Name: ex vivo preparations of the intact vomeronasal organ and accessory olfactory bulb
Uploaded: 2014-08-04T23:15:00
Description: Watch this Scientific Journal Video about Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb at JoVE.com

，F30 DC011673（GFH：NINDS，NIH）和得克萨斯大学西南的启动资金（JPM）：这项研究是由R00 DC011780（NINDS，NIH JPM）的支持。

所有实验均按照批准的得克萨斯大学西南机构动物护理和使用委员会的协议，并被选为以尽量减少应激，不适，疼痛经历的实验动物。 1，解剖商会自定义夹层室，小，薄塑料板所需要的最好的结果（ 图1）。提前尝试此协议的建造或取得该室。 2，解剖解决方案<ol><li>准备1升的标准人工脑脊液（ACSF）中的步骤3.22-5.4使用...

<ol>
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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reliable+sex+and+strain+discrimination+in+the+mouse+vomeronasal+organ+and+accessory+olfactory+bulb.">Reliable sex and strain discrimination in the mouse vomeronasal organ and accessory olfactory bulb.</a> The Journal of neuroscience : the official journal of the Society for Neuroscience. 33, 13903-13913 (2013).</li><li>Luo, M., Fee, M. S., Katz, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Encoding+pheromonal+signals+in+the+accessory+olfactory+bulb+of+behaving+mice.">Encoding pheromonal signals in the accessory olfactory bulb of behaving mice.</a> Science. 299, 1196-1201 (2003).</li><li>Binns, K. E., Brennan, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+electrophysiological+activity+in+the+accessory+olfactory+bulb+and+medial+amygdala+associated+with+mate+recognition+in+mice.">Changes in electrophysiological activity in the accessory olfactory bulb and medial amygdala associated with mate recognition in mice.</a> Eur J Neurosci. 21, 2529-2537 (2005).</li><li>Leszkowicz, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noradrenaline-induced+enhancement+of+oscillatory+local+field+potentials+in+the+mouse+accessory+olfactory+bulb+does+not+depend+on+disinhibition+of+mitral+cells.">Noradrenaline-induced enhancement of oscillatory local field potentials in the mouse accessory olfactory bulb does not depend on disinhibition of mitral cells.</a> Eur J Neurosci. 35, 1433-1445 (2012).</li><li>Ames, A., Gurian, B. 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P., Holy, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+ex+vivo+preparation+of+the+intact+mouse+vomeronasal+organ+and+accessory+olfactory+bulb.">An ex vivo preparation of the intact mouse vomeronasal organ and accessory olfactory bulb.</a> J Neurosci Methods. 177, 440-447 (2009).</li><li>Leinders-Zufall, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasensitive+pheromone+detection+by+mammalian+vomeronasal+neurons.">Ultrasensitive pheromone detection by mammalian vomeronasal neurons.</a> Nature. 405, 792-796 (2000).</li><li>Kato, H. K., Chu, M. W., Isaacson, J. S., Komiyama, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+sensory+representations+in+the+olfactory+bulb:+modulation+by+wakefulness+and+experience.">Dynamic sensory representations in the olfactory bulb: modulation by wakefulness and experience.</a> 76, 962-975 (2012).</li><li>Meeks, J. P., Arnson, H. A., Holy, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Representation+and+transformation+of+sensory+information+in+the+mouse+accessory+olfactory+system.">Representation and transformation of sensory information in the mouse accessory olfactory system.</a> Nature. 13, 723-730 (2010).</li></ol>

该VNO-AOB 体外在本协议中所述的制备是一个有用的替代体内5-7和急性现场片17实验AOB函数来麻醉。不像急性AOB切片实验，这也暴露了电路元件的电和光学记录，该制剂保留所有感觉传入和国内AOB连接。尽管这也可以在体内方法所述的麻醉剂，麻醉剂的存在不一定会改变神经功能，即兴奋性/抑制性的平衡，这是在这个和其他的嗅觉电路18，用于信息处理的...

在哺乳动物大脑感觉处理通常跨越多个往复连接的神经元回路，其中每个抽取特定的特征从感觉输入。在感觉通路，早期信息处理是正常的认知和行为至关重要。在附件嗅觉系统（AOS）的嗅球（AOB）是感官外围连接到该决定的荷尔蒙平衡1,2，3的侵略和觉醒4下游结构的主要神经回路。因此，该电路内的信息处理是密切相关的变化，动物的行为。 在嗅球位于小鼠和大鼠在主嗅球（MOB）下方的致密的背侧/尾/后一方面，血管化鼻腔鼻窦。该AOB接收传入的神经支配来自驻留在犁鼻器（VNO）的犁鼻周围感觉神经元（的VSN）的轴突，一个SMaL在口鼻前正上方的软腭升盲端管。这些轴突穿越隔组织在鼻腔的内侧边界的微妙片。使用麻醉小鼠5-7或可自由探索动物8一些研究探讨AOB的神经反应，以AOS的气味（如小鼠尿液） 在体内的来源。 在涉及（一）气管切开术，以确保麻醉过深，防止液体刺激5-7的心愿，（二）刺激交感型颈椎节6或犁鼻器5,7的直接插管引进非挥发性气味体内研究英勇麻醉以及（c）开颅手术带或不带额叶消融，以允许电极前进到AOB 6。清醒/行为研究微驱动8-10参与手术植入。总之，这些实验范式是强大的，但非常困难，往往需要麻醉。 体外 ）外取得了一些成功11-15。因为VNO和AOB之间的连接保持同侧，且由于中线室间隔组织可以在单个半球暴露于含氧的灌流，我们试图开发这样的单半球体外方法来分离这些结构，同时保持其功能性连接。最近，我们成功地实现了这一目标16。该制剂保持VNO和AOB活着和功能上连接至少4两 - 6小时，因为无论是轴突（沿中线软室间隔组织）和AOB较浅&lt;600μm的特征都可以访问的灌流充氧人工脑脊液（学联）。这VNO-AOB 体外制备允许引入控制刺激到VNO通过薄插管，并直接的视觉进入小AOB进行有针对性的电极放置和/或现场荧光显微镜。这种方法是有利的，如果一个人渴望学习这些电路在没有麻醉剂。因为这种方法切断离心连接，它不能很好地适合于查询到AOB功能的离心调制。该VNO-AOB 体外的准备是很难学，但一旦实现产生一个可靠的平台，探讨电路组织，信息处理和神经可塑性在这个强大的感测电路。

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			<td>Fisher Scientific</td>
			<td>3120030</td>
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			<td>Fisher Scientific</td>
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			<td>Fisher Scientific</td>
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			<td>Dissection Chamber</td>
			<td>Custom&#160;</td>
			<td>N/A</td>
			<td>See Fig. 1</td>
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			<td>Delrin plastic plank 0.6 cm x 1.5 cm x 0.1 cm</td>
			<td>Custom&#160;</td>
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			<td>Fisher Scientific</td>
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			<td>Fine Science Tools</td>
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			<td>Fine Science Tools</td>
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			<td>91150-20</td>
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			<td>Sigma-Aldrich</td>
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			<td>Sigma-Aldrich</td>
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			<td>Sigma-Aldrich</td>
			<td>various</td>
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			<td>myo-inositol</td>
			<td>Sigma-Aldrich</td>
			<td>various</td>
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			<td>Na-pyruvate</td>
			<td>Sigma-Aldrich</td>
			<td>various</td>
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			<td>Na-ascorbate</td>
			<td>Sigma-Aldrich</td>
			<td>various</td>
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			<td>HEPES buffer</td>
			<td>Sigma-Aldrich</td>
			<td>various</td>
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			<td>glucose</td>
			<td>Sigma-Aldrich</td>
			<td>various</td>
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ex vivo preparations of the intact vomeronasal organ and accessory olfactory bulb

鼠标附件嗅觉系统（AOS）是一个专门感觉传导通路检测非易失性社会异味，费洛蒙，和利它素。第一个神经回路在AOS途径，称为嗅球（AOB），它在建立性的典型行为，如领土的侵略和交配的重要作用。这个小（&lt;1 立方毫米）电路具有区分独特的行为状态，如性别，品系，并在同种的分泌物和排泄物化学感受线索压力的能力。虽然这个系统的紧凑型组织呈现为同时从电路的大部分记录了独特的机会，在AOB感觉处理的调查仍然充满挑战，主要是由于其大脑中的实验不利的位置。在这里，我们证明了多阶段夹层，消除了完好的AOB的前小鼠头盖骨的单个半球内，留下连接离子两者的周犁鼻感觉神经元（的VSN）和局部神经回路完好。该过程公开的AOB表面直接目视检查，便于在没有麻醉剂的电生理和从AOB电路元件的光学记录。在插入细导管进入犁鼻器（VNO），这家的VSN，可以直接在外围暴露于社会的气味和信息素，同时记录在AOB下游活动。此过程使控制查询到AOS信息处理，它可以揭示信息素连接变动风险行为机制的光。

取得成功，这种准备需要大量的实践，有几个步骤，在它可能会失败。每个人都应该想到需要很多的尝试取得成功之前。自定义剥离室需要成功完成该协议，并应事先在开始解剖的后期阶段获得。腔室的设计在图1中给出的是足以达到此目的，并且可以由相对廉价的塑料以最小的加工要求。如果一个人没有产生这样的腔容量，本地或在线加工企业可以进行协商，并可以构造室收费。 </p...

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Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

完整的犁鼻器和副嗅球的体外制剂

鼠标嗅球（AOB）已难以在感官编码的上下文中学习。在这里，我们表明，产生体外制剂，其中AOB神经元保持功能连接到他们的外设输入，便于研究的小鼠费洛蒙和利它素信息处理解剖。

The mouse accessory olfactory system (AOS) is a specialized sensory pathway for detecting nonvolatile social odors, pheromones, and kairomones. The first ...

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Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

Multielectrode Array Recordings of the Vomeronasal Epithelium

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Understanding neural circuits requires methods to record from many neurons simultaneously. For in vitro studies, one currently available technology is ...

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

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在小鼠嗅球成像气味诱发的活动，利用光学反射和自体荧光信号

In the brain, sensory stimulation activates distributed populations of neurons among functional modules which participate to the coding of the stimulus. ...

Imaging Neuronal Responses in Slice Preparations of Vomeronasal Organ Expressing a Genetically Encoded Calcium Sensor

在犁鼻器官的切片准备工作表示一个基因编码的钙传感器成像神经元的反应

The vomeronasal organ (VNO) detects chemosensory signals that carry information about the social, sexual and reproductive status of the individuals within ...

The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

嗅觉系统作为模型来研究轴突生长的模式和形态<em&gt;在体内</em

The olfactory system has the unusual capacity to generate new neurons throughout the lifetime of an organism. Olfactory stem cells in the basal portion of ...

Recording Temperature-induced Neuronal Activity through Monitoring Calcium Changes in the Olfactory Bulb of Xenopus laevis

Recording Temperature-induced Neuronal Activity through Monitoring Calcium Changes in the Olfactory Bulb of Xenopus laevis

通过的嗅球钙监控录像的变化温度引起的神经元电活动<em&gt;非洲爪蟾</em

The olfactory system, specialized in the detection, integration and processing of chemical molecules is likely the most thoroughly studied sensory system. ...

In-depth Physiological Analysis of Defined Cell Populations in Acute Tissue Slices of the Mouse Vomeronasal Organ

在小鼠犁鼻急性组织切片定义细胞群的深入生理学分析

In most mammals, the vomeronasal organ (VNO) is a chemosensory structure that detects both hetero- and conspecific social cues. Vomeronasal sensory ...

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 ...

Preparation of Rat Sciatic Nerve for Ex Vivo Neurophysiology

Preparation of Rat Sciatic Nerve for Ex Vivo Neurophysiology

大鼠坐骨神经用于 离体 神经生理学的制备

Ex vivo preparations enable the study of many neurophysiological processes in isolation from the rest of the body while preserving local tissue structure. ...