这项工作是由NSERC AHFMR支持。作者感谢玛丽亚姆Alaverdashvili和阿曼达Mauthe Kaddoura手稿和Hiroe山崎的意见，准备手术的帮助。

1。手术前准备<ol><li>适当的类型和剂量的麻醉药麻醉大鼠。在这里，我们用乌拉坦（1.5克/公斤，Sigma - Aldrich公司有限公司，圣路易斯，密苏里州） - 准备为20％溶液稀释的聚氨酯20克在100毫升磷酸缓冲液（PBS），即动物将收到7.5 intraperitonealy聚氨酯解决方案（IP）的毫升/公斤。由于其剂量反应曲线，聚氨酯的总剂量分为四个应用程序，每个相隔约30分钟。之前的前两个聚氨酯的应用，动物麻醉与异氟醚在3-4％的浓度（每分钟2升氧气）管理，并维持在2％（高达每分钟2升氧气），以防止动物从越来越强调通过注射。在手术过程中，isofluorane（2％浓度的2分钟）的额外的管理，如果需要的话，可以使用。 </li><li>剃须沿切口部位（头背侧）皮草，以确保皮草不会污染伤口和足够的面积，可消毒切口周围站点。 </li><li>防腐用有机碘或洗必泰肥皂和消毒剂（乙醇），准备手术部位。 </li><li>准备硅探针（NeuroNexus技术，安阿伯，MI）。探头可以使用70％的乙醇，用蒸馏水冲洗消毒。如果需要的话，探头可涂防腐准备（如染料我，Invitrogen公司有限公司）用荧光标记，稍后揭示探头的位置，在组织学分析。要做到这一点，我们用一个小的和非常柔软的油漆刷和微妙的接触与染料的硅探头的背面。此过程应该始终做了一个显微镜，以确保探头不被损坏或弯曲。经常检查，确保染料不会阻止任何渠道。探头上留下任何残余染料可以用酒精后取出，用清水冲洗。 </li><li>硅electrodES护理:由于其体积小，硅探针是微妙的，非常容易打破，即使使用具有相当的保健。因此，我们建议监测潜在的危险，在所有的时间，如:（一）保持在一个适当指定的框的探针;（二）防止崩溃或删除处理时的探头;（三）确保固定，立体造以防止任何意外;（四）录制时，周围组织和/或血液可以连接到探头。因此，重要的是录制后取出探头时要小心。强烈建议使用PBS或生理盐水溶液的流量恒定（V）始终清洁录制后探头。要清洁探头，强烈建议他们淹没在液体酶清洗剂（波士顿，博士伦）几分钟。始终用蒸馏水冲洗探针，并返回他们自己的专用盒。根据我们的经验，足够的照顾，确保硅探针10多个实验的良好品质的录音。如果通道"获取嘈杂"，在使用过程中，应排除后续分析。我们发现，通道之间的串扰是最小的;因此，嘈杂的通道通常有其他渠道的信号影响很小。 </li></ol> 2。外科<ol><li>任何手术操作前，涂抹于眼睛表面的眼软膏。 </li><li>管理地塞米松5毫克/毫升皮下注射，以减少脑部发炎。 </li><li>修复的动物，在整个手术的立体框架。 </li><li>管理2％盐酸利多卡因肾上腺素（2-4毫克/千克稀释至0.5％溶液）在切口部位皮下切口前，以减少伤口出血和局部麻醉的潜在风险。 </li><li>通过头皮中线2厘米的切口，横向和取代皮肤暴露颅骨表面。 </li><li>清除头皮背筋膜，钝性分离和控制microcautery或用骨蜡止血。 </li><li>两个钻孔，重视动物接地和参考的头骨枕骨的小螺丝钉。参考螺丝必须与硬脑膜接触。为了减少在录音电气噪声，这是建议保留在任何时候都湿了螺丝。要做到这一点，一个小的丙烯酸障碍，周围的螺丝可以建造或螺丝可以用琼脂覆盖。 </li><li>在坐标相应的皮质和/或皮质下面积（S）的利息5 1-2毫米直径的开颅手术。开颅手术（钻孔）期间需要使用压缩空气来消除骨粉。防止头骨温暖从钻井应用的PBS滴。第二次开颅手术，如果需要插入第二个探测器。在这段视频中，我们提出从两个皮质区同时录音，因此两个craniotomies。 </li><li>建立一个小的丙烯酸树脂屏障（如:郎牙科制造，有限责任公司，Wheeting，IL）的周边两个craniotomies为了保持PBS暴露硬脑膜上，防止它干燥。整个实验过程中不断应用PBS。 </li><li>打开硬脑膜问题都craniotomies。两面针（30半计）弯曲的提示（形成了钩），可用于该操作。 </li><li>将连接器的硅探针的立体manipulatiors。连接器连接到一个极点，才能有探头完全平行的立体操盘柄。这是至关重要的，以保持稳定的录音，并避免损坏探头。 </li><li>连接到硅探头的参考引脚的参考螺丝。为了减少电噪声，接地螺钉连接到:我）探头插头的接地;二）从录音设备的机箱接地;三）立体框架。 </li><li>将只是在软接触的硅探针的尖端。 </li><li>慢慢地引进日发送硅探头，并确保无阻力或弯曲，可以插入到大脑的硅探针的柄（图1及2）。 </li><li>仔细降低，直至达到所需的记录区域的探头。如果需要，加craniotomies顶部的PBS滴，以避免组织干燥。 </li><li>由于乌拉坦麻醉，只应使用终端程序，动物安乐死至少钠经口200mg/kg腹腔注射戊巴比妥稀释至不超过60毫克/毫升的浓度。其次是动物断头。 </li></ol> 3。代表性的成果: 局部场电位和扣球活动的代表录音如图3A所示。硅探头记录后执行穗排序（歧视尖峰来自不同的神经元;图3B，C; 6，7，8）提供调查涉及的神经元群活动的能力，我王等，以下过程:记忆和决策9 10，可塑性强，刺激的，自发的活动编码11 12和13个不同的药物的影响。 <img alt="图1" src="/files/ftp_upload/3282/3282fig1.jpg" /> 图1。答 :一个使用一个单一的柄的硅探头的例子是叠加在重建神经元（S. 坂田14提供）蒙太奇B.探头在每个8柄的四极配置的一个例子。C.原理图大鼠颅骨。绿色框表示在左，右的体感皮层的开颅手术的程度。 <img alt="图2" src="/files/ftp_upload/3282/3282fig2.jpg" /> 图2:与实验前额叶皮层和海马硅探针插入的一个例子。掉落的PBS覆盖了开颅手术，以保护暴露的BR泉干涸。以上位于小脑的两个螺丝将连接到地面和参考，分别。 <img alt="图3" src="/files/ftp_upload/3282/3282fig3.jpg" /> 图3。 A.图像显示500ms的有代表性的局部场电位（LFP）的电生理记录，从一个四极管（注:负电压绘制统筹）。插入显示-波形的尖峰B.二维单位从1四极管的功能空间集群的意见。每个集群从一个单一的单元（神经元）的尖峰。C.平均尖峰波形代表单位（颜色编码），在每一个单一的四极管的四个记录点 。D.波形能量（振幅），随着时间的推移峰值。注意:每个单元的相对恒定的能量值，这表明显示期的持续时间较稳定的记录。

<ol>
	<li>Buzsaki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+recording+of+neuronal+ensembles.">Large-scale recording of neuronal ensembles.</a> Nat. Neurosci. 7, 446-451 (2004).</li><li>Kipke, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advanced+neurotechnologies+for+chronic+neural+interfaces:+new+horizons+and+clinical+opportunities.">Advanced neurotechnologies for chronic neural interfaces: new horizons and clinical opportunities.</a> J. Neurosci. 28, 11830-11838 (2008).</li><li>Csicsvari, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Massively+parallel+recording+of+unit+and+local+field+potentials+with+silicon-based+electrodes.">Massively parallel recording of unit and local field potentials with silicon-based electrodes.</a> J. Neurophysiol. 90, 1314-1323 (2003).</li><li>Bartho, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+neocortical+principal+cells+and+interneurons+by+network+interactions+and+extracellular+features.">Characterization of neocortical principal cells and interneurons by network interactions and extracellular features.</a> J. Neurophysiol. 92, 600-608 (2004).</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. , (1998).</li><li>Lewicki, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+methods+for+spike+sorting:+the+detection+and+classification+of+neural+action+potentials.">A review of methods for spike sorting: the detection and classification of neural action potentials.</a> Network. 9, R53-R78 (1998).</li><li>Harris, K. D., Henze, D. A., Csicsvari, J., Hirase, H., Buzsaki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+tetrode+spike+separation+as+determined+by+simultaneous+intracellular+and+extracellular+measurements.">Accuracy of tetrode spike separation as determined by simultaneous intracellular and extracellular measurements.</a> J. Neurophysiol. 84, 401-414 (2000).</li><li>Luczak, A., Narayanan, N. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spectral+representation--analyzing+single-unit+activity+in+extracellularly+recorded+neuronal+data+without+spike+sorting.">Spectral representation--analyzing single-unit activity in extracellularly recorded neuronal data without spike sorting.</a> J. Neurosci. Methods. 144, 53-61 (2005).</li><li>Pastalkova, E., Itskov, V., Amarasingham, A., Buzsaki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Internally+generated+cell+assembly+sequences+in+the+rat+hippocampus.">Internally generated cell assembly sequences in the rat hippocampus.</a> Science. 321, 1322-1327 (2008).</li><li>Fujisawa, S., Amarasingham, A., Harrison, M. T., Buzsaki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavior-dependent+short-term+assembly+dynamics+in+the+medial+prefrontal+cortex.">Behavior-dependent short-term assembly dynamics in the medial prefrontal cortex.</a> Nat Neurosci. 11, 823-833 (2008).</li><li>Harris, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+do+neurons+work+together?+Lessons+from+auditory+cortex.">How do neurons work together? Lessons from auditory cortex.</a> Hear. Res. 271, 37-53 (2011).</li><li>Luczak, A., Bartho, P., Marguet, S. L., Buzsaki, G., Harris, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sequential+structure+of+neocortical+spontaneous+activity+in+vivo.">Sequential structure of neocortical spontaneous activity in vivo.</a> Proc. Natl. Acad. Sci. U.S.A. 104, 347-352 (2007).</li><li>Robbe, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cannabinoids+reveal+importance+of+spike+timing+coordination+in+hippocampal+function.">Cannabinoids reveal importance of spike timing coordination in hippocampal function.</a> Nat. Neurosci. 9, 1526-1533 (2006).</li><li>Sakata, S., Harris, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laminar+structure+of+spontaneous+and+sensory-evoked+population+activity+in+auditory+cortex.">Laminar structure of spontaneous and sensory-evoked population activity in auditory cortex.</a> Neuron. 64, 404-418 (2009).</li></ol>

没有利益冲突的声明。

本文演示了如何使用硅电极阵列记录从人口众多，同时在多个皮层区的神经元（&gt; 100）。为了在成功的手术和记录，应考虑以下问题: <ol><li>到所需区域的探头介绍:当插入脑组织中的探测器，它有可能造成相当大的损害。这可能会导致低质量的记录单位。为了避免这个问题，我们提出以下建议:（一）在一定程度的角度介绍了探头（推荐使用10度角）。通过这样做，可以减少所记录的神经元的树突状损害;（二）探针插入脑组织后，他们最初以更快的速度，每10-30秒（约50至100微米）降低，直到他们得到接近所需的记录。当探测目标区域约200微米，位置调整更慢（约imately 10微米，每2或3分钟）。 </li><li>稳定的录音:当探头在指定的区域是相当大的活​​动检测（在大多数的渠道鲜明尖峰）的，建议开始录制之前，等待约30分钟。这将使脑组织机械稳定的探针插入后，并确保一个更稳定的录音。 </li><li>脑脉动:偶尔可以看到大脑的脉动，可以显著降低录音的质量。在录音方面，一个小的开颅手术（只是足够的空间，以适应探头）可以减少大脑的脉动。如果有必要，枕大池可以被刺破。这样可以减少脑脊液压力和减少肿胀和脉动。 </li><li>硅探头配置:硅探头可以有多种形状和录制现场配置。例如，他们可以在不同的柄，柄的长度和厚度，并在记录点（;:www.neuronexustech.com看到如四极管与线性配置）的安排。一个特定的配置使用的探头选择取决于科学要回答的问题。例如，如果目标是记录在多个地点在一个特定的层（如在这些录音中提出）的神经元的人群，最好的选择是使用八柄和四极配置的探头。这允许多个单单位在每一个四极管和跨1.4毫米跨度8个地点的采样录音。另一个例子是，如果有人想研究在皮质层的活动传播，最好的选择是使用一个探头，一个柄定期分发的记录点，沿着这条柄，允许同时录制多个皮质层14。 </li></ol>

<table border="1"><tbody><tr><td> 名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td>聚氨酯</td><td> Sigma - Aldrich公司</td><td></td></tr><tr><td>磷酸盐缓冲液（PBS） </td><td></td><td></td></tr><tr><td>异氟醚</td><td>森</td><td> 02241315 </td></tr><tr><td>硅探针</td><td> NeuroNexus技术</td><td>采用4 × 2 - 春节 - 5MM - 150 - 200 - 312 </td></tr><tr><td>染料我（18（3）介质隔离集成电路） </td><td> Invitrogen公司有限公司</td><td> 617830 </td></tr><tr><td>地塞米松5 </td><td> Vetoquinol </td><td> 0A0640B </td></tr><tr><td>眼膏</td><td> Vetoquinol </td><td> 005165414 </td></tr><tr><td>利多卡因盐酸肾上腺素的2％ </td><td> Bimeda - MTC </td><td> 00141984 </td></tr><tr><td>立体定向框架</td><td> KOPF仪器</td><td> 1430 - B </td></tr><tr><td>电极操纵器和支架</td><td> KOPF仪器</td><td> 960 </td></tr><tr><td>鼠耳，酒吧</td><td> KOPF仪器</td><td> 1955年</td></tr><tr><td>大鼠适配器</td><td> KOPF仪器</td><td> 920 </td></tr><tr><td>麻醉面罩</td><td> KOPF仪器</td><td> 906 </td></tr><tr><td>解剖刀</td><td> Roboz手术仪器</td><td> RS - 9843 </td></tr><tr><td>手术刀片</td><td>模范</td><td> 0086 </td></tr><tr><td>微解剖剪刀</td><td> Roboz手术仪器</td><td> RS - 5910 </td></tr><tr><td>螺丝</td><td>古尔德Pasteners有限公司</td><td> 14084701 </td></tr><tr><td>止血钳</td><td>抢劫盎司手术仪器</td><td> RS - 7130 RS - 7211 </td></tr><tr><td> 30 ½表针</td><td>流式细胞 - 迪金森</td><td> 305115 </td></tr><tr><td>微电机控制</td><td> Foredom </td><td> HP4 - 917 </td></tr><tr><td>解剖钳</td><td> Roboz手术仪器</td><td> RS - 5237 </td></tr><tr><td>骨蜡</td><td> Lukens </td><td> M263500 </td></tr><tr><td>压缩空气瓶</td><td>猎鹰安全产品，公司</td><td> 50 - 10521C </td></tr><tr><td>酶法清洁</td><td>波士顿，博士伦</td><td> BAUSCH497628 </td></tr><tr><td>蒸馏水</td><td></td><td></td></tr><tr><td>牙科丙烯酸粉末</td><td>郎牙科</td><td> 1220 </td></tr><tr><td>牙科丙烯酸液体</td><td>郎牙科</td><td> 1404 </td> &lt;/ TR&gt; <tr><td>数字山猫SX系统</td><td> Neuralynx公司</td><td> 4SX - Z400 </td></tr><tr><td>猎豹Softwere </td><td> Neuralynx公司</td><td></td></tr><tr><td>鳄鱼电缆</td><td></td><td></td></tr></tbody></table>

recording large-scale neuronal ensembles with silicon probes in the anesthetized rat

从神经合奏的大型电生理记录提供了机会，以研究大脑如何协调各种各样的扣球从它的神经元活动的行为。同时监察扣球从大量的神经元在多个地方的神经回路活动的一个最有效的方法是通过使用硅电极阵列1-3。 动作电位产生大量的跨膜电压的变化，在细胞somata附近。这些输出信号可以测量导体放置在一个神经元的接近。如果有许多活动（扣球）在针尖附近的神经元，电极记录其中，单个神经元的贡献是"电气距离"加权相结合的信号。硅探头是理想的记录电极监测，由于大量的记录点（64）和体积小的多个神经元。 Furthermo再次，多个站点可以被安排在一毫米的距离，从而使神经元的活动，同时在不同的皮质层或多个皮层柱的录音（图1）。更重要的是，记录点的几何精确分布，还可以隔离单个神经元的4的空间关系的决心。在这里，我们描述的一种急性，大规模的左，右前肢同时在硅探针（图2）麻醉大鼠体感皮层神经元记录。

Watch this Scientific Journal Video about Recording Large-scale Neuronal Ensembles with Silicon Probes in the Anesthetized Rat at JoVE.com

Recording Large-scale Neuronal Ensembles with Silicon Probes in the Anesthetized Rat

神经元的活动外的录音使用的麻醉大鼠硅探针进行说明。这种技术允许跨多个脑区的信息，来自100多个神经元同时获得。它提供了有关单细胞神经合奏在多个本地电路动态决议的信息。

在麻醉大鼠的硅探头录制的大型神经元合唱

Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of ...

recording-large-scale-neuronal-ensembles-with-silicon-probes

Research

JoVE Journal

Neuroscience

24.8K 次观看.  University of Lethbridge. 神经元的活动外的录音使用的麻醉大鼠硅探针进行说明。这种技术允许跨多个脑区的信息，来自100多个神经元同时获得。它提供了有关单细胞神经合奏在多个本地电路动态决议的信息。

视频: Recording Large-scale Neuronal Ensembles with Silicon Probes in the Anesthetized Rat

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects1-11. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design.
Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (<a href="http://www.neuronexustech.com/" >http://www.neuronexustech.com/</a>; <a href="http://www.sbmicrosystems.com/">http://www.sbmicrosystems.com/</a>; <a href="http://www.acreo.se/">http://www.acreo.se/</a>) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

移动硅探针的神经行为鼠害的大型记录

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

科研

神经科学

A Fully Automated and Highly Versatile System for Testing Multi-cognitive Functions and Recording Neuronal Activities in Rodents

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be investigated in a single task sequence. The unique feature of this system is a custom-made, acoustically transparent chamber that eliminates many of the issues associated with auditory cue control in most commercially available chambers. The ease with which operant devices can be added or replaced makes this system quite versatile, allowing for the implementation of a variety of auditory, visual, and olfactory behavioral tasks. Automation of the system allows fine temporal (10 ms) control and precise time-stamping of each event in a predesigned behavioral sequence. When combined with a multi-channel electrophysiology recording system, multiple cognitive brain functions, such as motivation, attention, decision-making, patience, and rewards, can be examined sequentially or independently.

In this report, we present a fully automated and highly versatile system capable of simultaneously testing multi-cognitive behaviors and recording neuronal activities for rodents.

一个多认知功能测试，并记录在啮齿动物的神经元活动的完全自动化和高度灵活的系统

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be ...

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of the dendritic tree, which eventually leads to neuronal output of action potentials at the axon. The influence of diverse spatial and temporal parameters of specific synaptic input on neuronal output is currently under investigation, e.g. the distance-dependent attenuation of dendritic inputs, the location-dependent interaction of spatially segregated inputs, the influence of GABAergig inhibition on excitatory integration, linear and non-linear integration modes, and many more.
With fast micro-iontophoresis of glutamate and GABA it is possible to precisely investigate the spatial and temporal integration of glutamatergic excitation and GABAergic inhibition. Critical technical requirements are either a triggered fluorescent lamp, light-emitting diode (LED), or a two-photon scanning microscope to visualize dendritic branches without introducing significant photo-damage of the tissue. Furthermore, it is very important to have a micro-iontophoresis amplifier that allows for fast capacitance compensation of high resistance pipettes. Another crucial point is that no transmitter is involuntarily released by the pipette during the experiment.
Once established, this technique will give reliable and reproducible signals with a high neurotransmitter and location specificity. Compared to glutamate and GABA uncaging, fast iontophoresis allows using both transmitters at the same time but at very distant locations without limitation to the field of view. There are also advantages compared to focal electrical stimulation of axons: with micro-iontophoresis the location of the input site is definitely known and it is sure that only the neurotransmitter of interest is released. However it has to be considered that with micro-iontophoresis only the postsynapse is activated and presynaptic aspects of neurotransmitter release are not resolved. In this article we demonstrate how to set up micro-iontophoresis in brain slice experiments.

In this article we introduce fast micro-iontophoresis of neurotransmitters as a technique to investigate integration of postsynaptic signals with high spatial and temporal precision.

谷氨酸和GABA快速微离子导入:一个有用​​的工具，调查突触整合

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of ...

Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to monitor the disease course in the follow-up period. Changes in the VEP parameters closely correlate with pathological damage in the optic nerve. This protocol provides a detailed description about the rodent model of optic nerve microinjection, in which a partial demyelination lesion is produced in the optic nerve. VEP recording techniques are also discussed. Using skull implanted electrodes, we are able to acquire reproducible intra-session and between-session VEP traces. VEPs can be recorded on individual animals over a period of time to assess the functional changes in the optic nerve longitudinally. The optic nerve demyelination model, in conjunction with the VEP recording protocol, provides a tool to investigate the disease processes associated with demyelination and remyelination, and can potentially be employed to evaluate the effects of new remyelinating drugs or neuroprotective therapies.

Focal demyelination is induced in the optic nerve using lysolecithin microinjection. Visual evoked potentials are recorded via skull electrodes implanted over the visual cortex to examine the signal conduction along the visual pathway in vivo. This protocol details the surgical procedures underlying electrode implantation and optic nerve microinjection.

在实验视神经脱髓鞘模型大鼠视觉诱发电位记录

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to ...

Extracellular Recording of Neuronal Activity Combined with Microiontophoretic Application of Neuroactive Substances in Awake Mice

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and awake animals. Therefore, methods allowing the manipulation of synaptic systems in awake animals are required in order to determine the contribution of synaptic inputs to neuronal processing unaffected by anesthetics. Here, we present methodology for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. By combining these procedures, we performed microiontophoretic injections of gabazine to selectively block GABAA receptors in neurons of the inferior colliculus of head-restrained mice. Gabazine successfully modified neural response properties such as the frequency response area and stimulus-specific adaptation. Thus, we demonstrate that our methods are suitable for recording single-unit activity and for dissecting the role of specific neurotransmitter receptors in auditory processing.
The main limitation of the described procedure is the relatively short recording time (~3 hr), which is determined by the level of habituation of the animal to the recording sessions. On the other hand, multiple recording sessions can be performed in the same animal. The advantage of this technique over other experimental procedures used to manipulate the level of neurotransmission or neuromodulation (such as systemic injections or the use of optogenetic models), is that the drug effect is confined to the local synaptic inputs to the target neuron. In addition, the custom-manufacture of electrodes allows adjustment of specific parameters according to the neural structure and type of neuron of interest (such as the tip resistance for improving the signal-to-noise ratio of the recordings).

We present methods for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. This technique allows the detailed analysis of putative local synaptic inputs to the neuron of interest.

神经元电活动外记录与神经活性物质在清醒小鼠微电泳应用相结合

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and ...

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

The olfactory system, specialized in the detection, integration and processing of chemical molecules is likely the most thoroughly studied sensory system. However, there is piling evidence that olfaction is not solely limited to chemical sensitivity, but also includes temperature sensitivity. Premetamorphic Xenopus laevis are translucent animals, with protruding nasal cavities deprived of the cribriform plate separating the nose and the olfactory bulb. These characteristics make them well suited for studying olfaction, and particularly thermosensitivity. The present article describes the complete procedure for measuring temperature responses in the olfactory bulb of X. laevis larvae. Firstly, the electroporation of olfactory receptor neurons (ORNs) is performed with spectrally distinct dyes loaded into the nasal cavities in order to stain their axon terminals in the bulbar neuropil. The differential staining between left and right receptor neurons serves to identify the &#947;-glomerulus as the only structure innervated by contralateral presynaptic afferents. Secondly, the electroporation is combined with focal bolus loading in the olfactory bulb in order to stain mitral cells and their dendrites. The 3D brain volume is then scanned under line-illumination microscopy for the acquisition of fast calcium imaging data while small temperature drops are induced at the olfactory epithelium. Lastly, the post-acquisition analysis allows the morphological reconstruction of the thermosensitive network comprising the &#947;-glomerulus and its innervating mitral cells, based on specific temperature-induced Ca2+ traces. Using chemical odorants as stimuli in addition to temperature jumps enables the comparison between thermosensitive and chemosensitive networks in the olfactory bulb.

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

Here we describe a protocol for measuring and analyzing temperature responses in the olfactory bulb of Xenopus laevis. Olfactory receptor neurons and mitral cells are differentially stained, after which calcium changes are recorded, reflecting a sensitivity of some neural networks in the bulb to temperature drops induced at the nose.

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. ...

Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats

The electroretinogram (ERG) and visual evoked potential (VEP) are commonly used to assess the integrity of the visual pathway. The ERG measures the electrical responses of the retina to light stimulation, while the VEP measures the corresponding functional integrity of the visual pathways from the retina to the primary visual cortex following the same light event. The ERG waveform can be broken down into components that reflect responses from different retinal neuronal and glial cell classes. The early components of the VEP waveform represent the integrity of the optic nerve and higher cortical centers. These recordings can be conducted in isolation or together, depending on the application. The methodology described in this paper allows simultaneous assessment of retinal and cortical visual evoked electrophysiology from both eyes and both hemispheres. This is a useful way to more comprehensively assess retinal function and the upstream effects that changes in retinal function can have on visual evoked cortical function.

This protocol describes simultaneous measurement of electroretinogram and visual evoked potentials in anesthetized rats.

电图同时记录和麻醉大鼠视觉诱发电位

The electroretinogram (ERG) and visual evoked potential (VEP) are commonly used to assess the integrity of the visual pathway. The ERG measures the ...

Implantation of Chronic Silicon Probes and Recording of Hippocampal Place Cells in an Enriched Treadmill Apparatus

An important requisite for understanding brain function is the identification of behavior and cell activity correlates. Silicon probes are advanced electrodes for large-scale electrophysiological recording of neuronal activity, but the procedures for their chronic implantation are still underdeveloped. The activity of hippocampal place cells is known to correlate with an animal&#39;s position in the environment, but the underlying mechanisms are still unclear. To investigate place cells, here we describe a set of techniques which range from the fabrication of devices for chronic silicon probe implants to the monitoring of place field activity in a cue-enriched treadmill apparatus. A micro-drive and a hat are built by fitting and fastening together 3D-printed plastic parts. A silicon probe is mounted on the micro-drive, cleaned, and coated with dye. A first surgery is performed to fix the hat on the skull of a mouse. Small landmarks are fabricated and attached to the belt of a treadmill. The mouse is trained to run head-fixed on the treadmill. A second surgery is performed to implant the silicon probe in the hippocampus, following which broadband electrophysiological signals are recorded. Finally, the silicon probe is recovered and cleaned for reuse. The analysis of place cell activity in the treadmill reveals a diversity of place field mechanisms, outlining the benefit of the approach.

We describe the diverse steps to implant chronic silicon probes and to record place cells in mice that are running head-fixed on a cue-enriched treadmill apparatus.

富氧运动器械中慢性硅探针的植入及海马位细胞的记录

An important requisite for understanding brain function is the identification of behavior and cell activity correlates. Silicon probes are advanced ...

Chronic Transcranial Electrical Stimulation and Intracortical Recording in Rats

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a closed-loop event-triggered manner. Although many studies are focusing on the possible benefits and side-effects of TES in healthy and pathologic brains, there are still many fundamental open questions regarding the mechanism of action of the stimulation. Therefore, there is a clear need for a robust and reproducible method to test the acute and the chronic effects of TES in rodents. TES can be combined with regular behavioral, electrophysiological, and imaging techniques to investigate neuronal networks in vivo. The implantation of transcranial stimulation electrodes does not impose extra constraints on the experimental design while it offers a versatile, flexible tool to manipulate brain activity. Here we provide a detailed, step-by-step protocol to fabricate and implant transcranial stimulation electrodes to influence brain activity in a temporally constrained manner for months.

This detailed protocol describes transcranial stimulation electrode placement on the temporal bone in order to investigate the short- and long-term effects of transcranial electrical stimulation in freely moving rats.

大鼠慢性经颅电刺激及 Intracortical 记录

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a ...

Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats

Monitoring the activity patterns of a large population of neurons over many days in awake animals is a valuable technique in the field of systems neuroscience. One key component of this technique consists of the precise placement of multiple electrodes into desired brain regions and the maintenance of their stability. Here, we describe a protocol for the construction of a 3D-printable hyperdrive, which includes eighteen independently adjustable tetrodes, and is specifically designed for in vivo extracellular neural recording in freely behaving rats. The tetrodes attached to the microdrives can either be individually advanced into multiple brain regions along the track, or can be used to place an array of electrodes into a smaller area. The multiple tetrodes allow for simultaneous examination of action potentials from dozens of individual neurons, as well as local field potentials from populations of neurons in the brain during active behavior. In addition, the design provides for simpler 3D drafting software that can easily be modified for differing experimental needs.

We present the construction of a 3D-printable hyperdrive with eighteen independently adjustable tetrodes. The hyperdrive is designed to record brain activity in freely behaving rats over a period of several weeks.