Name: odorant-induced responses recorded from olfactory receptor neurons using the suction pipette technique
Uploaded: 2012-04-05T12:00:00
Description: Watch this Scientific Journal Video about Odorant-induced Responses Recorded from Olfactory Receptor Neurons using the Suction Pipette Technique at JoVE.com

这项工作是由美国国立卫生研究院DC009613，人类前沿科学计划和莫雷护理奖学金（JR）的支持。

1。录像设置的记录室被安装在尼康TE2000U Eclipse的倒置显微镜相衬嵌合的光学上的气动工作台和电屏蔽使用一个法拉第笼。有机玻璃录音室包括两个部分，由阻挡层部分分离，并粘附到硅烷化的载玻片。该室的部分之一是用来解决细胞，而另一种是用于在录制过程中，以尽量减少刺激曝光过早曝光的结算，但尚未使用的单元格，以加臭剂。的记录设置包括的吸入移液管（见下文...

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	<li>Reisert, J., Zhao, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perspectives+on:+Information+and+coding+in+mammalian+sensory+physiology:+Response+kinetics+of+olfactory+receptor+neurons+and+the+implications+in+olfactory+coding.">Perspectives on: Information and coding in mammalian sensory physiology: Response kinetics of olfactory receptor neurons and the implications in olfactory coding.</a> J. Gen. Physiol. 138, 303-310 (2011).</li><li>Kaupp, U. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+signalling+in+vertebrates+and+insects:+differences+and+commonalities.">Olfactory signalling in vertebrates and insects: differences and commonalities.</a> Nat. Rev. Neurosci. 11, 188-200 (2010).</li><li>Tirindelli, R., Dibattista, M., Pifferi, S., Menini, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+pheromones+to+behavior.">From pheromones to behavior.</a> Physiol. Rev. 89, 921-956 (2009).</li><li>Kleene, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+electrochemical+basis+of+odor+transduction+in+vertebrate+olfactory+cilia.">The electrochemical basis of odor transduction in vertebrate olfactory cilia.</a> Chem. Senses. 33, 839-859 (2008).</li><li>Baylor, D. A., Lamb, T. D., Yau, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responses+of+retinal+rods+to+single+photons.">Responses of retinal rods to single photons.</a> J. Physiol. 288, 613-634 (1979).</li><li>Wang, J., Kefalov, V. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-cell+Suction+Recordings+from+Mouse+Cone+Photoreceptors.">Single-cell Suction Recordings from Mouse Cone Photoreceptors.</a> J. Vis. Exp. (35), e1681 (2010).</li><li>Lowe, G., Gold, G. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+spatial+distributions+of+odorant+sensitivity+and+odorant-induced+currents+in+salamander+olfactory+receptor+cells.">The spatial distributions of odorant sensitivity and odorant-induced currents in salamander olfactory receptor cells.</a> J. Physiol. 442, 147-168 (1991).</li><li>Reisert, J., Matthews, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Na+-dependent+Ca2++extrusion+governs+response+recovery+in+frog+olfactory+receptor+cells.">Na+-dependent Ca2+ extrusion governs response recovery in frog olfactory receptor cells.</a> J. Gen. Physiol. 112, 529-535 (1998).</li><li>Reisert, J., Matthews, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adaptation+of+the+odour-induced+response+in+frog+olfactory+receptor+cells.">Adaptation of the odour-induced response in frog olfactory receptor cells.</a> J. Physiol. 519, 801-813 (1999).</li><li>Matthews, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+compact+modular+flow+heater+for+the+superfusion+of+mammalian+cells.">A compact modular flow heater for the superfusion of mammalian cells.</a> J. Physiol. 518P, 13 (1999).</li><li>Reisert, J., Matthews, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+recording+of+receptor+current+and+intraciliary+Ca2++concentration+in+salamander+olfactory+receptor+cells.">Simultaneous recording of receptor current and intraciliary Ca2+ concentration in salamander olfactory receptor cells.</a> J. Physiol. 535, 637-645 (2001).</li></ol>

<p class="jove_content"&gt;抽吸移液器技术是一个电的方法，该方法被用于记录气味诱导慢受体电流和快速的双相动作电位同时从ORN。由于细胞的质膜不违反，此方法离开确保没有改变的加臭剂的反应，由于细胞质离子浓度或稀释的细胞内因素变化的细胞内环境不受干扰。细胞可以记录从较长的持续时间（在小鼠在青蛙和1小时至4小时）。这种技术的另一个优点是，因为在单元主体内的记录移液器，即纤毛和树突状旋钮?...

<table border="1"><tbody><tr><td> 的材料的名称 </td><td> 类型 </td><td> 公司 </td><td> 目录/ 型号 </td><td> 评论 </td></tr><tr><td>航空表</td><td>设备</td><td>新港</td><td></td><td></td></tr><tr><td>气泵</td><td>设备</td><td>新港</td><td> ACGP </td><td></td></tr><tr><td>用移液管拔轮器</td><td>设备</td><td>萨特</td><td> P-97 </td><td></td></tr><tr><td>高硼硅玻璃</td><td>设备</td><td> WPI </td><td> 1B150-4 </td><td></td></tr><tr><td>尼康的Eclipse倒置显微镜</td><td>设备</td><td>尼康</td><td> TE2000U </td><td>配备汞灯，GFP过滤器和目标的20倍和5倍，至少</td></tr><tr><td>放大器的PC-501A </td><td>设备</td><td>华纳</td><td> 64-0008 </td><td>探头1GΩ </td></tr><tr><td>钻石刀</td><td>设备</td><td>定做</td><td></td><td></td></tr><tr><td> A / D数字化仪Mikro1401 </td><td>设备</td><td>剑桥电子设计</td><td></td><td></td></tr><tr><td>过滤单元3382 </td><td>设备</td><td>克罗恩海特公司</td><td></td><td></td></tr><tr><td>信号</td><td>软件</td><td>剑桥电子设计</td><td></td><td></td></tr><tr><td>成型的Ag / AgCl颗粒</td><td>设备</td><td> WPI </td><td> 64-1297 </td><td></td></tr><tr><td>吸管架</td><td>设备</td><td>华纳</td><td> 64-0997 </td><td>自定义修改，以适应 探头</td></tr><tr><td>录音室</td><td>设备</td><td>定做</td><td></td><td></td></tr><tr><td>微操作机器人 MP85-1028 </td><td>设备</td><td>萨特仪器</td><td>微操作机器人 MP85-1028 </td> &lt;TD&gt; </td></tr><tr><td>矿物油</td><td>解</td><td>西格玛</td><td> 330779-1L </td><td></td></tr><tr><td>示波器TDS 1001 </td><td>设备</td><td>泰克公司</td><td></td><td></td></tr><tr><td>三管方玻璃管</td><td>设备</td><td>华纳</td><td> 64-0119 </td><td> 0.6 mm内径，长5厘米</td></tr><tr><td>阀</td><td>设备</td><td>利公司</td><td></td><td></td></tr><tr><td> Valvelink 8.2 </td><td>设备</td><td>自动化科学</td><td></td><td></td></tr><tr><td> SF-77B灌注快一步</td><td>设备</td><td>华纳</td><td></td><td></td></tr></tbody></table>

odorant-induced responses recorded from olfactory receptor neurons using the suction pipette technique

动物采样的臭味通过化学感受系统位于鼻腔周围环境。化学感受信号的复杂行为的影响，如食物的选择，捕食，同种和配偶识别和其他社会有关的线索。位于嗅觉受体神经元（ORNs）中嵌入鼻腔的嗅觉上皮细胞背侧部分。这些双极神经元的轴突发送到嗅球（见图1，Reisert钊1，最初发表于普通生理学杂志“）和扩展的单枝晶的纤毛辐射到的粘液覆盖的嗅觉上皮细胞的边界上皮细胞。的纤毛包含的信号转导机制，最终导致兴奋性通过睫状转导通道，环核苷酸门控通道（CNG）的Ca 2 +活化的Cl -的信道（ 图1）流入的电流。随之而来的depola三丙触发在电池主体2-4的动作电位的产生。 在这个视频中，我们描述了使用的“吸吸液管技术”，记录的气味引起的反应ORNs。该方法最初发展到记录从杆感光体5和jove.com修改，以记录从鼠标锥感光体6，可以发现在该方法的一个变种。后来被改编吸入吸液管技术也记录ORNs 7,8。简要地说，在解离的嗅觉上皮细胞和细胞分离后，整个细胞的ORN体被吸入到的记录吸管的前端。枝晶和纤毛仍然暴露于沐浴液，从而解决方案的变化，例如，使气味或药物受体阻滞剂的应用。在此配置中，还没有进入到细胞内环境获得（的没有全细胞电压钳）和细胞内的电压保持自由变化。这一切都OWS受体电流缓慢，起源于发射的细胞体9纤毛和快动作电位的同时记录。动力学这两个信号之间的差异在允许它们被分开使用不同的过滤器设置。这种技术可以用在任何野生型或基因敲除小鼠，或选择性地从记录ORNs也表达GFP标记的特定子集ORNs，例如表示一个给定的加臭剂的受体或离子通道。

Watch this Scientific Journal Video about Odorant-induced Responses Recorded from Olfactory Receptor Neurons using the Suction Pipette Technique at JoVE.com

Odorant-induced Responses Recorded from Olfactory Receptor Neurons using the Suction Pipette Technique

嗅觉受体神经元（ORNs）气味信号转换成一个受体的电流，从而触发动作电位，被输送到第二个神经元在嗅球中。这里，我们描述的吸入移液管的技术，同时记录从小鼠ORNs电流和动作电位的加臭剂诱导的受体。

从使用吸吸液管技术的嗅觉受体神经元的气味引起的反应记录

Animals sample the odorous environment around them through the chemosensory systems located in the nasal cavity. Chemosensory signals affect complex ...

Research

JoVE Journal

Neuroscience

Analyzing Responses of Mouse Olfactory Sensory Neurons Using the Air-phase Electroolfactogram Recording

鼠标采用的空气相Electroolfactogram录制的嗅觉感官神经元响应分析

Animals depend on olfaction for many critical behaviors, such as finding food sources, avoiding predators, and identifying conspecifics for mating and ...

科研

神经科学

Application of a NMDA Receptor Conductance in Rat Midbrain Dopaminergic Neurons Using the Dynamic Clamp Technique

一个NMDA受体电导的大鼠中脑多巴胺能神经元的动态钳技术中的应用

Neuroscientists study the function of the brain by investigating how neurons in the brain communicate. Many investigators look at changes in the ...

Isolating Nasal Olfactory Stem Cells from Rodents or Humans

隔离啮齿类动物或人类的鼻嗅干细胞

The olfactory mucosa, located in the nasal cavity, is in charge of detecting odours. It is also the only nervous tissue that is exposed to the external ...

Targeting Olfactory Bulb Neurons Using Combined In Vivo Electroporation and Gal4-Based Enhancer Trap Zebrafish Lines

Targeting Olfactory Bulb Neurons Using Combined In Vivo Electroporation and Gal4-Based Enhancer Trap Zebrafish Lines

针对嗅球神经元采用组合在体内电穿孔和GAL4基于增强陷阱斑马鱼线

In vivo electroporation is a powerful method for delivering DNA expression plasmids, RNAi reagents, and morpholino anti-sense oligonucleotides to specific ...

Flash Photolysis of Caged Compounds in the Cilia of Olfactory Sensory  Neurons

闪光光解笼状化合物在嗅感觉神经元的纤毛

Photolysis of caged compounds allows the production of rapid and localized increases in the concentration of various physiologically active compounds1. ...

Imaging pHluorin-tagged Receptor Insertion to the Plasma Membrane in Primary Cultured Mouse Neurons

成像pHluorin标记插入到质膜受体原代培养的小鼠神经元

A better understanding of the mechanisms governing receptor trafficking between the plasma membrane (PM) and intracellular compartments requires an ...

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 ...

Perforated Patch-clamp Recording of Mouse Olfactory Sensory Neurons in Intact Neuroepithelium: Functional Analysis of Neurons Expressing an Identified Odorant Receptor

鼠标嗅觉感官神经元的完整神经上皮穿孔膜片钳记录：神经元表达鉴定的气味受体功能分析

Analyzing the physiological responses of olfactory sensory neurons (OSN) when stimulated with specific ligands is critical to understand the basis of ...

Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

使用单一感器录音来检测臭虫的嗅觉神经元的反应，以信息化学物质

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house ...

In Vivo Three-Dimensional Two-Photon Microscopy to Study Conducted Vascular Responses by Local ATP Ejection Using a Glass Micro-Pipette

在 Vivo 三维双光子显微镜中,利用玻璃微移器研究本地 ATP 喷射的血管反应

Maintenance of normal brain function requires a sufficient and efficient supply of oxygen and nutrition by a complex network of vessels. However, the ...