Name: neonatal subventricular zone electroporation
Uploaded: 2013-02-11T13:00:00
Description: Watch this Scientific Journal Video about Neonatal Subventricular Zone Electroporation at JoVE.com

这项工作是由国防部（理念发展奖，W81XWH-10-1-0041，AB），CT干细胞补助金（AB），和国立健康NRSA 10668225（DMF）的补助金。本发明的材料的基础工作部分由康涅狄格州的支持下，康涅狄格干细胞研究资助计划。它的内容是完全的责任作者的观点，并不一定代表国家的康涅狄格州，康涅狄格州或CT创新公共健康部的官方意见，公司的资助者不参与研究设计，数据收集分析，决定，公布，或准备的稿子。

本程序是在按照与耶鲁IACUC要求。科学家应确保IACUC准则的批准，随后根据其机构的要求。 1。第1节。制备DNA，解决方案和玻璃吸管<ol><li>生成高纯度（OD 260/280&gt; 1.80），高浓度（微克/微升&gt; 2.5）内毒素的DNA。 </li><li>准备0.9％的生理盐水溶液和无菌过滤器，通过一个22微米的过滤器。 </li><li>将10厘米火抛光的高硼硅玻璃毛细管（OD：1.5毫米，1.10毫米）到一个模型中PP-830成?...

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在这里，我们详细新生儿SVZ电的技术，这种技术能够迅速和有力的标记和操作SVZ干细胞和它们的后代。有几个优点，在与其它技术相比，电穿孔具有。首先，考虑的重点标记的细胞，一个是能分辨细胞自主和非细胞自主的影响。其次，使用诱导系统的遗传操作，允许一个用来比较的效果之前或之后突触融合。此外，人们可以绕过使用多个质粒与含有loxP位点的质粒通过电穿孔CRE表达小鼠。第四，报...

<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>重抛光的高硼硅管</td><td>萨特仪器</td><td> BF150-110-10 </td><td></td></tr><tr><td>双级玻璃微普勒</td><td>成重</td><td> PC-10H </td><td></td></tr><tr><td>固绿</td><td>菲舍尔科学</td><td> 0521192205 </td><td></td></tr><tr><td> ECM 830方波脉冲发生器</td><td>哈佛设备</td><td> 45-0052 </td><td></td></tr><tr><td> Tweezertrodes </td><td>哈佛设备</td><td> 45-0488 </td><td></td></tr><tr><td>光纤光源</td><td> Fisher Scientific则</td><td> 12-562-36 </td><td></td></tr><tr><td>卤钨灯灯</td><td> USHIO美国公司</td><td> 1002247 </td><td></td></tr><tr><td> Picospritzer II </td><td>帕克工具</td><td> 052-0312-900 </td><td></td></tr></tbody></table>

neonatal subventricular zone electroporation

神经干细胞（NSCs）线的产后侧脑室，并产生多种细胞类型，包括神经元，星形胶质细胞和室管膜细胞。了解，利用其独特的修复潜力更好地理解大脑和中枢神经系统紊乱的分子通路的负责NSC的自我更新，承诺和分化是至关重要的。往前哺乳动物系统操纵的方法需要耗费时间和昂 ​​贵的基因工程的努力在整个动物的第2级。因此，绝大多数研究探讨NSC分子在体外或在无脊椎动物中的功能。 在这里，我们展示了技术操作简单，快速的新生儿的NPC，被称为新生儿脑室下区（SVZ）电。十年前制定类似的技术来研究胚胎神经干细胞，并资助研究cortical开发3,4。最近研究产后的啮齿动物前脑5-7。这种技术的结果SVZ神经干细胞在强大的标签和他们的后代。因此，产后SVZ电哺乳动物国科会基因工程提供了成本和时间效益的替代方案。

新生儿脑室下区电在几乎所有的放射状胶质连续的背，背外侧和侧脑室下区继“清扫”运动的镊子电极（ 图1E）的标签。然而，电可定制的要求，在各自的实验，但不扫电极，并使用特定的位置和方向，详细的视频。例如，由于背侧局部放射状胶质不同于那些衬心室的横向部分，一个可以选择只有一个单一的区域9,10 electroporate。更多特殊性的也可以使用较小的电极。 <p clas...

Watch this Scientific Journal Video about Neonatal Subventricular Zone Electroporation at JoVE.com

Neonatal Subventricular Zone Electroporation

我们展示出一种微创技术，新生儿脑室下区电。该技术包括新生幼崽的侧脑室注射质粒DNA和通电时提供神经干细胞和基因操纵

新生儿脑室下区电穿孔

Neural stem cells (NSCs) line the postnatal lateral ventricles and give rise to multiple cell types which include neurons, astrocytes, and ependymal ...

Research

JoVE Journal

Neuroscience

The Subventricular Zone En-face: Wholemount Staining and Ependymal Flow

脑室下区恩的脸：Wholemount染色和室管膜流量

The walls of the lateral ventricles contain the largest germinal region in the adult mammalian brain. The subventricular zone (SVZ) in these walls is an ...

科研

神经科学

In vivo Electroporation of Developing Mouse Retina

In vivo Electroporation of Developing Mouse Retina

在发展中国家小鼠视网膜体内电穿孔

The functional characterization of genes expressed during mammalian retinal development remains a significant challenge.  Gene targeting to generate ...

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection

在子宫内电穿孔：鼠标控制时空基因Transefection

In order to understand the function of genes expressed in specific region of the developing brain, including signaling molecules and axon guidance ...

Preparation of Acute Subventricular Zone Slices for Calcium Imaging

钙成像的急性下区片的制备

The subventricular zone (SVZ) is one of the two neurogenic zones in the postnatal brain. The SVZ contains densely packed cells, including neural ...

Neonatal Pial Surface Electroporation

新生儿软脑膜表面电穿孔

Over the past several years the pial surface has been identified as a germinal niche of importance during embryonic, perinatal and adult neuro- and ...

Subtype-selective Electroporation of Cortical Interneurons

皮质的interneurons的亚型选择性电穿孔

The study of central nervous system (CNS) maturation relies on genetic targeting of neuronal populations. However, the task of restricting the expression ...

Cell Sorting of Neural Stem and Progenitor Cells from the Adult Mouse Subventricular Zone and Live-imaging of their Cell Cycle Dynamics

神经干细胞和祖细胞来自成年小鼠脑室下区细胞分选和实时成像的细胞周期动力学

Neural stem cells (NSCs) in the subventricular zone of the lateral ventricles (SVZ) sustain olfactory neurogenesis throughout life in the mammalian brain. ...

Isolation and Culture of Adult Neural Stem Cells from the Mouse Subcallosal Zone

分离和成人神经干细胞的培养从鼠标Subcallosal区

Adult neural stem cells (aNSCs) can be used for the regeneration of damaged brain tissue. NSCs have the potential for differentiation and proliferation ...

Spinal Cord Neurons Isolation and Culture from Neonatal Mice

脊髓神经元从新生小鼠中分离和培养

We present a protocol for the isolation and culture of spinal cord neurons. The neurons are obtained from neonatal C57BL/6 mice and are isolated on ...

Continuous Video Electroencephalogram during Hypoxia-Ischemia in Neonatal Mice

新生小鼠缺氧缺血期间的连续视频脑电图

Hypoxia ischemia is the most common cause of neonatal seizures. Animal models are crucial for understanding the mechanisms and physiology underlying ...