Name: in ovo electroporation in embryonic chick retina
Uploaded: 2012-02-05T13:00:00
Description: Watch this Scientific Journal Video about In Ovo Electroporation in Embryonic Chick Retina at JoVE.com

我们要感谢他的高清摄像头（炮VIXIA HFS100）使用玛斯恩维尔先生。这项工作是支持的，一部分由来自NIH的“（EY018738），新泽西州的脊髓研究委员会（08-3074-可控硅 - E-0和10-3091-SCR-E 0），布希生物医学研究奖补助。

由于该协议的某些部分进行轻微修改先前在其他的朱庇特的视频和论文3所述，我们不会在这里详细讨论。 1。鸡蛋处理和针的制备<ol><li>鸡蛋可以储存在13℃左右的葡萄酒在凉爽长达1周。如果温度过高，胚胎开始正常发展，而较低的温度下会导致高死亡率。一旦你准备好孵化从酒柜中取出的鸡蛋和鸡蛋与较大的一端设置垂直向上。保持至少2个小时之前，他们在37.5℃...

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	<li>Doh, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+retinal+cell+development+in+chick+embryo+by+immunohistochemistry+and+in+ovo+electroporation+techniques.">Analysis of retinal cell development in chick embryo by immunohistochemistry and in ovo electroporation techniques.</a> BMC. Dev. Biol. 10, (2010).</li><li>Blank, M. C., Chizhikov, V., Millen, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+Ovo+Electroporations+of+HH+Stage+10+Chicken+Embryos.">In Ovo Electroporations of HH Stage 10 Chicken Embryos.</a> J. Vis. Exp. (9), e408-e408 (2007).</li><li>Nakamura, H., Funahashi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Introduction+of+DNA+into+chick+embryos+by+in+ovo+electroporation.">Introduction of DNA into chick embryos by in ovo electroporation.</a> Methods. 24, 43-48 (2001).</li><li>Hamburger, V., Hamilton, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+series+of+normal+stages+in+the+development+of+the+chick+embryo.+1951.">A series of normal stages in the development of the chick embryo. 1951.</a> Dev. Dyn.. 195, 231-272 (1992).</li><li>Hamburger, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+stage+series+of+the+chick+embryo.">The stage series of the chick embryo.</a> Dev. Dyn. 195, 273-275 (1992).</li></ol>

有针对性的视网膜注射，并在 E4类阶段OVO在胚胎视网膜电针对视网膜祖细胞，导致在单细胞水平上的所有六个主要的视网膜细胞类型的能力，可视化。 在OVO电在HH10（E1.5）针对视泡是能够转染细胞的发展，形成了眼。然而，这些细胞具有非常高的营业额，在这个时候，这种方法是不特定的视网膜细胞。这可能是高的细胞周转率，防止持续稳定表达。由E4时，胚胎不够发达，眼?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>受精卵无病原体（SPF）的白来航鸡蛋</td><td>朝阳农场（卡茨基尔，纽约） </td><td></td><td></td></tr><tr><td>鸡蛋孵化器</td><td> GQF制造，大草原，GA </td><td>型号-1550 </td><td>设置湿度60％和37.5°C。 </td></tr><tr><td>玻璃毛细管</td><td>世界精密仪器公司</td><td> TW150F-4 </td><td></td></tr><tr><td> 0.1毫升注射器</td><td>汉密尔顿有限公司，内华达州里诺</td><td>气密1710 </td><td>另外1ml注射器，以及可用于</td></tr><tr><td>硅的Masterflex（双氧水）管</td><td>科尔 - 帕默</td><td> HV-96400-13 </td&gt; <td>附加玻璃针注射器剪下一小块（1厘米） </td></tr><tr><td>镊子</td><td>杜蒙</td><td>机管局</td><td></td></tr><tr><td>微操作机器人</td><td>世界精密仪器公司</td><td> M3301-M3 </td><td></td></tr><tr><td>质粒DNA </td><td></td><td></td><td> pCAG-GFP是借来的，从康妮Cepko博士</td></tr><tr><td>固绿FCF </td><td> Sigma-Aldrich公司</td><td> F7252 </td><td>稀释用PBS为0.025％ </td></tr><tr><td>脉冲发生器</td><td>哈佛仪器，马</td><td> A型肉毒毒素流脑830 </td><td>方波发生器</td></tr><tr><td>电极</td><td>哈佛仪器，马</td><td> A型肉毒毒素模型514 </td><td>我们的电极相距3-5毫米间距</td></tr><tr><td>单色数码相机</td><td>德国蔡司， </td><td>一xiocam的MRM </td><td></td></tr><tr><td>荧光解剖显微镜</td><td>徕卡公司，德国</td><td>徕卡MZ16FA </td><td></td></tr><tr><td>直立荧光显微镜</td><td>德国蔡司， </td><td>蔡司Axio上成像仪A1 </td><td></td></tr></tbody></table>

in ovo electroporation in embryonic chick retina

鸡胚胎的视网膜是一个很好的工具来研究在高等脊椎动物的视网膜发育。由于庞大的规模和外部的发展，它是相对非常容易操纵的鸡胚视网膜，利用重组DNA / RNA技术。胚胎视网膜电DNA / RNA的结构将有一个很大的优势，研究视网膜发育过程中视网膜干/祖细胞的基因调控。如检测报告基因检测，基因过度表达，基因敲除（shRNA的）等不同类型，可以使用电穿孔技术。该视频演示了有针对性的视网膜注射到鸡胚视网膜OVO在汉堡和汉密尔顿阶段22-23，这是关于胚胎第4天（E4），电。在这里，我们表明在OVO电技术的快速和便捷，使质粒DNA的表达绿色荧光蛋白（GFP）作为标记直接传递到鸡胚视网膜下的空间和通过电脉冲，以促进视网膜干/祖细胞的DNA吸收。视网膜注射和电E4类的新方法，让所有的视网膜细胞类型的可视化，包括后期新生神经元，这一直是注射和电的传统方法难以在E1.5 2。

Watch this Scientific Journal Video about In Ovo Electroporation in Embryonic Chick Retina at JoVE.com

In Ovo Electroporation in Embryonic Chick Retina

该视频的总体目标是显示如何执行有针对性的视网膜注射<em&gt;在OVO</em&gt; DNA / RNA的结构，在汉堡包和汉密尔顿22-23阶段，这是关于胚胎第4天（E4），成鸡胚胎视网膜电。这种技术是非常有用的，以研究基因表达，基因调控，并在发展中国家小鸡视网膜的形态学变化。

在OVO在鸡胚视网膜电穿孔

Chicken embryonic retina is an excellent tool to study retinal development in higher vertebrates. Because of large size and external development, it is ...

Research

JoVE Journal

Biology

In Ovo Electroporation in Embryonic Chick Retina

In Ovo Electroporations of HH Stage 10 Chicken Embryos

在OVO的HH阶段10个鸡胚Electroporations

Large size and external development of the chicken embryo have long made it a valuable tool in the study of developmental biology.  With the advent of ...

科研

生物学

Method for Whole Mount Antibody Staining in Chick

整装在鸡的抗体染色方法

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ...

Chick ex ovo Culture and ex ovo CAM Assay: How it Really Works

Chick ex ovo Culture and ex ovo CAM Assay: How it Really Works

小鸡前OVO文化前OVO CAM含量：它是如何真正起作用

Chicken eggs in the early phase of breeding are between in vitro and in vivo systems and provide a vascular test environment not only to study ...

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

电子显微镜对胚胎的制备果蝇胚胎心管

The morphogenesis of the Drosophila  embryonic heart tube has emerged as a valuable model system for studying cell migration, cell-cell adhesion and cell ...

Primary Culture and Plasmid Electroporation of the Murine Organ of Corti. 

原代培养及质粒电穿孔尔蒂小鼠器官。

In all mammals, the sensory epithelium for audition is located along the spiraling organ of Corti that resides within the conch shaped cochlea of the ...

Deciphering Axonal Pathways of Genetically Defined Groups of Neurons in the Chick Neural Tube Utilizing in ovo Electroporation

Deciphering Axonal Pathways of Genetically Defined Groups of Neurons in the Chick Neural Tube Utilizing in ovo Electroporation

利用鸡胚神经管破译基因定义的组的神经元轴突途径在OVO电穿孔

Employment of enhancer elements to drive expression of reporter genes in neurons is a widely used paradigm for tracking axonal projection. For tracking ...

Video Bioinformatics Analysis of Human Embryonic Stem Cell Colony Growth

视频人类胚胎干细胞克隆生长的生物信息学分析

Because video data are complex and are comprised of many images, mining information from video material is difficult to do without the aid of computer ...

Single Cell Transfection in Chick Embryos

单细胞转染鸡胚

A central theme in developmental biology is the diversification of lineages and the elucidation of underlying molecular mechanisms. This entails a ...

Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes

Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes

膜片钳和灌注技术研究离子通道的表达爪蟾卵母细胞

The protocol presented here is designed to study the activation of the large conductance, voltage- and Ca2+-activated K+ (BK) channels. The protocol may ...

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

在体内电穿孔Morpholinos到成年斑马鱼视网膜

Many devastating inherited eye diseases result in progressive and irreversible blindness because humans cannot regenerate dying or diseased retinal ...