Name: whole mount preparation of the adult drosophila ventral nerve cord for giant fiber dye injection
Uploaded: 2011-06-04T14:00:00
Description: Watch this Scientific Journal Video about Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection at JoVE.com

描述该项目是从国家儿童健康和人类发展研究所的支持标记的内容完全是作者的责任，并不一定代表的国家儿童健康和人类发展或研究所的官方意见，赠款数R01HD050725美国国立卫生研究院。我们感谢他们的手稿和视频输入Godenschwege实验室以及芭芭拉Schreader成员。

1。腹神经索的剖析可在室温下进行解剖和染料的GF灌装。对于所有清扫步骤冷（4℃）生理盐水应使用减慢细胞代谢和维持神经元存活更长的时间。 <ol><li>麻醉与CO 2成蝇（4天或以上），或阴寒他们，直到他们被固定在冰上。麻醉用冰块将需要更多的时间（最多30分钟），但将保持更长的时间固定的苍蝇。 </li><li>放置一个飞在Sylgard涂层的培养皿。剥离需要使用高倍率...

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	<li>Power, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+thoracico-abdominal+nervous+system+of+an+adult+insect+Drosophila+melanogaster.">The thoracico-abdominal nervous system of an adult insect Drosophila melanogaster.</a> J. Comp. Neurol. 88, 347-409 (1948).</li><li>Allen, M. J., Godenschwege, T. A., Tanouye, T. A., Phelan, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Making+an+escape:+development+and+function+of+the+Drosophila+giant+fibre+system.">Making an escape: development and function of the Drosophila giant fibre system.</a> Semin. Cell Dev. Biol. 17, 31-41 (2006).</li><li>Boerner, J., Duch, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Average+shape+standard+atlas+for+the+adult+Drosophila+ventral+nerve+cord.">Average shape standard atlas for the adult Drosophila ventral nerve cord.</a> J. Comp. Neurol. 518, 2437-2455 (2010).</li><li>Gu, H., O'Dowd, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cholinergic+synaptic+transmission+in+adult+Drosophila+Kenyon+cells+in+sity.">Cholinergic synaptic transmission in adult Drosophila Kenyon cells in sity.</a> J. Neurosci. 26, 265-272 (2006).</li><li>Boerner, J., Godenschwege, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+for+the+Drosophila+ventral+nerve+cord+standard+in+neuronal+circuit+reconstruction+and+in-depth+analysis+of+mutant+morphology.">Application for the Drosophila ventral nerve cord standard in neuronal circuit reconstruction and in-depth analysis of mutant morphology.</a> J. Neurogent. 24, 158-1567 (2010).</li><li>Godenschwege, T. A., Kristiansen, L. V., Uthaman, S. B., Hortsch, M., Murphey, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+conderved+role+for+Drosophila+Neuroglian+and+human+L1-CAM+in+central-synapse+formation.">A conderved role for Drosophila Neuroglian and human L1-CAM in central-synapse formation.</a> Curr. Biol. 16, 12-23 (2006).</li><li>Uthaman, S. B., Godenschwege, T. A., Murphey, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mechanism+distinct+from+highwire+for+the+Drosophila+ubiquitin+conjugase+bendless+in+synaptic+growth+and+maturation.">A mechanism distinct from highwire for the Drosophila ubiquitin conjugase bendless in synaptic growth and maturation.</a> J. Neurosci. 28, 8615-8623 (2008).</li><li>Storkebaum, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dominant+mutations+in+the+tyrosyl-tRNA+synthetase+gene+recapitulate+in+Drosophila+features+of+human+Charcot-Marie-Tooth+neuropathy.">Dominant mutations in the tyrosyl-tRNA synthetase gene recapitulate in Drosophila features of human Charcot-Marie-Tooth neuropathy.</a> Proc. Natl. Acad. Sci. USA. 106, 11782-11787 (2009).</li><li>Allen, M. J., Godenschwege, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+recordings+from+the+Drosophila+giant+fiber+system+(GFS).">Electrophysiological recordings from the Drosophila giant fiber system (GFS).</a> Cold Spring Harb. Protoc. 2010, pdb.prot5453-pdb.prot5453 (2010).</li><li>Augustin, H., Allen, M. J., Partridge, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+Recordings+from+the+Giant+Fiber+Pathway+of+D.+melanogaster.">Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster.</a> J. Vis. Exp. , (2011).</li></ol>

没有从果蝇体内提取的神经系统，可以解剖。这有两个好处，第一，夹层导致神经系统的损害不大，并保持连接第二，大多数神经肌肉和感觉器官。表演解剖描述的那样，准备向前伸直政府飞行服务队的染料标记的样品。为方便起见，政府飞行服务队的运动神经元突触后留附着的肌肉。轴突，因此，没有损坏，保持神经元的存活更长的时间。此外，对神经系统的损害是可能扰乱间隙连接功能，从?...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论</th></tr><tr><td> Sylgard 184有机硅弹性体套件</td><td>道康宁</td><td></td><td></td></tr><tr><td>解剖镜下</td><td> AmScope </td><td> SM - 2TZ </td><td></td></tr><tr><td> Vannas剪刀超细</td><td>学术仪器</td><td> VS1023 </td><td></td></tr><tr><td>杜蒙钳Dumontstar 55 </td><td>精细科学的工具</td><td> 11295-51 </td><td></td></tr><tr><td>奥斯特利茨昆虫引脚</td><td>精细科学的工具</td><td> 26002-10 </td><td>直径0.1毫米</td></tr><tr><td>硼硅玻璃电极</td><td>世界精密仪器</td><td> 1B100F - 4 </td><td></td></tr><tr><td>垂直移液器拖轮700C </td><td>大卫Kopf仪器</td><td>型号700C </td><td></td></tr><tr><td>路西法黄河CH达丽星盐</td><td>西格玛</td><td> L0259 </td><td> 1％H 2 O </td></tr><tr><td>固定舞台直立显微镜和相机</td><td>尼康</td><td> FN - 1和DS - U2的相机</td><td></td></tr><tr><td>计划Fluor公司10X空气目的</td><td>尼康</td><td>原讼法庭计划Fluor公司10X NA 0.3的WD16毫米</td><td></td></tr><tr><td>福陆公司40X水浸渍目的</td><td>尼康</td><td>原讼法庭福陆公司40X W NA 0.80的WD2.0毫米</td><td></td></tr><tr><td> 3D液压微操作机器人</td><td> Narishige国际</td><td> MW0 - 3型</td><td></td></tr><tr><td>放大器</td><td>入门仪器公司</td><td>型号5A </td><td></td></tr></tbody></table>

whole mount preparation of the adult drosophila ventral nerve cord for giant fiber dye injection

为了分析神经元轴突和树突状的形态，这是必须获得准确的神经元结构的标签。准备以及标记的样品几乎没有组织损伤，使我们能够分析细胞形态和相互比较个别样品，从而使突变异常的识别。 在证明清扫方法的神经系统仍然主要是里面的成人飞。通过背侧切口，腹部和胸部开了，大部分内脏器官都将被删除。只有背侧，腹侧神经线（VNC）和宫颈结缔组织（CVC）的巨型纤维（GFS）1大轴突外露，而大脑含有的GF胞体和树突仍然完整的头2 。在这个准备，最神经的VNC应保持连接到他们的肌肉。 剥离后，细胞内的巨型纤维（GF）用荧光染料填充证明。 CVC GF轴突位于背表面，从而可以很容易地与微分干涉对比（DIC）的光学显微镜下可视化。这使得在此网站与染料的GF轴突注射标签VNC的轴突和他们的终端包括整个的GF。该方法结果可靠和强大的政府飞行服务队的允许填充啶显微镜后，立即进行成像神经元染色。另外，使用的标准的免疫组织化学程序 3适用于高分辨率的共聚焦显微镜，荧光信号可增强。

Watch this Scientific Journal Video about Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection at JoVE.com

Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection

一个<em&gt;在体内</em&gt;成人解剖<em&gt;果蝇</em&gt;腹神经索（VNC）是证明。这种特殊的解剖方法会导致损伤小到VNC允许的高分辨率成像的荧光染料的巨纤维神经元的后续标签。

成人整装准备果蝇巨纤维染料注射腹侧神经索

To analyze the axonal and dendritic morphology of neurons, it is essential to obtain accurate labeling of neuronal structures. Preparing well labeled ...

Research

JoVE Journal

Neuroscience

Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection

Visualization of the Embryonic Nervous System in Whole-mount Drosophila Embryos

Visualization of the Embryonic Nervous System in Whole-mount Drosophila Embryos

在全安装的胚胎神经系统的可视化果蝇胚胎

The Drosophila embryo is an attractive model system for investigating the cellular and molecular basis of neuronal development. Here we describe the ...

科研

神经科学

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

从巨纤维通路的电生理记录 D。果蝇</em

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" ...

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

电检测屏幕的配对Nanoinjection和生物活性化合物的使用果蝇巨人纤维系统

Screening compounds for in vivo activity can be used as a first step to identify candidates that may be developed into pharmacological agents1,2. We ...

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 ...

DiI-Labeling of DRG Neurons to Study Axonal Branching in a Whole Mount Preparation of Mouse Embryonic Spinal Cord

DII的标签DRG神经元的研究轴突分支在小鼠胚胎脊髓整装准备

Here we  present a technique to label the trajectories of small groups of DRG neurons  into the embryonic spinal cord by diffusive staining using the ...

Dissection of the Transversus Abdominis Muscle for Whole-mount Neuromuscular Junction Analysis

Dissection of the Transversus Abdominis Muscle for Whole-mount Neuromuscular Junction Analysis

全山神经肌肉交界分析的 横向解剖阿布多米尼斯 肌肉

Analysis of neuromuscular junction morphology can give important insight into the physiological status of a given motor neuron. Analysis of thin flat ...

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

嗅觉受体神经元在全山免疫标记<em&gt;果蝇</em&gt;天线

Odorant molecules bind to their target receptors in a precise and coordinated manner. Each receptor recognizes a specific signal and relays this ...

An In Vitro Adult Mouse Muscle-nerve Preparation for Studying the Firing Properties of Muscle Afferents

An In Vitro Adult Mouse Muscle-nerve Preparation for Studying the Firing Properties of Muscle Afferents

一<em&gt;在体外</em&gt;成年小鼠肌肉，神经准备留学肌肉传入放电的放电特性

Muscle sensory neurons innervating muscle spindles and Golgi tendon organs encode length and force changes essential to proprioception. Additional ...

Whole Mount Dissection and Immunofluorescence of the Adult Mouse Cochlea

全山解剖和成年小鼠耳蜗的免疫

The organ of Corti, housed in the cochlea of the inner ear, contains mechanosensory hair cells and surrounding supporting cells which are organized in a ...

The Preparation of Oblique Spinal Cord Slices for Ventral Root Stimulation

斜脊髓切片用于神经前根刺激的制备

Electrophysiological recordings from spinal cord slices have proven to be a valuable technique to investigate a wide range of questions, from cellular to ...