Name: a procedure for implanting organized arrays of microwires for single-unit recordings in awake, behaving animals
Uploaded: 2014-02-14T14:15:00
Description: Watch this Scientific Journal Video about A Procedure for Implanting Organized Arrays of Microwires for Single-unit Recordings in Awake, Behaving Animals at JoVE.com

这项研究是由美国国家药物滥用研究所的支持给予DA 006886（MOW）和DA 032270（DJB）。

悉心照顾，必须采取保持无菌条件下（如在指南中描述的实验动物9的护理和使用），同时准备和进行以下步骤。以下协议符合本指南实验动物的护理和使用，并已被批准的机构动物护理和使用委员会，罗格斯大学。据估计，在后续的步骤将需要3-6小时来完成。 植入微丝阵列： <ol><li>在麻醉状态下的动物的地方使用50毫克/千克戊巴比妥钠（IP）和管理硝酸甲基阿托品10...

<ol>
	<li>Carter, M., Shieh, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Electrophysiology In: Guide to research techniques in neuroscience. , (2009).</li><li>Aston-Jones, G., Siggins, G. R., Kupfer, D., Bloom, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Electrophysiology. In: Psychopharmacology: The Fourth Generation of Progress. , (1995).</li><li>Root, D. H., 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=Differential+roles+of+ventral+pallidum+subregions+during+cocaine+self-administration+behaviors.">Differential roles of ventral pallidum subregions during cocaine self-administration behaviors.</a> J. Comp. Neurol. 521 (3), 558-588 (2013).</li><li>Cardin, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dissecting+local+circuits+in+vivo:+integrated+optogenetic+and+electrophysiology+approaches+for+exploring+inhibitory+regulation+of+cortical+activity.">Dissecting local circuits in vivo: integrated optogenetic and electrophysiology approaches for exploring inhibitory regulation of cortical activity.</a> (3-4), 106-103 (2012).</li><li>Ma, S., 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=Amphetamine's+dose-dependent+effects+on+dorsolateral+striatum+sensorimotor+neuron+firing.">Amphetamine's dose-dependent effects on dorsolateral striatum sensorimotor neuron firing.</a> Behav. Brain Res. , (2013).</li><li>Ghitza, U. 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=Persistent+cue-evoked+activity+of+accumbens+neurons+after+prolonged+abstinence+from+self-administered+cocaine.">Persistent cue-evoked activity of accumbens neurons after prolonged abstinence from self-administered cocaine.</a> J. Neurosci. 23 (19), 7239-7245 (2003).</li><li>Tang, C., 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=Changes+in+activity+of+the+striatum+during+formation+of+a+motor+habit.">Changes in activity of the striatum during formation of a motor habit.</a> Eur. J. Neurosci. 25 (4), 1212-1227 (2007).</li><li>Tang, C., 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=Dose+and+rate-dependent+effects+of+cocaine+on+striatal+firing+related+to+licking.">Dose and rate-dependent effects of cocaine on striatal firing related to licking.</a> J. Pharmacol. Exp. Ther. 324 (2), 701-713 (2008).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=National+Research+Council.">National Research Council.</a> Guide for the Care and Use of Laboratory Animals: Eighth Edition. , (2011).</li><li>Fabbricatore, A. 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=Electrophysiological+evidence+of+mediolateral+functional+dichotomy+in+the+rat+accumbens+during+cocaine+self-administration:+tonic+firing+patterns.">Electrophysiological evidence of mediolateral functional dichotomy in the rat accumbens during cocaine self-administration: tonic firing patterns.</a> Eur. J. Neurosci. 30 (12), 2387-2400 (2009).</li><li>Root, D. H., 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=Slow+phasic+and+tonic+activity+of+ventral+pallidal+neurons+during+cocaine+self-administration.">Slow phasic and tonic activity of ventral pallidal neurons during cocaine self-administration.</a> Synapse. 66 (2), 106-127 (2012).</li><li>Root, D. H., 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=Rapid-phasic+activity+of+ventral+pallidal+neurons+during+cocaine+self-administration.">Rapid-phasic activity of ventral pallidal neurons during cocaine self-administration.</a> Synapse. 64 (9), 704-713 (2010).</li><li>Tang, C. C., 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=Decreased+firing+of+striatal+neurons+related+to+licking+during+acquisition+and+overtraining+of+a+licking+task.">Decreased firing of striatal neurons related to licking during acquisition and overtraining of a licking task.</a> J. Neurosci. 29 (44), 12952-12961 .</li><li>Paxinos, G., Watson, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Rat Brain in Stereotaxic Coordinates. , (1997).</li></ol>

细胞外记录代表一个强大的实验技术，它可以被纳入几乎所有的实验准备在神经科学。已经植入在有组织的阵列线可被跟踪为它们的轴穿过大脑并进入其靶区域（ 图5A）。当在非绝缘微丝尖端创建一个小的，实验后病变从不锈钢线创建一个小的铁矿床，可以使用溶液精确地标记非绝缘微丝尖端的位置（其中，被记录的单单元） 5％亚铁氰化钾和10％的HCl（ 图5A和5B;</s...

电生理记录让科学家研究生物细胞的电特性。在中枢神经系统中，其中电脉冲作为信号传导机制，这些录音是为了解神经功能1-2特别重要的。在单一单元中记录行为的动物，其已被插入到大脑中的微电极能够在一段时间内神经元的产生动作电位的变化进行记录。 虽然许多技术允许一个记录大脑活动，单机电是最精确的方法，允许的分辨率在单个神经元的水平之一。当空间特异性程度高是需要的，微丝可以用于靶向离散子细胞核或brain3内细胞的合奏。单机录音也受益于高时间分辨率的录音是准确的在微秒级。并且， 在体内唤醒录音允许完整电路的相互作用，与传入和传出突起，全身化学和激素的影响，和生理参数的天然环境。神经信号从感觉输入，运动行为，认知过程，神经化学/药理学，或者某种组合。因此，感觉，运动，认知和化学影响的偏析就必须周密的实验，有效的突发事件和控制，可以允许每个上述影响的评估。总而言之，在录音动物行为让实验者观察多个信息源的集成功能电路内，并推导出电路功能更全面的模型。 单单元记录也具有许多缺点的，其中任何实验者应该意识到受损。首先，录音可能难以进行。事实上，日的性质Ë探头放大器和植入微丝，使空间和时间特异性，这些记录也使得录音易受外来电信号（ 即电气“噪声”）的影响。因此，在一个电系统解决问题的能力，就必须电生理原理和装置的发达技术的理解。同样重要的是要注意，在某些情况下，在细胞外记录记录的电信号可代表多个神经信号的总和。此外，单单元活动的目标区域内的人口活动的普遍性通常可以通过细胞异质性的目标区域内的程度的限制（但参阅卡丁4）。例如，电极可能会偏向于记录高振幅输出神经元，以代替其他细胞。单单元记录的可解释性增加通过组合记录与其它技术包括，但不限于，电气（顺或逆向），化学（ 例如，离子电渗疗法或设计师受体）或光遗传学刺激4，临时神经inactivations，感觉检查5，断开的程序，或免疫组织化学3。 在下面，我们将列举必需的材料和步骤，植入大鼠的组织微丝阵列（尽管该协议可以适于在其他物种中使用）的协议。在我们的实验室中使用的固定阵列的过程和风格已被证明可靠的纵向录音，可以在一个月的时间内维持6-8相同的神经元记录的。这使得这个过程非常适用于研究阶段性反应实验的刺激，塑料改变神经反应，或者学习和动机的机制。

<table>
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr>
			<td>Table 1. List of Surgical Materials</td>
		</tr>
		<tr>
			<td>Gauze</td>
			<td>Fisher (MooreBrand)</td>
			<td>19-898-144</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton Swabs</td>
			<td>Fisher (Puritan)</td>
			<td>S304659</td>
			<td></td>
		</tr>
		<tr>
			<td>Nembutal (Pentobarbital)</td>
			<td>Sigma Aldrich</td>
			<td>P3761</td>
			<td></td>
		</tr>
		<tr>
			<td>Atropine Methyl Nitrate</td>
			<td>Sigma Aldrich</td>
			<td>A0382</td>
			<td></td>
		</tr>
		<tr>
			<td>Baytril (Enrofloxacin)</td>
			<td>Butler Shein (Bayer)</td>
			<td>1040007</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine Hydrochloride</td>
			<td>Butler Shein</td>
			<td>SKU# 023061</td>
			<td></td>
		</tr>
		<tr>
			<td>Betadine (Povidone-Iodine)</td>
			<td>Fisher (Perdue)</td>
			<td>19-066452&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotax</td>
			<td>Kopf</td>
			<td>Model 900</td>
			<td></td>
		</tr>
		<tr>
			<td>Cauterizing Tool</td>
			<td>Stoelting</td>
			<td>59017</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting Microscope</td>
			<td>Nikon</td>
			<td>SMZ445</td>
			<td></td>
		</tr>
		<tr>
			<td>Dental Drill</td>
			<td>Buffalo</td>
			<td>37800</td>
			<td></td>
		</tr>
		<tr>
			<td>Bacteriostatic Saline</td>
			<td>Bulter Schein</td>
			<td>8973</td>
			<td></td>
		</tr>
		<tr>
			<td>Jewlers Skrews</td>
			<td>Stoelting</td>
			<td>51457</td>
			<td></td>
		</tr>
		<tr>
			<td>Microwire Array</td>
			<td>Microprobes</td>
			<td>Custom (Flexible)</td>
			<td></td>
		</tr>
		<tr>
			<td>Ground Wire</td>
			<td>Omnetics</td>
			<td>Custom Plug</td>
			<td></td>
		</tr>
		<tr>
			<td>Dental Acrylic</td>
			<td>Fisher (BAS)</td>
			<td>50-854-402</td>
			<td></td>
		</tr>
		<tr>
			<td>Absorbable Sutures</td>
			<td>Fisher (Ethicon)</td>
			<td>NC0258473</td>
			<td></td>
		</tr>
		<tr>
			<td>Puralube (Opthalamic Ointment/Lubricant)</td>
			<td>Fisher (Henry Schein)</td>
			<td>008897</td>
			<td></td>
		</tr>
	</tbody>
</table>
<table>
	<tbody>
		<tr>
			<td>Table 2. List of Surgical Instruments</td>
		</tr>
		<tr>
			<td>2x Microforceps</td>
			<td>George Tiemann &#38; Co.</td>
			<td>#160-57</td>
			<td>Multi-use (e.g. clearing debris in skull window)</td>
		</tr>
		<tr>
			<td>2x Forceps</td>
			<td>George Tiemann &#38; Co.</td>
			<td>#160-93</td>
			<td>Multi-use (e.g. tying sutures)</td>
		</tr>
		<tr>
			<td>6x Hemostats</td>
			<td>George Tiemann &#38; Co.</td>
			<td>#105-1125</td>
			<td>Clamp and open incision</td>
		</tr>
		<tr>
			<td>1x Small scissors</td>
			<td>George Tiemann &#38; Co.</td>
			<td>#105-411</td>
			<td>Cut sutures after tying</td>
		</tr>
		<tr>
			<td>1x Tissue forceps</td>
			<td>George Tiemann &#38; Co.</td>
			<td>#105-222</td>
			<td>Holding tissue while suturing</td>
		</tr>
		<tr>
			<td>1x Needle holder</td>
			<td>George Tiemann &#38; Co.</td>
			<td>#105-1259</td>
			<td>Holding suture needle</td>
		</tr>
		<tr>
			<td>1x Scalpel holder (with #11 blade)</td>
			<td>George Tiemann &#38; Co.</td>
			<td>#105-80 (w/ #105-71 blade)</td>
			<td>Making skull incision</td>
		</tr>
		<tr>
			<td>1x # 22 Scalpel blade</td>
			<td>George Tiemann &#38; Co.</td>
			<td># 160-381</td>
			<td>Shaving scalp</td>
		</tr>
		<tr>
			<td>1x Surgical Spatula</td>
			<td>George Tiemann &#38; Co.</td>
			<td>#160-718</td>
			<td>Scraping skull to clear tissue on skull</td>
		</tr>
		<tr>
			<td>Machine/Jewelers Screws</td>
			<td>Various</td>
			<td>N/A</td>
			<td>0/80 x 1/8&#8221;</td>
		</tr>
	</tbody>
</table>
<table>
	<tbody>
		<tr>
			<td>Table 3. List of Equipment for Recording Electrophysiological Signals</td>
		</tr>
		<tr>
			<td>Microwire Array &#38; Connector</td>
			<td>Micro Probe, Inc. (Gaithersburg, MD)</td>
			<td>&#160;N/A</td>
			<td>Cranially implanted in target recording region. Arrays are customized based on desired wire spacing, length, etc.</td>
		</tr>
		<tr>
			<td>(Part No. Based on array characteristics)</td>
		</tr>
		<tr>
			<td>Unity-Gain Harness/Headstage</td>
			<td>M.B. Turnkey Designs (Hillsborough, NJ)</td>
			<td>Proj 1200</td>
			<td>Initial amplification of neural signal; allows for propagation of small neural signals.</td>
		</tr>
		<tr>
			<td>Commutator (&#38; Optional Fluid Swivel)</td>
			<td>Plastics One, Inc. (Roanoke, VA)</td>
			<td>SL18C</td>
			<td>Allows animals to freely rotate while propagating electrical signal to preamp</td>
		</tr>
		<tr>
			<td>Pre-Amplifier</td>
			<td>M.B. Turnkey Designs (Hillsborough, NJ)</td>
			<td>Proj 1198</td>
			<td>Differentially amplifies neural signals against a reference electrode.</td>
		</tr>
		<tr>
			<td>Filter &#38; Amplifier</td>
			<td>M.B. Turnkey Designs (Hillsborough, NJ)</td>
			<td>Proj 1199</td>
			<td>Band-pass filters and further amplifies the differentially amplified signal.</td>
		</tr>
		<tr>
			<td>Acquisition Computer</td>
			<td>EnGen (Phoenix, AZ)</td>
			<td>N/A (Custom Build)</td>
			<td>Runs software and hardware for behavioral and neural data acquisition.</td>
		</tr>
		<tr>
			<td>A/D Card&#160;</td>
			<td>Data Translation (Marlboro, MA)</td>
			<td>DT-3010</td>
			<td>Digitizes neural signals for computer sampling.</td>
		</tr>
		<tr>
			<td>Digital I/O Card</td>
			<td>Measurement Computing (Norton, MA)</td>
			<td>PCI CTR-05</td>
			<td>Acquires behavioral inputs and outputs</td>
		</tr>
	</tbody>
</table>

a procedure for implanting organized arrays of microwires for single-unit recordings in awake, behaving animals

体内电生理记录在清醒，行为的动物提供了强有力的方法对理解神经信号在单细胞水平。该技术允许实验者在时间和区域研究具体的放电模式，以提供持续的行为相关记录的动作电位。此外，单个单元的记录可以与为了产生神经功能的全面解释了大量的其他技术相结合。在这篇文章中，我们描述了麻醉，准备微丝植入。随后，我们列举了必要的设备和手术步骤，准确地插入微丝数组转换为目标结构。最后，我们简要地描述用于从阵列中的每个单独的电极来记录设备。所述的固定微丝阵列非常适合于慢性注入并允许神经数据的纵向记录在几乎任何行为编写，上。我们讨论了追踪电极轨道进行三角微丝的位置，以及如何以植入微丝结合免疫组化技术，以提高记录结果的解剖特异性。

一个用本实验室记录电生理信号设备的列表可以在表3中可以找到。以下从手术恢复，单单元通过插入一个单位增益探头插入植入连接器记录。此探头通过电缆连接到一个换向器，其能够自由旋转而不在电生理记录符通过使用电滑环。换向器允许受试者在行为，这是该制剂的原理优势之一在录制过程中自由移动。信号，然后通过前置放大器（10倍的增益），用于差分放大信号对所选择?...

Watch this Scientific Journal Video about A Procedure for Implanting Organized Arrays of Microwires for Single-unit Recordings in Awake, Behaving Animals at JoVE.com

A Procedure for Implanting Organized Arrays of Microwires for Single-unit Recordings in Awake, Behaving Animals

植入微丝的组织阵列中单个单元的电生理记录的使用存在许多技术上的挑战。方法来执行这种技术和必要的设备描述。此外，有益利用有组织的微丝阵列由不同的神经次区域具有高空间选择性记录进行了讨论。

一个程序植入微丝的有组织阵列的单机录音清醒，举止动物

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell ...

Research

JoVE Journal

Neuroscience

Electrode Fabrication and Implantation in Aplysia californica for Multi-channel Neural and Muscular Recordings in Intact, Freely Behaving Animals

Electrode Fabrication and Implantation in Aplysia californica for Multi-channel Neural and Muscular Recordings in Intact, Freely Behaving Animals

电极的制作及植入海兔californica完整的，自由的行为的动物多通道的神经和肌肉的录音

Recording from key nerves and muscles of Aplysia during feeding behavior allows us to study the patterns of neural control in an intact animal.  ...

科研

神经科学

Single-unit In vivo Recordings from the Optic Chiasm of Rat

Single-unit In vivo Recordings from the Optic Chiasm of Rat

单机在体内</em从大鼠视交叉>录音

Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic ...

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

逆行荧光染料标记允许有针对性的外单细胞记录技术鉴定神经元<em&gt;在体内</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Design and Assembly of an Ultra-light Motorized Microdrive for Chronic Neural Recordings in Small Animals

设计和组装的超轻型电动微型硬盘（Microdrive）慢性神经录音的小动物

The ability to chronically record from populations of neurons in freely behaving animals has proven an invaluable tool for dissecting the function of ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

双突触诱发的星形胶质细胞和神经元的反应，在急性海马脑片电生理记录

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

Simultaneous Long-term Recordings at Two Neuronal Processing Stages in Behaving Honeybees

在两个神经元处理阶段在做人蜜蜂同时长期录音

In both mammals and insects neuronal information is processed in different higher and lower order brain centers. These centers are coupled via convergent ...

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

一个程序，植入脊柱商会纵<em&gt;在体内</em&gt;鼠标脊髓成像

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by ...

Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice

Optogenetic 在行为小鼠海马θ振荡中的作用

Extensive data on relationships of neural network oscillations to behavior and organization of neuronal discharge across brain regions call for new tools ...

Investigating Object Representations in the Macaque Dorsal Visual Stream Using Single-unit Recordings

用单单元记录法研究猕猴背视流中的对象表示

Previous studies have shown that neurons in parieto-frontal areas of the macaque brain can be highly selective for real-world objects, disparity-defined ...

Chronic Implantation of Multiple Flexible Polymer Electrode Arrays

多柔性聚合物电极阵列的慢性植入

Simultaneous recordings from large populations of individual neurons across distributed brain regions over months to years will enable new avenues of ...