Name: morphological analysis of drosophila larval peripheral sensory neuron dendrites and axons using genetic mosaics
Uploaded: 2011-11-07T12:00:00
Description: Watch this Scientific Journal Video about Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics at JoVE.com

作者感谢理化学研究所的资金。我们也感谢Cagri Yalgin，加Delandre，和周杰伦帕里什讨论遗传和免疫组化协议。

1.Preparation试剂<ol><li>准备钙离子免费HL3.1生理盐水28。 <ol><li> （毫米）：70氯化钠，氯化钾，氯化镁2 20 10 3，碳酸氢钠5 HEPES，115蔗糖，和5海藻糖; pH值7.2。过滤消毒，并储存于4 ° C。 注意：CA + +免费的解决方案可以防止肌肉收缩，在清扫。 </li></ol></li><li>设为聚- L -赖氨酸（PLL）盖玻片。 <ol><li> 4.2毫升水溶解100毫克PLL和300μL分装在Eppendorf管中，并冻?...

<ol>
	<li>Grueber, W. B., Jan, L. Y., Jan, Y. 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果蝇幼虫DA神经元模型提供了一个优良的遗传系统调查机制，控制神经元的形态和电路形成。 MARCM是一般用于标签和产生突变的DA神经元克隆。 MARCM，我们使用一个泛神经（ 如Gal4 的 C155）或DA神经元特异性司机。使用泛神经的驱动程序，它可以直接使用公共储备中心广泛使用的几个股票。然而，使用DA神经元的特定的驱动程序可以是有利的，因为标记克隆不会产生在中?...

<table border="1"><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td> SZX16荧光解剖镜下（GFPHQ过滤） </td><td>奥林巴斯</td><td> SZX16 </td><td></td></tr><tr><td>活昆虫钳</td><td>福斯特</td><td> 26030-10 </td><td></td></tr><tr><td> 26毫米x 76毫米抑郁症的载玻片</td><td> Toshinriko有限公司</td><td> T8 - R004 </td><td></td></tr><tr><td> Sylgard 184（或Silpot 184） </td><td>道康宁</td><td> 3097358-1004 </td><td></td></tr><tr><td>聚- L -赖氨酸</td><td>西格玛</td><td> P - 1524 </td><td>该产品已被证明最有效的</td></tr><tr><td> DPX安装培养​​基</td><td>西格玛</td><td> 44581 </td><td></td></tr><tr><td>兔抗GFP </td><td> Invitrogen公司</td><td> A - 11122 </td><td>稀释1:500 </td></tr><tr><td>大鼠抗- CD8 </td><td> Caltag </td><td> 5H10 </td><td>稀释1:200 </td></tr><tr><td>小鼠抗CD2 </td><td>抗体ð serotec </td><td> MCA443R </td><td>稀释1:700 </td></tr><tr><td>鼠抗- Fasciclin2 </td><td> DSHB </td><td> 1D4 </td><td>稀释1:10 </td></tr></table>

morphological analysis of drosophila larval peripheral sensory neuron dendrites and axons using genetic mosaics

神经系统的发育需要的神经元​​的地位和身份，准确的神经元类的特定的树突状发展和轴突布线，正确规范。最近的树突状树枝状（DA） 的果蝇幼虫周围神经系统（PNS）的感觉神经元已成为强大的遗传模型，在其中阐明神经元分化的一般和特定类的机制。有四个主要的DA神经元类（I - IV）1 。他们被命名为了增加枝蔓乔木的复杂性，并在其分化2-10的基因控制类特定的差异。 DA感官系统是一个实际的模型研究的树突状形态 11-13的控制背后的分子机制，因为：1）它可以利用强大的基因在果蝇中的可用工具，2）的优势，DA神经元枝蔓乔木展开在下方的光骑士尺寸只有2AR幼虫角质层很容易地可视化体内的高分辨率，3）类特定的树突形态的多样性有利于比较分析，发现控制简单与高度分支的树突树形成的关键要素，以及4）树突状乔木的定型不同的DA能神经元的形状有利于形态的统计分析。 DA神经元活动的修改输出的一个幼虫的运动中枢模式发生器 14-16 。不同的DA神经元类有不同的感觉方式，和它们的激活会引起不同的行为反应14,16-20。此外，不同类别发送到果蝇幼虫中枢神经系统的腹 ​​神经索（VNC） 的 21轴索预测刻板。这些预测终止两个DA神经元感觉方式和位置在体壁的树突状领域7,22地形意见书23。因此，DA的轴突预测的检查可用于澄清的内在机制， 地形测绘7,22,23，以及一个简单的电路调节幼虫运动14-17的接线。 我们在座的生成和分析遗传马赛克24标记的DA通过MARCM（马赛克Repressible细胞标记分析）1,10,25和FLP - 22,26,27技术（在图1总结）神经元的实用指南。

Watch this Scientific Journal Video about Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics at JoVE.com

Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics

感觉神经元的树突分枝<em&gt;果蝇</em&gt;幼虫周围神经系统是有用的模型，以澄清一般和神经元类特定的神经细胞分化的机制。我们提出了一个实用指南生成和分析树突状树枝状的神经元遗传马赛克。

形态分析果蝇幼虫外周感觉神经元树突和轴突使用遗传马赛克

Nervous system development requires the correct specification of neuron position and identity, followed by accurate neuron class-specific dendritic ...

Research

JoVE Journal

Neuroscience

Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics

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

Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall

Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall

免疫组化标记在感觉神经元树突的微管，气管，并在肌肉果蝇幼虫体壁

To understand how differences in complex cell shapes are achieved, it is important to accurately follow microtubule organization. The Drosophila larval ...

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

可视化和遗传操作在小鼠大脑皮层和使用海马树突和棘<em&gt;在子宫内</em&gt;电穿孔

In utero electroporation (IUE) has become a powerful technique to study the development of different regions of the embryonic nervous system 1-5. To date ...

Voltage-sensitive Dye Recording from Axons, Dendrites and Dendritic Spines of Individual Neurons in Brain Slices

电压敏感染料记录单个神经元在大脑切片的轴突，树突和树突棘

Understanding the biophysical properties and functional organization of single neurons and how they process information is fundamental for understanding ...

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 ...

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 ...

Production and Isolation of Axons from Sensory Neurons for Biochemical Analysis Using Porous Filters

生产和轴突隔离从感觉神经元的生化分析使用多孔过滤器

Neuronal axons use specific mechanisms to mediate extension, maintain integrity, and induce degeneration. An appropriate balance of these events is ...

Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells

Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells

去除<em&gt;果蝇</em从幼虫鱼片的感官神经元和表皮细胞的免疫荧光分析&gt;肌肉组织

Drosophila larval dendritic arborization (da) neurons are a popular model for investigating mechanisms of neuronal morphogenesis. Da neurons develop in ...

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 ...

A Microbiomechanical System for Studying Varicosity Formation and Recovery in Central Neuron Axons

中枢神经元轴突静脉曲张形成和恢复的 Microbiomechanical 系统研究

Axonal varicosities are enlarged structures along the shafts of axons with a high degree of heterogeneity. They are present not only in brains with ...