Name: paired nanoinjection and electrophysiology assay to screen for bioactivity of compounds using the drosophila melanogaster giant fiber system
Uploaded: 2012-04-15T14:00:00
Description: Watch this Scientific Journal Video about Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System at JoVE.com

我们要承认在马里实验室和Godenschwege的实验室，特别是艾琳米泽成员，征求意见，并帮助这个协议。这项工作是由国立神经疾病研究所和中风授予R21NS06637 FM和TAG; AB公司是由国家科学基金会奖号码082925，URM的资助：为未来的研究人员结合生物学。

1。电钻机设置<ol><li>详细描述了为电钻机设置所需的设备由奥古斯丁等 。在该杂志14。为所需的电生理仪器的详细解释，请参阅本文。 </li><li>加入了第六的微操作机器人，其持有的nanoinjector修改先前所描述的电钻机设置14。为方便应置于两者之间的刺激电极显微操作，在图2所示的动物的头。 </li><li>在实验开始前，确保你有一个旋转轴与所有显微操作的舒适?...

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这里介绍的nanoinjection /电生物活性，可快速筛选的化合物在果蝇的神经系统。这是一个小说在体内的技术，需要少量的化合物，在一个特点的神经电路引起的各种分子靶点的作用。这种方法可以用来测试不同的化合物的生物活性，从未知毒素市售药物制剂。 在这里，我们证明了我们的检测功能，使用工作重点，其中有巨人光纤系统（GFS）的果蝇（ 图5）效?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>录音玻璃电极：硼硅玻璃毛细管</td><td>世界精密仪器</td><td> 1B100F-4 </td><td> 1.0毫米外径，0.58毫米编号</td></tr><tr><td>刺激</td><td>草仪器</td><td>型号S48 </td><td></td></tr><tr><td>放大器</td><td>获得仪器公司</td><td>型号5A </td><td></td></tr><tr><td>数据采集​​软件：Digidata </td><td>分子器件</td><td>型号1440A </td><td></td></tr><tr><td>数据采集​​软件：pCLAMP </td><td>分子器件</td><td>第10版</td><td></td></tr><tr><td>体视显微镜瓦特第i个光纤显微镜环形灯</td><td> AmScope </td><td> SM-4T 型号HL250-AR </td><td></td></tr><tr><td>解剖的安装范围</td><td> AmScope </td><td> SM-2TZ </td><td></td></tr><tr><td>风筝手动微操作手和倾斜基地</td><td>世界精密仪器</td><td>型号：M3301 风筝：＃风筝M3-L的型号</td><td></td></tr><tr><td> 果蝇野生的10E基因型（野生型菌株） </td><td>布卢明顿联合中心</td><td>股票＃3892 </td><td></td></tr><tr><td>立式吸管拔</td><td>大卫Kopf仪器</td><td>型号700C </td><td></td></tr><tr><td>注射的玻璃micropipettes：硼硅玻璃毛细管</td><td>世界精密仪器</td><td>目录＃4878 </tD&gt; <td> 1.14毫米外径，内径0.5mm </td></tr><tr><td>硅油</td><td>费舍尔</td><td>目录＃S159-500 </td><td></td></tr><tr><td>倒角</td><td>萨特仪器有限公司</td><td>克拉的棕色类型 型号＃BV-10 </td><td></td></tr><tr><td> nanoliter2000 </td><td>世界精密仪器</td><td>目录＃B203XVY </td><td></td></tr><tr><td>蓝色食用色素</td><td>麦考密克</td><td> N / A </td><td>配料：水，丙烯乙二醇，FD＆C蓝色1和0.1％丙酯（防腐剂）。 </td></tr><tr><td> methyllycaconitine柠​​檬酸（工作重点） </td><td> tocris生命科学</td><td>目录＃1029 </td><td></td></tr><tr><td>塑料蜡棒</td><td> ？生公司 （阿克伦美国俄亥俄州） </td><td></td><td></td></tr></tboDY&gt; </table>

paired nanoinjection and electrophysiology assay to screen for bioactivity of compounds using the drosophila melanogaster giant fiber system

体内活性筛选化合物可以用来作为第一步，以确定可能到1,2药物制剂开发的候选人。我们开发了一种新型的nanoinjection /电检测，使检测的神经电路介导的在果蝇 3,4逃生响应功能的生物活性化合物的调节作用。我们在体内实验中，使用果蝇巨纤维系统（GFS， 图1）允许不同类型的化合物，如小分子肽的筛选，只需要最小数量引起的效果。此外，果蝇的飞行服务队提供了一个潜在的分子靶点神经或肌肉的种类繁多。 （GFS）巨纤维突触电（峡路口）以及化学（胆碱）到外围的中间神经突触（PSI）和“Tergo Trochanteral肌肉神经元（TTMn）5 </SUP&gt;。背侧纵肌神经元DLMn（）连接的PSI是依赖Dα7烟碱乙酰胆碱受体（nAChRs）的6。最后，神经肌肉接头（NMJ）的TTMn和DLMn跳（TTM）和飞行肌（DLM），是谷氨酸7-12。在这里，我们演示了如何注入化合物的纳升数量级，而获得巨人光纤系统，以及如何监察该电路的功能复合的影响的电生理细胞内记录。我们显示枸橼酸methyllycaconitine（MLA）的，胆碱受体拮抗剂，破坏PSI的DLMn连接，但不能连接或跳跃或飞行肌肉的功能NMJ的GF TTMn的的的特异性。 在这部影片开始之前，它是至关重要的，你仔细地观察和熟悉的朱庇特的视频名为“电生理记录，从四melan巨纤维通路“ogaster从奥古斯丁等 7，视频这里的目的是为这一现有技术的扩展。在这里，我们使用的细节，只对另外的的配对nanoinjections和监测技术的电生理记录方法和重点。

Watch this Scientific Journal Video about Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System at JoVE.com

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

迅速<em&gt;在体内</em&gt;分析测试neuromodulatory使用巨人光纤系统（GFS）的化合物<em&gt;果蝇</em&gt;描述。 nanoinjections动物的头，随着政府飞行服务队的电生理记录可以揭示生物活​​性化合物对神经或肌肉。

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 ...

Research

JoVE Journal

Neuroscience

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

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" ...

科研

神经科学

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

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

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 ...

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

标记的单细胞在中枢神经系统<em&gt;果蝇</em

In this  article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the ...

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster

在自发活动的测定<em&gt;果蝇</em

Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such ...

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster

简单的检测方法的社交回避的量化<em&gt;果蝇</em

Drosophila melanogaster is an emerging model to study different aspects of social interactions. For example, flies avoid areas previously occupied by ...

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

在气味引导行为的高分辨率定量<em&gt;果蝇</em&gt;使用<em&gt; Flywalk</em&gt;范式

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To ...

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster

一个行为分析对机械感觉MARCM为基础的克隆<em&gt;果蝇</em

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense ...

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster

高通量检测，以检查个体产蛋首<em&gt;果蝇</em

Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. ...

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

一个简单的方法来衡量在改建悬赏寻求行为使用<em&gt;果蝇</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

毛细管馈线分析测量的摄食<em&gt;果蝇</em

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...