Name: visualization of larval segmental nerves in 3rd instar drosophila larval preparations
Uploaded: 2010-09-29T17:00:00
Description: Watch this Scientific Journal Video about Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations at JoVE.com

SG是从美国纽约州立大学布法罗分校和约翰R. Oishei基金会的资金支持。

1。试剂的制备<ol><li>准备1X夹层缓冲区使用128毫米氯化钠，4MM，4MM 氯化钙氯化镁2，2毫米氯化钾，5MM HEPES和36mm的蔗糖。 PH值至7.2的解决方案和过滤消毒。 </li><li>准备使用1:1稀释16％的甲醛和1个夹层缓冲固定液。 </li><li>准备1X PBT的使用1X的磷酸盐缓冲液（PBS），PH值在7.2和Triton X - 100的一个比例为1:100。 </li><li>准备5％牛血清白蛋白（BSA）的0.01％，钠叠氮化物在1X PBS。 </li></ol><p cla...

<ol>
	<li>Gunawardena, S., Goldstein, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disruption+of+Axonal+Transport+and+Neuronal+Viability+by+Amyoid+Precursor+Protein+Mutations+in+Drosophila.">Disruption of Axonal Transport and Neuronal Viability by Amyoid Precursor Protein Mutations in Drosophila.</a> Neuron. 32 (2), 389-401 (2001).</li><li>Gunawardena, S., Her, L. S., Brusch, R. G., Laymon, R. A., Niesman, I. R., Gordesky-Gold, B., Sintasath, L., Bonini, N. M., Goldstein, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disruption+of+axonal+transport+by+loss+of+huntingtin+or+expression+of+pathogenic+polyQ+proteins+in+Drosophila.">Disruption of axonal transport by loss of huntingtin or expression of pathogenic polyQ proteins in Drosophila.</a> Neuron. 40 (1), 1-2 (2003).</li><li>Hurd, D. D., Saxton, W. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinesin+mutations+cause+motor+neuron+disease+phenotypes+by+disrupting+fast+axonal+transport+in+Drosophila.">Kinesin mutations cause motor neuron disease phenotypes by disrupting fast axonal transport in Drosophila.</a> Genetics. 144 (3), 1075-1085 (1996).</li><li>Zinsmaier, K. E., Eberle, K. K., Buchner, E., Walter, N., Benzer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Paralysis+and+early+death+in+cysteine+string+protein+mutants+of+Drosophila.">Paralysis and early death in cysteine string protein mutants of Drosophila.</a> Science. 263, 977-980 (1994).</li></ol>

果蝇幼虫节段性神经是一个功能强大的系统研究轴突运输的机制。如果正确执行， 第三龄幼虫清扫的中枢神经系统，百佳，和相关的组织，如肌肉和NMJs，的所有组件都可以在一个单一的幼虫研究。我们已经适应了从赫德和萨克斯顿（1996年）出版的一本议定书。该协议还可以适应测试其他神经元抗体，但抗体的优化应该做。小学和中学的抗体孵育时间可以改变，可以添加或阻塞的?...

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visualization of larval segmental nerves in 3rd instar drosophila larval preparations

果蝇作为研究的发展和神经系统的功能强大的模型系统，特别是由于其方便的遗传学和完全测序的基因组，新兴。此外，幼虫的神经系统是一个理想的模型系统研究轴突运输机制，幼虫节段性神经束包含其细胞位于大脑和身体的长度沿结束他们的神经末梢内的机构轴突。在这里，我们描述幼虫节段性神经突触囊泡蛋白的可视范围内的程序。如果做得正确，对神经系统的所有组件，连同相关的组织，如肌肉和NMJs，保持完好，无破损，并准备要可视化。 第三龄幼虫携带各种突变解剖，固定，与突触囊泡抗体孵育，可视化和比较野生型幼虫。此过程可适应几种不同的突触或神经抗体和多种蛋白质分布的变化，可以很容易地观察到幼虫节段性神经内。

Watch this Scientific Journal Video about Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations at JoVE.com

Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations

果蝇幼虫提供了一个理想的模型系统调查内幼虫节段性神经轴突运输的机制。使用此过程中， 第三龄幼虫携带各种突变可以比野生型幼虫。

3幼虫节段性神经的可视化 RD龄果蝇幼虫的筹备工作

Drosophila melanogaster is emerging as a powerful model system for studying the development and function of the nervous system, particularly because of ...

Research

JoVE Journal

Neuroscience

Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

In vivo Visualization of Synaptic Vesicles Within Drosophila Larval Segmental Axons

在果蝇幼虫分部轴突在体内的突触小泡的可视化

Elucidating the mechanisms of axonal transport has shown to be very important in determining how defects in long distance transport affect different ...

科研

神经科学

Mapping and Application of Enhancer-trap Flippase Expression in Larval and Adult Drosophila CNS

Mapping and Application of Enhancer-trap Flippase Expression in Larval and Adult Drosophila CNS

幼虫和成人测绘和增强子陷阱Flippase表达中的应用果蝇中枢神经系统

The Gal4/ UAS binary method is powerful for gene and neural circuitry manipulation in Drosophila. For most neurobiological studies, however, Gal4 ...

Dissection and Immunohistochemistry of Larval, Pupal and Adult Drosophila Retinas

Dissection and Immunohistochemistry of Larval, Pupal and Adult Drosophila Retinas

幼虫，蛹和成人的解剖和免疫组化<em&gt;果蝇</em&gt;视网膜

The compound eye of Drosophila melanogaster consists of about 750 ommatidia (unit eyes). Each ommatidium is composed of about 20 cells, including ...

Forebrain Electrophysiological Recording in Larval Zebrafish

斑马鱼幼虫的前脑的电生理记录

Epilepsy affects nearly 3 million people in the United States and up to 50 million people worldwide. Defined as the occurrence of spontaneous unprovoked ...

Functional Analysis of the Larval Feeding Circuit in Drosophila

Functional Analysis of the Larval Feeding Circuit in Drosophila

在幼虫馈电电路的功能分析果蝇</i

The serotonergic  feeding circuit in Drosophila  melanogaster larvae can be used to investigate neuronal substrates of  critical importance during the ...

Live Imaging of Drosophila Larval Neuroblasts

Live Imaging of Drosophila Larval Neuroblasts

的实时成像<em&gt;果蝇</em&gt;幼虫神经母细胞

Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given ...

Why Quantification Matters: Characterization of Phenotypes at the Drosophila Larval Neuromuscular Junction

Why Quantification Matters: Characterization of Phenotypes at the Drosophila Larval Neuromuscular Junction

为什么量化事项：表型的表征在<em&gt;果蝇</em&gt;幼虫神经肌肉接头

Most studies on morphogenesis rely on qualitative descriptions of how anatomical traits are affected by the disruption of specific genes and genetic ...

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

Using Linear Agarose Channels to Study Drosophila Larval Crawling Behavior

用线性琼脂糖渠道研究<em&gt;果蝇</em&gt;幼虫爬行行为

Drosophila larval crawling is emerging as a powerful model to study neural control of sensorimotor behavior. However, larval crawling behavior on flat ...

Single-cell Resolution Fluorescence Live Imaging of Drosophila Circadian Clocks in Larval Brain Culture

Single-cell Resolution Fluorescence Live Imaging of Drosophila Circadian Clocks in Larval Brain Culture

单细胞分辨荧光活成像在幼虫脑培养中的果蝇昼夜节律时钟

The circadian pacemaker circuit orchestrates rhythmic behavioral and physiological outputs coordinated with environmental cues, such as day/night cycles. ...

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons

跟踪果蝇幼虫行为对嗅神经元 Optogenetic 刺激的反应

The ability of insects to navigate toward odor sources is based on the activities of their first-order olfactory receptor neurons (ORNs). While a ...