Name: axoplasm isolation from rat sciatic nerve
Uploaded: 2010-09-24T17:00:00
Description: Watch this Scientific Journal Video about Axoplasm Isolation from Rat Sciatic Nerve at JoVE.com

此协议允许轴浆隔离与胶质细胞和血管组织污染最小化。该方法的产量约为70-100微克每8-10周龄Wistar大鼠共轴浆蛋白质。 1。解剖坐骨神经神经<ol><li>安乐死两个大鼠颈椎脱位的CO 2吸入。触诊缺乏心跳开始清扫前确认死亡。 </li><li>拭清扫面积70％的乙醇。 </li><li>切开皮肤，单独的肌肉和隔离坐骨神经使用不破坏血管的剪刀和镊子仔细。解剖坐骨神经两个（每个约1.5厘...

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	<li>Hanz, S., Perlson, E., Willis, D., Zheng, J. Q., Massarwa, R., Huerta, J. J., Koltzenburg, M., Kohler, M., van-Minnen, J., Twiss, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axoplasmic+importins+enable+retrograde+injury+signaling+in+lesioned+nerve.">Axoplasmic importins enable retrograde injury signaling in lesioned nerve.</a> Neuron. 40, 1095-1104 (2003).</li><li>Michaelevski, I., Medzihradszky, K. F., Lynn, A., Burlingame, A. L., Fainzilber, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axonal+transport+proteomics+reveals+mobilization+of+translation+machinery+to+the+lesion+site+in+injured+sciatic+nerve.">Axonal transport proteomics reveals mobilization of translation machinery to the lesion site in injured sciatic nerve.</a> Mol Cell Proteomics. 9 (5), 976-987 (2010).</li><li>Rishal, I., Michaelevski, I., Rozenbaum, M., Shinder, V., Medzihradszky, K. F., Burlingame, A. L., Fainzilber, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Axoplasm+isolation+from+peripheral+nerve.">Axoplasm isolation from peripheral nerve.</a> Dev Neurobiol. 70, 126-133 (2010).</li><li>Twiss, J. L., Fainzilber, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ribosomes+in+axons--scrounging+from+the+neighbors.">Ribosomes in axons--scrounging from the neighbors.</a> Trends Cell Biol. 19, 236-243 (2009).</li></ol>

生化和蛋白质组学分析已确认，此过程中降低血清和神经胶质细胞污染3相比，轴浆隔离前面描述的方法，通过机械挤压 1 。我们已经用这个轴浆隔离协议，探讨坐骨神经损伤后 2的动力蛋白为基础的逆行信号，并期望它有许多其他进程的探索，在成人末梢神经轴突的神经胶质细胞相互作用4实用。

<ul><li>雄性Wistar大鼠（8-10周龄） </li><li> PBS 0.2X和PBS 1X解决方案包含蛋白酶抑制剂（Roshe）2片，每50毫升</li><li> Eppendorfs为1.5毫升</li><li>消毒塑料菜35毫米</li><li>解剖工具包括：剪刀和精细镊子</li><li>双目和台灯</li></ul>抗体鼠抗GFAP阳性克隆GA - 5是从Sigma（G6171）。兔抗白蛋白是从Cedarlane（CLAG5140）。鼠抗-微管蛋白β3和兔抗gERK自Sigma（分别为T2200和M5670）。

axoplasm isolation from rat sciatic nerve

纯周围神经轴突胞浆（轴浆）的分离是许多生物过程的生化研究的关键。在这篇文章中，我们展示和描述基于以下步骤从成年大鼠坐骨神经神经轴浆隔离协议：（1）清扫的神经分册和结缔组织分离;（2）潜伏期短段神经分册低渗介质释放和裂解髓鞘轴突非结构;（3）提取其余的轴突丰富的材料。该制剂的蛋白质组学和生化特性，已证实为轴突组成部分的高度浓缩。

Watch this Scientific Journal Video about Axoplasm Isolation from Rat Sciatic Nerve at JoVE.com

Axoplasm Isolation from Rat Sciatic Nerve

我们展示基于神经分册，并在低渗介质孵化剥离释放髓鞘裂解非轴索结构，其余轴突丰富的材料提取成年大鼠坐骨神经神经轴浆隔离协议。

从大鼠坐骨神经轴浆隔离

Isolation of pure axonal cytoplasm (axoplasm) from peripheral nerve is crucial for biochemical studies of many biological processes. In this article, we ...

Research

JoVE Journal

Neuroscience

Reproducible Mouse Sciatic Nerve Crush and Subsequent Assessment of Regeneration by Whole Mount Muscle Analysis

重复性小鼠坐骨神经挤压和再生的后续评估整装肌肉分析

Regeneration in the peripheral nervous system (PNS) is widely studied both for its relevance to human disease and to understand the robust regenerative ...

科研

神经科学

In Vivo Electrophysiological Measurements on Mouse Sciatic Nerves

In Vivo Electrophysiological Measurements on Mouse Sciatic Nerves

在对小鼠坐骨神经体内的电测量

Electrophysiological studies allow a rational classification of various neuromuscular diseases and are of help, together with neuropathological ...

Minimally-invasive Technique for Injection into Rat Optic Nerve

微创技术注入到老鼠视神经

The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central ...

Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination

在实验视神经脱髓鞘模型大鼠视觉诱发电位记录

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to ...

In Vivo Gene Transfer to Schwann Cells in the Rodent Sciatic Nerve by Electroporation

In Vivo Gene Transfer to Schwann Cells in the Rodent Sciatic Nerve by Electroporation

体内基因转移到雪旺细胞在啮齿动物坐骨神经电穿孔

The formation of the myelin sheath by Schwann cells (SCs) is essential for rapid conduction of nerve impulses along axons in the peripheral nervous ...

In Vivo Electrophysiological Measurement of the Rat Ulnar Nerve with Axonal Excitability Testing

In Vivo Electrophysiological Measurement of the Rat Ulnar Nerve with Axonal Excitability Testing

在体内轴突兴奋性试验对大鼠尺神经的电生理测量

Electrophysiology enables the objective assessment of peripheral nerve function in vivo. Traditional nerve conduction measures such as amplitude and ...

Analysis of Immune Cells in Single Sciatic Nerves and Dorsal Root Ganglion from a Single Mouse Using Flow Cytometry

应用流式细胞仪分析单侧坐骨神经和背根神经节的免疫细胞

Nerve-resident immune cells in the peripheral nervous system (PNS) are essential to maintaining neuronal integrity in a healthy nerve. The immune cells of ...

Facial Nerve Surgery in the Rat Model to Study Axonal Inhibition and Regeneration

大鼠模型中的面部神经外科研究同氧抑制与再生

This protocol describes consistent and reproducible methods to study axonal regeneration and inhibition in a rat facial nerve injury model. The facial ...

Use of In Vivo Single-fiber Recording and Intact Dorsal Root Ganglion with Attached Sciatic Nerve to Examine the Mechanism of Conduction Failure

使用 Vivo 单纤维记录和带附骨神经的固定背道根结条，以检查传导失败机制

Single-fiber recording has been a classical and effective electrophysiological technique over the last few decades because of its specific application for ...

3D Kinematic Analysis for the Functional Evaluation in the Rat Model of Sciatic Nerve Crush Injury

坐骨神经粉碎损伤大鼠模型中功能评估的3D运动学分析

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of ...