Name: laser capture microdissection of drosophila peripheral neurons
Uploaded: 2010-05-24T17:00:00
Description: Watch this Scientific Journal Video about Laser Capture Microdissection of Drosophila Peripheral Neurons at JoVE.com

我们感谢博士。裕农扬和韦斯Grueber提供援助，带LCM飞的股票，在本研究中使用的，弗吉尼亚Espina，伊曼纽尔Petricoin博士和博士兰斯利奥塔。作者承认这项研究的支持（DNC）和乔治梅森大学教务长办公室（EPRI）的托马斯F和凯特米勒Jeffress纪念信托。

对果蝇 LCM的外周神经元的一般意见允许来自6个小时，一个星期或液晶显示模块不再取决于组织类型和所需的细胞数量。 所有的程序都进行严格的无RNase条件以下的标准程序。幼虫表达无论是21 - 7 - GAL4，无人机mCD8：：GFP或PPK - GAL4 UAS - mCD8：：GFP转基因记者线，这些实验中使用。 1。准备幼虫<ol><li>收集30-40年龄相?...

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本协议描述了我们对果蝇通过LCM的外周神经元的隔离的优化方法。虽然这LCM协议旨在为特定的隔离单，类特定或多个果 ​​蝇的大神经元从3龄幼虫阶段的发展，稍作修改的协议可以很容易地适用于其他果蝇的细胞类型的捕捉从所有发展使用不同的GAL4的阶段 ， 记者UAS - mCD8 - GFP转基因标签的细胞类型或类型的利益。我们的研究结果表明，激光捕获微解剖DA?...

设备： <ul><li>低温恒温器</li><li> 50 ml锥形管幻灯片固定，漂洗，胰蛋白酶处理和乙醇/二甲苯脱水</li><li> -80 ° C冰箱</li><li>孵化器</li><li> PixCell IIe上LCM仪器与福陆公司的300 epifluorescence光学优化EGFP（分子器件，分子器件） </li></ul>

laser capture microdissection of drosophila peripheral neurons

树突状树枝状（DA）神经元的果蝇周围神经系统（PNS）在调查的基础类特定的树突形态发生 1,2的分子机制提供一个极好的的模型系统。为了方便类特定的DA神经元发展的分子生物学分析，至关重要的是在一个纯粹的人口获得这些细胞。虽然一系列不同的细胞和组织特异性的RNA分离技术中存在的果蝇细胞，包括基于磁珠细胞净化3,4，荧光激活细胞分选（FACS）5-8，和RNA结合蛋白为基础的战略9 ，概无这些方法可以很容易地利用空间精度与高度隔离的单个或多个类特定的果蝇DA能神经元。激光捕获显微切割（LCM）已成为一个非常强大的工具，可以用来从组织切片中分离出特定的细胞类型，具有高度的空间分辨率和精度。然后，可以从分离的细胞获得的RNA用于包括在一个给定的细胞类型10-16的QRT - PCR和基因芯片表达谱分析。迄今为止，液晶显示模块没有被广泛应用在果蝇的组织和细胞17,18，包括发展第三龄幼虫阶段的DA能神经元的分析。 在这里，我们目前对果蝇的大神经元使用的红外（IR）类LCM的隔离，我们优化的协议。这种方法允许单一，类特定或多个的DA能神经元具有高特异性和空间分辨率的捕捉。年龄相匹配的三龄幼虫表达的UAS - mCD8：绿色荧光蛋白 19基因的控制下的IV级DA神经元具体PPK - GAL4 20驱动程序或泛DA神经元特异性21 - 7 - GAL4 21驱动程序是用于这些实验。从孤立的DA能神经元获得的RNA是非常高的质量，并可以直接用于下游应用，包括QRT - PCR或芯片分析。此外，这种液晶显示模块协议，可以方便地用于捕捉到其他果蝇的细胞类型的不同阶段的发展依赖于具体，GAL4驱动 GFP的表达模式的细胞类型的。

Watch this Scientific Journal Video about Laser Capture Microdissection of Drosophila Peripheral Neurons at JoVE.com

Laser Capture Microdissection of Drosophila Peripheral Neurons

在这个视频文章中，我们目前的方法隔离单个或多个<em&gt;果蝇</em&gt; DA神经元使用红外线（IR）的捕获激光捕获显微切割（LCM）的类三龄幼虫。从孤立的神经元获得的RNA可以很容易地用于下游应用，包括QRT - PCR或芯片分析。

激光捕获显微切割技术的果蝇外周神经元

The dendritic arborization (da) neurons of the Drosophila peripheral nervous system (PNS) provide an excellent model system in which to investigate the ...

Research

JoVE Journal

Neuroscience

Laser Capture Microdissection of Drosophila Peripheral Neurons

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

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

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 ...

科研

神经科学

Non-Laser Capture Microscopy Approach for the Microdissection of Discrete Mouse Brain Regions for Total RNA Isolation and Downstream Next-Generation Sequencing and Gene Expression Profiling

Non-Laser Capture Microscopy Approach for the Microdissection of Discrete Mouse Brain Regions for Total RNA Isolation and Downstream Next-Generation Sequencing and Gene Expression Profiling  |

总RNA的分离和下游下一代测序和基因表达谱的非激光捕获显微切割离散小鼠脑地区显微镜方法

As technological platforms, approaches such as next-generation sequencing, microarray, and qRT-PCR have great promise for expanding our understanding of ...

Preparation of Drosophila Central Neurons for in situ Patch Clamping

Preparation of Drosophila Central Neurons for in situ Patch Clamping

制备<em&gt;果蝇</em&gt;中央神经元<em在原位</em&gt;膜片钳

Short generation times and facile genetic techniques make the fruit fly Drosophila melanogaster an excellent genetic model in fundamental neuroscience ...

Laser Capture Microdissection of Enriched Populations of Neurons or Single Neurons for Gene Expression Analysis After Traumatic Brain Injury

激光捕获显微切割的丰富种群的创伤性脑损伤后神经元或单个神经元的基因表达分析

Long-term cognitive disability after TBI is associated with injury-induced neurodegeneration in the hippocampus-a region in the medial temporal lobe that ...

Laser Capture Microdissection of Neurons from Differentiated Human Neuroprogenitor Cells in Culture

神经元的分化的人类Neuroprogenitor细胞培养激光捕获显微切割

Neuroprogenitor cells (NPCs) isolated from the human fetal brain were expanded under proliferative conditions in the presence of epidermal growth factor ...

Production of RNA for Transcriptomic Analysis from Mouse Spinal Cord Motor Neuron Cell Bodies by Laser Capture Microdissection

通过激光捕获微分路，从小鼠脊髓运动神经细胞体中生成RNA进行转录分析

Preparation of high-quality RNA from cells of interest is critical to precise and meaningful analysis of transcriptional differences among cell types or ...

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 ...

Olfactory Neurons Obtained through Nasal Biopsy Combined with Laser-Capture Microdissection: A Potential Approach to Study Treatment Response in Mental Disorders

通过鼻腔活检获得联合激光捕获显微切割的嗅觉神经元：一种可能的方法来研究治疗反应的精神障碍

Bipolar disorder (BD) is a severe neuropsychiatric disorder with poorly understood pathophysiology and typically treated with the mood stabilizer, lithium ...

Laser Capture Microdissection - A Demonstration of the Isolation of Individual Dopamine Neurons and the Entire Ventral Tegmental Area

激光捕获显微切割 - 个人多巴胺神经元的分离和整个腹侧被盖区的示范

Laser capture microdissection (LCM) is used to isolate a concentrated population of individual cells or precise anatomical regions of tissue from tissue ...

Visualization of the Axonal Projection Pattern of Embryonic Motor Neurons in Drosophila

Visualization of the Axonal Projection Pattern of Embryonic Motor Neurons in Drosophila

胚胎运动神经元轴突投影模式的可视化<em&gt;果蝇</em

The establishment of functional neuromuscular circuits relies on precise connections between developing motor axons and target muscles. Motor neurons ...