Name: a molecular readout of long-term olfactory adaptation in c. elegans
Uploaded: 2012-12-22T14:00:00
Description: Watch this Scientific Journal Video about A Molecular Readout of Long-term Olfactory Adaptation in C. elegans at JoVE.com

我们想感谢斯科特·哈密尔顿，奥哈洛伦实验室的成员，仔细阅读这篇稿子。我们也感谢匿名审稿人很好的建议和有见地的意见。

1。建设GFP标记EGL-4表达动物<ol><li> ODR-3基因（使用2678 bp的直接上游的起始密码子）的启动子下克隆的翻译融合GFP :: EGL-4：（对）ODR-3 :: GFP :: EGL-4。在化学传感器的神经元对ODR-3启动子驱动表达：AWA; AWB; AWC，以及在ASH弱。 </li><li>进样的质粒（对）ODR-3 :: GFP :: EGL-4成野生型（N2）的动物在50纳克/微升的共注射的标记物，使用标准的种系的转化技术8：（对）OFM-1?...

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这里描述的气味诱导核项目的GFP标记的EGL-4分子提供了一个强大的分子C.嗅觉适应中读出的线虫 。气味引起的核易位检测很简单，只需要几天的准备时间。 pyIs500，我们已经建立了这些实验动物，表达了标记，这说明了的AWC神经元以及表达GFP标记的EGL-4蛋白。因此，我们认为，实验者可以非常迅速，几乎没有工作经验这一类的神经元（也许是经检查几十动物）后开始感觉很舒服?...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> Bacto琼脂</td><td> Difco公司</td><td> DF0140-07-4 </td><td> NGM板</td></tr><tr><td>氯化钠</td><td>费舍尔化工</td><td> S671-10 </td><td> NGM板</td></tr><tr><td>细菌用蛋白胨</td><td> Difco公司</td><td> DF0118-07-2 </td><td> NGM板</td></tr><tr><td>磷酸氢二钾</td><td>费舍尔化工</td><td> S375-500 </td><td> S-基底缓冲和NGM板</td></tr><tr><td>磷酸二氢钾</td><td>费舍尔化工</td><td> P285-500 </td><td> S-基底缓冲和NGM板</td></tr><tr><td> Kimwipes - 小</td><td> Kimberly-Clark公司</td><td> LS2770 </td><td></td></tr><tr><td>乙醇100％ </td><td>金盾化工有限公司有限公司</td><td> 43196-115 </td><td>稀释气味的趋化实验</td></tr><tr><td>氯化钙</td><td> Sigma-Aldrich公司</td><td> C8106-500G </td><td> NGM板</td></tr><tr><td>硫酸镁</td><td> MP Biomedicals </td><td> 150136-500G </td><td> NGM板</td></tr><tr><td>叠氮化钠99％ </td><td> Fisher Scientific则</td><td> ICN10289180 </td><td>麻醉剂</td></tr><tr><td>琼脂糖 - 超</td><td> Invitrogen公司</td><td> 16500-500 </td><td>琼脂糖凝胶垫</td></tr><tr><td>苯甲醛</td><td> Sigma-Aldrich公司</td><td> B1334-100G </td><td> AWC气味</td></tr><tr><td>丁酮，ACS级</td><td> Sigma-Aldrich公司</td><td> 360473-500ML </td><td> AWC气味</td></tr><tr><td>微量离心管 - 1.5毫升彩色</td><td>丹维尔</td><td> LS8147 </td><td></td></tr><tr><td>巴斯德移液管一次性玻璃5-3/4“ </td><td> Fisher Scientific则</tD&gt; <td> 13-678-20B </td><td></td></tr><tr><td> Stratalinker </td><td> Stratagene公司</td><td> Stratalinker 2400 </td><td> UV整合</td></tr><tr><td>过滤器真空瓶 - 500毫升</td><td>的Nalgene </td><td> 09-740-25B </td><td></td></tr></tbody></table>

a molecular readout of long-term olfactory adaptation in c. elegans

在持续的刺激感官的神经元减少其对信号的灵敏度，以适应他们的反应。适应的反应有助于形状的重视和保护细胞过度刺激。 C.适应内的嗅觉电路线虫最早是由Colbert和巴格曼1,2。在这里，作者定义的参数的嗅觉适应的范例，这是他们设计一个隔离的能力，以适应挥发性气味的化学传感器翼细胞C型（AWC）感觉神经元检测缺陷的突变体的遗传筛选。当野生型C.线虫动物暴露于一个有吸引力的AWC-检测到的气味3为30分钟，他们会适应他们的气味的响应，然后将忽略适应中的趋化行为〜1小时测定气味。当野生型C.线虫动物暴露于一个有吸引力的AWC-〜1小时检测到的气味，他们将忽略在交流的适应气味hemotaxis行为分析〜3小时。这两个阶段的嗅觉适应性C.线虫被描述为短期嗅觉适应30分钟后，气味暴露引起的，长期的嗅觉适应60分钟后，气味暴露引起的。 L&#39;Etoile酒店等。后来的工作，发现了一种蛋白激酶G（PKG）EGL-4的短期和长期的嗅觉适应性AWC神经元所需要的。 EGL-4蛋白含有核定位序列，有必要为长期的嗅觉适应性反应，但可有可无的短期嗅觉适应性反应的AWC 4。绿色荧光蛋白标记EGL-4，这是可能的可视化本地化的EGL-4在AWC暴露在长时间的气味。使用此功能齐全的GFP标记的EGL-4（GFP :: EGL-4）分子，我们已经能够开发出分子读出长期的嗅觉适应性AWC 5。使用这个米olecular读出的嗅觉适应，我们已经能够进行正向和反向遗传筛选，以确定突变的动物表现出有缺陷的亚细胞定位模式GFP :: EGL-AWC 6,7。在这里，我们描述：1）建造的GFP :: EGL-4表达动物; 2）的协议，用于培养动物长期臭味诱导核易位测定;和3）的得分长期臭味诱导核易位核GFP :: EGL-4状态的事件和恢复（重新过敏）。

本地化模式的GFP :: EGL-4英寸的AWC长时间气味曝光之前和之后的一个例子示出在图2中。在此之前长时间的气味曝光，GFP :: EGL-4定位于细胞质中的AWC（图2B），80分钟后，气味暴露GFP :: EGL-4定位于细胞核内的AWC（图2D）。在行为层面上，与胞内GFP :: EGL-4 AWC动物被吸引到一个点源的气味（ 图2C图形代表的趋化实验结果的不适应动物-注意的趋化指数接近1?...

Watch this Scientific Journal Video about A Molecular Readout of Long-term Olfactory Adaptation in C. elegans at JoVE.com

A Molecular Readout of Long-term Olfactory Adaptation in C. elegans

在这里，我们描述了一个长期的嗅觉适应性的分子读出<em&gt;秀丽隐杆线虫</em&gt;。蛋白激酶G，EGL-4，稳定的适应措施是必要的，在初级感觉神经元对AWC。在长时间的气味暴露EGL-4转位从细胞质到细胞核的AWC。

长期的嗅觉适应性的分子读数<em&gt; C.线虫</em

During sustained stimulation most sensory neurons will adapt their response by decreasing their sensitivity to the signal. The adaptation response helps ...

Research

JoVE Journal

Neuroscience

A Molecular Readout of Long-term Olfactory Adaptation in C. elegans

C. elegans Positive Butanone Learning, Short-term, and Long-term Associative Memory Assays

C. elegans Positive Butanone Learning, Short-term, and Long-term Associative Memory Assays

线虫的积极丁酮学习，短期，长期联想记忆检测

The memory of experiences and learned information is critical for organisms to make choices that aid their survival. C. elegans navigates its environment ...

科研

神经科学

In vivo Laser Axotomy in C. elegans

In vivo Laser Axotomy in C. elegans

在 C 在体内激光干切断线虫

Neurons communicate with other cells via axons and dendrites, slender membrane extensions that contain pre- or post-synaptic specializations. If a neuron ...

Automated Quantification of Synaptic Fluorescence in C. elegans

Automated Quantification of Synaptic Fluorescence in C. elegans

突触荧光自动定量<em&gt; C。线虫</em

Synapse strength refers to the amplitude of postsynaptic responses to presynaptic neurotransmitter release events, and has a major impact on overall ...

In vivo Neuronal Calcium Imaging in C. elegans

In vivo Neuronal Calcium Imaging in C. elegans

C.在体内神经元钙成像技术线虫

The nematode worm C. elegans is an ideal model organism  for relatively simple, low cost neuronal imaging in vivo. Its small transparent body and simple, ...

Long-term Behavioral Tracking of Freely Swimming Weakly Electric Fish

自由游泳弱电鱼的长期行为跟踪

Long-term behavioral tracking can capture and quantify natural animal behaviors, including those occurring infrequently. Behaviors such as exploration and ...

Long-term Time Lapse Imaging of Mouse Cochlear Explants

老鼠耳蜗外植体的长期时间推移成像

Here we present a method for long-term time-lapse imaging of live embryonic mouse cochlear explants. The developmental program responsible for building ...

Agarose Microchambers for Long-term Calcium Imaging of Caenorhabditis elegans

Agarose Microchambers for Long-term Calcium Imaging of Caenorhabditis elegans

琼脂糖微室的长期钙成像<em&gt;秀丽隐杆线虫</em

Behavior is controlled by the nervous system. Calcium imaging is a straightforward method in the transparent nematode Caenorhabditis elegans to measure ...

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

的自动分析<em&gt;℃。线虫</em&gt;游泳行为使用天书软件

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has ...

Quadruple Immunostaining of the Olfactory Bulb for Visualization of Olfactory Sensory Axon Molecular Identity Codes

嗅球的四次免疫染色用于嗅觉感觉轴突分子识别码的可视化

The mouse olfactory system is often used to study mechanisms of neural circuit formation&#12288;because of its simple anatomical structure. An Olfactory ...

Using an Adapted Microfluidic Olfactory Chip for the Imaging of Neuronal Activity in Response to Pheromones in Male C. Elegans Head Neurons

Using an Adapted Microfluidic Olfactory Chip for the Imaging of Neuronal Activity in Response to Pheromones in Male C. Elegans Head Neurons

应用一种适应微流控嗅觉芯片的神经活性成像对雄性线虫神经元的信息素反应

The use of calcium indicators has greatly enhanced our understanding of neural dynamics and regulation. The nematode Caenorhabditis elegans, with its ...