美国国家药物滥用研究所，美国国立卫生研究院授予5R44DA026220-03

这里使用的OPAD（Stoelting公司，伊利诺斯Wood Dale）中描述的一个示例实验中使用辣椒素的一般术语。操作员具有自由编程，有许多选项和疼痛模型，虽然无数次的实验。例如，镇痛药减少伤害措施6,3,4,5,7和其他疼痛模型，如慢性缩窄性损伤农产品和炎症伤害性的行为增加3,9。这些模型很容易适应以下协议。 对于所有的实验中，雄性Sprague-SD大鼠（250-300克，查尔斯河，罗利，NC）。这些被安置在对22℃的温度和31％的湿度控制的房间与一个正常的12小时的光/暗周期（早上6时至下午6时亮起），都可以免费获得食物和水，但禁食时。行为会议进行过程中亮过相。这些设施AAALAC认证和批准了所有程序bŸ佛罗里达IACUC大学。 1.Training和基线会话<ol><li>食品快速啮齿动物每个OPAD会话的前一天晚上（如:15 + / - 1小时这个实验）。 </li><li>鼠害必须先进行培训，直到观察到一致的行为非厌恶的温度（例如33-37℃）。一般来说，大约六次会议（每周三次两星期）足以培养小鼠或大鼠舔约600-1,000或2000次会议，分别。 </li></ol> 2。测试前的准备和辣椒素治疗<ol><li>无毛老鼠是最好的所有的操作性程序，如果这是不可能的，啮齿动物必须有他们的脸毛（颊毛，而不是只的触须垫/胡须，因为这有一个对啮齿动物的导航效果）进行精确测试前1-2天去除记录的行为。 </li><li>颊毛作为温度绝缘体，使热和冷痛觉。要除去毛，麻醉大鼠，加眼膏，脸颊的头发剃光，用指甲刀，应用脱毛霜，等待2-4分钟，然后用清水洗净。见诺伊贝特等3完整的方法论。 </li><li>在当天的测试中，麻醉动物（如1-2.5％异氟醚吸入）和双眼，以防止他们从晒出任何外用药物治疗，并保持在眼睛越来越兽医opthalamic软膏。 </li><li>应用辣椒素霜（0.1％），双侧脸颊，用无菌棉签暴露。等待5分钟。擦去奶油纱布浸泡在温水（40℃左右）。用酒精棉签擦拭脸颊，并设置一个计时器，30分钟。 </li><li>允许啮齿动物有足够的时间从麻醉中恢复，然后返回它的笼子。等待，直到它可以提高其头部在胸骨的姿势在为了防止笼床上用品的愿望。 </li></ol> 3。 OPAD系统编程协议和实验<ol><li> Ø创新的关键f OPAD行为测试系统是颜面部的软件，任何迷宫（Stoelting有限公司，伊利诺斯Wood Dale）驱动系统，允许用户程序并创建新的实验。 </li><li>下面提供了如何编写一个简单的实验是一个通用的例子，但有更多的选择的温度斜坡协议和疼痛模型辣椒素以外。 </li><li>实验，可以在任何点设计和保存。打开对OPADs和开放软件。打开白噪声控制环境噪声。在"文件"，选择新实验。在副标题"议定书"的协议名称，并选择是否被蒙蔽。 </li><li>根据"OPAD笼"中选择"新OPAD笼"，然后"添加所有连接OPAD笼。"在"输出"中选择"温度控制器"，然后"热元素"。开始温度（例如，一个中性33-37℃）进行调整。 </li><li>如果需要，调整斜坡"OPAD温度循环下的温度。"为了改变它的温度从中立到热冷做出调整。 </li><li>对于"1"，"设定温度（°C）"= 45，"斜坡持续时间"= 30秒，和"留值"= 3分钟。对于"2"，33次，30秒和3分钟。对于"3"，7，60秒和3分钟。为"4"，60秒，33分钟和3分钟。检查框"后一段时间:"，选择3分钟。 </li><li>在"字段"添加任何额外票据有关科目。例如，使一个地区的动物ID，选择"新领域"，将名称更改为"动物标识。"然后选择"动物"，"文字"，"动物ID使用此字段。" </li><li> OPAD在"舞台"，将自动设置的"第一阶段"。设置测试期间的持续时间和命名的阶段，如果需要。对于这个斜坡会话，设置到18分钟。 </li><li>阶段的注意事项:对于大多数实验，行为会不会持续时间超过10-20分钟。在那之后，鼠类酒足饭饱。在数天的实验中，附加的阶段可以添加的每一天，使数据分析更简单。 </li&gt; <li>在"计算"，选择"新的计算"，并将它命名为"L / F"舔/面对比。在标示为"输入计算在该地区下面的"调整说"舔:激活/联系方式:激活"。 </li><li>要创建一个简单的数据分析的时间段选择"分析"和"新的时间段"。名称"33°C"选择框"这段时间在各个阶段是一样的"，对于"开始:"写0"完:"放3分钟。 </li><li>重复上述步骤，为每个时间段。产品名称:"斜坡33-45°C。"开始于:3分钟。结束于:3.5分钟。 "45°C"，3.5分钟，6.5分钟。 "斜坡45-33°C"，6.5分，7分。 "33°C"，7分钟，10分钟。 "斜坡33-7°C"，10分钟，11分钟。 "12°C"，11分钟，14分钟。 "斜坡7-33°C，14分钟，15分钟。 "33°C"，15分钟，18分钟。 </li><li>保存并命名的协议。注意:在新的实验中，已储存的协议可以被重新使用。 </li></ol> 4。运行试验<ol><li>准备一个房间TE温度奖励混合物OPAD瓶。比例为2:1的水:甜炼乳工作得很好，虽然也可使用蔗糖或糖精的解决方案。 </li><li>将液体捕捉托盘，有机玻璃笼，金属地板炉排OPAD机。将接线笼。 </li><li>地方奖励瓶站和调整，使壶嘴可以达到的啮齿动物。起初，瓶子可以放在进一步回在笼子里，然后撤回，以产生更好的面部接触。 </li><li>将实验到OPAD软件。添加要测试的动物的数量。在副标题"实验"添加"标题"，然后添加治疗组（辣椒素和控制）。添加各组动物的数量。 </li><li>在副标题为"测试"，选择"动物治疗和数据"。添加字母（A，B，C等）治疗和动物ID。框现在应该有指定的动物在其屏幕上的ID。 </li><li>按OPAD盒上的按钮。这将将热压调整到合适的温度。当指示灯变为橙色，放置灭鼠里面，并再次按下按钮。在这个实验中，大鼠将开始30分钟后的辣椒素乳膏擦去。 </li><li>绿灯将打开。距离调整为瓶子是从包装盒中，因此必须使大鼠颊，鼻毛，区域可以舔的热压接触。适当的调整，将导致在一个坚实的红色方块上面舔概述橙色盒子接触。 </li><li>妥善安置的奖励瓶的检测至关重要。如果瓶子太靠近大鼠舔未做接触或触摸鼻子的奖励瓶，使许多舔出现只有一个轮廓的红色方块表示，而不是实心方块舔。 </li><li>一旦测试会话是，OPAD在实验者的口气提醒。返回啮齿类动物的笼子里。如果另一个啮齿动物进行测试后框将籼稻TE动物标识。重复步骤4.5-4.7，根据需要，直到实验结束。 </li></ol> 5。分析，绘图和的与OPAD软件数据统计分析<ol><li>在副标题"结果"选择是否看到一个文本，图形，或统计分析报告。 </li><li>在"图形报告设置"框中，选择变量检查。例如，根据"计算结果"检查"L / F"框中。 </li><li>指定"在x轴秀"的"时间段"和"显示不同的系列"作为"治疗"。选择"查看报告"。保存，打印，复印，或通过电子邮件发送的报告可以做到在这个时候。某些数据点可能会被排除在下面的框中分组步骤，如果需要的话。 </li><li>在副标题"数据"是一个可调上市以电子表格的形式，如果需要从实验的原始数据。注:现在所有的数据被保存，并在以后的时间可以被操纵和分析。 </li></ol> 6。清理<ol><li>关掉机器，并除去笼布线。清洗和消毒炉排，盒，瓶，液体纸盒。这些组件可以通过手或洗碗机洗涤。 </li></ol>

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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+operant+assay+of+thermal+pain+in+conscious,+unrestrained+rats.">An operant assay of thermal pain in conscious, unrestrained rats.</a> J. Neurosci. Methods. 97, 19-29 (2000).</li><li>Vierck, C. J., Kline, R., Wiley, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+operant+escape+and+innate+reflex+responses+to+nociceptive+skin+temperatures+produced+by+heat+and+cold+stimulation+of+rats.">Comparison of operant escape and innate reflex responses to nociceptive skin temperatures produced by heat and cold stimulation of rats.</a> Behav. Neurosci. 118, 627-635 (2004).</li><li>Nolan, T. A., Caudle, R. M., Neubert, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+caloric+and+non-caloric+sweet+reward+solutions+on+thermal+facial+operant+conditioning.">Effect of caloric and non-caloric sweet reward solutions on thermal facial operant conditioning.</a> Behav. Brain Res. 216, 723-725 (2011).</li><li>Taha, S. A., Katsuura, Y., Noorvash, D., Seroussi, A., Fields, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Convergent,+not+serial,+striatal+and+pallidal+circuits+regulate+opioid-induced+food+intake.">Convergent, not serial, striatal and pallidal circuits regulate opioid-induced food intake.</a> Neuroscience. 161, 718-733 (2009).</li></ol>

理查德·米尔斯和克里斯·劳埃德Stoelting公司雇员生产和自由访问这篇文章是由Stoelting有限公司

OPAD系统是一个易于使用的，高通量的检测，能够检测在啮齿动物的疼痛感知的变化。该系统的高吞吐量的性质意味着，许多动物可以由一个人在一天内进行测试。这是由于OPAD软件系统，因为它允许多达16个盒，在一台计算机上同时运行。这意味着，在初始安装后的时间，约48操作性运行（每运行18分钟），可以进行一个小时，甚至更如果会话时间设置为每级的更短的时间。这允许在数百种动物每天疼痛测试。用最传统的止痛检测，这种量的测试是不实际。 与我们以前的工作相一致，啮齿动物的行为是痛苦的条件下改变。在非有毒期间，啮齿动物通常具有很长的饮用水较量中，他们保持接触与热压。在厌恶45°C或1°C的条件下，啮齿动物有更短的较量，因为他们不能维持很长一段时间的接触。因此，舔/面对比（舔数除以面部接触的数量在一个会话）改变疼痛。辣椒素的敏感性增加热痛表现出较低的L / F比在处理与未处理的啮齿类动物在45°C温度。止痛药可以返回舔/面对比水平类似于非痛苦的条件3。动物模型的临床相关的深神经组织疼痛，如三叉神经痛疼痛的条件，很容易产生皮肤上（如应用辣椒素霜）虽然是最简单的检测方法，此法对疼痛，也可以改变行为操作性颜面部检测9 。两者合计这些数据支持的证据表明，OPAD改建在热和冷疼痛，疼痛阈值，除了操作性的有害化学剂，如辣椒素敏感颜面部疼痛检测的能力疼痛和镇痛6,3,8,17,10,5,12,18,11,9检测许多其他条件。 OPAD测量疼痛的系统是一个更多的临床相关的，有意义的，人道的方法检测疼痛比反射为基础的措施。这些传统的措施，像爪子用von Frey细丝19和甩尾实验20撤出痛觉已经使用了一个多世纪，但他们只能测量实验者造成刺激反应。动物难以控制，和"痛觉"主要是本地化到脊髓。对于人类来说，主观疼痛的经验也很重要，因为人们只是要求报告自己的主观程度的疼痛。动物自我报告他们的痛苦在基于操作性的程序的能力将是一个基本的疼痛研究的突破，为1。 OPAD，动物们在AP的选择是否响应的刺激ainful或不。如果实在是太痛苦，动物只是减少他们的企图达到的奖励，从而限制他们接触到疼痛。这是一个更人性化，更少的压力检测时相比，很多基于反射的动物往往有自己的运动受到限制，无法控制他们所接触到的金额疼痛刺激措施。需要摆脱疼痛是一种内在的驱动器在所有的动物和OPAD采用这种行为，而不是补偿，它像其他伤害性检测。运动远离疼痛的反射为基础的措施操作性任务正变得越来越普遍的领域。其他团体使用非反射为基础的措施，如检查顿饭持续时间21,22,23和热痛逃生范式24（评论其他止痛措施，我们建议我们的第一参考 1）。这些元素组合成一个统一的措施，舔/面对比，WH OPADICH检查食物的摄入和需要，以摆脱痛苦的刺激。另一个好处是，该法是能够测量在很长一段时间（1-2个月）疼痛不失灵敏度7,9。由于自己的优势，通过反射为基础的测试，很好地适应了这种压力较小，更人性化的检测测量啮齿动物伤害行为的长期变化。 阿片类药物的剂量效应和疼痛阈值时相比，反射为基础的措施操作性疼痛措施往往得到不同的结果。虽然基于反射的措施，通常用于高剂量的阿片类药物25的一些研究表明，较低的剂量所需要的响应的操作式的检测26,27,28。高剂量药物，也可以干扰操作性的措施，但这些都是OPAD 6检测。其他的研究也表明，逃离痛苦的刺激的阈值是不同的操作性versu的Ş反射为基础的措施29,2,30暗示的动物对疼痛的感觉，而他们的脊髓反射的速度之间的主要区别。 OPAD的一个好处是啮齿动物可以选择是否要执行任务，这允许灭鼠表达逃避或回避行为。这种复杂的行为，需要决策皮质伤害性的量来控制啮齿动物感觉14,29,15,30。虽然逃生和避税行为可以干扰与反射为基础的措施，这些疼痛行为的OPAD是一个不可分割的组成部分。疼痛阈值和低剂量的阿片类药物操作性检测所需的差异表明疼痛和镇痛更高的灵敏度比传统的反射为基础的措施。 虽然OPAD可以比传统的检测，一些实验条件和药物直接测量疼痛可能有不利的影响，在此试验中，必须控制。改造s的食欲动机可以改变这个实验的行为。这可以反映由在奖励本身31或奖励的动机6的差。必须小心，以确保动物的动机的奖励是恒定的，因为许多药物可干扰动机。例如，高剂量的吗啡和其他阿片类药物可能会导致贪食为甜，脂肪物质32将改变响应操作性颜面部检测6。虽然这并不表明，这种基于操作性的奖励冲突范式使用，包括焦虑和成瘾领域具有广泛的影响（ 即在一个给定的疼痛刺激的存在改变的奖励方面）重要的是要控制食欲改变疼痛测试会话。这些改变的动机不会出现在这些临床相关的低剂量的镇痛作用仍然完好3。方法之一CONTROL这种可能的混淆，以确保给定的药物的剂量不会增加在中性温度（33-37℃）的行为。测试药物与非药物组在一个中立的温度应该是第一步加入疼痛组件之前。此外，由于一些基线会话是可能的在一个测试会话使用OPAD这些问题可以被检测到，并且可以控制在一个单一的行为会议。由于空腹时间表可以改变这个实验的动机，重要的是要保持一致。我们通常做一个通宵快，但其他可能的时间表。例如，我们已经尝试了每天的短前禁食6小时（结果未发表）。这允许每天测试，而不是每隔一天。此外，unfasted大鼠也回应测定9。无论空腹技术，用它主要是重要的保持一致，在整个测试控制激励因素。 如甩尾，von Frey细丝）既不是软件驱动的，也不高通量。软件驱动系统提供了一个显着的进步行为研究如何设计，如何对数据进行收集和分析，在这个实验中的使用将允许增加基本疼痛研究在未来更多的临床翻译。该系统预计将有一个显着的影响ADVA通过实践与疼痛有关的未来研究，因为这些操作性的行为学研究提供必要的链接了解高阶结构上整体的疼痛行为的影响。

慢性的，不受控制疼痛仍然是一个重大的公共卫生问题和新颖的镇痛治疗往往不能从替补席翻译床头。这种缺乏成功的部分原因是由于使用的低效行为分析在疼痛反射为基础的措施不一定完全模拟人类的痛苦条件1,2，具体而言，缺乏一个可靠的，高通量，市售在大鼠和小鼠体内疼痛评估测定。我们在座的高吞吐量，易于使用的版本，我们基于操作性的痛觉测定。这个新的系统是基于我们以前的操作性颜面部疼痛检测已被证明是检测不同的止痛方式，包括冷，热，机械3,4,5敏感。从这些措施中，各种各样的领域都进行了研究，包括止痛药6,3,4,5,7，疼痛，如炎症，痛觉过敏和异常疼痛<s升&gt; 3,8，三叉神经痛，周围痛觉调制辣椒素，辣椒素类似物，薄荷，和icillin 8,10,5,11 通过 TRP通道。心理影响，如焦虑引起的疼痛12调制和安慰剂效应13，也已经证明，颜面部的操作性测试，暗示它可能是适合用于测量充分体验痛苦，而不是简单的痛觉。 颜面部疼痛评估设备（OPAD）使用的奖励/冲突检测，这使得啮齿动物之间作出选择接收加强奖励或逃避厌恶刺激，从而控制疼痛的感觉在会话期间14,15。鼠害首先训练他们的脸按到温度控制热压为了获得食品瓶含有液体的奖励。训练结束后，刺激温度可以被加热或冷却和不同的重对应可以表明动物感知痛觉或镇痛水平。 OPAD也能在温度急剧变化，允许在一个单一的测试环节在高温和低温的基准测试和评估疼痛。在这里，我们提出了一个简单的协议，它突出OPAD疼痛的变化引起的热，冷的检测能力，并且TRPV1激动剂辣椒素16。下面使用辣椒素作为热增敏剂，因为它有几个好处，因为它是该测定非组织破坏和先前已被证明诱导面部异常性疼痛和痛觉过敏中的啮齿类动物模型8。 ，我们将演示如何，OPAD软件也能迅速获得，分析，图表，对啮齿动物的行为数据进行统计分析。

<table border="1">
 <tbody>
 <tr>
 <td>Name of Reagent/Material</td>
 <td>Company</td>
 <td>Catalog Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Sweetended Condensed Milk</td>
 <td>Borden</td>
 <td>5272910108</td>
 <td></td>
 </tr>
 <tr>
 <td>Capzasin-HP 0.1%</td>
 <td>Chattem, Inc.</td>
 <td>0032648-02</td>
 <td></td>
 </tr>
 <tr>
 <td>Isopropyl alcohol</td>
 <td>CVS</td>
 <td>5042826245</td>
 <td></td>
 </tr>
 <tr>
 <td>Isoflurane</td>
 <td>Piramal Healthcare</td>
 <td>66794-013-25</td>
 <td></td>
 </tr>
 <tr>
 <td>Opthalamic vet ointment</td>
 <td>Dechra</td>
 <td>17033-211-38</td>
 <td></td>
 </tr>
 <tr>
 <td>Hair remover lotion</td>
 <td>Church and Dwight Co., Inc</td>
 <td>NRLBB-22339-04</td>
 <td></td>
 </tr>
 <tr>
 <td>OPAD System</td>
 <td>Stoelting</td>
 <td>67500</td>
 <td></td>
 </tr>
 <tr>
 <td>Additional OPAD cages</td>
 <td>Stoelting</td>
 <td>67501</td>
 <td></td>
 </tr>
 <tr>
 <td>Granulated cylinder</td>
 <td>Cole-Parmer</td>
 <td>EW-34512-11</td>
 <td></td>
 </tr>
 <tr>
 <td>Paper towels</td>
 <td>ANY</td>
 <td>ANY</td>
 <td></td>
 </tr>
 <tr>
 <td>Cotton tipped applicators</td>
 <td>Fisher</td>
 <td>23-400-101</td>
 <td></td>
 </tr>
 <tr>
 <td>Fluotec 4 Vaporizer</td>
 <td>Ohmeda</td>
 <td>39711</td>
 <td></td>
 </tr>
 <tr>
 <td>Hair clippers</td>
 <td>Oster</td>
 <td>78005-010</td>
 <td></td>
 </tr>
	 </tbody>
 </table>

use of the operant orofacial pain assessment device (opad) to measure changes in nociceptive behavior

我们提出了一个操作性疼痛清醒的，有意识的啮齿动物的检测系统。颜面部疼痛评估设备（OPAD）评估疼痛行为在更多的临床相关的方式，不依靠痛觉反射为基础的措施。食物禁食，无毛（或剃了毛的）啮齿动物放入一个有机玻璃室，它有两个基于珀尔帖的热压，可以进行编程之间的任何温度7℃和60℃。训练啮齿动物接触这些以访问的奖励瓶。在会话过程中，一些行为疼痛结果会自动记录并保存。这些措施包括奖励瓶激活的数量（舔）和脸部接触刺激（面接触），但舔/面部比（总数每会话/总数接触舔）也可以创建自定义等措施。刺激的温度可以设置为一个单一的温度或在一个会话内的多个温度。 OPAD是一种高通量，易于使用的操作性试验，这将导致在未来更好的翻译疼痛研究，因为它包含了皮质的输入，而不是依靠脊髓反射的伤害性检测。

典型的结果示OPAD在图1A-D中的一个单一的啮齿类动物的行为。舔会话的每个环节，在中性33°C的温度，但厌恶的低（45℃和7°C） 图1A所示。 图1B表明，长期接触较量是在33°C是典型的非伤害性刺激的温度。的持续时间的减小，接触期间的温度是痛苦的增加的数量。 图1C是图斜坡协议OPAD被编程，以用于所有的测试会话， 图1D显示随着时间的推移以克摄入的总量奖励。同样地舔的数量，动物喜欢的中性温度超过痛苦的。舔/ OPAD进行了计算，并说明在面对比（L / F）为基准会话图1E。这个比例远高于在三无痛苦33°C会议（20-46的舔每面接触）的痛苦会议45°C（3舔每面接触）和7比°C（1舔每面接触）。重复测量单因素方差分析有显着性意义（F（4,52）= 6.2182，P &lt;0.001）L / F比温度的效​​果。邦费罗尼的测试进行比较时，33°C与7°C（P &lt;0.05），45°C和33°C（2）（P &lt;0.01），33℃（2）与7°C（ P &lt;0.01）。 N = 16适用于所有温度。在图1F辣椒素处理啮齿类动物（N = 8）从幼稚大鼠（N = 8）在任何中性33°C的温度差异不显着。辣椒素治疗的啮齿动物确实有一个显着低于L / F的比例，在45°C（T-测试，T（13）= 2.9350，P = 0.012）。辣椒素组有较高的L / F比在7°C，但并不显着。 图1。接触测量痛觉与OPAD。OPAD单一啮齿动物的行为是绘制Å）数量舔， 乙），Ç）温度的热压在会议期间，和D）奖励的摄入量以克为单位。ê）的舔/面对比例是三无痛苦的33°C会议期间的痛苦会显着降低45°C和12°C（重复测量单因素方差分析，F（4,52）= 6.2182，P &lt;0.001，邦费罗尼测试33°C与7°C（P &lt;0.05，＃），45°C与33°C（2）（P &lt;0.01，**），33°C（2） - 7°C （P &lt;0.01，＃＃）F）辣椒素治疗的啮齿动物有一个显着较低的L / F比在45℃（T检验，t（13）= 2.9350，P = 0.012），但在没有中性气温。 1E和天真1F辣椒素和N = 8 N = 8，N = 16。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/50336/50336fig1large.jpg" target="_blank">点击此处查看大图</a> 。

Watch this Scientific Journal Video about Use of the Operant Orofacial Pain Assessment Device OPAD to Measure Changes in Nociceptive Behavior at JoVE.com

Use of the Operant Orofacial Pain Assessment Device OPAD to Measure Changes in Nociceptive Behavior

我们提出了一个用户友好的，高通量的操作性在清醒的，有意识的啮齿动物的疼痛行为的评价体系。颜面部疼痛评估设备（OPAD）可以评估疼痛通过奖励/冲突范式，从而提供了一种更人性化的方式测试。该协议将产生更多的临床相关数据和平移啮齿动物。

使用操作性口面疼痛评估设备（OPAD）测量伤害行为的变化

We present an operant  system for the detection of pain in awake, conscious rodents. The Orofacial  Pain Assessment Device (OPAD) assesses pain behaviors ...

use-operant-orofacial-pain-assessment-device-opad-to-measure-changes

Nanyang Technological University

Research

JoVE Journal

Behavior

Use of the Operant Orofacial Pain Assessment Device (OPAD) to Measure Changes in Nociceptive Behavior

21.1K 次观看.  University of Florida College of Dentistry. 我们提出了一个用户友好的，高通量的操作性在清醒的，有意识的啮齿动物的疼痛行为的评价体系。颜面部疼痛评估设备（OPAD）可以评估疼痛通过奖励/冲突范式，从而提供了一种更人性化的方式测试。该协议将产生更多的临床相关数据和平移啮齿动物。

视频: Use of the Operant Orofacial Pain Assessment Device OPAD to Measure Changes in Nociceptive Behavior

The Crossmodal Congruency Task as a Means to Obtain an Objective Behavioral Measure in the Rubber Hand Illusion Paradigm

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden hand is touched. If the viewed and felt touches are given at the same time then this is sufficient to induce the compelling experience that the rubber hand is one's own hand. The RHI can be used to investigate exactly how the brain constructs distinct body representations for one's own body. Such representations are crucial for successful interactions with the external world. To obtain a subjective measure of the RHI, researchers typically ask participants to rate statements such as "I felt as if the rubber hand were my hand". Here we demonstrate how the crossmodal congruency task can be used to obtain an objective behavioral measure within this paradigm.
The variant of the crossmodal congruency task we employ involves the presentation of tactile targets and visual distractors. Targets and distractors are spatially congruent (i.e. same finger) on some trials and incongruent (i.e. different finger) on others. The difference in performance between incongruent and congruent trials - the crossmodal congruency effect (CCE) - indexes multisensory interactions. Importantly, the CCE is modulated both by viewing a hand as well as the synchrony of viewed and felt touch which are both crucial factors for the RHI.
The use of the crossmodal congruency task within the RHI paradigm has several advantages. It is a simple behavioral measure which can be repeated many times and which can be obtained during the illusion while participants view the artificial hand. Furthermore, this measure is not susceptible to observer and experimenter biases. The combination of the RHI paradigm with the crossmodal congruency task allows in particular for the investigation of multisensory processes which are critical for modulations of body representations as in the RHI.

We demonstrate how an objective measure can be employed in the widely employed rubber hand illusion paradigm. This measure is obtained by modifying the well-established crossmodal congruency task. This task allows the investigation of multisensory processes which are critical for modulations of body representations as in the rubber hand illusion.

一致性的任务作为一种手段来获取目标行为的措施中的橡胶手错觉范式Crossmodal

The rubber hand illusion (RHI) is a popular experimental paradigm. Participants view touch on an artificial rubber hand while the participants' own hidden ...

科研

行为学

Meal Duration as a Measure of Orofacial Nociceptive Responses in Rodents

A lengthening in meal duration can be used to measure an increase in orofacial mechanical hyperalgesia having similarities to the guarding behavior of humans with orofacial pain. To measure meal duration unrestrained rats are continuously kept in sound attenuated, computerized feeding modules for days to weeks to record feeding behavior. These sound-attenuated chambers are equipped with chow pellet dispensers. The dispenser has a pellet trough with a photobeam placed at the bottom of the trough and when a rodent removes a pellet from the feeder trough this beam is no longer blocked, signaling the computer to drop another pellet. The computer records the date and time when the pellets were taken from the trough and from this data the experimenter can calculate the meal parameters. When calculating meal parameters a meal was defined based on previous work and was set at 10 min (in other words when the animal does not eat for 10 min that would be the end of the animal&#39;s meal) also the minimum meal size was set at 3 pellets. The meal duration, meal number, food intake, meal size and inter-meal interval can then be calculated by the software for any time period that the operator desires. Of the feeding parameters that can be calculated meal duration has been shown to be a continuous noninvasive biological marker of orofacial nociception in male rats and mice and female rats. Meal duration measurements are quantitative, require no training or animal manipulation, require cortical participation, and do not compete with other experimentally induced behaviors. These factors distinguish this assay from other operant or reflex methods for recording orofacial nociception.

A lengthening in meal duration represents orofacial nociceptive behavior in rodents similar to the guarding behavior of humans with orofacial pain. Eating is a behavior that requires no training or animal manipulation, requires cortical participation, and is not competing with other experimentally induced behaviors, distinguishing this assay from alternative reflex or operant measurements.

膳食持续时间作为衡量啮齿动物的奥体非感知反应的指标

A lengthening in meal duration can be used to measure an increase in orofacial mechanical hyperalgesia having similarities to the guarding behavior of ...

Assessment of Murine Exercise Endurance Without the Use of a Shock Grid: An Alternative to Forced Exercise

Using laboratory mouse models, the molecular pathways responsible for the metabolic benefits of endurance exercise are beginning to be defined. The most common method for assessing exercise endurance in mice utilizes forced running on a motorized treadmill equipped with a shock grid. Animals who quit running are pushed by the moving treadmill belt onto a grid that delivers an electric foot shock; to escape the negative stimulus, the mice return to running on the belt. However, avoidance behavior and psychological stress due to use of a shock apparatus can interfere with quantitation of running endurance, as well as confound measurements of postexercise serum hormone and cytokine levels. Here, we demonstrate and validate a refined method to measure running endurance in na&#239;ve C57BL/6 laboratory mice on a motorized treadmill without utilizing a shock grid. When mice are preacclimated to the treadmill, they run voluntarily with gait speeds specific to each mouse. Use of the shock grid is replaced by gentle encouragement by a human operator using a tongue depressor, coupled with sensitivity to the voluntary willingness to run on the part of the mouse. Clear endpoints for quantifying running time-to-exhaustion for each mouse are defined and reflected in behavioral signs of exhaustion such as splayed posture and labored breathing. This method is a humane refinement which also decreases the confounding effects of stress on experimental parameters.

A method to assess exercise endurance in laboratory mice without the use of a shock grid is demonstrated. This method is a humane refinement that can decrease the confounding effects of stress on experimental parameters.

小鼠运动耐力的评估没有一个冲击电网的使用:另一种强迫运动

Using laboratory mouse models, the molecular pathways responsible for the metabolic benefits of endurance exercise are beginning to be defined. The most ...

Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of the most commonly used platforms to measure behaviors in animal models. It is a fast and relatively easy test that provides a variety of behavioral information ranging from general ambulatory ability to data regarding the emotionality of the subject animal. As it relates to rodent models, the procedure allows the study of different strains of mice or rats both laboratory bred and wild-captured. The technique also readily lends itself to the investigation of different pharmacological compounds for anxiolytic or anxiogenic effects. Here, a protocol for use of the open field maze to describe mouse behaviors is detailed and a simple analysis of general locomotor ability and anxiety-related emotional behaviors between two strains of C57BL/6 mice is performed. Briefly, using the described protocol we show Wild Type mice exhibited significantly less anxiety related behaviors than did age-matched Knock Out mice while both strains exhibited similar ambulatory ability.

A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

使用开放式现场迷宫来衡量小鼠的运动能力和焦虑样行为

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of ...

A Protocol of Manual Tests to Measure Sensation and Pain in Humans

Numerous qualitative and quantitative techniques can be used to test sensory nerves and pain in both research and clinical settings. The current study demonstrates a quantitative sensory testing protocol using techniques to measure tactile sensation and pain threshold for pressure and heat using portable and easily accessed equipment. These techniques and equipment are ideal for new laboratories and clinics where cost is a concern or a limiting factor. We demonstrate measurement techniques for the following: cutaneous mechanical sensitivity on the arms and legs (von-Frey filaments), radiant and contact heat sensitivity (with both threshold and qualitative assessments using the Visual Analog Scale (VAS)), and mechanical pressure sensitivity (algometer, with both threshold and the VAS). The techniques and equipment described and demonstrated here can be easily purchased, stored, and transported by most clinics and research laboratories around the world. A limitation of this approach is a lack of automation or computer control. Thus, these processes can be more labor intensive in terms of personnel training and data recording than the more sophisticated equipment. We provide a set of reliability data for the demonstrated techniques. From our description, a new laboratory should be able to set up and run these tests and to develop their own internal reliability data.

The goal of this procedure is to demonstrate a battery of quantitative techniques for sensory and pain measurement in humans. The equipment and techniques described are commonly found in pain clinics or are easy to obtain.

手动测试的协议来衡量感觉和疼痛人类

Numerous qualitative and quantitative techniques can be used to test sensory nerves and pain in both research and clinical settings. The current study ...

Simultaneous Recordings of Cortical Local Field Potentials and Electrocorticograms in Response to Nociceptive Laser Stimuli from Freely Moving Rats

Electrocortical responses, elicited by laser heat pulses that selectively activate nociceptive free nerve endings, are widely used in many animal and human studies to investigate the cortical processing of nociceptive information. These laser-evoked brain potentials (LEPs) consist of several transient responses that are time-locked to the onset of laser stimuli. However, the functional properties of the LEP responses are still largely unknown, due to the lack of a sampling technique that can simultaneously record neural activities at the surface of the cortex (i.e., electrocorticogram [ECoG] and scalp electroencephalogram [scalp EEG]) and inside the brain (i.e., local field potential [LFP]). To address this issue, we present here an animal protocol using freely moving rats. This protocol is composed of three main procedures: (1) animal preparation and surgical procedures, (2) a simultaneous recording of ECoG and LFP in response to nociceptive laser stimuli, and (3) data analysis and feature extraction. Specifically, with the help of a 3D-printed protective shell, both ECoG and LFP electrodes implanted on the rat&#39;s skull were securely held together. During data collection, laser pulses were delivered on the rat&#39;s forepaws through gaps in the bottom of the chamber when the animal was in spontaneous stillness. Ongoing white noise was played to avoid the activation of the auditory system by the laser-generated ultrasounds. As a consequence, only nociceptive responses were selectively recorded. Using the standard analytical procedures (e.g., band-pass filtering, epoch extraction, and baseline correction) to extract stimulus-related brain responses, we obtained results showing that LEPs with a high signal-to-noise ratio were simultaneously recorded from ECoG and LFP electrodes. This methodology makes the simultaneous recording of ECoG and LFP activities possible, which provides a bridge of electrocortical signals at the mesoscopic and macroscopic levels, thereby facilitating the investigation of nociceptive information processing in the brain.

We developed a technique that simultaneously records both electrocorticography and local field potentials in response to nociceptive laser stimuli from freely moving rats. This technique helps establish a direct relationship of electrocortical signals at the mesoscopic and macroscopic levels, which facilitates the investigation of nociceptive information processing in the brain.

在自由运动大鼠的反应中, 皮质局部电位和电皮质图谱的同时记录

Electrocortical responses, elicited by laser heat pulses that selectively activate nociceptive free nerve endings, are widely used in many animal and ...

Multi-Modal Signals for Analyzing Pain Responses to Thermal and Electrical Stimuli

The assessment of pain relies mostly on methods that require a person to communicate. However, for people with cognitive and verbal impairments, existing methods are not sufficient as they lack reliability and validity. To approach this problem, recent research focuses on an objective pain assessment facilitated by parameters of responses derived from physiology, and video and audio signals. To develop reliable automated pain recognition systems, efforts have been made in creating multimodal databases in order to analyze pain and detect valid pain patterns. While the results are promising, they only focus on discriminating pain or pain intensities versus no pain. In order to advance this, research should also consider the quality and duration of pain as they provide additional valuable information for more advanced pain management. To complement existing databases and the analysis of pain regarding quality and length, this paper proposes a psychophysiological experiment to elicit, measure, and collect valid pain reactions. Participants are subjected to painful stimuli that differ in intensity (low, medium, and high), duration (5 s / 1 min), and modality (heat / electric pain) while audio, video (e.g., facial expressions, body gestures, facial skin temperature), and physiological signals (e.g., electrocardiogram [ECG], skin conductance level [SCL], facial electromyography [EMG], and EMG of M. trapezius) are being recorded. The study consists of a calibration phase to determine a subject&#8217;s individual pain range (from low to intolerable pain) and a stimulation phase in which pain stimuli, depending on the calibrated range, are applied. The obtained data may allow refining, improving, and evaluating automated recognition systems in terms of an objective pain assessment. For further development of such systems and to investigate pain reactions in more detail, additional pain modalities such as pressure, chemical, or cold pain should be included in future studies. Recorded data of this study will be released as the &#8220;X-ITE Pain Database&#8221;.

This article focuses on the experimental elicitation of pain through heat (thermal) and electrical stimulation while recording physiological, visual, and paralinguistic responses. It aims at collecting valid multimodal data for analyzing pain based on its intensity, quality, and duration.&#160;

分析疼痛对热和电刺激反应的多模态信号

The assessment of pain relies mostly on methods that require a person to communicate. However, for people with cognitive and verbal impairments, existing ...

Chronic Post-Ischemia Pain Model for Complex Regional Pain Syndrome Type-I in Rats

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical condition. However, the underlying mechanisms of CRPS-I have yet to be revealed. It is known that ischemia/reperfusion is one of the leading factors that causes CRPS-I. By means of prolonged ischemia and reperfusion of the hind limb, the rat chronic post-ischemia pain (CPIP) model has been established to mimic CRPS-I. The CPIP model has become a well-recognized animal model for studying the mechanisms of CRPS-I. This protocol describes the detailed procedures involved in the establishment of the rat model of CPIP, including anesthesia, followed by ischemia/reperfusion of the hind limb. Characteristics of the rat CPIP model are further evaluated by measuring the mechanical and thermal hypersensitivities of the hind limb as well as the nocifensive responses to acute capsaicin injection. The rat CPIP model exhibits several CRPS-I-like manifestations, including hind limb edema and hyperemia in the early stage after establishment, persistent thermal and mechanical hypersensitivities, and increased nocifensive responses to acute capsaicin injection. These characteristics render it a suitable animal model for further investigation of the mechanisms involved in CRPS-I.

Provided here is a protocol that details steps to establish an animal model of chronic post-ischemia pain (CPIP). This is a well-recognized model mimicking human complex regional pain syndrome type-I. Mechanical and thermal hypersensitivities are further evaluated, as well as capsaicin-induced nocifensive behaviors observed in the CPIP rat model.

大鼠复杂区域疼痛综合征I型慢性缺血后疼痛模型

Complex regional pain syndrome type-I (CRPS-I) is a neurological disease that causes severe pain among patients and remains an unresolved medical ...

Operant Conditioning Task to Measure Song Preference in Zebra Finches

An operant conditioning paradigm is used to test the song preference of female zebra finches. Finches are placed in a two-chambered cage with a connecting opening and indicate their preference for a song by landing on a perch within each chamber. By interrupting the infrared beam from a photoelectric sensor above each perch, the bird activates the playback of a song through a speaker located on each side of the cage. Freely available software is used to trigger the song playback from each perch. To determine the song preference of each animal, her chamber preference is first identified by triggering no song playback when she lands on each perch. This chamber preference is then compared to her song preference. A minimum activity threshold is set to ensure the preference is real. Using this method, we show that paired females prefer the song of their partner. This method was used to understand the contribution of dopamine to the formation and maintenance of song preference.

We describe a technique to evaluate song preference in zebra finches. Females are placed in a two-chambered cage and song preference is measured by the number of times she triggers the playback of one song by landing on a perch within one chamber, compared with triggering a different song in the second chamber. Perch landings are counted using infrared sensors.

操作调节任务，以测量斑马芬奇中的歌曲首选项

An operant conditioning paradigm is used to test the song preference of female zebra finches. Finches are placed in a two-chambered cage with a connecting ...

The Use of Traditional Fear Tests to Evaluate Different Emotional Circuits in Cattle

Animal temperament is complex and has implications for productivity and economic profitability. Quantifying an animal&#39;s response to differing stimuli may facilitate breeding selections and identify animals that are better suited to specific management strategies. Multiple tests have been developed to evaluate cattle temperament (e.g., exit velocity, chute score, pen score, open field test, startle test, bovine zero maze), but each of these tests evaluates the animal&#39;s response to different stimuli (e.g., isolation, novel environment, startle, willingness to enter an enclosed area). Cattle temperament has been observed to be relatively stable over time. However, the evaluation of temperament has the potential to be influenced by current conditions, previous experiences, and observer bias. Many of these temperament tests have been improperly categorized as fear tests and have also been criticized for being subjective. This paper provides a framework for standardizing behavioral tests for cattle and suggests that these different evaluations assess different aspects of the animal&#39;s overall temperament.

Here, we present a protocol to conduct a variety of behavioral tests in cattle that have been designed to evaluate emotions. A battery of behavioral tests (open field test, startle test, bovine zero maze, exit velocity, pen score, and chute score) were conducted to evaluate different components of animal temperament.

使用传统恐惧测试来评估牛的不同情绪回路

Animal temperament is complex and has implications for productivity and economic profitability. Quantifying an animal&#39;s response to differing stimuli ...