Name: a general method for evaluating deep brain stimulation effects on intravenous methamphetamine self-administration
Uploaded: 2016-01-22T13:00:00.000+00:00
Duration: 9 min 16 s
Description: Watch this Scientific Journal Video about A General Method for Evaluating Deep Brain Stimulation Effects on Intravenous Methamphetamine Self-Administration at JoVE.com

Supported by the 2014-2015 Neurosurgery Research and Education Foundation (NREF) award and a 2014 Grant-In-Aid Award from Louisiana State University Shreveport School of Medicine (J.A.W.). We thank S. Harold and C.M. Keller for their invaluable technical assistance and teaching.

所有程序由LSUHSC机构动物护理和使用委员会批准并进行按照美国国立卫生研究院"的实验动物护理的原则"。 1.啮齿动物驯化和限食<ol><li>使用成年Wistar大鼠是3个月的实验开始老。房子大鼠单独使用在配备有层流单元和空气过滤器在温度和湿度控制，AAALAC认可的动物护理设施上的扭转12小时亮/暗循环笼（在0600小时的灯关）。 </li><li>提供水和标准啮齿动物食物自由，直到体重大约是380 - 400克随后，食物限制大鼠，并在85至90％的自由喂养的体重在实验会议保持，以便采集响应于甲基苯丙胺和维护。 </li><li>从到达整个实验处理每天在家里笼室啮齿类动物为了会议记录体重及调整日常食物分配。 </li><li>一旦目标的权重都实现了，准备手术植入患有慢性留置颈静脉导管和颅内刺激电极每鼠。 </li></ol> 2.颈内静脉穿刺置管<ol><li>导管准备<ol><li>制备硅橡胶管的一个13厘米长为0.012 x 0.025内径&#39;，创建一个硅氧烷球4厘米使用额外硅胶管和电灼管的一端，并允许风干。 </li><li>浸在来自于柑橘几分钟一苎烯系溶剂的管的另一端，并允许膨胀。连接膨胀管连接到一个不锈钢导向导管弯曲成直角。 </li><li>锚不锈钢引导套管用牙科丙烯酸水泥生物相容性网格的1"方的弯曲基地。 </li><li>内外冲洗导管，用乙醇和蒸馏水:河注入空气通过油管消除残余液滴。晾干O / N。 </li></ol></li><li>无菌技术<ol><li>执行使用无菌手术技术在一个专用的动物外科套件中的所有手术。灭菌用高压灭菌器械和植入物，并通过将防水纸张上或桌子铺上无菌巾（S）准备一个无菌区。 </li><li>戴无菌手套，并保持所有的仪器，植入物及手术纱布在无菌区的过程中。擦拭每台仪器用酒精棉签其次是20秒，程序间的珠灭菌时多次手术执行。 </li></ol></li><li>麻醉<ol><li>预处理大鼠阿托品（硫酸盐）（0.04毫克/公斤，平方），然​​后加入戊巴比妥（20 - 50毫克/ kg，腹腔）实现麻醉。 </li><li>注射用无菌青霉素G普鲁卡因悬浮液动物（75000单位，即时通讯）和镇痛药（卡洛芬，5 - 10毫克/公斤，SC或KEToprofen，2 - 5毫克/公斤，SC）之前立即手术，以减少围手术期的感染和疼痛，分别。 </li><li>检查是否有足够的麻醉通过提供一个适度的脚趾捏的动物，如果没有反应发生，然后继续。敷眼润滑剂两眼。 </li></ol></li><li>手术现场准备<ol><li>剃对大鼠的背上一个2×2厘米背补丁只是后侧到连接肩胛骨的线。剃一个1×1厘米腹侧补丁在颚骨和胸骨之间的右颈部区域。 </li><li>用酒精擦拭垫随后聚维酮碘溶液剃光领域。将大鼠在无菌巾，并让优碘，然后再继续干。 </li></ol></li><li>导管植入<ol><li>使平行连接肩胛骨在midscapular区域上使用10-刃刀背面的线的切口。使用止血钳到皮肤从底层结缔组织分开来创建为一个平面网状导管基地。灌溉用无菌生理盐水的区域，并盖上之前转动到老鼠背部无菌纱布。 </li><li>作出正确的颚骨和使用10-刃刀胸骨之间的对角切口。使用止血钳到皮肤从底层结缔组织分开来定位颈静脉。注:颈静脉呈白色/银色和光泽，是更大的雄性大鼠高于女性。 </li><li>放置一个抹刀静脉下方，轻轻扎暴露静脉的4-0丝线缝合周围的顶（近部）和静脉推回颈部。 </li><li>分离结缔组织下，下侧壁颈部皮肤，但高于肌肉来创建一个肤浅的口袋里。由隧道背臂和向上后面推从颈部切口到midscapular切口消毒套管针。通过插入刚性导丝向上穿过套管针从颈部端并进入远侧导管运行从背面到颈部的导管。拉通过对颈切口和所附远侧导管将跟随导丝回来。 </li><li>通过在先前放置近端缝线拉隔离右颈静脉过刮刀。使用一球的剪刀，使对静脉的上部小部分切割。 </li><li>插入一个弯钳进入静脉切开打开它。保持镊子隔开但到位，通过导管末端在钳尖之间并进入静脉约2 - 3厘米那里它将终止右心房外面。 </li><li>通过把4-0丝线绕远端静脉固定导管。领带的近端和远端缝合在一起的"盒子"结添加额外的稳定性。 </li><li>冲洗导管肝素化无菌生理盐水并提请回血，确认成功植入。修剪缝合1 - 2毫米以上的节，掖下颈部皮肤剩余近端导管。关闭与您选择的适当的缝合/方法。如果非absor巴布尔缝合线或U形钉时，它们必须在全身麻醉下10-14天被除去。覆盖切口用无菌棉签抗生素软膏。 </li><li>锚远侧导管/导向导管/网格组件使用可吸收缝线皮下背面组织在两个网状基体的相对的角落。关闭背切口周围使用中断，非反相吸收缝合线的引导套管。覆盖切口用无菌棉签抗生素软膏。 </li></ol></li><li>术后护理<ol><li>用3毫升注射器冲洗用肝素化的0.9％无菌盐水的溶液导管和放置一个闭孔到导向套管，以防止堵塞。 </li><li>冲洗，每天每只鼠的导管以保持开放。 </li><li>紧接着的步骤，将在其家笼老鼠在一个加热垫的外科手术室，观察到的意识和自发运动的回报。</li><li>返回回收的老鼠的菌落室和允许五至七天到颅内手术之前通过。称重，处理和每日评估其一般状况，包括检查对感染和评价动物行为，外观和活动水平。 </li><li>咨询动物资源的兽医，如果出现任何问题，并按照任何推荐的治疗方案。注入动物止痛剂（卡洛芬，5 - 10毫克/公斤，SC或酮洛芬，2 - 5毫克/公斤，SC）根据需要治疗的围手术期疼痛。 </li></ol></li></ol> 3.颅内电极放置<ol><li>手术准备<ol><li>如在2.2节执行无菌条件下，所有的手术。 </li></ol></li><li>麻醉<ol><li>放置动物在麻醉诱导室和提供异氟烷气体流入腔在1000 - 2000毫升/分钟的蒸发器设定为5％。 </li><li>一旦动物是横卧，从室中删除在第二鼻锥放在软垫立体定向的操作平台。从切换室气流鼻锥和跑气保用蒸发器设置在2 - 3％。 </li><li>当需要维持稳定的呼吸和手术过程中对刺激无反应调节汽化器。 </li><li>注入大鼠用无菌青霉素G普鲁卡因悬浮液（75,000单位，即时通讯）和镇痛药（丁丙诺啡0.05至0.5mg /千克SC）之前立即手术，以减少围手术期的感染和疼痛，分别。 </li><li>检查是否有足够的麻醉通过提供一个适度的脚趾捏的动物，如果没有反应发生，然后继续。敷眼润滑剂两眼。 </li></ol></li><li>手术现场准备<ol><li>剃鼠的头的顶部和放置在鼠耳棒的过程中，以保持其头部固定。用酒精擦拭垫随后聚维酮碘溶液剃光面积。让优碘，然后再继续干。 </li></ol></li><li>电极植入<ol><li>抓头皮之间，略先于耳钳，并用剪刀整个基地削减。这manuveur将删除在中期颅骨皮肤的1.5×1厘米的区域。 </li><li>使用10刀片，以便通过骨膜圆周切口向下的头骨和一个弯钳刮，并取出脑髓。灌溉用无菌生理盐水的区域，轻拍过量血液和盐用纱布，并允许颅骨完全干燥所以骨性标志，包括前囟，可以清楚地看到。 </li><li>对于双侧手术，安装两个双极铂铱电极，一成的立体操作平台的两边各电极座。移动第一电极就位〜1毫米以上前囱和写下立体定向坐标前后（AP）和内侧 - 外侧（ML）的位置，这将在所述数字显示器上显示。实际上并不触摸电极尖端的S​​KULL因为电极将不再起作用。重复此过程的另一个电极。 </li><li>计算最终的AP和ML坐标基于感兴趣的靶结构。移动电极到该位置与尖端刚好颅骨上方获得初始背腹（DV）的数字显示坐标。计算基于感兴趣的靶结构的最后的DV深度。 注意:伏隔核壳是针对在本实施例给出其已知参与使用以下立体定向药物圆满行为8的坐标相对于前囟: AP进入坐标= [数字显示的前囟坐标] + 1.6 ML入口坐标= [数字显示的前囟坐标]±2.4左/右 DV深度= [数字显示在在AP / ML进入颅骨表面坐标] - 8.5 </li><li>标记用永久性毫安每个电极的颅骨的表面上的突出的项目位置rker。不要磕碰或者这个动作过程中触摸电极的尖端。 </li><li>使用圆球状金刚石涂层毛刺钻1.4毫米的孔，​​在每个标记。要小心，不要通过头骨投身到颅内跳马的高速钻头。使用弯钳刺破硬脑膜后颅骨已经钻出了。 </li><li>使用圆球状金刚石涂层毛刺钻0.7毫米孔的颅骨螺钉放置电极条目后面附加的四个位置。使用手动螺丝刀放置4 0.8（直径）×3.2（长）mm的不锈钢螺钉插入颅骨，两个在中线的每一侧。紧紧保护这些螺丝高达约一半的长度入颅，因为他们是主要的基础设施，将举行的地方颅盖在未来几周和几个月。 </li><li>通过手动操作使管理该旋钮通过其burrhole进入大脑所计算的DV深度小心地将第一电极的Z坐标电极支架。转动旋钮的速度大致等于每秒1/2圈以防止对电极的前端不应有的损失。确保电极尖端进入大脑时，不接触burrhole的骨边缘。 </li><li>安全使用超级胶水层叠在burrhole和后螺钉，随后牙科水泥第一电极。一旦这种结构已完全干燥，从支架上取下电极。重复的插入和固井方法，所述第二电极。应用牙科用粘固一直到皮肤边缘，但不与皮肤重叠，因为这松开颅水泥帽长期。 </li></ol></li><li>术后护理<ol><li>将两个防尘帽在电极底座，防止堵塞。 </li><li>紧接着的步骤，将在其家笼老鼠在一个加热垫的外科手术室，观察到的意识和自发运动的回报。 </li><li>返回老鼠恢复为thË殖民地的房间，让五天开始实验前通过。称重，处理和每日评估大鼠的一般状态，包括检查对感染和评价动物行为，外观和活动水平。 </li><li>咨询动物资源的兽医，如果出现任何问题，并按照任何推荐的治疗方案。注入动物止痛剂（丁丙诺啡0.05至1毫克/千克SC），为需治疗围手术期疼痛。颅内出血可配合使用非甾体类止痛药更普遍（卡布洛芬，5 - 10毫克/公斤，SC或酮洛芬，2 - 5毫克/公斤，SC），所以使用丁丙诺啡围手术期颅内手术。 </li></ol></li></ol> 4.操作性设备<ol><li>使用包含声音衰​​减外壳内的塑料和不锈钢操作性条件反射腔运行行为实验。 </li><li>与排风扇提供通风和白色NOI装备每个机柜SE掩盖无关的声音。 </li><li>使用一台个人电脑和一个行为软件接口系统进行编程的程序，并收集实验数据。 </li><li>普通安装<ol><li>装备各实验室安装在一个墙壁与位于每个杠杆上方的刺激光室的两个响应杠杆。指定杠杆之一的"活性"杠杆，以便它被按下时产生一个编程的结果。 </li><li>程序中的积极响应杆上面直接位于每操作性会议期间照亮刺激光，说明药物的有效性。 </li><li>拥有超过2.8秒伴随着的房子光在对面的墙上持续了5秒，刺激光去输液交付甲基苯丙胺（0.05毫克/公斤/输液100微升0.9％氯化钠）对主动杆结果的响应关在30秒的超时。 </li><li>计数响应在主动杆，但他们应该没有凉城计划在30秒的超时时间quences。对于完成后，在不活动的杠杆记录反应，但他们应该没有计划的后果。 </li></ol></li></ol> 5.静脉注射（IV）甲基苯丙胺的自我管理程序<ol><li>一般准备<ol><li>加载老鼠进入操作室应尽快平静，尽量减少行为的假象。附加一个不锈钢弹簧拴住于导向套管上的啮齿动物的背部，并具有防漏流体回转悬浮操作性室的上方。 </li><li>确保连接管道的从旋转中的电机驱动泵位于声音衰减外壳外部20毫升药物注射器的完整性。要做到这一点，推塑料连接管的至少四分之一英寸到金属旋转尖端和药物注射器针尖，直到它不会与中度拉滑落。此消彼长的旋转和皮带装配允许相对奔放米ovement动物。 </li><li>至周五每天周一进行操作性会话在大约相同的时间。 </li></ol></li><li>获得<ol><li>为了便于快速采集四甲基苯丙胺自我管理的，运行大鼠每日6小时会话四到五个连续天。 </li><li>在一个固定的比例reinforcementduring的FR-1 + 30秒的时间表，只收到一个输液四甲基苯丙胺的每按在活动杆之后是30秒超时举办这些会议（ 例如，没有提示或奖励的后果发生压无论杆）。注意:此初始延长和"容易"的访问将导致在大多数鼠类获取显著吸毒行为在小于或等于一周（ 图3）。 </li></ol></li><li>保养<ol><li>在培训的第二个星期，至周五运行大鼠每天2小时的会议周一到维护和完善IV威百亩基安非他明的自我管理。 </li><li>上加强件的固定比率的FR-1 + 30秒超时时间表进行会话。 </li><li>记录稳定，激烈时的甲基苯丙胺展示跨越每个会话的总数为三个连续会话（ 图4）和输注的跨过第一30分钟的累积数量大于输注期间的累积数的变化小于10％的响应第二30分钟（ 图5）。注意:该标准确保了大鼠开发一种载药模式在会话指示成瘾行为19而不是简单地随便使用的开始。 </li></ol></li><li>会后<ol><li>在每届会议结束时，断开啮齿动物的背部皮带。冲洗导管与0.1毫升含有800单位链激酶，以防止血块0.9％盐溶液中。插入阻塞器到每一个引导套管，防止堵塞返回在R之前的ats到饲养笼。 </li><li>每个实验会议结束周三在整个实验过程中后立即测试导管的通畅。 <ol><li>准备3毫升注射器，用22克针，肝素含有抑菌盐水来测试导管通畅。附加一个4到6英寸长的塑料管，以针和另一端，以对动物的背部的导管插管组件的金属柱的一端。 </li><li>注入0.1至0.2ml生理盐水，以确保清晰的流程，然后绘制注射器活塞回来。如果导管是专利，它应该既冲洗容易且拉回血，将在管道中可见。释放活塞和注入另一0.2 ml至刷新所有血液回通过导管。 </li><li>如果血液不能被撤回，然后取出3毫升注射器和油管的金属柱。 </li><li>准备1毫升注射器，用22克针，包括美索比妥钠，速效麻醉剂，以进一步检验导管通畅。附加一个4到6英寸长的塑料管，以针和另一端，以对动物的背部的导管插管组件的金属柱的一端。 </li><li>注入1.5毫克，并迅速从对动物的背面的金属交取出1毫升注射器和油管。重新连接3毫升注射器充满肝素抑菌生理盐水注入0.1 - 0.2毫升如果动物失去肌肉张力在3秒，然后将导管专利和实用。 </li></ol></li></ol></li><li>请参阅补充文件"常见错误"的缺陷部分，解决不良反应的甲基苯丙胺，无法获得甲基苯丙胺的自我管理和大鼠的提取难度。 </li></ol> 6.大脑刺激装置<ol><li>使用10至12丛玻璃箱（12×18×18）（宽x高x）运行DBS实验。盖与坚硬不透明外每箱纸覆盖盒子的背面和侧面，以防止大鼠查看或彼此交互。离开清晰的前面板上发现这样考官可以在刺激会议观看动物。 </li><li>盖的箱子的顶部用一个半透板，防止老鼠从同时允许气流逸出。使用该面板，以支持位于上述各框，以方便啮齿动物头帽和刺激系统之间的电连接的换向器。 </li><li>使用刺激系统，可以提供稳定的电流，以多个同时畜为DBS实验。使用一个系统，该系统包括一个用户可编程数字信号处理器/通信接口的，刺激剂，刺激器的电池组，信道分离器箱，及附带的软件（见材料说明书）。 </li><li>使用自定义长度的电缆连接到刺激的通道端口连接到每个commutat优越的电子底座要么。注意:需要将取决于单个实验室的长度。这些电缆的动物区之外，无需被覆盖在不锈钢弹簧。 </li><li>换向器的劣质电子底座连接到使用覆盖着不锈钢弹簧16的电缆啮齿动物的头帽植入电极座。确保电缆足够长的时间，让自由移动到机箱的每一个领域，而不在头帽显著紧张。注:电缆两端大致相当于老鼠的头会站在自己的四条腿，当是通常是足够的。 </li><li>脑刺激编程<ol><li>使用个人计算机和编程软件的刺激参数 （例如，波形，频率，脉冲宽度，刺激间延迟，电流幅度），并收集实验数据进行编程。 </li><li>使用可视化编程语言，指定功能的每个设备将执行以满足experi精神的端点和哪些数据将被存储和/或预计的用于查看实时性。运行这个特定的项目的命令表明在图1中。 </li><li>指定所需的频率，脉冲宽度，并进入视觉控制面板（图2）在实验前的起始振幅。高频刺激大鼠的典型参数是类似于在临床人类脑深部刺激用于:频率为130〜180赫兹，60至90毫秒的脉冲宽度，及为100的电流幅度至250微安4,8-10。注:低电流用于在啮齿动物，由于其尺寸减小相比，灵长类动物。 </li></ol></li></ol> 7.深部脑刺激过程<ol><li>要加载到老鼠的箱子，从换向器装上不锈钢弹簧线的头帽每个电极座。通过运行5μA电流的频率测试每个电极的阻抗1000赫兹，持续2秒。 <ol><li>如果阻抗等于或小于125千欧，然后继续进行实验，因为电极是能够提供治疗刺激。如果阻抗大于125千欧，考虑从实验除去动物因为电极的高电阻可能截断电流以潜在亚治疗水平。 </li></ol></li><li>通过一个或两个在此期间，它们将被连接到电极线（多个），但是未接收到任何活性治疗习惯模拟会话中运行的影响。模拟测试将消除任何非特异性行为效应。紧随每个模拟会话，运输老鼠操作性盒四甲基苯丙胺自我管理的每日2小时会议。 </li><li>此消彼长的大鼠分成两组，活性刺激和假刺激队列，使得基线药物摄入不组之间​​显著不同。 </li><li>执行日常DBS会议S上啮齿动物的队列，在此期间他们接受无论是主动脑电刺激或无刺激3小时，这取决于它们的组分配5天。紧随每个DBS会议上，交通老鼠操作性盒四甲基苯丙胺自我管理的每日2小时会议。 <ol><li>仔细在每个DBS会话的至少一部分观察动物，以确保刺激不会造成明显的改变动物的行为。如果在刺激后/出现的任何异常行为时，一定要记录这些意见。注:作者在这篇文章中描述的实验过程中有没有注意到显著行为改变或改变食物/饮水量。 </li></ol></li><li>改变DBS治疗，电参数，和DBS会话，并根据需要取决于假设操作性会话之间的时间的长度。 </li></ol>

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The authors have no competing financial interests with regards to this methodology.

虽然深部脑刺激的确切机制尚未完全表征，DBS功效为马达和精神病症，可能会导致从电治疗和各种皮质和皮质脑区域随时间6,22-26运作之间的动态相互作用。而甲基苯丙胺递送至鼠类非偶然的方法是公描述27,28，这些方法最适合于药物的药物动力学和神经化学效应27-29离散调查。操作性静脉注射毒品的自我管理，通过将动力药物的元素，非常适合怎样的电疗法，如星展银行与病理行为，随着时间的推移相互作用的研究。我们描述的步骤检查DBS的特遣队使用甲基苯丙胺一个环境的影响，在不同的环境。 还有在我们的四甲基苯丙胺自ADMINIST三个关键步骤配给模式:快速采集和药物摄入升级1）在长时间访问会话感应，药物摄入量稳定，高速率的2）在随后的简短访问会话维护和吸毒前装模式3）发展。这个范例可以实现在一个2至3个星期时限与10 - 12只大鼠的实验中，这是既具有成本效益和非常适合于测试的DBS定头帽中使用心理兴奋剂啮齿动物的潜在有限寿命的效果。这个过程中，这样的结合一段长的存取19,30,31其他范式合理模拟物质使用障碍的某些方面;它展示了使用和高激励的两个升级来获得药物早期会话"载药"，这是人类的依赖与娱乐使用19,30的重要方面。谁早就进入暴露在四甲基苯丙胺鼠害也表明认知缺陷32的，不同的反应，药物治疗33，药物动力学34和神经化学改变35更类似于人类患慢性甲基苯丙胺使用障碍比啮齿类动物仅用短访问曝光。 同样，也有在我们的脑深部刺激过程三个关键步骤:1）习惯化的DBS环境，包括头系绳连接，一个或两个"模拟"课程，2）日，间歇交付使用商业系统积极刺激， 3）DBS断开和后续运输毒品的设置。这种模式被设计为模仿非侵入性疗法，如TMS而不是传统的连续的DBS的过程。完全植入，可编程DBS系统，如那些用于常见的运动障碍3将患者从几个上述原因精神兴奋药的依赖患有轻微可行的。 IntermittenT电治疗策略，不涉及高风险的手术和调养，像TMS，可以更好地适应这一患者群体。我们所描述的方法将允许调查人员发展和完善的治疗策略，可以修改与毒品有关的行为，同时该药物环境之外在限制的时间内交付。有越来越多的证据认为是特定神经生理缺陷23后形成图案或联合全身药物治疗36使出持久精神病和药物相关的行为的积极影响数周治疗后已停止瞬态颅内电刺激。 初始优秀的手术技术和持续的关心的在激烈的药物使用多个手术部位的需求是这种方法的主要限制。如果两个静脉导管或DBS电极成为非经营性和/或感染，鼠须退出研究。在严格的无菌技术颈静脉导管和颅内电极位置最好从以前的独立启动这些程序经验丰富的调查经验。 此过程是适合于若干修改和将来调查，包括检查:1）交替刺激参数（例如， -刺激波形，脉冲宽度，频率，幅度），2）其它潜在的治疗脑的目标（如， -伏隔核。核心，内侧前额叶皮质，脑，缰核），3）不同的DBS交付模式 （例如， -每日星展交货，周星展交货，星展银行在不同的时间间隔操作性会议上，星展银行收购之前），以及或许是最令人兴奋之前， 4）短时DBS的组合和模仿选择性的途径光遗传学刺激，发挥持久的行为修改36药剂。

甲基苯丙胺是产生强烈和长时间的兴奋由于急性增加突触单胺，尤其是多巴胺精神刺激。甲基苯丙胺的依赖是估计有25 3400万全球用户，并没有有效的治疗1,2流行的健康问题。有大量需要制定甲基苯丙胺依赖新的治疗策略。深部脑刺激（DBS）是一种使用大脑"起搏器"正常化发生在某些疾病破坏性神经元放电模式，包括帕金森氏症，肌张力障碍，和特发性震颤3一个神经外科手术。最近人类病例报告表明，DBS也可以是一种有效的治疗酒精和药物依赖，但关于精神兴奋剂的临床前证据（例如，可卡因，甲基苯丙胺）被限制4-8。 连续深部脑刺激，因为它是currentlŸ实践，需要从患者和他/她的家人卓越的合作。细致的伤口护理和个人卫生需要保护下面的起搏器硬件，这是容易受到感染，即使在谁不使用静脉注射药物产生的菌血症患者。定期随访的DBS装置的也是必要的，因为该系统的开环设计;有经验的医师改变现代DBS的设置，在日常门诊预约3降低目标的症状。这种处理的模式将在可卡因和脱氧麻黄碱用户由于他们的具有挑战性的心理情况所限制。通过检查DBS效果当治疗期间在药物使用的环境9-11可卡因自身给药程序连续递送几啮齿动物研究已经模仿此不切实际范例。 非侵入性连续技术不需要留置硬件，例如经颅磁刺激（TMS），其可以用于治疗物质使用障碍12的一个更好的选择。 TMS递送非侵入性地用外部headcoil生成期间每日在特定脑靶电场，间歇性治疗。 ħ线圈或"深"TMS让更深大脑结构被刺激，除了皮质部位，扩大其潜在用途13,14的最近问世。这两种疗法在一系列会议连续发表在不同的环境比主药的使用，并已在人类和啮齿类动物对药物的依赖13,15-17试验表明承诺。治疗甲基苯丙胺依赖患者的窗口很可能会在清醒的时期，如法院规定的康复，而不是在街头狂欢时，他们可以体验暴力或不稳定行为18。因此，本文的目的是描述电刺激的递送在时间上和空间上与药物使用环境中，为进一步接近什么是可能在人类中，为第IV甲基苯丙胺依赖治疗分开。

<table><tbody><tr><td>Rodent operant chambers</td><td>Med Associates, Inc</td><td>ENV-008CT</td><td>Med Associates Inc. PO Box 319 St. Albans, Vermont 05478 USA Phone: (802) 527-2343</td></tr><tr><td>Kopf Small Animal Stereotaxic Instrument with Digital Display Console</td><td>Kopf Instruments</td><td>Model 940</td><td>Kopf Phone: 1-877-352-3275 Fax: 1-818-352-3275 Email: sales@kopfinstruments.net</td></tr><tr><td>Z-Series 3-DSP Bioamp Processor</td><td>Tucker Davis Technologies</td><td>RZ5D</td><td>Tucker-Davis Technologies&amp;nbsp;11930 Research Circle Alachua, FL 32615&amp;nbsp;USA Ph: 386-462-9622&amp;nbsp;www.tdt.com</td></tr><tr><td>Z-Series 32-Channel Stimulator</td><td>Tucker Davis Technologies</td><td>IZ2-32</td><td>Software is accompanied by a manual that discusses how to program experiments using the OpenEx platform, which can be accessed here: http://www.tdt.com/files/manuals/OpenEx_User_Guide.pdf</td></tr><tr><td>48 Volt LI-ION Battery Pack for IZ2 Stimulator</td><td>Tucker Davis Technologies</td><td>LZ48-200</td><td></td></tr><tr><td>32-Channel Splitter Box for PZ5</td><td>Tucker Davis Technologies</td><td>S-BOX_PZ5</td><td></td></tr><tr><td>OpenEx Ext Software Package for Multi-Channel Neural Recording</td><td>Tucker Davis Technologies</td><td>OpenEx</td><td></td></tr><tr><td>Platinum-iridium stimulating electrodes</td><td>Plastics One Inc</td><td>MS303/8-B/SPC ELECT PT 2C TW .005&quot;</td><td>Plastics One Inc P.O.Box 21465, S.W. Roanoke, VA 24018, PH 540-772-7950</td></tr><tr><td>2-channel cables between stimulator and commutator</td><td>Plastics One Inc</td><td>305-441/2 W/ Spring</td><td></td></tr><tr><td>2-channel cables between commutator and electrode pedestal</td><td>Plastics One Inc</td><td>305-305 W/ Spring</td><td></td></tr><tr><td>4-channel commutators</td><td>Plastics One Inc</td><td>SL2+2C and SL2+SC/SB</td><td></td></tr></tbody></table>

a general method for evaluating deep brain stimulation effects on intravenous methamphetamine self-administration

Substance use disorders, particularly to methamphetamine, are devastating, relapsing diseases that disproportionally affect young people. There is a need for novel, effective and practical treatment strategies that are validated in animal models. Neuromodulation, including deep brain stimulation (DBS) therapy, refers to the use of electricity to influence pathological neuronal activity and has shown promise for psychiatric disorders, including drug dependence. DBS in clinical practice involves the continuous delivery of stimulation into brain structures using an implantable pacemaker-like system that is programmed externally by a physician to alleviate symptoms. This treatment will be limited in methamphetamine users due to challenging psychosocial situations. Electrical treatments that can be delivered intermittently, non-invasively and remotely from the drug-use setting will be more realistic. This article describes the delivery of intracranial electrical stimulation that is temporally and spatially separate from the drug-use environment for the treatment of IV methamphetamine dependence. Methamphetamine dependence is rapidly developed in rodents using an operant paradigm of intravenous (IV) self-administration that incorporates a period of extended access to drug and demonstrates both escalation of use and high motivation to obtain drug.

以下的IV颈静脉导管和颅内DBS电极的放置，啮齿动物可成功地在短暂的复苏期过后训练自我管理四甲基苯丙胺。 图3显示了大鼠将获得并在2天后的扩展访问升级甲基苯丙胺自我施用药物与由4天，平均每个会话168±12注入。 大鼠然后转移到操作性训练每天2小时的时间表的原因有两个:1），以防止甲基苯丙胺毒性和严重的行为变化的情况与持久性，长时间的访问和2），以建立响应该相对稳定的速率可以通过各种操纵治疗性干预。 图4表明，每个短访问会话总输注在第二周的平均数量为75±8和一般天的日常变化小于10％。 图5表明，大鼠devel软件包OP增加的动力服药相比，天如由摄入的"前装"模式的出现，通过培训6天1.一旦开发，其效果在很大程度上维持在随后的会话（数据未显示） 。 图6显示了双边DBS在非药物环境递送导致操作性四甲基苯丙胺自我施用的三个5天相比于假手术组刺激的显着降低。 （ - 8.5，ML±2.4 AP + 1.6，DV），伏隔核的外壳是针对使用下面的立体坐标中相对于前囟门鉴于其已知的参与毒品圆满的行为8。刺激参数和持续时间分别宽松地基于以前出版的经验与DBS对精神疾病8,20,21的治疗，但可以根据实验者的需要进行调整。误差棒是适度的，而不是所有的日子意图通道意义表示的，可以在行为评估尽管明确的治疗效果可以看出响应的范围内。增加大鼠每个实验的数量可以帮助弥补这一自然的变化。 11只动物最初用于这个实验。一个动物安乐死的拒食手术后，一个动物被排除因癫痫发作，一个动物被排除由于DBS电极故障留给我们一共有8只动物（N = 4深水; N = 4个活动DBS）。一般开始用约10 - 12只为每个实验将允许其成功完成。 <img alt="图1" src="/files/ftp_upload/53266/53266fig1.jpg" /> 图1. 可视编程语言。研究者使用可视编程语言，如这里所示的例子中，设计一个程序，它可以提供脑刺激到多个灵魂LS同时在用户输入的参数， <a href="https://www.jove.com/files/ftp_upload/53266/53266fig1large.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/53266/53266fig2.jpg" /> 图2. 视觉控制面板。在此之前的实验开始，研究者指定所希望的频率，脉冲宽度，并在一个可视控制面板的左侧幅度。在这里，刺激参数为:电流强度200μA;脉冲宽度61毫秒;脉冲频率130赫兹。一旦刺激开始，显示在右侧的有功电流输送的波形。 <a href="https://www.jove.com/files/ftp_upload/53266/53266fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 图3. 收购四甲基苯丙胺的自我管理的总操作性响应（360分钟）的数据采用重复测量方差分析与定义为重复测量的日常会话分析。所有的分析都认为P &lt;0.05被认为是显著。数据是平均值±标准差。在结束前四天操作性培训的日常6小时操作性会议共注射甲基苯丙胺+ P &lt;0.05相比，会话1和2， <a href="https://www.jove.com/files/ftp_upload/53266/53266fig3large.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/53266/53266fig4.jpg" /> 图4。使用重复测量方差分析与定义为重复测量的日常会话维护四甲基苯丙胺的自我管理的总操作性响应（120分钟）的数据进行了分析。所有的分析都认为P &lt;0.05被认为是显著。数据是平均值±标准差。在过操作性的培训，展示稳定而强烈的吸毒行为，第二周每天2小时操作性会议总甲基苯丙胺输液。 <a href="https://www.jove.com/files/ftp_upload/53266/53266fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/53266/53266fig5.jpg" /> 图5. 发展动因服药。操作性作出答复的是总量为每15分钟第一个小时和数据进行了分析使用重复我asures ANOVA与定义为重复测量每15分钟象限。所有的分析都认为P &lt;0.05被认为是显著。数据是平均值±标准差。 "前装"的模式是不存在的操作性培训的第一天，但​​在第二周的发展，显示出强大的动力服药。 + P &lt;0.05相比，30，45和60分钟，++ P &lt;0.05相比，45和60分钟，+++ P &lt;0.05相比，60分钟。 <a href="https://www.jove.com/files/ftp_upload/53266/53266fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图6" src="/files/ftp_upload/53266/53266fig6.jpg" /> 图 6. 四甲基苯丙胺的自我管理DBS的影响，总操作性在第一个小时响应（60分钟）的数据是使用一个混合方差分析与betwe分析EN治疗（DBS VS深水）除变量和日常会话的重复测量。所有的分析都认为P &lt;0.05被认为是显著。数据是平均值±标准差。双边预操作性深部脑刺激甲基苯丙胺注射的次数显著减少了操作性治疗3天，4响应的第一个60分钟，7 + P &lt;0.05相比，假手术组和基线响应。响应返回给每日治疗结束后的基线水平。 <a href="https://www.jove.com/files/ftp_upload/53266/53266fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about A General Method for Evaluating Deep Brain Stimulation Effects on Intravenous Methamphetamine Self-Administration at JoVE.com

A General Method for Evaluating Deep Brain Stimulation Effects on Intravenous Methamphetamine Self-Administration

This article describes the delivery of intracranial electrical stimulation that is temporally and spatially separate from the drug-use environment for the treatment of IV methamphetamine dependence.

一般方法评估对静脉注射甲基苯丙胺的自我管理深部脑刺激的影响

Substance use disorders, particularly to methamphetamine, are devastating, relapsing diseases that disproportionally affect young people.  There is a need ...

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Research

JoVE Journal

Behavior

15.1K 次观看.  Louisiana State University. This article describes the delivery of intracranial electrical stimulation that is temporally and spatially separate from the drug-use environment for the treatment of IV methamphetamine dependence.

视频: A General Method for Evaluating Deep Brain Stimulation Effects on Intravenous Methamphetamine Self-Administration

Novel Apparatus and Method for Drug Reinforcement

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction. Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in animal research to model various components of drug reinforcement, consumption, and addiction in humans. For this study, we used a novel approach to studying drug reinforcement in rats by combining traditional CPP and self-administration methodologies. We assembled an apparatus using two Med Associate operant chambers, sensory stimuli, and a Plexiglas-constructed neutral zone. These modifications allowed our experiments to encompass motivational aspects of drug intake through self-administration and drug-free assessment of drug/cue conditioning strength with the CPP test. In our experiments, rats self-administered cocaine (0.75 mg/kg/inj, i.v.) during either four (e.g., the "short-term") or eight (e.g., the "long-term") alternating-day sessions in an operant environment containing distinctive sensory cues (e.g., olfactory and visual). On the alternate days, in the other (differently-cued) operant environment, saline was available for self-infusion (0.1 ml, i.v.). Twenty-four hours after the last self-administration/cue-pairing session, a CPP test was conducted. Consistent with typical CPP findings, there was a significant preference for the chamber associated with cocaine self-administration. In addition, in animals undergoing the long-term experiment, a significant positive correlation between CPP magnitude and the number of cocaine-reinforced lever responses. In conclusion, this apparatus and approach is time and cost effective, can be used to examine a wide array of topics pertaining to drug abuse, and provides more flexibility in experimental design than CPP or self-administration methods alone.

Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in research to model various components of drug reinforcement, consumption, and addiction in humans. In this report, we combined traditional CPP and self-administration methods as a novel approach to studying drug reinforcement and addiction in rats.

新型仪器和药物加固方法

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction.  Operant drug ...

科研

神经科学

A Novel Approach to Assess Motor Outcome of Deep Brain Stimulation Effects in the Hemiparkinsonian Rat: Staircase and Cylinder Test

Deep brain stimulation of the subthalamic nucleus is an effective treatment option for Parkinson&#39;s disease. In our lab we established a protocol to screen different neurostimulation patterns in hemiparkinsonian (unilateral lesioned) rats. It consists of creating a unilateral Parkinson&#39;s lesion by injecting 6-hydroxydopamine (6-OHDA) into the right medial forebrain bundle, implanting chronic stimulation electrodes into the subthalamic nucleus and evaluating motor outcomes at the end of 24 hr periods of cable-bound external neurostimulation. The stimulation was conducted with constant current stimulation. The amplitude was set 20% below the individual threshold for side effects. The motor outcome evaluation was done by the assessment of spontaneous paw use in the cylinder test according to Shallert and by the assessment of skilled reaching in the staircase test according to Montoya. This protocol describes in detail the training in the staircase box, the cylinder test, as well as the use of both in hemiparkinsonian rats. The use of both tests is necessary, because the staircase test seems to be more sensitive for fine motor skill impairment and exhibits greater sensitivity to change during neurostimulation. The combination of the unilateral Parkinson model and the two behavioral tests allows the assessment of different stimulation parameters in a standardized way.

Deep brain stimulation (DBS) is an effective treatment option for Parkinson's disease. We established a study design to screen novel stimulation paradigms in rats. The protocol describes the use of the staircase test and cylinder test for motor outcome assessment in DBS treated hemiparkinsonian rats.

一种新的方法，以评估在帕金森病大鼠脑深部电刺激效应的运动结果:楼梯和缸测试

Deep brain stimulation of the subthalamic nucleus is an effective treatment option for Parkinson&#39;s disease. In our lab we established a protocol to ...

行为学

Methods for Intravenous Self Administration in a Mouse Model

Animal models have been developed to study the reinforcing effects of drugs, including the intravenous self-administration (IVSA) paradigm. The advantages of using an IVSA paradigm to study the reinforcing properties of drugs of abuse such as cocaine include the fact that the drug is self-administered instead of experimenter-administered, the schedule of reinforcement can be altered, and accurate measurement of the quantities of drug consumed as well as the timing and pattern of IV injections can be obtained. Furthermore, the intravenous route of administration avoids potential confounds related to first pass metabolism or taste, and produces rapid increases in blood and brain drug levels. As outlined in this video, intravenous self-administration can be obtained without prior food restriction or prior drug training following careful catheter placement during surgery and meticulous daily catheter flushing and maintenance. Experimental procedures outlined in this paper include a description of animal housing and acclimation methods, operant training using sweetened milk solutions, and catheter implantation surgery.

The intravenous self-administration (IVSA) paradigm is considered to be the gold standard in examining the reinforcing properties of drugs of abuse in rodents. This manuscript outlines the experimental procedures and surgical techniques necessary to obtain reliable IVSA data. In particular, meticulous catheter implantation and maintenance are highlighted.

在小鼠模型中静脉自我管理的方法

Animal models have been developed to study the reinforcing effects of drugs, including the intravenous self-administration (IVSA) paradigm. The advantages ...

Microdialysis of Ethanol During Operant Ethanol Self-administration and Ethanol Determination by Gas Chromatography

Operant self-administration methods are commonly used to study the behavioral and pharmacological effects of many drugs of abuse, including ethanol. However, ethanol is typically self-administered orally, rather than intravenously like many other drugs of abuse. The pharmacokinetics of orally administered drugs are more complex than intravenously administered drugs. Because understanding the relationship between the pharmacological and behavioral effects of ethanol requires knowledge of the time course of ethanol reaching the brain during and after drinking, we use in vivo microdialysis and gas chromatography with flame ionization detection to monitor brain dialysate ethanol concentrations over time. 
Combined microdialysis-behavioral experiments involve the use of several techniques. In this article, stereotaxic surgery, behavioral training and microdialysis, which can be adapted to test a multitude of self-administration and neurochemical centered hypotheses, are included only to illustrate how they relate to the subsequent phases of sample collection and dialysate ethanol analysis. Dialysate ethanol concentration analysis via gas chromatography with flame-ionization detection, which is specific to ethanol studies, is described in detail. Data produced by these methods reveal the pattern of ethanol reaching the brain during the self-administration procedure, and when paired with neurochemical analysis of the same dialysate samples, allows conclusions to be made regarding the pharmacological and behavioral effects of ethanol.

A method to determine the time course of ethanol concentration in the brains of rats during operant ethanol self-administration is described. Gas chromatography with flame ionization detection is used to quantify ethanol in the dialysate samples, because it has the sensitivity required for the small volumes that are generated.

微透析的乙醇操作性乙醇的自我管理和乙醇的测定气相色谱

Operant self-administration methods are commonly used to study the behavioral and pharmacological effects of many drugs of abuse, including ethanol.  ...

A Procedure for Implanting Organized Arrays of Microwires for Single-unit Recordings in Awake, Behaving Animals

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell level. The technique allows experimenters to examine temporally and regionally specific firing patterns in order to correlate recorded action potentials with ongoing behavior. Moreover, single-unit recordings can be combined with a plethora of other techniques in order to produce comprehensive explanations of neural function. In this article, we describe the anesthesia and preparation for microwire implantation. Subsequently, we enumerate the necessary equipment and surgical steps to accurately insert a microwire array into a target structure. Lastly, we briefly describe the equipment used to record from each individual electrode in the array. The fixed microwire arrays described are well-suited for chronic implantation and allow for longitudinal recordings of neural data in almost any behavioral preparation. We discuss tracing electrode tracks to triangulate microwire positions as well as ways to combine microwire implantation with immunohistochemical techniques in order to increase the anatomical specificity of recorded results.

Implanting organized arrays of microwires for use in single-unit electrophysiological recordings presents a number of technical challenges.&#160;Methods for performing this technique and the equipment necessary are described.&#160;Also, the beneficial use of organized microwire arrays to record from distinct neural subregions with high spatial selectivity is discussed.

一个程序植入微丝的有组织阵列的单机录音清醒，举止动物

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

深部脑刺激与同步功能磁共振成像在鼠害

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

Microelectrode Guided Implantation of Electrodes into the Subthalamic Nucleus of Rats for Long-term Deep Brain Stimulation

Deep brain stimulation (DBS) is a widely used and effective therapy for several neurologic disorders, such as idiopathic Parkinson&#8217;s disease, dystonia or tremor. DBS is based on the delivery of electrical stimuli to specific deep anatomic structures of the central nervous system. However, the mechanisms underlying the effect of DBS remain enigmatic. This has led to an interest in investigating the impact of DBS in animal models, especially in rats. As DBS is a long-term therapy, research should be focused on molecular-genetic changes of neural circuits that occur several weeks after DBS. Long-term DBS in rats is challenging because the rats move around in their cage, which causes problems in keeping in place the wire leading from the head of the animal to the stimulator. Furthermore, target structures for stimulation in the rat brain are small and therefore electrodes cannot easily be placed at the required position. Thus, a set-up for long-lasting stimulation of rats using platinum/iridium electrodes with an impedance of about 1 M&#937; was developed for this study. An electrode with these specifications allows for not only adequate stimulation but also recording of deep brain structures to identify the target area for DBS. In our set-up, an electrode with a plug for the wire was embedded in dental cement with four anchoring screws secured onto the skull. The wire from the plug to the stimulator was protected by a stainless-steel spring. A swivel was connected to the circuit to prevent the wire from becoming tangled. Overall, this stimulation set-up offers a high degree of free mobility for the rat and enables the head plug, as well as the wire connection between the plug and the stimulator, to retain long-lasting strength.

A method for implanting electrodes into the subthalamic nucleus (STN) of rats is described. Better localization of the STN was achieved by using a microrecording system. Furthermore, a stimulation set-up is presented that is characterized by long-lasting connections between the head of the animal and the stimulator.

微电极引导电极植入到大鼠丘脑底核的长期深部脑刺激

Deep brain stimulation (DBS) is a widely used and effective therapy for several neurologic disorders, such as idiopathic Parkinson&#8217;s disease, ...

A Protocol for Measuring Cue Reactivity in a Rat Model of Cocaine Use Disorder

Cocaine use disorder (CUD) follows a trajectory of repetitive self-administration during which previously neutral stimuli gain incentive value. Cue reactivity, the sensitivity to cues previously linked with the drug-taking experience, plays a prominent role in human craving during abstinence. Cue reactivity can be assessed as the attentional orientation toward drug-associated cues that is measurable as appetitive approach behavior in both preclinical and human studies. Herein describes an assessment of cue reactivity in rats trained to self-administer cocaine. Cocaine self-administration is paired with the presentation of discrete cues that act as conditioned reinforcers (i.e., house light, stimulus light, infusion pump sounds). Following a period of abstinence, lever presses in the cocaine self-administration context accompanied by the discrete cues previously paired with cocaine infusion are measured as cue reactivity. This model is useful to explore neurobiological mechanisms underlying cue reactivity processes as well as to assess pharmacotherapies to suppress cue reactivity and therefore, modify relapse vulnerability. Advantages of the model include its translational relevance, and its face and predictive validities.&#160;The primary limitation of the model is that the cue reactivity task can only be performed infrequently and must only be used in short duration (e.g., 1 hour), otherwise rats will begin to extinguish the pairing of the discrete cues with the cocaine stimulus. The model is extendable to any positively reinforcing stimulus paired with discrete cues; though particularly applicable to drugs of abuse, this model may hold future applications in fields such as obesity, where palatable food rewards can act as positively reinforcing stimuli.

Cue reactivity is conceptualized as sensitivity to cues linked with drug-taking experiences that contribute to craving and relapse in abstinent humans. Cue reactivity is modeled in rats by measuring attentional orientation toward drug-associated cues that results in appetitive approach behavior in a cue reactivity test following self-administration and forced abstinence.

可卡因使用障碍大鼠模型中提示反应性测定的一种协议

Cocaine use disorder (CUD) follows a trajectory of repetitive self-administration during which previously neutral stimuli gain incentive value. Cue ...

An Invasive Method for the Activation of the Mouse Dentate Gyrus by High-frequency Stimulation

Electrical high-frequency stimulation (HFS), using implanted electrodes targeting various brain regions, has been proven as an effective treatment for various neurological and psychiatric disorders. HFS in the deep region of the brain, also named deep-brain stimulation (DBS), is becoming increasingly important in clinical trials. Recent progress in the field of high-frequency DBS (HF-DBS) surgery has begun to spread the possibility of utilizing this invasive technique to other situations, such as treatment for major depression disorder (MDD), obsessive-compulsive disorder (OCD), and so on. 
Despite these expanding indications, the underlying mechanisms of the beneficial effects of HF-DBS remain enigmatic. To address this question, one approach is to use implanted electrodes that sparsely activate distributed subpopulations of neurons by HFS. It has been reported that HFS in the anterior nucleus of the thalamus could be used for the treatment of refractory epilepsy in the clinic. The underlying mechanisms might be related to the increased neurogenesis and altered neuronal activity. Therefore, we are interested in exploring the physiological alterations by the detection of neuronal activity as well as neurogenesis in the mouse dentate gyrus (DG) before and after HFS treatment. 
In this manuscript, we describe methodologies for HFS to target the activation of the DG in mice, directly or indirectly and in an acute or chronic manner. In addition, we describe a detailed protocol for the preparation of brain slices for c-fos and Notch1 immunofluorescent staining to monitor the neuronal activity and signaling activation and for bromodeoxyuridine (BrdU) labeling to determine the neurogenesis after the HF-DBS induction. The activation of the neuronal activity and neurogenesis after the HF-DBS treatment provides direct neurobiological evidence and potential therapeutic benefits. Particularly, this methodology can be modified and applied to target other interested brain regions such as the basal ganglia and subthalamic regions for specific brain disorders in the clinic.

This protocol shows how to set up a reliable HFS method in mice. Neurons throughout the hippocampal dentate gyrus are electrically stimulated by HFS directly and indirectly in vivo. Neuronal activity and molecular signaling are examined by c-fos and Notch1 immunofluorescent staining, respectively; neurogenesis is quantified by bromodeoxyuridine labeling assay.

高频刺激激活小鼠齿状回的侵袭方法

Electrical high-frequency stimulation (HFS), using implanted electrodes targeting various brain regions, has been proven as an effective treatment for ...

Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry (CIS-FSCV) to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine

Phasic dopamine (DA) release from the ventral tegmental area (VTA) to the nucleus accumbens plays a pivotal role in reward processing and reinforcement learning. Understanding how the diverse neuronal inputs into the VTA control phasic DA release can provide a better picture of the circuitry that controls reward processing and reinforcement learning. Here, we describe a method that combines intra-VTA cannula infusions of pharmacological agonists and antagonists with stimulation-evoked phasic DA release (combined infusion and stimulation, or CIS) as measured by in vivo fast-scan cyclic voltammetry (FSCV). Using CIS-FSCV in anesthetized rats, a phasic DA response can be evoked by electrically stimulating the VTA with a bipolar electrode fitted with a cannula while recording in the nucleus accumbens core. Pharmacological agonists or antagonists can be infused directly at the stimulation site to investigate specific VTA receptors&#39; roles in driving phasic DA release. A major benefit of CIS-FSCV is that VTA receptor function can be studied in vivo, building on in vitro studies.

Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry CIS-FSCV to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine

The goal of this protocol is to directly manipulate ventral tegmental area receptors to study their contribution to subsecond dopamine release.

联合输注和刺激与快速扫描循环伏安法（CIS-FSCV）评估腹侧被盖区域受体对相位多巴胺的调节

Phasic dopamine (DA) release from the ventral tegmental area (VTA) to the nucleus accumbens plays a pivotal role in reward processing and reinforcement ...