联合穿梭盒子训练与电的Cortex记录和刺激作为一种工具来研究感知和学习
Summary
穿梭箱回避学习是行之有效的行为神经。本协议描述了如何穿梭盒子学习啮齿类动物可以与特定地点的电气皮层内微刺激（ICMS） 和体内录音作为研究学习和感知的多个方面的工具的同时慢性相结合。
Abstract
穿梭箱回避学习是行为神经科学和实验设置一个行之有效的方法是传统的定制;必要的设备现在可以通过一些商业公司。该协议提供了一个双向穿梭箱回避学习范式的啮齿类动物的详细说明（在这里长爪沙鼠， 长爪沙鼠 ）的结合位点特异性电气皮层内微刺激（ICMS），并同时慢性电在体内录音。详细的协议，适用于研究在不同的啮齿类动物的学习行为和感知的多个方面。
听觉皮层电路如下条件刺激的位点特异性ICMS作为工具来测试特定的传入，传出和皮质内连接的感知相关。不同的活动模式可以通过使用不同的刺激电极ARR被诱发AYS的地方，层依赖ICMS或远处ICMS网站。利用行为信号检测分析可以判断哪个刺激策略是最有效的用于引发一个行为上可检测的和显着信号。此外，使用不同的电极设计（表面电极，深部电极等）并行多信道-录音允许对这种学习过程的时间过程研究神经元观测。这将是讨论的行为设计的变化是如何能提高认知的复杂性（ 如探测，识别，逆转学习）。
Introduction
行为神经科学的一个基本目标是建立神经元的结构和功能特性，学习和认知之间的具体联系。与感知和学习有关的神经活性可以通过在多个位点的动作电位并局部场电位的各种大脑结构电生理记录进行研究。尽管电生理记录提供的神经活动和行为之间的相关协会，直接电皮层内微刺激（ICMS）一个多世纪以来一直是最直接的方法神经元的兴奋的人群和他们的行为和感知效果1的测试因果关系– 3。许多研究表明，动物是能够利用电刺激的知觉的任务范围内，例如初级视4吨取决于刺激位置的各种空间和时间属性onotopic 5，或躯体6区皮层。在皮层电诱发活动的传播主要是通过轴索纤维及其分布的突触连接2，在皮质，显然是一层依赖7的布局。得到的多突触活化ICMS诱发是今后更广泛的比电场2,8,9的直接影响。这就解释了为什么感性效应阈值由皮质内微刺激诱 发可以强烈地层依赖8,10,11和站点相关的9。最近的一项研究证明了细节，上层的刺激产生的主要supragranular层corticocortical电路更广泛的激活，而皮质导致更深层的一个焦点，反复corticoefferent intracolumnar激活刺激。并行行为实验表明，后者具有低得多的听觉检测苏氨酸esholds 8。因此，位点特异性ICMS作为条件刺激的优点是利用与电生理记录结合因果涉及特定皮质电路激活8学习和感知在穿梭箱行为的措施。
双向穿梭箱范式是研究回避学习12一个完善的实验室设备。梭箱包括2个舱室的一道坎或门口分开。甲条件刺激（CS），其通过适当的信号象光或声音来表示，是偶然随后厌恶无条件刺激（US），如例如一个足部电击在金属网格地板。受试者可以学习，以避免与美国通过从一个穿梭箱隔室中响应于所述CS穿梭到其他。穿梭箱学习涉及区分学习阶段的序列13,14：首先，科学预测，美国从CS由经典条件和从美国逃出用仪器调节，因为美国是在穿梭终止。在下一阶段，科目学习完全避免美国通过穿梭于应对美国发病前的CS（ 回避反应）。一般情况下，穿梭箱学习包括经典性条件反射，有助于调理，以及根据学习阶段14的目标导向的行为。
梭箱程序，可以方便地建立，一般经过几次的日常训练课15产生强大的行为– 17。除了简单的逃避性（检测），穿梭箱可进一步用于通过采用GO / NOGO范式研究刺激歧视。这里，训练动物，以避免与美国由一个条件反应（CR）（去行为;梭成相对隔室）响应于<STRONG>去刺激（CS +）和NOGO行为 （留在当前车厢;无CR）响应NOGO刺激（CS-）并行微刺激和记录神经活动的高密度多电极阵列允许学习。的生理机制成功的学习标的。几个技术细节，根本的穿梭盒子训练，ICMS和平行电成功的组合，将被讨论。
Protocol
Representative Results
Discussion
此协议通过使用双向厌恶足部电击控制穿梭盒子系统描述同时位点特异性ICMS和多通道电生理记录在一个学习的动物的方法。协议强调技术的关键概念为这样的组合，并指出只有通过它的共同接地电极接地的动物，留下gridfloor在浮动电压的重要性。在这里，听梭箱学习应用于长爪沙鼠的学习相关的塑料重组这些动物的听觉皮层中得到了广泛的8,12,14,15,21,22研究。然而，所描述的协议可以适于…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作是由来自Deustche研究联合会DFG和莱布尼茨研究所神经生物学的资助。我们感谢玛丽亚 – 滨海Zempeltzi和凯萨琳·奥尔的技术援助。
Materials
| Teflon-insulated stainless steel wire | California Fine Wire | diam. 50µm w/ isolation | |
| Pin connector system | Molex Holding GmbH | 510470200 | 1.25 mm pitch PicoBlade |
| TEM grid Quantifoil | Science Services | EQ225-N27 | |
| Dental acrylic Paladur | Heraeus Kulzer | 64707938 | |
| Hand-held drill OmniDrill35 | WPI | 503599 | |
| Ketamine 500mg/10ml | Ratiopharm GmbH | 7538837 | |
| Rompun 2%, 25ml | Bayer Vital GmbH | 5066.0 | |
| Sodium-Chloride 0.9%, 10ml | B.Braun AG | PRID00000772 | |
| Lubricant KY-Jelly | Johnson & Johnson | ||
| Shuttle-box E10-E15 | Coulbourn Instruments | H10-11M-SC | |
| Stimulus generator MCS STG 2000 | Multichannel Systems | ||
| Plexon Headstage cable 32V-G20 | Plexon Inc. | HSC/32v-G20 | |
| Plexon Headstage 32V-G20 | Plexon Inc. | HST/32v-G20 | |
| PBX preamplifier 32 channels | Plexon Inc. | 32PBX box | |
| Multichannel Acquisition System | Plexon Inc. | MAP 32/HLK2 | |
| Cryostate CM3050 S | Leica Microsystems GmbH | ||
| Signal processing Card Ni-Daq | National Instruments | ||
| Lab StandardTM Stereotaxic Instruments | Stoelting Co. | ||
| Audio attenator g.pah | g.pah Guger technologies | ||
| Cresyl violet acetate | Roth GmbH | 7651.2 | |
| Roticlear | Roth GmbH | A538.1 | |
| Sodium acetate trihydrate | Roth GmbH | 6779.1 | |
| Potassium hexacyanoferrat(II) trihydrate | Roth GmbH | 7974.2 | |
| Di-sodium hydrogen phospahte dihydrate | Merck | 1,065,801,000 | |
| ICM Impedance Conditioning Module | FHC | 55-70-0 | |
| Animal Temperarture Controler | World Precision Instruments | ATC2000 |
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