井的轨道（显示数据表的注意书）的建议。 另外，最好在一天的同一时间做了一定的实验重复。例如，如果你开始的3个小时的...

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a behavioral assay to measure responsiveness of zebrafish to changes in light intensities 

视动反射（OKR）是一个基本的视觉反射展出，大部份脊椎动物和眼视觉场景相对稳定起着重要作用。然而，OKR需要检测的动物移动的条纹，它可能是鱼类，未能表现出一种OKR未必完全失明。斑马鱼突变体，没有视动反应彗星（NRC）在任何光线条件下测试，并报告完全失明没有OKR。此前，我们已经表明，OFF神经节细胞的活性，可在这些突变体的记录。为了确定是否与没有NRC的突变体，如OKR突变鱼可以检测到简单的轻的递增和递减，我们开发的视觉电机行为检测器（VMR）。在这个实验中，单斑马鱼幼虫放置在每个允许同时使用自动视频跟踪系统的幼虫监测的96孔板以及。每个幼虫的30分钟指示灯亮起，指示灯熄灭30分钟的运动反应记录和量化。聚鱼后灯被称为惊跳反应，随后返回到低比基线活动电机活动的一个简短的扣球，被称为冻结。聚鱼也大幅增加他们的自发活动紧随灯光熄灭，只有逐步在几分钟内恢复到基线自发活动。 NRC的突变体的回应同样光聚鱼关，但表现出他们的平均活动为WT鱼相比略有减少。电机响应活动光NRC突变延迟和缓慢。有一个NRC的突变响应缓慢的上升时间，光相比WT光响应。结果表明，NRC鱼是不完全失明。因为硬骨鱼类可以检测到通过非视网膜组织中，我们确认使用chokh（CHK）的突变体，完全没有眼睛，从发展的最初阶段，直接的行为反应的光强度的变化需要的完整的眼睛。在我们的VMR检测，CHK突变体表现出没有惊吓反应，无论是轻或关，表明横向眼睛调解这种行为。 VMR检测形容这里补充行之有效的OKR检测，不测试斑马鱼幼虫的能力以应对光强的变化。此外，VMR检测的自动化，适合于高通量筛选驱动的视觉反应的光强度的缺陷。

Watch this Scientific Journal Video about A Behavioral Assay to Measure Responsiveness of Zebrafish to Changes in Light Intensities at JoVE.com

A Behavioral Assay to Measure Responsiveness of Zebrafish to Changes in Light Intensities

我们开发的视觉运动反应，反应轻的递增和递减定量幼虫斑马鱼的电机输出。我们还研究了斑马鱼的视觉突变，包括没有视动反应（NRC）的突变体，它被认为是完全失明的另一个视力检测，视动反射测试时。

斑马鱼的响应行为的检测，以测量光强的变化

The optokinetic reflex (OKR) is a basic visual reflex exhibited by most vertebrates and plays an important role in stabilizing the eye relative to the ...

Research

JoVE Journal

Biology

A Behavioral Assay to Measure Responsiveness of Zebrafish to Changes in Light Intensities 

Intravenous Microinjections of Zebrafish Larvae to Study Acute Kidney Injury

斑马鱼幼虫的研究急性肾损伤的静脉Microinjections

In this video article we describe a zebrafish model of AKI using gentamicin as the nephrotoxicant.  The technique consists of intravenous microinjections ...

科研

生物学

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结合免疫荧光和三维保存的间期核荧光原位杂交DNA研究在3D核组织的变化

Fluorescent in situ hybridization using DNA probes  on 3-dimensionally preserved nuclei followed by 3D confocal microscopy (3D DNA  FISH) represents the ...

Analysis of Apoptosis in Zebrafish Embryos by Whole-mount Immunofluorescence to Detect Activated Caspase 3

Whole-mount immunofluorescence to detect activated Caspase 3 (Casp3 assay) is useful to identify cells undergoing either intrinsic or extrinsic apoptosis ...

In vitro Methylation Assay to Study Protein Arginine Methylation

In vitro Methylation Assay to Study Protein Arginine Methylation

在体外甲基化分析研究蛋白质精氨酸甲基化

Protein arginine methylation is one of the most abundant post-translational modifications in the nucleus. Protein arginine methylation can be identified ...

Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes

定量免疫检测，以衡量在中心体的变异蛋白水平

Centrosomes are small but important organelles that serve as the poles of mitotic spindle to maintain genomic integrity or assemble primary cilia to ...

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

对于波长和红/近红外光治疗对氧化应激强度的范围影响的评估方法<em&gt;在体外</em

Red/near-infrared light therapy (R/NIR-LT), delivered by laser or light emitting diode (LED), improves functional and morphological outcomes in a range of ...

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使用拉姆齐试验来测量液体分泌和离子流率在<em&gt;果蝇</em&gt;马氏管

Modulation of renal epithelial ion transport allows organisms to maintain ionic and osmotic homeostasis in the face of varying external conditions. The ...

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体外试验来测量磷脂酰乙醇胺甲基转移酶活

Phosphatidylethanolamine methyltransferases are biosynthetic enzymes that catalyze the transfer of one or more methyl group(s) from ...

Nephrotoxin Microinjection in Zebrafish to Model Acute Kidney Injury

肾毒素显微注射斑马鱼到模型急性肾损伤

The kidneys are susceptible to harm from exposure to chemicals they filter from the bloodstream. This can lead to organ injury associated with a rapid ...