Funding for this project was provided by the National Health and Medical Research Council (NHMRC) 1046203 (BVB, AJV) and Melbourne Neuroscience Institute Fellowship (CTN).

所有的实验过程是根据实践的照顾和动物用于科学目的的使用澳大利亚代码，设置了由国家健康与医学研究委员会在澳大利亚进行。来自墨尔本大学，理学院，动物伦理委员会（批准文号0911322.1）获得伦理关。 慢性VEP电极1.植入前注:如果同时ERG和VEP信号要收集的动物必须VEP通过手术植入电极至少1周的信号收集前。 <ol><li>通过用氯己定（在70％乙醇0.5％）清洗消毒前实验手术台上。使用前高压灭菌所有的手术设备。盖上无菌手术单的动物。确保所有实验者戴上医用口罩，隔离衣和消毒手套。 </li><li>以3升/分钟的流速的3.5％的异氟烷与O 2 -诱发麻醉3。保持烷sthesia在整个手术1.5％和2升/分。由缺少爪子捏反射的保证麻醉足够的深度。 </li><li>在角膜上施加1％羧甲基纤维素钠，以防止眼睛干燥。 </li><li>剃30毫米×30mm的区域中的前额，后对眼睛和前到耳朵。 </li><li>放置在热垫（37℃）的动物，以保持体温并用立体框架稳定动物的头。 </li><li>消毒用10％聚维酮碘的三倍剃区域。避开眼睛附近的区域用含酒精的消毒剂，是与实践的标准一致列明外科技师协会。 </li><li>让头部中位数矢状切口用手术刀和真皮组织的这种消费一个约20毫米直径的圆，露出颅骨。 </li><li>刮磨和用纱布干燥，露出冠状面和矢状颅缝卸下底层的骨膜。 </li><li>我们ING连接到一个钻牙毛刺，在立体坐标环钻两个孔（0.7毫米直径，深度约1毫米），通过两个半球颅骨:7毫米尾鳍前囟3毫米的横向中线。 </li><li>螺钉在不锈钢螺丝（直径0.7mm时，长度为3毫米，具洗必泰灭菌）为两个预先制作孔高达约1毫米（2毫米螺钉的暴露）的深度，以允许牢固锚固。此接触硬膜，而不会损坏底层的皮层组织。 </li><li>在4〜8点0缝线 - 干燥用纱布颅骨，并用两个3缩回松弛的皮肤准备牙科用汞合金手术区。 </li><li>传播的牙科用汞合金在暴露的头骨到位，以固定（​​步骤1.10描述不锈钢螺丝）螺丝电极。保证〜螺丝1.5毫米保持暴露记录。 </li><li>卸下回缩缝线。 </li><li>注入0.5％卡洛芬皮下（5毫克/千克）镇痛和盐水（0.9％氯化钠，10.5毫升）皮下注射补液。 </li><li>让动物在笼子里单独恢复。直到它已经恢复了足够的意识，以保持胸骨斜卧不要让动物无人值守。 </li><li>不要返回动物公司其他动物的，直到它已完全手术（最少5天）恢复。 </li><li>继续管理0.5％，每天一次，镇痛卡洛芬皮下（5毫克/千克）4天。 </li><li>记录ERG和VEP1周手术。 </li></ol> 2. ERG和VEP记录<ol><li>数据收集准备<ol><li>使用计算机软件来同时触发的刺激，并根据下推荐的设置采集数据2。 <ol><li>放大信号3（ERG:×1000，VEP:x10,000）与由分离的前置放大器和放大器内部设置的增益，并与两个眼睛阻抗相匹配。 </li><li>设置采样率的ERG到4 kHz超过650毫秒记录窗口（2,560点），要做到这一点，单击在数据采集软件（名称和材料的软件，请参阅表的版本）"时基"选项卡，选择"2,560"为样本，而"500毫秒"时间它会返回一个650毫秒的记录窗口。 <ol><li>使用相同的方法，为VEP​​采样速率在250毫秒划时代设置为10千赫。允许为ERG和VEP录音10毫秒刺激前基线。要做到这一点，点击"设置"选项卡;选择"刺激"，弹出一个新的对话窗口;在该窗口中选择从下拉列表中的"模式"，"把脉";而对于"延迟"的值设置为"10毫秒"。 </li></ol></li><li>设置ERG带通滤波，以0.3 - 1,000赫兹（ - 3分贝）。这可以通过点击"生物放大器"中的数据采集软件来完成。然后设置为"高通"到"0.3Hz"的价值，并为"低通"到"1 kHz的价值"。 </li><li> 2.1.1.3在使用上述方法，设置VEP带通设置，以0.1 -作为视觉（符合ISCEV）的临床电推荐的国际社会为人类VEP录音6 - 100赫兹（3 dB为单位）。 </li></ol></li></ol></li><li>电极的制备<ol><li>分别由2银线或鳄鱼夹连接到电极引线，定制使ERG有效/无效和VEP有效/无效电极。商购获得的接地电极。 </li><li>对于4定做电极，切断从电极引线延长男性的结束。用手术刀刀片确保内钢丝未损坏除去1厘米外聚四氟乙烯绝缘涂层的。 </li><li>预方式通过切割银线（厚度0.3mm）的70毫米的长度，并形成一个环〜直径8mm包围鼠眼ERG的非活动电极。通过成形于1毫升枪头环路制备均匀圆。 </li><lI&gt;通过削减白银70毫米的长度，并形成一个椭圆〜8mm的长度方向的直径钩住大鼠门齿预时尚VEP无活性电极。 </li><li>预时尚切割银线的30毫米长，形成了一个小环轻轻接触大鼠角膜的ERG主动电极（〜1-2毫米直径） </li><li>牢固地缠绕在一起与暴露的内部钢丝的银附着的电极（2 ERG活性，2 ERG不活动，1 VEP不活动）的电极引线。 </li><li>多余的绝缘暴露的金属用胶带，以降低光伏文物。 </li><li>在ERG非活动电极棒一小片钩 - 环紧固件的（〜5毫米×20毫米），遮蔽胶带，以使稳定附着到啮齿动物颈带。 </li><li>鳄鱼夹附着在电极的内导线引出使VEP有源电极。 </li><li>之前的录音，电镀银导线的暴露表面（ 即，不活动环和ACTI使用9伏直流电源20秒，以改善信号传导已经尖）与酰氯。 <ol><li>要做到这一点，将其浸入ERG电极丝（充当主单元的阳极）到生理盐水的银尖;此电极导线的另一端​​连接到一个9伏电池的正端子。 </li><li>另一线（阴极）连接到该电池的负极端子，并浸入另一端插入盐水为好。 20秒后，断开并观察ERG电极丝的条子尖被以白色为主色调均匀涂抹。 注意:每个实验会议准备新的ERG电极（〜最多8小时），以确保氯化物涂层的通畅。 </li></ol></li></ol></li><li>动物准备<ol><li>在防光室暗适应录音前的动物过夜（≥8小时）。通过关闭室内灯光，关闭所有门和百叶窗确保最大的暗适应。通过将不透光材料，尽量减少周围漏光外面厚厚的黑色窗帘门/窗和放置电脑屏幕路口。 </li><li>进行动物准备在暗室内昏暗的红色发光二极管的帮助（LED; 17.4 cd.m -2，λ 最大 = 600纳米），以维持杆灵敏度。 </li><li> （5毫克/公斤60）肌内注射氯胺酮/赛拉嗪麻醉大鼠。由于没有一个爪子捏反射的确认麻醉足够的深度。 </li><li>为了保持镇静，如果需要的话经过50分钟施用麻醉的进一步剂量（首次剂量的50％）。 </li><li>有关其他表面麻醉适用0.5％proxymetacaine一滴每只眼睛，和眨眼掉多余的液体。 </li><li>对于瞳孔扩大适用0.5％托给每只眼睛一滴，然后擦干多余的液体。 </li></ol></li><li> ERG和VEP电极定位<ol><li>将动物ERG平台，在Ganzfeld碗坐落在法拉第笼门前。避免使用电热垫，因为它Ç一个引入电噪音进入电生理记录。注意:该平台被连接到一个循环加热的水平台以维持体温。 </li><li>安全动物平台牢牢但没有放置紧紧围绕颈背钩 - 环紧固件的带。 </li><li>钩围绕麻醉大鼠的底部门齿惰性VEP电极。 </li><li>通过眼部周围的环绕赤道巩膜环非侵入性的ERG不活动的电极位置。通过附加电极周围的颈背的钩 - 环紧固件条稳定这一点。重复对侧眼。 </li><li>法尔胜VEP通过鳄鱼夹连接到不锈钢螺丝头骨预先植入的电极活跃。 </li><li>放置1％羧甲基纤维素钠对角膜ERG的活性电极的放置之前一小滴，以改善信号质量。注:粘胶液还有助于保持整个实验到微型角膜水化迈兹干燥型白内障的啮齿动物7的形成。 </li><li>放在下门牙1％羧甲基纤维素钠的一小滴以改善VEP非活动电极的接触，从而信号质量。 </li><li>定位ERG主动电极使用连接到一个定制的立体手臂显微轻触中央角膜表面。 </li><li>插入2 - 接地针电极（不锈钢）皮下到尾5毫米。 </li><li>如有必要，干任何多余的液体从下眼睑的记录，以改善信号质量的前向。 </li><li>滑动平台接近Ganzfeld碗确保动物的眼睛与碗的开口对齐，以使甚至两个视网膜照度（见步骤2.4.1）。 </li><li>关闭法拉第笼，以减少多余的噪音。 </li></ol></li><li>数据采集<ol><li>使用昏暗的测试闪光（ - 0.52日志cd.sm -2），以评估是否电极placem耳鼻喉科令人满意2。在控制条件下，这将导致在〜800μV的ERG振幅和不大于10％的眼间的变异。如果需要重新定位电极。 </li><li>继测试闪光让动物暗适应了录音之前完全黑暗的10分钟。 </li><li>的使用Ganzfeld碗，同时收集ERG和VEP光刺激目前闪烁超过〜500毫秒的时间窗口同步信号。从调光器进展到更亮，以保持足够的暗适应特定的波形的光的水平。 </li><li>收集在一系列的发光能量信号引发STR，b波和ERG的A / B波的波形。平均多个信号的调光照明水平（20重复），少在明亮发光的能量（1重复）。逐渐拉长刺激间间隔为1〜180秒，从最暗到亮的光级。请参阅表1为例协议。 </li><li>为ISO后期ERG杆和视锥反应，利用配对闪光模式8。启动四次闪烁在1.52日志cd.sm -2，500毫秒刺激间隔2之间。数字减去从混合波形（1 次闪光）的锥形波形（3 次或4 次闪光）以导出推定杆响应。 </li><li>以记录VEP信号，平均20重复在明亮发光的能量（ 即 ， - 0.52〜1.52日志cd.sm -2，5秒刺激间间隔）。注意，在该序列中的第一闪光灯返回常规暗适应ERG应答。 </li><li>允许1 - 用于重新适应3分钟（20）VEP下一亮ERG步骤之前扫过之后，这取决于光能量。 </li><li>数据采集​​完成后，安乐死用戊巴比妥钠（325毫克/毫升，3ml）中的心内注射麻醉动物。 </li></ol></li></ol><table border="1" fo:keep-together.within-page="1" fo:保持与 - next.within页="总是"&gt; <tbody><tr><td> 波形 </td><td> 刺激光能（日志cd.sm -2） </td><td> 重复次数 </td><td> 刺激间隔（秒） </td></tr><tr><td> STR </td><td> -6.24 </td><td> 20 </td><td> 2 </td></tr><tr><td> STR </td><td> -5.93 </td><td> 20 </td><td> 2 </td></tr><tr><td> STR </td><td> -5.6 </td><td> 20 </td><td> 2 </td></tr><tr><td> STR </td><td> -5.33 </td><td> 20 </td><td> 2 </td></tr><tr><td>杆b波</td><td> -4.99 </td><td> 10 </td><td> 2 </td></tr><tr><td>杆b波</td><td> -4.55 </td><td> 10 </td><td> 2 </td></tr><tr><td>杆b波</td><td> -4.06 </td><td>五</td><td>五</td></tr><tr><td>杆b波</td><td> -3.51 </td><td>五</td><td>五</td></tr><tr><td>杆b波</td><td> -3.03 </td><td> 1 </td><td> 1五</td></tr><tr><td>杆b波</td><td> -2.6 </td><td> 1 </td><td> 15 </td></tr><tr><td>杆b波</td><td> -1.98 </td><td> 1 </td><td> 15 </td></tr><tr><td>混合A- / b波</td><td> -1.38 </td><td> 1 </td><td>三十</td></tr><tr><td>混合A- / b波</td><td> -0.94 </td><td> 1 </td><td>三十</td></tr><tr><td rowspan="2">闪光灯1:20的混合A- / B波平均:VEP </td><td rowspan="2"> -0.52 </td><td rowspan="2"> 20 </td><td>五</td></tr><tr><td> （接下来的前90秒） </td></tr><tr><td rowspan="2">闪光灯1:20的混合A- / B波平均:VEP </td><td rowspan="2"> 0.04 </td><td rowspan="2"> 20 </td><td>五</td></tr><tr><td> （接下来的前120秒） </td></tr><tr><td rowspan="2">闪光灯1:20的混合A- / B波平均:VEP </td><td rowspan="2"> 0.58 </td><td rowspan="2"> 20 </td><td>五</td></tr><tr><td> （接下来的前180秒） </td></tr><tr><td rowspan="2"> FLA文件H 1:20的混合A- / B波平均:VEP </td><td rowspan="2"> 1.2 </td><td rowspan="2"> 20 </td><td>五</td></tr><tr><td> （接下来的前180秒） </td></tr><tr><td rowspan="2">闪光灯1:20的混合A- / B波平均:VEP </td><td rowspan="2"> 1.52 </td><td rowspan="2"> 20 </td><td>五</td></tr><tr><td> （接下来的前180秒） </td></tr><tr><td>锥A- / b波</td><td> 1.52 </td><td> 4 </td><td> 0.5 </td></tr></tbody></table> 表1. ERG和VEP记录协议使用激励能源的范围。从暗淡（顶）的经济刺激演示进度亮（底部）闪烁，有足够的刺激间隔，以保证暗适应。在协议结束时，呈现四个闪烁以短的时间间隔重复以引起锥体介导的应答。 3. ERG波形分析注:ERG和VEP分析先前详细描述3,9,10的。以下部分提供了简要概述。 <ol><li>数字电压 - 时间格式的电子表格软件进行数据分析导出的信号。 </li><li>罗德photoreceptoral功能<ol><li>建模一个波PIII的前缘与延迟高斯（等式1）11。 PIII（I，T）= PIII室∙[1 - EXP（ - 我 ∙-S（T - T 的 D）2）]在t&gt; T D（公式1） </li><li>优化配合了两个亮的发光能量的合奏12,13（ 即 1.22和1.52日志cd.sm -2）。 </li><li>模型到a-波幅度的90％，以避免后期receptoral入侵14。 注意:该模型返回photoreceptoral响应的饱和幅度（罗PIII，μV），灵敏度（S，M 2 .CD -1 .s的-3）和延迟（叔D，毫秒）。 </li></ol></li><li>杆双极细胞FUNCT离子<ol><li>数字减去PIII模型（见上文）从混合波形与覆振荡电位返回该混合PII。 </li><li>提取混合PII杆PII，数字减去从混合PII的视锥细胞反应（ 第三或1.52日志cd.sm 4 次闪光-2）（1.52日志cd.sm 1 日闪-2）。 </li><li>然后，应用一个低通滤波器的波形（46.9赫兹，-3分贝，布莱克曼窗）以除去振荡电位。剩余的波形是在杆的PII响应10。 </li><li>提取杆PII峰值和（1.52日志cd.sm -2低于-2日志cd.sm -2和杆隔离PII）10绘制它反对一切刺激强度。 </li><li>模型使用双曲函数（公式2），其中提供内视网膜细胞的完整性的量度这些数据。 V（I）= V MAX（I N /（I N+ K N））（式2） 注意:此方程返回最大PII响应（Ⅴ 最大值 ，μV），1 /灵敏度（K，日志cd.sm-2）和功能（n）的15的斜率。 </li></ol></li><li>锥双极细胞功能 注:由于视锥细胞反应是采取在一个单一的强度（1.52日志cd.sm -2）的幅度和时间在这个光照水平的重新调整。 <ol><li>提取最大锥PII回应2,16。 </li><li>提取到这个最大响应对应2,16隐含的时间。 </li></ol></li><li>神经节细胞的功能<ol><li>由于STR是小信号，应用一个低通滤波器具有50Hz的缺口的波形，以消除高频和线路噪音（46.9赫兹，-3分贝，布莱克曼窗）。 </li><li>提取最大PSTR响应3.17。 </li><li>提取到这个最大响应对应3.17隐含的时间。 </li></ol></li></ol> 4.安娜VEP波形的裂解<ol><li>提取VEP（P1，N1和P2）的最大和最小分量。详细见参考文献3,6。 </li><li>明示的振幅为从它们的前峰或槽（P1N1和N1P2）3,6-槽-峰振幅。 </li><li>提取隐含的时间（ 它 ）到这个最大响应对应（P1 了 ，N1 它 ， 它 P2）3,6。 </li></ol>

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The authors have no disclosures relevant to this work.

在ERG和VEP分别从视网膜和皮质，视觉功能的目标的措施。同时记录的优点是，整个视觉通路的更全面的观点得到。具体地，从它们的并发评估中的补充信息可在视觉通路提供损伤部位的一个更清楚的划分（ 例如，对于具有重叠ERG但不同VEP表现18，病症当视神经病变可以与初级脑萎缩19共存， 20时，或VEP的损失可能会受到伤病的表现在多个地点在视觉通路21,22?...

电图（ERG）和视觉诱发电位（VEP）的测量提供视觉通路的完整性有用的定量评估。该ERG测量视网膜对光刺激的电反应，而VEP措施而相同的光从事件视网膜初级视觉皮层的视觉通路的相应功能的完整性。该原稿描述了在一个通常使用的实验室模型，大鼠的记录和ERG和VEP响应分析的协议。 该ERG通过量化来一道闪光视网膜的总电反应提供了许多关键的视网膜细胞类的功能完整性的指标。一个协调的一系列离子通量的光引发发病和偏移，产生可使用放置在眼外表面的电极来测量电压检测的变化。所得波形表示的本身在组合明确定义的部件的里斯，在幅度，定时和频率不同。的研究有相当体已经表明这些分量在许多脊椎动物视网膜并且该组件可从彼此分离相对保守。通过审慎地选择刺激（闪光刺激，背景，刺激间隔）的条件下和在选择合成波形的特定特性来分析一个可以确信返回视网膜细胞1,2的一组特定的量度。这些特征背后的效用，因此ERG的广泛应用，视网膜功能的非侵入性的措施。此手稿着重于方法用于测量ERG和分析其功能返回有关一些在视网膜的主要细胞类，即光感受器（PIII的成分），双极细胞（PII中成分）和视网膜神经节细胞的信息（正暗视阈值反应或PSTR）。 3的区域V1的视辐射串行通信。在啮齿类动物中，多数-从每只眼睛交互对4视神经纤维的（90 95％）和支配对侧中脑。不像ERG，它是作为还不可能将VEP的不同部件归因于特定的细胞类，从而5改变任意位置沿视觉通路可能影响VEP波形。尽管如此，VEP是视觉性能和视觉通路的完整性的有用的非侵入性的措施。的VEP，与ERG一起使用时，可以提供视觉系统（ 即，视网膜/视觉通路）的更完整的评价。 ERG和VEP录音可以在隔离或组合来进行，这取决于APPLI阳离子。本文介绍的方法可以从两个眼睛视网膜和大脑皮质视觉诱发电生理和麻醉大鼠两个半球同时评估。这是为了更全面地评估视网膜功能和上游的效果，在视网膜功能变化可能对视觉诱发皮质功能的有效方法。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Alligator clip</td>
			<td>generic brand</td>
			<td>HM3022</td>
			<td>Stainless steel 26 mm clip for connecting VEP screw electrodes to cables</td>
		</tr>
		<tr>
			<td>Bioamplifier</td>
			<td>ADInstruments</td>
			<td>ML 135</td>
			<td>For amplifying ERG and VEP signals</td>
		</tr>
		<tr>
			<td>Carboxymethylcellulose sodium 1.0%</td>
			<td>Allergan</td>
			<td>CAS 0009000-11-7</td>
			<td>Viscous fluid for improving signal quality of the active ERG electrode</td>
		</tr>
		<tr>
			<td>Carprofen 0.5%</td>
			<td>Pfizer Animal Health Group</td>
			<td>CAS 53716-49-7</td>
			<td>Proprietary name: Rimadyl injectable (50 mg/mL). For post-surgery analgesia, diluted to 0.5% (5 mg/mL) in normal saline</td>
		</tr>
		<tr>
			<td>Chlorhexadine 0.5%</td>
			<td>Orion Laboratories</td>
			<td>27411, 80085</td>
			<td>For disinfecting surgical instruments</td>
		</tr>
		<tr>
			<td>Circulating water bath</td>
			<td>Lauda-K&#246;nigshoffen</td>
			<td>MGW Lauda</td>
			<td>For maintaining body temperature of the anesthetized animal during surgery and electrophysiological recordings</td>
		</tr>
		<tr>
			<td>Dental amalgam</td>
			<td>DeguDent GmbH</td>
			<td>64020024</td>
			<td>For encasing the electrode-skull assembly to make it more robust</td>
		</tr>
		<tr>
			<td>Dental burr</td>
			<td>Storz Instruments, Bausch and Lomb</td>
			<td>#E0824A</td>
			<td>A miniature drill head of ~0.7mm diameter for making a small hole in the skull over each hemisphere to implant VEP screws</td>
		</tr>
		<tr>
			<td>Drill</td>
			<td>Bosch</td>
			<td>Dremel 300 series</td>
			<td>An automatic drill for trepanning</td>
		</tr>
		<tr>
			<td>Electrode lead</td>
			<td>Grass Telefactor&#160;</td>
			<td>F-E2-30</td>
			<td>Platinum cables for connecting silver wire electrodes to the amplifier</td>
		</tr>
		<tr>
			<td>Faraday Cage</td>
			<td>custom-made</td>
			<td></td>
			<td>Ensures light proof to maintain dark adaptation. Encloses the Ganzfeld setup to improve signal to noise ratio</td>
		</tr>
		<tr>
			<td>Gauze swabs</td>
			<td>Multigate Medical Products Pty Ltd</td>
			<td>57-100B</td>
			<td>For drying the surgical incision and exposed skull surface during surgery</td>
		</tr>
		<tr>
			<td>Ganzfeld integrating sphere</td>
			<td>Photometric Solutions International</td>
			<td></td>
			<td>Custom designed light stimulator: 36 mm diameter, 13 cm aperture size</td>
		</tr>
		<tr>
			<td>Velcro</td>
			<td>VELCRO Australia Pty Ltd</td>
			<td>VELCRO Brand Reusable Wrap</td>
			<td>Hook-and-loop fastener to secure the electrodes and the animal on the recording platform</td>
		</tr>
		<tr>
			<td>Isoflurane 99.9%</td>
			<td>Abbott Australasia Pty Ltd</td>
			<td>CAS 26675-46-7</td>
			<td>Proprietary Name: Isoflo(TM) Inhalation anaaesthetic. Pharmaceutical-grade inhalation anesthetic mixed with oxygen gas for VEP electrode implant surgery</td>
		</tr>
		<tr>
			<td>Ketamine&#160;</td>
			<td>Troy Laboratories</td>
			<td>Ilium Ketamil</td>
			<td>Proprietary name: Ketamil Injection, Brand: Ilium. Pharmaceutical-grade anesthetic for electrophysiological recording</td>
		</tr>
		<tr>
			<td>Luxeon LEDs</td>
			<td>Phillips Lighting Co.</td>
			<td></td>
			<td>For light stimulation twenty 5 watt and one 1 watt LEDs.</td>
		</tr>
		<tr>
			<td>Micromanipulator</td>
			<td>Harvard Apparatus</td>
			<td>BS4 50-2625</td>
			<td>Holds the ERG active electrode during recordings</td>
		</tr>
		<tr>
			<td>Needle electrode</td>
			<td>Grass Telefactor&#160;</td>
			<td>F-E2-30</td>
			<td>Subcutaneously inserted in the tail to serve as the ground electrode for both the ERG and VEP</td>
		</tr>
		<tr>
			<td>Phenylephrine 2.5% minims&#160;</td>
			<td>Bausch and Lomb</td>
			<td>CAS 61-76-7</td>
			<td>Instilled with Tropicamide to achieve maximal dilation for ERG recording</td>
		</tr>
		<tr>
			<td>Povidone iodine 10%</td>
			<td>Sanofi-Aventis</td>
			<td>CAS 25655-41-8</td>
			<td>Proprietory name: Betadine, Antiseptic to prepare the shaved skin for surgery 10%, 500 mL</td>
		</tr>
		<tr>
			<td>Powerlab data acquisition system</td>
			<td>ADInstruments</td>
			<td>ML 785</td>
			<td>Controls the LEDs</td>
		</tr>
		<tr>
			<td>Proxymetacaine 0.5%</td>
			<td>Alcon Laboratories&#160;</td>
			<td>CAS 5875-06-9</td>
			<td>For corneal anaesthesia during ERG recordings</td>
		</tr>
		<tr>
			<td>Saline solution</td>
			<td>Gelflex</td>
			<td></td>
			<td>Non-injectable, for electroplating silver wire electrodes</td>
		</tr>
		<tr>
			<td>Scope Software</td>
			<td>ADInstruments</td>
			<td>version 3.7.6</td>
			<td>Simultaneously triggers the stimulus via the Powerlab system and collects data</td>
		</tr>
		<tr>
			<td>Silver (fine round wire)</td>
			<td>A&#38;E metal</td>
			<td>0.3 mm</td>
			<td>Used to make active and inactive ERG electrodes, and the inactive VEP electrode</td>
		</tr>
		<tr>
			<td>Stainless streel screws&#160;</td>
			<td>MicroFasterners</td>
			<td></td>
			<td>0.7 mm shaft diameter, 3 mm in length to be implanted over the primary visual cortex and serve as the active VEP electrodes</td>
		</tr>
		<tr>
			<td>Stereotaxic frame</td>
			<td>David Kopf</td>
			<td>Model 900</td>
			<td>A small animal stereotaxic instrument for locating the primary visual cortices according to Paxinos &#38; Watson&#39;s 2007 rat brain atlas coordinates</td>
		</tr>
		<tr>
			<td>Surgical blade</td>
			<td>Swann-Morton Ltd.</td>
			<td>0206</td>
			<td>For incising the area of skin overlaying the primary visual cortex to implant the VEP electrodes</td>
		</tr>
		<tr>
			<td>Suture</td>
			<td>Shanghai Pudong Jinhuan Medical Products Co.,Ltd</td>
			<td></td>
			<td>3-0 silk braided suture non-absorbable, for skin retraction during VEP electrode implantation surgery</td>
		</tr>
		<tr>
			<td>Tobramycine eye ointment 0.3%</td>
			<td>Alcon Laboratories&#160;</td>
			<td>CAS 32986-56-4</td>
			<td>Proprietary name: Tobrex. Prophylactic antibiotic ointment applied around the skin wound after surgery</td>
		</tr>
		<tr>
			<td>Tropicamide 0.5%</td>
			<td>Alcon Laboratories&#160;</td>
			<td>CAS 1508-75-4</td>
			<td>Proprietary name: 0.5% Mydriacyl eye drop, Instilled to achieve mydriasis for ERG recording</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Troy Laboratories</td>
			<td>Ilium Xylazil-100</td>
			<td>Pharmaceutical-grade anesthetic for electrophysiological recording</td>
		</tr>
		<tr>
			<td>Pipette tip&#160;</td>
			<td>Eppendorf Pty Ltd</td>
			<td>0030 073.169</td>
			<td>Eppendorf epTIPS 100 - 5000 mL, for custom-made electrodes</td>
		</tr>
		<tr>
			<td>Microsoft Office Excel</td>
			<td>Microsoft</td>
			<td>version 2010</td>
			<td>spreadsheet software for data analysis</td>
		</tr>
		<tr>
			<td>Lethabarb Euthanazia Injection</td>
			<td>Virbac (Australia) Pty Ltd</td>
			<td>LETHA450</td>
			<td>325 mg/mL pentobarbital sodium for rapid euthanazia</td>
		</tr>
	</tbody>
</table>

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.

ERG的a波（&gt; -1.38日志cd.sm -2），B波（&gt; - 4.99日志cd.sm -2）个STR（&lt; - 4.99日志cd.sm -2）和VEPs（&gt; - 0.52登录cd.sm -2）同时（ 图1和记录3）。在非常暗淡闪烁，一个正STR（PSTR）被认为在约闪光后110毫秒，并且在大约220毫秒的负STR（NSTR）（ 图1和2）。具有大b波的ERG，50到100毫秒的中等闪光可以以其PII响?...

Watch this Scientific Journal Video about Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats at JoVE.com

Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats

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 ...

simultaneous-recording-electroretinography-visual-evoked-potentials

Research

JoVE Journal

Neuroscience

12.2K 次观看.  University of Melbourne. This protocol describes simultaneous measurement of electroretinogram and visual evoked potentials in anesthetized rats. 

视频: Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats

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

Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of behaviors from the spiking activity of its neurons. One of the most effective methods to monitor spiking activity from a large number of neurons in multiple local neuronal circuits simultaneously is by using silicon electrode arrays1-3.
Action potentials produce large transmembrane voltage changes in the vicinity of cell somata. These output signals can be measured by placing a conductor in close proximity of a neuron. If there are many active (spiking) neurons in the vicinity of the tip, the electrode records combined signal from all of them, where contribution of a single neuron is weighted by its 'electrical distance'. Silicon probes are ideal recording electrodes to monitor multiple neurons because of a large number of recording sites (+64) and a small volume. Furthermore, multiple sites can be arranged over a distance of millimeters, thus allowing for the simultaneous recordings of neuronal activity in the various cortical layers or in multiple cortical columns (Fig. 1). Importantly, the geometrically precise distribution of the recording sites also allows for the determination of the spatial relationship of the isolated single neurons4. Here, we describe an acute, large-scale neuronal recording from the left and right forelimb somatosensory cortex simultaneously in an anesthetized rat with silicon probes (Fig. 2).

Extracellular recordings of neuronal activity using silicon probes in the anesthetized rat will be described. This technique allows information to be obtained across multiple brain areas from more than 100 neurons simultaneously. It provides information with single cell resolution about neuronal ensembles dynamics in multiple local circuits.

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 ...

科研

神经科学

Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation

One of the central tasks in retinal neuroscience is to understand the circuitry of retinal neurons and how those connections are responsible for shaping the signals transmitted to the brain. Photons are detected in the retina by rod and cone photoreceptors, which convert that energy into an electrical signal, transmitting it to other retinal neurons, where it is processed and communicated to central targets in the brain via the optic nerve. Important early insights into retinal circuitry and visual processing came from the histological studies of Cajal1,2 and, later, from electrophysiological recordings of the spiking activity of retinal ganglion cells - the output cells of the retina3,4.
A detailed understanding of visual processing in the retina requires an understanding of the signaling at each step in the pathway from photoreceptor to retinal ganglion cell. However, many retinal cell types are buried deep in the tissue and therefore relatively inaccessible for electrophysiological recording. This limitation can be overcome by working with vertical slices, in which cells residing within each of the retinal layers are clearly visible and accessible for electrophysiological recording.
Here, we describe a method for making vertical sections of retinas from larval tiger salamanders (Ambystoma tigrinum). While this preparation was originally developed for recordings with sharp microelectrodes5,6, we describe a method for dual whole-cell voltage clamp recordings from photoreceptors and second-order horizontal and bipolar cells in which we manipulate the photoreceptor's membrane potential while simultaneously recording post-synaptic responses in horizontal or bipolar cells. The photoreceptors of the tiger salamander are considerably larger than those of mammalian species, making this an ideal preparation in which to undertake this technically challenging experimental approach. These experiments are described with an eye toward probing the signaling properties of the synaptic ribbon - a specialized synaptic structure found in a only a handful of neurons, including rod and cone photoreceptors, that is well suited for maintaining a high rate of tonic neurotransmitter release7,8 - and how it contributes to the unique signaling properties of this first retinal synapse.

We describe the preparation of thin retinal slices from aquatic tiger salamanders (Ambystoma tigrinum) and explain how we use these slices to study synaptic processing in the retina by obtaining dual whole-cell voltage clamp recordings from photoreceptors and second-order horizontal and bipolar cells.

同时光感受器和二阶神经元在两栖类动物的视网膜切片制备全细胞记录

One of the central tasks in retinal neuroscience  is to understand the circuitry of retinal neurons and how those connections are  responsible for shaping ...

Simultaneous Electrophysiological Recording and Calcium Imaging of Suprachiasmatic Nucleus Neurons

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in regulating the intracellular calcium concentration. Changing environmental conditions, such as the light-dark cycle, can modify neuronal and neural network activity and the expression of a family of circadian clock genes within the suprachiasmatic nucleus (SCN), the location of the master circadian clock in the mammalian brain. Excitatory synaptic transmission leads to an increase in the postsynaptic Ca2+ concentration that is believed to activate the signaling pathways that shifts the rhythmic expression of circadian clock genes. Hypothalamic slices containing the SCN were patch clamped using microelectrodes filled with an internal solution containing the calcium indicator bis-fura-2. After a seal was formed between the microelectrode and the SCN neuronal membrane, the membrane was ruptured using gentle suction and the calcium probe diffused into the neuron filling both the soma and dendrites. Quantitative ratiometric measurements of the intracellular calcium concentration were recorded simultaneously with membrane potential or current. Using these methods it is possible to study the role of changes of the intracellular calcium concentration produced by synaptic activity and action potential firing of individual neurons. In this presentation we demonstrate the methods to simultaneously record electrophysiological activity along with intracellular calcium from individual SCN neurons maintained in brain slices.

Procedures are described to perform simultaneous recordings of membrane potential or current and changes of intracellular calcium concentration. Suprachiasmatic nucleus neurons are filled with the calcium indicator bis-fura-2 using a patch clamp electrode in the whole cell patch clamp configuration.

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in ...

Extracting Visual Evoked Potentials from EEG Data Recorded During fMRI-guided Transcranial Magnetic Stimulation

Transcranial Magnetic Stimulation (TMS) is an effective method for establishing a causal link between a cortical area and cognitive/neurophysiological effects. Specifically, by creating a transient interference with the normal activity of a target region and measuring changes in an electrophysiological signal, we can establish a causal link between the stimulated brain area or network and the electrophysiological signal that we record. If target brain areas are functionally defined with prior fMRI scan, TMS could be used to link the fMRI activations with evoked potentials recorded. However, conducting such experiments presents significant technical challenges given the high amplitude artifacts introduced into the EEG signal by the magnetic pulse, and the difficulty to successfully target areas that were functionally defined by fMRI. Here we describe a methodology for combining these three common tools: TMS, EEG, and fMRI. We explain how to guide the stimulator&#39;s coil to the desired target area using anatomical or functional MRI data, how to record EEG during concurrent TMS, how to design an ERP study suitable for EEG-TMS combination and how to extract reliable ERP from the recorded data. We will provide representative results from a previously published study, in which fMRI-guided TMS was used concurrently with EEG to show that the face-selective N1 and the body-selective N1 component of the ERP are associated with distinct neural networks in extrastriate cortex. This method allows us to combine the high spatial resolution of fMRI with the high temporal resolution of TMS and EEG and therefore obtain a comprehensive understanding of the neural basis of various cognitive processes.

This paper describes a method for collecting and analyzing electroencephalography (EEG) data during concurrent transcranial magnetic stimulation (TMS) guided by activations revealed with functional magnetic resonance imaging (fMRI). A method for TMS artifact removal and extraction of event related potentials is described as well as considerations in paradigm design and experimental setup.

提取脑电数据视觉诱发电位记录在功能磁共振成像引导下经颅磁刺激

Transcranial Magnetic Stimulation (TMS) is an effective method for establishing a causal link between a cortical area and cognitive/neurophysiological ...

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

An In vivo electroretinogram (ERG) signal is composed of several overlapping components originating from different retinal cell types, as well as noise from extra-retinal sources. Ex vivo ERG provides an efficient method to dissect the function of retinal cells directly from an intact isolated retina of animals or donor eyes. In addition, ex vivo ERG can be used to test the efficacy and safety of potential therapeutic agents on retina tissue from animals or humans. We show here how commercially available in vivo ERG systems can be used to conduct ex vivo ERG recordings from isolated mouse retinas. We combine the light stimulation, electronic and heating units of a standard in vivo system with custom-designed specimen holder, gravity-controlled perfusion system and electromagnetic noise shielding to record low-noise ex vivo ERG signals simultaneously from two retinas with the acquisition software included in commercial in vivo systems. Further, we demonstrate how to use this method in combination with pharmacological treatments that remove specific ERG components in order to dissect the function of certain retinal cell types.

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

Ex vivo ERG can be used to record electrical activity of retinal cells directly from isolated intact retinas of animals or humans. We demonstrate here how common in vivo ERG systems can be adapted for ex vivo ERG recordings in order to dissect the electrical activity of retinal cells.

同时<em&gt;体外</em两个由视网膜&gt;功能测试<em&gt;在体内</em&gt;电图系统

An In vivo electroretinogram (ERG) signal is composed of several overlapping components originating from different retinal cell types, as well as noise ...

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 ...

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the pathophysiological basis of these diseases, it is necessary to precisely characterize in which way the processing of information is disturbed between the different neuronal parts of the circuit. Using extracellular in vivo electrophysiological recordings, it is possible to accurately delineate neuronal activity within a neuronal network. The application of this method has several advantages over alternative techniques, e.g., functional magnetic resonance imaging and calcium imaging, as it allows a unique temporal and spatial resolution and does not rely on genetically engineered organisms. However, the use of extracellular in vivo recordings is limited since it is an invasive technique that cannot be universally applied. In this article, a simple and easy to use method is presented with which it is possible to simultaneously record extracellular potentials such as local field potentials and multiunit activity at multiple sites of a network. It is detailed how a precise targeting of subcortical nuclei can be achieved using a combination of stereotactic surgery and online analysis of multi-unit recordings. Thus, it is demonstrated, how a complete network such as the hyperdirect cortico-basal ganglia loop can be studied in anesthetized animals in vivo.

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

In this study, the methodology is presented on how to perform multi-site in vivo electrophysiological recordings from the hyperdirect pathway under urethane anesthesia.

急性<em&gt;体内</em&gt;麻醉大鼠超导途径局部场电位和多单位活动的电生理记录

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

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 ...

Visual Evoked Potential Recordings in Mice Using a Dry Non-invasive Multi-channel Scalp EEG Sensor

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable because of the plunger-spring-barrel structural facet and mechanical strengths resulting from metal materials. The whole process for acquiring the VEP responses in vivo from a mouse consists of four steps: (1) sensor assembly, (2) animal preparation, (3) VEP measurement, and (4) signal processing. This paper presents representative measurements of VEP responses from multiple mice with a submicro-voltage signal resolution and sub-hundred millisecond temporal resolution. Although the proposed method is safer and more convenient compared to other previously reported animal EEG acquiring methods, there are remaining issues including how to enhance the signal-to-noise ratio and how to apply this technique with freely moving animals. The proposed method utilizes easily available resources and shows a repetitive VEP response with a satisfactory signal quality. Therefore, this method could be utilized for longitudinal experimental studies and reliable translational research exploiting non-invasive paradigms.

We designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable. This paper describes the whole process from manufacturing the proposed EEG electrode to signal processing of visual evoked potential (VEP) signals measured on a mouse scalp using a dry non-invasive multi-channel EEG sensor.

干无创多通道头皮脑电传感器在小鼠视觉诱发电位记录中的应用

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable ...

Stimulus-specific Cortical Visual Evoked Potential Morphological Patterns

This paper presents a methodology for the recording and analysis of cortical visual evoked potentials (CVEPs) in response to various visual stimuli using 128-channel high-density electroencephalography (EEG). The specific aim of the described stimuli and analyses is to examine whether it is feasible to replicate previously reported CVEP morphological patterns elicited by an apparent motion stimulus, designed to simultaneously stimulate both ventral and dorsal central visual networks, using object and motion stimuli designed to separately stimulate ventral and dorsal visual cortical networks.&#160; Four visual paradigms are presented: 1. Randomized visual objects with consistent temporal presentation. 2. Randomized visual objects with inconsistent temporal presentation (or jitter).&#160; 3. Visual motion via a radial field of coherent central dot motion without jitter.&#160; 4. Visual motion via a radial field of coherent central dot motion with jitter.&#160; These four paradigms are presented in a pseudo-randomized order for each participant.&#160; Jitter is introduced in order to view how possible anticipatory-related effects may affect the morphology of the object-onset and motion-onset CVEP response.&#160; EEG data analyses are described in detail, including steps of data exportation from and importation to signal processing platforms, bad channel identification&#160;and removal, artifact rejection, averaging, and categorization of average CVEP morphological pattern type based upon latency ranges of component peaks. Representative data show that the methodological approach is indeed sensitive in eliciting differential object-onset and motion-onset CVEP morphological patterns and may, therefore, be useful in addressing the larger research aim. Given the high temporal resolution of EEG and the possible application of high-density EEG in source localization analyses, this protocol is ideal for the investigation of distinct CVEP morphological patterns and the underlying neural mechanisms which generate these differential responses.&#160; &#160; &#160;&#160;

In this paper, we present a protocol to investigate differential cortical visual evoked potential morphological patterns through stimulation of ventral and dorsal networks using high-density EEG. Visual object and motion stimulus paradigms, with and without temporal jitter, are described. Visual evoked potential morphological analyses are also outlined.&#160;&#160;

刺激特定的皮质视觉引发潜在形态模式

This paper presents a methodology for the recording and analysis of cortical visual evoked potentials (CVEPs) in response to various visual stimuli using ...