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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</ol>

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混淆）。通过测量ERG和VEP同时，视网膜和皮质响应之间的增益的指数也可以导出。这可以提供，以检测细微病变的有用工具。目前的协议允许ERG和VEP测量实验室常用的作用，但可以很容易地广告艾普特其他哺乳动物物种23-25。啮齿动物ERG和VEP波形提供人眼观察到26-28响应的临床合理替代。 通过设计一个特定的刺激协议，可在单个记录会话期间获得既ERG和VEP响应。 表1示出光水平与连续闪光之间的恢复时间适当考虑一个进展。这个协议提供了必要最大化信号对噪声的特性，并通过氯胺酮的单剂量提供的麻药窗口内限制记录时间之间的平衡:赛拉嗪。因此，该技术可能是用于研究基本生理和疾病的视觉功能的客观定量测量是有用的。 视觉系统的全面评估可以通过同时评估双边视网膜反应和视觉诱发皮层反应来实现。然而，每个技术也可以在隔离进行和单眼代替双眼，以简化程序。目前的协议描述选择给老鼠有一杆为主的视网膜杆通路分离暗ERG和VEP信号。如果光适于响应是对研究更感兴趣的，但是也可以通过预适应进行明视视网膜电图和VEP信号到背景光。 这种技术的一个主要限制是有必要进行麻醉的条件下的步骤，以使稳定的电极放置。不过，这种方法提供了可靠的信号与噪声的特性使细微的改变治疗检测。 由于STR及其光适应灵敏度的小振幅，几个步骤需要密切观察，以确保这种反应的成功记录。首先，充分暗适应需要被实现，其包括在昏暗的灯光红暗隔夜适应（≥8小时），放置电极（17.4 cd.m -2，λ 最大 = 600纳米），并重新暗适应下一个昏暗的测试闪光（10分钟为- 0.52日志CD。 SM-2）。 （ 即，20个信号）搜集与短刺激间的间隔（ 即 ，2秒）另外，STR的信号-噪声特性可以通过多个信号平均来得到改善。一双眼和皮质的这种综合评估的优点是使比较对侧记录3。因此，应特别注意在制作电极（ 即 ，相同的尺寸和形状的电极）采取，以确保最小的跨眼睛和跨皮质变性。 鉴于双方ERG和VEP技术在视觉通路及其与疾病相关的流程体内的措施，提供广泛的使用，它会整理等具体途径-P是有用的rotocols（ 例如 ，开/关或锥形子类型特异性），并同时执行ERG / VEP录音用不同的刺激模式（ 例如 ，闪烁，图案，锯齿），以在临床诊断扩展该技术的应用。在未来的这个应用程序的另一种合乎逻辑的步骤也将是从意识的同时29记录ERG和VEP，可自由移动的动物，以避免对神经生理学30麻药的影响。

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><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&amp;ouml;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 ~ 0.7 mm 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&amp;nbsp;</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&amp;nbsp;</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 W and one 1 W 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&amp;nbsp;</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&amp;nbsp;</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&amp;nbsp;</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&amp;amp;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&amp;nbsp;</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 &amp;amp; Watson&#039;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&amp;nbsp;</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&amp;nbsp;</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</td><td>Eppendorf Pty Ltd</td><td>0030 073.169</td><td>Eppendorf epTIPS 100 - 5,000 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响应被分析开始后之间的峰（ 图1和2）。在这种刺激的能量，峰值前的负a波是微不足道的。在明亮的发光能量的负偏转波变成能够与PIII响应（ 图2）来量化更为突出。暗视VEP波形显示了一个否定的答复（P1N1; 15 - 70毫秒窗口）后跟<html>一个正偏转（N1P2; 30 - 100毫秒）（ 图3和4）。 <img alt="图1" src="/files/ftp_upload/54158/54158fig1.jpg" /> 图1.组平均ERG波形:ERG改变随刺激强度。号码波形左侧标明用来引出波形发光曝光。注意每个面板的不同幅度和时间尺度。在调光器的发光能量的暗视阈响应的正和负分量可以引起（PSTR，NSTR）。由于刺激能量变亮，a和b波的反应可用来测定，而配对的闪光模式可以测量锥回应。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54158/54158fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> PLOAD / 54158 / 54158fig2.jpg"/&gt; 图2. ERG分析。（A）杆感光功能，可以通过使用PIII到A波模型进行检测。 A-波在1.22和1.52日志cd.sm -2（空心圆，○）特别适合作为与PIII合奏（灰线，方程1）到返回室PIII最低的90％（饱和幅度，μV） S（灵敏度，米2 .CD -1 .s的-3）和TD（定时延迟，毫秒）的参数。（B）的杆双极细胞功能（平均值±SEM）可由强度响应系列杆的PII建模来测定（空心圆○）与中落桨功能（灰线）。这将返回2V 以下 （饱和幅度，μV），K（1 /灵敏度，登录CD SM -2）和N（斜率）。（C）视网膜神经节细胞的功能是在昏暗的光能量检测和PSTR峰值量化（PSTR AMP）和时间（ 它 PSTR </sUB&gt;）。（D）锥双极细胞功能引起与锥PII峰值（锥PII 安培 ）和时间（锥PII 吧 ）量化配对闪光模式。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54158/54158fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/54158/54158fig3.jpg" /> 图3.平均水平VEP波形，视觉诱发电位波形的形状改变随刺激能量。号码波形左侧标明用来引出波形发光曝光。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54158/54158fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/54158/54158fig4.jpg" /> Figu4.重新分析VEP和强度响应函数。 （一）VEP振幅分析作为峰谷（P1N1）和谷峰（N1P2）幅度。这些反应的隐含时间（ 它 ）也返回（P1 了 ，N1 它 ，P2 它 ）。（二）VEP P1N1幅度（平均值±SEM）随着经济刺激能量增加而增加。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54158/54158fig4large.jpg" target="_blank">请点击此处查看大图这个数字。</a>

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

Nanyang Technological University

Research

JoVE Journal

Neuroscience

12.3K 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

Implantation and Recording of Wireless Electroretinogram and Visual Evoked Potential in Conscious Rats

The full-field electroretinogram (ERG) and visual evoked potential (VEP) are useful tools to assess retinal and visual pathway integrity in both laboratory and clinical settings. Currently, preclinical ERG and VEP measurements are performed with anesthesia to ensure stable electrode placements. However, the very presence of anesthesia has been shown to contaminate normal physiological responses. To overcome these anesthesia confounds, we develop a novel platform to assay ERG and VEP in conscious rats. Electrodes are surgically implanted sub-conjunctivally on the eye to assay the ERG and epidurally over the visual cortex to measure the VEP. A range of amplitude and sensitivity/timing parameters are assayed for both the ERG and VEP at increasing luminous energies. The ERG and VEP signals are shown to be stable and repeatable for at least 4 weeks post surgical implantation. This ability to record ERG and VEP signals without anesthesia confounds in the preclinical setting should provide superior translation to clinical data.

We show surgical implantation and Recording procedures to measure visual electrophysiological signals from the eye (electroretinogram) and brain (visual evoked potential) in conscious rats, which is more analogous to the human condition where recordings are conducted without anesthesia confounds.

植入和无线电图记录和视觉诱发电位清醒大鼠

The full-field electroretinogram (ERG) and visual evoked potential (VEP) are useful tools to assess retinal and visual pathway integrity in both ...

科研

行为学

Using the Electroretinogram to Assess Function in the Rodent Retina and the Protective Effects of Remote Limb Ischemic Preconditioning

The ERG is the sum of all retinal activity. The ERG is usually recorded from the cornea, which acts as an antenna that collects and sums signals from the retina. The ERG is a sensitive measure of changes in retinal function that are pan-retinal, but is less effective for detecting damage confined to a small area of retina. In the present work we describe how to record the &#8216;flash&#8217; ERG, which is the potential generated when the retina is exposed to a brief light flash. We describe methods of anaesthesia, mydriasis and corneal management during recording; how to keep the retina dark adapted; electrode materials and placement; the range and calibration of stimulus energy; recording parameters and the extraction of data. We also describe a method of inducing ischemia in one limb, and how to use the ERG to assess the effects of this remote-from-the-retina ischemia on retinal function after light damage. A two-flash protocol is described which allows isolation of the cone-driven component of the dark-adapted ERG, and thereby the separation of the rod and cone components. Because it can be recorded with techniques that are minimally invasive, the ERG has been widely used in studies of the physiology, pharmacology and toxicology of the retina. We describe one example of this usefulness, in which the ERG is used to assess the function of the light-damaged retina, with and without a neuroprotective intervention; preconditioning by remote ischemia.

The electroretinogram (ERG) is an electrical potential generated by the retina in response to light. This paper describes how to use the ERG to assess retinal function, in dark-adapted rats, and how it can be can be used to assess a neuroprotective intervention, in the present case remote ischemic preconditioning.

使用电生理评估功能，在啮齿动物的视网膜和远程肢体缺血预适应的保护作用

The ERG is the sum of all retinal activity. The ERG is usually recorded from the cornea, which acts as an antenna that collects and sums signals from the ...

神经科学

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 ...

Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the neurogenesis of EEG is still very limited. Local field potentials (LFPs) recorded via a multi-laminar microelectrode can provide a more detailed account of simultaneous neural activity across different cortical layers in the neocortex, but the technique is invasive. Combining EEG and LFP measurements in a pre-clinical model can greatly enhance understanding of the neural mechanisms involved in the generation of EEG signals, and facilitate the derivation of a more realistic and biologically accurate mathematical model of EEG. A simple procedure for acquiring concurrent and co-localized EEG and multi-laminar LFP signals in the anesthetized rodent is presented here. We also investigated whether EEG signals were significantly affected by a burr hole drilled in the skull for the insertion of a microelectrode. Our results suggest that the burr hole has a negligible impact on EEG recordings.

This protocol describes a simple method for concurrent recording of co-localized electroencephalography (EEG) and multi-laminar local field potential in an anesthetized rat. A burr hole drilled in the skull for the insertion of a microelectrode is shown to produce negligible distortion of the EEG signal.

啮齿动物联合局部脑电图和局部电位的同步记录

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the ...

Electroretinogram Recording for Infants and Children under Anesthesia to Achieve Optimal Dark Adaptation and International Standards

Electroretinogram (ERG) is the only clinical objective test available to assess retinal function. Full-field ERG (ffERG) measures the panretinal rod and cone photoreceptor function as well as inner retinal function and is an important measure in the diagnosis and management of inherited retinal diseases as well as inflammatory, toxic, and nutritional retinopathies. Adhering to international standards and maintaining retinal dark adaptation are critical to acquire valid and reliable dark-adapted (scotopic) and light-adapted (photopic) ffERG responses. Performing ffERG in infants and children is challenging and often requires general anesthesia in the operating room. However, maintaining retinal dark adaptation in the operating room is becoming increasingly difficult given the numerous light sources from anesthesiology monitoring systems and other equipment. A practical and widely applicable method for ffERG testing is described in the operating room that optimizes retinal dark adaptation. The method reduces operating room time by dark-adapting the patient before general anesthesiology is instituted. The operating room is modified for dark adaptation and any remaining light source in the darkened operating room is minimized with the use of a modified portable foldable darkroom that encloses the patient&#8217;s head and the ERG examiner during ffERG scotopic recordings. The simple method adheres to ffERG international standards and provides valid reliable scotopic and photopic ffERG recordings that are critical to assess objective retinal function in this young age group where subjective assessment of visual function such as visual acuity and visual fields are not possible. Furthermore, the ffERG is the gold standard clinical test in detecting early onset inherited retinal diseases including Leber congenital amaurosis where approved gene therapy has become available. In sedated conditions, very low amplitude ffERG signals can be detected due to minimal orbicularis muscle activity interference, which is particularly relevant in patients after gene therapy to detect improved amplitude responses.

Adhering to international standards and maintaining retinal dark adaptation are critical to acquire valid full-field electroretinogram responses in the diagnosis and management of inherited retinal diseases. A practical protocol using a portable darkroom is provided to obtain full-field&#160;electroretinogram for infants and children under sedation or general anesthesia in the operating room setting.

麻醉下婴儿和儿童电雷电图记录，实现最佳黑暗适应和国际标准

Electroretinogram (ERG) is the only clinical objective test available to assess retinal function. Full-field ERG (ffERG) measures the panretinal rod and ...

医学

Full-Circle Cauterization of Limbal Vascular Plexus for Surgically Induced Glaucoma in Rodents

Glaucoma, the second leading cause of blindness worldwide, is a heterogeneous group of ocular disorders characterized by structural damage to the optic nerve and retinal ganglion cell (RGC) degeneration, resulting in visual dysfunction by interrupting the transmission of visual information from the eye to the brain. Elevated intraocular pressure is the most important risk factor; thus, several models of ocular hypertension have been developed in rodents by either genetic or experimental approaches to investigate the causes and effects of the disease. Among those, some limitations have been reported such as surgical invasiveness, inadequate functional assessment, requirement of extensive training, and highly variable extension of retinal damage. The present work characterizes a simple, low-cost, and efficient method to induce ocular hypertension in rodents, based on low-temperature, full-circle cauterization of the limbal vascular plexus, a major component of aqueous humor drainage. The new model provides a technically easy, noninvasive, and reproducible subacute ocular hypertension, associated with progressive RGC and optic nerve degeneration, and a unique post-operative clinical recovery rate that allows in vivo functional studies by both electrophysiological and behavioral methods.

The goal of this protocol is to characterize a novel model of glaucomatous neurodegeneration based on 360&#176; thermic cauterization of limbal vascular plexus, inducing subacute ocular hypertension.

角膜缘血管丛全圆烧灼治疗啮齿动物手术诱发的青光眼

Glaucoma, the second leading cause of blindness worldwide, is a heterogeneous group of ocular disorders characterized by structural damage to the optic ...

Preparing a Rat Model to Assess Visual Pathway Integrity Using an Electrophysiology Setup

Source: Nguyen, C. T. et al., <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/54158">Simultaneous Recording of Electroretinography and Visual Evoked Potentials in Anesthetized Rats.</a>&#160;J. Vis. Exp. (2016)
This video demonstrates the preparation of a rat model to study retinal health and visual pathway integrity. Visual evoked potential and electroretinography reference and active electrodes are connected to the rat, followed by aligning the platform with the rat in front of the Ganzfeld bowl for even retinal illumination and closing the Faraday cage to avoid external interference.

准备大鼠模型以使用电生理学设置评估视觉通路完整性

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

JoVE Encyclopedia of Experiments

Neurophysiology

Electrophysiology Techniques

Direct-Coupled Electroretinogram Recording of the Retinal Pigment Epithelium in a Mouse

Source: Miyagishima, K. J., et al. <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/61491">Direct-Coupled Electroretinogram (DC-ERG) for Recording the Light-Evoked Electrical Responses of the Mouse Retinal Pigment Epithelium.</a> J. Vis. Exp. (2020).
This video demonstrates the recording of a direct-coupled electroretinogram (DC-ERG) on a dark-adapted, anesthetized mouse retinal pigment epithelium (RPE). After applying dilating drops, the animal is placed in a recording setup, and the capillary electrode filled with a salt buffer is placed on the cornea. After establishing the baseline, DC-ERG is recorded.

小鼠视网膜色素上皮的直接耦合视网膜电图记录

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Mouse ...

Imaging and Optical Techniques

Recording of Visual Evoked Potentials via the Skull Electrodes in a Rat Model

Source: You, Y., et al. <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/52934">Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination.</a> J. Vis. Exp. (2015).
This video demonstrates the process of measuring visual evoked potentials or VEPs in a dark-adapted, anesthetized rat with pre-implanted skull electrodes, which record the electrical signals from the brain&#39;s visual cortex in response to light stimuli.

通过大鼠模型中的颅骨电极记录视觉诱发电位

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. VEP ...

Concurrent Electroencephalographic and Local Field Potential Recordings in an Anesthetized Rat

Source: Kang, S., et al. <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/56447">Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent.</a> J. Vis. Exp. (2017).
This video demonstrates the procedure for concurrent recordings of co-localized electroencephalography (EEG) and local field potentials (LFPs) in an anesthetized rat. The process involves preparing the rat and attaching stimulating electrodes to its whisker pads. An EEG spider electrode is placed on the rat&#39;s skull for surface EEG measurements. At the same time, a multi-channel microelectrode is inserted into a specific brain region through a burr hole in the skull to record neural activity from a localized brain area.