这项研究是由美国国立卫生研究院/ NIAAA（AA11852和AA007471）的补助金。

1。立体定向手术<ol><li>所有程序遵循美国国立卫生研究院实验动物护理和使用指南，机构动物管理和使用委员会批准。 </li><li>利用立体定位仪，以及处理Long-Evans大鼠，用异氟烷麻醉，植入了一个21号套管（后microdilaysis探针插入）（塑料件，罗阿诺克，VA）的大脑区域的利益之上，和系绳螺栓被内置到头部阶段（用于支持微透析设备）。 </li><li>术中和术后监测大鼠仔细。确保所有的动物获得了至少一个星期的术后护理和恢复，并开始执行以下步骤之前，健康。一个朱庇特的视频的立体定向手术方法是可用的。 </li></ol> 2。操作性培训<ol><li>一个星期后，手术后的恢复，动物训练杆按10％的蔗糖搜索解决方案n中的MED Associates的操作性室配备一个的lickometer，伸缩杆，的吸管（如前所述霍华德等人，2009年）（MedAssociates，公司，佛蒙特州，美国）2从MedAssociates软件。被用来设计所有操作性计划（MedAssociates公司，美国佛蒙特州）。 </li><li>一旦培训，以应对蔗糖，开始作用于一个适当的训练计划，在此期间，乙醇逐渐加入到多个喝酒聚会的解决方案。 </li><li>例如，我们的实验室目前使用了8天的训练计划，其中乙醇褪色的解决方案，最终在10％乙醇/ 10％蔗糖饮水​​解决方案。控制动物只收到10％的蔗糖或不喝酒的解决方案。要使用的特定类型的操作性时刻表可以很宽的范围内变化，但仍然可以进行下面所述的一般方法，微透析采样3 </li></ol> 3。前微透析过程：绑定<ol&gt; <li>第二到最后训练（在我们的例子中，这是第 7 届培训课程）的前一天晚上，习惯于动物的纽带弹簧，连接的纽带螺栓在他们的头阶段。弹簧连接到微透析旋转和反的平衡杠杆臂，这是暂停笼旁边的一个环形支架和夹子，使动物可以自由移动，在它的笼子里。动物系留过夜，在操作性房间，在自己的笼子里自由采食食物和水，习惯于他们的网络共享设置。 </li><li>翌日，大鼠完成其操作性的程序与系绳到位，习惯于动物该系绳的感觉，而执行其行为的任务。束缚装置安装的操作性腔室的顶部附近，以允许悬浮液的系绳与旋转的中心上方的车顶的操作性室的壁。所有这一切都被置于内的声音衰减腔。达产后R的会话，返回其家笼子里的操作性房间，等待microdiaysis探针插入大鼠。 </li></ol> 4。前的微透析程序：插入微透析探针微透析实验的前一天，大鼠后的一天完成行为训练<ol><li>微透析实验的前一天，在鼠已完成操作性的会话，而系留，与异氟醚麻醉大鼠，并取出导管闭孔。慢慢地（超过5分钟）微透析探针插入，灌注人工脑脊液（ACSF），通过的颅引导插管插入的大脑区域的利益。我们利用实验室构建的探头和微程序仿照Doyon等人，2003年和佩蒂特和正义，1991年3，4 </li><li>使用前面讨论的圈养装置暂停线以上的动物探头的入口和出口。 </li><li>打开的p长袍下降到0.2μl/ min的流速。再次，老鼠度过夜在操作性房间。 </li></ol> 5。微透析过程：样品采集与食欲和消费型相分离的自我管理会议<ol><li>在实验开始前至少2小时，转了探头，其工作流量。我们使用1.0或2.0μl/ min的不同的大脑区域。检查探头的流量是一致的，和至少90％的通过使用Hamilton注射器测量体积随着时间的推移的设定流量。 </li><li>透析样品（5或7分钟）之前，期间和之后的操作性会议。采样间隔取决于大脑区域，神经递质被分析，透析液浓度的被分析物，和用于分析的分析化学设备的灵敏度。的行为阶段如下： <ol><li> 基线：在实验初期，在动物的饲养笼（4例），收集透析样本。 </li><li> 转让后的家庭笼基线样品的采集，传输的老鼠的操作性室的。栓大鼠的传输需要格外小心，确保微透析液传输线不纠结，和，大鼠保持平静。紧随转让后，激活操作性的程序，因为你从基线/转让样品首先等待样品。 </li><li> 等待：继续收集大鼠等待杆延伸到所述腔室（3-4取决于大脑区域的样品）的样品。 </li><li> 饮酒：杆被提出并加压后，饮用溶液一瓶（10％sucrose/10％乙醇或10％蔗糖）可为大约20分钟（3-4样品）的大鼠。 </li><li> 后饮料：饮料期后，瓶子收缩，但老鼠仍然是我n的操作性室，约20分钟（3-4个样品）。 </li></ol></li></ol> 6。的微透析过程：微透析样品的制备乙醇分析<ol><li>前两个样品动物自我管理后，被评估为乙醇浓度的乙醇溶液中，所有样品。吸移管可以是1或2微升等分从感兴趣的样品成2毫升玻璃小瓶。气密隔（9毫米红色保利螺丝帽，PTFE / SIL隔垫，安捷伦科技），然后密封的小瓶。乙醇分析等分试样（1或2微升）的体积取决于总的样品体积，和所需的任何附加的分析样品的体积。 </li><li>如果样本将用于购买的神经化学分析，存储在适当的后移液等分的乙醇定量样品。 </li><li>例如，我们的实验室多巴胺的样本进行分析。要做到这一点，在干冰上放置样品，在实验过程中然后，储存在-80℃下的样品后的实验。我们使用2微升等份5分钟用流速为2.0微升/分钟的探针对伏隔核中下收集样品乙醇分析。这使得至少7微升，其余为以后的分析多巴胺的高效液相色谱 - 电化学检测。 </li><li>对于样本，其中有前额叶皮层细胞外多巴胺浓度要低得多，我们使用取1μL乙醇分析样品1.0μl/ min的流速为7分钟。 </li></ol> 7。微透析后的程序<ol><li>的微透析实验结束后，麻醉大鼠，并取下探头。更换闭孔，如果动物还没有立即安乐死。在三天之内的实验，收集大脑。否则，可视化的探针道可能是不可能的。 </li><li>大脑应删除我根据批准的动物使用的协议。我们用戊巴比妥钠（150毫克/公斤，ip）可过量，随后用盐水洗涤，然后福尔马林在盐水通过心脏灌注。淹没大脑前至少12小时切片福尔马林盐溶液。 </li><li>冠部分为100微米片的大脑。甲酚紫染色切片，并检查正确的探头位置，5，6 </li></ol> 8。乙醇浓度的样本进行分析<ol><li>所收集的样品，以及作为外部标准（0.3125 - 乙醇的20mM）我们的气相色谱仪，用火焰电离检测系统上运行。此系统包括一台Varian CP 3800带有火焰离子化检测器的气相色谱仪，在Bruker（瓦里安）8400顶空自动进样器加热至50℃，和一个HP INNOWAX毛细管柱（30米×0.53毫米×1.0微米膜厚），与氦气作为流动相。 </li><li>色谱图的记录和与chromatogr分析aphy软件如瓦里安星色谱工作站软件，将被在这里具体讨论。 </li><li>要准备系统对乙醇的分析，使用循环水浴加热自动进样器板，然后打开另外两个煤气瓶（空气和氢，氦始终保持运行状态保存的蜡柱）。 </li><li>液化气罐的压力，以及的样品你打算运行，使你能保持纤维和隔计数的次数已使用，让他们在适当的时候可以改变（注射纤维500;隔记录-100穿刺）。 </li><li>启动的启动的方法，该方法准备系统运行样品（在表2中的程序参数）。等待系统报告说，它是“准备就绪”。 </li><li>启动一个20分钟的“烧”，准备通过使其经受高温解吸任何污染，它吸收的纤维试样分析而处于静止状态。 </li><li>标准是由稀释与水的95％的乙醇，至终体积为100 mL，请使用的容积的在一个化学品的烧瓶中通风柜价的238μl的。这将创建一个40 mM的的乙醇溶液。我们使用一个1:1的的串行，，以创建产品稀释20，10，5星，2.5，1.25时，0.625，0.3125 mM的的标准的。如果在的标准每个浓度中，请使用相同的的的体积将会采取何种从透析样本的等分试样，。用移液管成一个2毫升的玻璃管形瓶中的等分试样，，并然后密封该有一个空气的紧隔垫的样品瓶。 </li><li>所有的乙醇样本顷中的自动进样器加热，，直到的整个的液体等分试样已被汽化的。 ，我们加热我们的的的的样品为约30 min。 </li><li>本节描述了星工作站的的软件，我们与我们的的气体色​​谱仪中中使用。其他软件可能会可以使用，但下面的描述中可能不适用的。 </li><li>要运行样品，创建一个示例列表，并记下哪些样品是在其中自动进样器插槽，，，，并如何许多的倍的样品应被刺破的。请务必，，以的路线的数据的人文件到您所选择的的倍呃。然后，选择您的样品的首选方法，并开始运行。我们使用在表2中指出的运行参数。 </li><li>我们透析液的乙醇计划吸收样品3分钟和解吸入氦气流，送入蜡列，1分钟。但是，用户可以编写程序，以满足您的特定需求。 </li><li>在蜡中的列分开的样品组分，然后通过氢火焰离子化检测器的含碳化合物，如乙醇，燃烧和释放离子。在创建一个电流，它被转换为电压和记录导致像一个如下所示的（图1）在色谱从检测器的阳极到阴极的电子流增加这样的结果。电压的变化是正比于通过检测器的量的碳传递跨time.7 </li><li>系统完成后运行这些示例，您将需要检查每个峰的整合。星工作站软件，蓝峰在工具栏上的图标上单击。每个峰的颜色，点击并拖动箭头来调整基线。重新整合，每一个峰值，然后再移动到下一组峰。峰分析软件可以是任何常规色谱软件。 </li><li>星工作站软件中，单击“批处理报表在您的工具栏图标。从您指定的文件夹，将每个样本的批次报告打印样本报告。最常见的色谱软件系统软件可定制的报告。 </li><li>报告可以进行编程，以显示您所选择的信息。我们目前使用的峰高，但也可以进行编程，以峰面积的报告。 </li><li>要关闭系统，启动关机方法（参数在表2），关闭水浴，并关闭氢气和空气罐时，柱箱温度达到30°C。 </li></ol> 9。酒精数据<ol><li>绘制的峰值高度为每个已知的外部标准的乙醇浓度的函数（图2A）。使用由这些点给出的线性函数，从峰的高度来计算的乙醇浓度，由每个透析液样品给出。 </li><li>通过绘制随时间的透析液中的乙醇的浓度，我们可以看到在我们的行为会议期间大脑中的感兴趣区域的图案的乙醇水平。实施例的数据，如下所示（图2B），乙醇的浓度的透析液中的自我管理会话期间跨越时间被表示为。 </li></ol> 10。一般维护：纤维应改为每500穿刺，并且隔膜每100穿刺要改变纤维<ol><li>气相色谱仪键盘按菜单，选择8400，按回车键，选择更改注射器，按回车键。自动进样器会自动定位到允许的纤维（SPME纤维大会，75微米，Supelco公司分析，Carboxen-PDMS BellefoNTE，PA）被删除。 </li><li>打开门，涵盖了纤维。拧松锁紧螺母，并将闩锁，纤维集合体被删除。以光纤组件。保存在装配体中的纤维，然后拧下螺母拧纤维。 </li><li>用一个新的更换旧的纤维，和重新组装的纤维集合体。自动进样器更换组件，重新锁定和装​​配拧到位。当您完成后，按“更改”键盘上，和自动进样器将准备自己使用。 </li></ol>要改变隔<ol><li>首先，拧开盖子覆盖在隔（温度.450直径一般空调的隔垫，瓦里安），然后删除旧的隔垫。座新的隔垫入接头。重新拧紧瓶盖，并用扳手拧紧。然后，使用注射针来刺穿隔垫，使纤维不会打破首次被刺穿隔垫。 </li></oL&gt; 11。代表性的成果图1示出了例如三种不同浓度的乙醇的标准和大鼠透析液乙醇自我管理会话结束时收集的样品的色谱。乙醇峰应该是相对对称，具有一致的滞留时间，以及信号噪声比&gt; 10。如果未能满足这些标准，您的系统需要维护。质量色谱正确制备的标准产生一个线性的标准曲线（R2≥0.99;图2A），用于计算从乙醇中的乙醇浓度的透析液样品收集自给药动物超过他们的自我管理会话的过程中（图2B） 。 <img src="/files/ftp_upload/4142/4142fig1.jpg" alt="图1" /> 图1实施例的色谱图。一微升乙醇标准或透析液样品被加载到一个气体Chromatograph小瓶和分析如文中所述。 A）的峰值为2.5，5.0和10毫米乙醇标准的覆盖。 B）的峰值从大鼠具有自我管理的乙醇产生的透析液样品。 <img src="/files/ftp_upload/4142/4142fig2.jpg" alt="图2" /> 图2的图形从例如图1中所示的实验结果。 A）乙醇的标准曲线。 B）的时间过程中，整个透析液乙醇浓度的乙醇自我管理会话。 <table border="1"><tbody><tr><td> 硬件 </td><td> 参数 </td><td> 开始设置 </td><td> 运行设置 </td><td> 休息设置 </td></tr><tr><td colspan="5"> 8400布鲁克（瓦里安）自动进样器 </td></tr><tr><td></td><td>进样方式</td><td>固相微萃取</td><td>固相微萃取</td><td> N / A </td></tr> &lt;TR&gt; <td></td><td>溶剂穿透深度</td><td> 0％ </td><td> 20％ </td><td> N / A </td></tr><tr><td></td><td>样品的穿透深度</td><td> 20％ </td><td> 20％ </td><td> N / A </td></tr><tr><td></td><td>吸收时间</td><td> 0.01分钟</td><td> 3分钟</td><td> N / A </td></tr><tr><td></td><td>解吸时间</td><td> 19分钟</td><td> 1分钟</td><td> N / A </td></tr><tr><td></td><td>清洁模式溶剂源</td><td>我</td><td>我</td><td> N / A </td></tr><tr><td></td><td>清洁模式吸附，解吸时间</td><td> 0.01分钟</td><td> 0.01分钟</td><td> N / A </td></tr><tr><td></td><td>水浴加热自动进样器* </td><td> 50°C </td><td> 50°C </td><td>离</td></tr><tr><td colspan="5"> CP-3800瓦里安气相色谱仪 </td></tr><tr><td> 喷油器炉 </TD&gt; <td>烤箱电源</td><td>上</td><td>上</td><td>上</td></tr><tr><td></td><td>烤箱温度</td><td> 250°C </td><td> 220°C </td><td> 30°C </td></tr><tr><td> 柱温箱 </td><td>稳定时间</td><td> 0.10分钟</td><td> 0.10分钟</td><td> 0.10min </td></tr><tr><td></td><td>温度</td><td> 65°C </td><td> 65°C </td><td> 30°C </td></tr><tr><td> 柱 </td><td>流动相流速</td><td> 8.5毫升/分钟</td><td> 8.5毫升/分钟</td><td> N / A </td></tr><tr><td></td><td>柱压</td><td> 〜6磅</td><td> 〜6磅</td><td> ≥0.1磅</td></tr><tr><td> FID检测器 </td><td>烤箱电源</td><td>上</td><td>上</td><td>上</td></tr><tr><td></td><td>温度</td><td> 220°C </td><td> 220℃ </td><td> 120°C </td></tr&gt; <tr><td></td><td>电子产品</td><td>上</td><td>上</td><td>离</td></tr><tr><td></td><td>时间常数</td><td>快</td><td>快</td><td>快</td></tr><tr><td></td><td>范围</td><td> 11 </td><td> 11 </td><td> 11 </td></tr><tr><td></td><td>自动调零</td><td>是</td><td>是</td><td>是</td></tr></tbody></table> 表2中。气相色谱仪，用火焰电离检测系统中的参数。此表显示的参数，用来准备的三个方案（启动设置），运行（运行设置）和维护系统，而不是使用（休息设置）的。

<ol>
	<li>Geiger, B. M., Frank, L. E., Caldera-Siu, A. D., Pothos, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Survivable+stereotaxic+surgery+in+rodents.">Survivable stereotaxic surgery in rodents.</a> J. Vis. Exp. (20), e880 (2008).</li><li>Howard, E. C., Schier, C. J., Wetzel, J. S., Gonzales, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+dopamine+response+in+the+nucleus+accumbens+core-shell+border+differs+from+that+in+the+core+and+shell+during+operant+ethanol+self-administration.">The dopamine response in the nucleus accumbens core-shell border differs from that in the core and shell during operant ethanol self-administration.</a> Alcohol Clin. Exp. Res. 33, 1355-1365 (2009).</li><li>Doyon, W. M., York, J. L., Diaz, L. M., Samson, H. H., Czachowski, C. L., Gonzales, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dopamine+activity+in+the+nucleus+accumbens+during+consummatory+phases+of+oral+ethanol+self-administration.">Dopamine activity in the nucleus accumbens during consummatory phases of oral ethanol self-administration.</a> Alcohol Clin. Exp. Res. 27, 1573-1582 (2003).</li><li>Pettit, H. O., Justice, J. B. <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+dose+on+cocaine+self-administration+behavior+and+dopamine+levels+in+the+nucleus+accumbens.">Effect of dose on cocaine self-administration behavior and dopamine levels in the nucleus accumbens.</a> Brain Res. 539, 94-102 (1991).</li><li>Paxinos, G., Kus, L., Ashwell, K. W. S., Watson, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Chemoarchitectonic Atlas of The Rat Forebrain. , (1998).</li><li>Paxinos, G., Watson, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Rat Brain in Stereotaxic Coordinates. , (1986).</li><li>Skoog, D. A., Holler, F. J., Nieman, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Principles of Instrumental Analysis. , (1998).</li><li>Doyon, W. M., Anders, S. K., Ramachandra, V. S., Czachowski, C. L., Gonzales, R. A. <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+operant+self-administration+of+10%+ethanol+plus+10%+sucrose+on+dopamine+and+ethanol+concentrations+in+the+nucleus+accumbens.">Effect of operant self-administration of 10% ethanol plus 10% sucrose on dopamine and ethanol concentrations in the nucleus accumbens.</a> J Neurochem. 93, 1469-1481 (2005).</li><li>Carrillo, J., Gonzales, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+single+exposure+to+voluntary+ethanol+self-administration+produces+adaptations+in+ethanol+consumption+and+accumbal+dopamine+signaling.">A single exposure to voluntary ethanol self-administration produces adaptations in ethanol consumption and accumbal dopamine signaling.</a> Alcohol. 45, 559-5566 (2011).</li><li>Howard, E. C., Schier, C. J., Wetzel, J. S., Duvauchelle, C. L., Gonzales, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+shell+of+the+nucleus+accumbens+has+a+higher+dopamine+response+compared+with+the+core+after+non-contingent+intravenous+ethanol+administration.">The shell of the nucleus accumbens has a higher dopamine response compared with the core after non-contingent intravenous ethanol administration.</a> Neuroscience. 154, 1042-1053 (2008).</li><li>Robinson, D. L., Lara, J. A., Brunner, L. J., Gonzales, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+Ethanol+Concentrations+in+the+Extracellular+Fluid+of+the+Rat+Brain.">Quantification of Ethanol Concentrations in the Extracellular Fluid of the Rat Brain.</a> J. Neurochem. 75, 1685-1693 (2000).</li><li>Gonzales, R. A., McNabb, J., Yim, H. J., Ripley, T., Bungay, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+Microdialysis+of+Ethanol+in+Rat+Striatum.">Quantitative Microdialysis of Ethanol in Rat Striatum.</a> Alcohol Clin. Exp. Res. 22, 858-867 (1998).</li></ol>

这篇文章是由美国医学协会，

应用程序和限制在啮齿类动物模型药物成瘾的药物自我管理。许多药物的滥用，以这种方式建模，可静脉给药，其中的药物被直接传送到车厢中部。这允许剂量的自我管理会话进行密切监测。由于乙醇通常是口服自我的管理，它是更难监测血药浓度，由于吸收和代谢的个体差异。通过使用微透析样品的大脑区域的利益，我们不仅能够监控模式到达该地区的乙醇，但我们还可以在同一区域同时监测神经递质的变化随着时间的推移，在每一个自我管理阶段。 大脑中的神经化学改变及药物引起的反应与药物滥用和依赖，因此，能够同时测量确保神经化学和药物浓度在特定的自我管理阶段提供了一个非常强大而独特的工具。记住相​​关的透析液浓度被测物的行为是一个问题的微透析管道的物理特性。具体而言，所花费的时间从微透析探针的内腔的流体被转移到收集管中是至关重要的，和更短的时间是更好的。在我们的实验室中，我们构建了探头，这样的时间是90秒左右的。使用这些条件下，我们已经发现在起始的乙醇消耗，饮料和饮料后的期间，作为透析液的乙醇浓度的增加而降低的过程，有一个增加accumbal多巴胺。2,3,8,9这些实验，当与药理学研究中的数据相结合，使我们能够解析出严格的药理作用乙醇和自我管理相关的环境因素通道法兰神经递质的浓度。 应该指出的是，这个特殊的联合行为的微透析技术的应用是适合我们的实验室目前的研究兴趣。它的目的是评估的时空格局乙醇，达到大脑中的神经递质的变化在同一区域的模式相比，这样我们就可以同步的自我管理行为与这些措施。派生的透析液中的乙醇浓度在体内探针恢复不纠正，只有一小部分的大脑乙醇的组织浓度。如果需要定量微量渗析乙醇，乙醇提取馏分，探头从细胞外空间分成扩散应通过实验确定。从我们的实验室方法，并进一步讨论，请参阅以前的出版物。10,11,12 虽然这个协议说明了如何使用的气体通道romatography随着固相微萃取乙醇从微透析样品的顶空，可以使用其他的方法进行分析的微透析样品的乙醇含量。然而，替代方法可能会遭受一些缺点。较不敏感的分析方法，例如，可能需要更大的采样次数大于在这里示出的5-7分钟的样品体积，有必要。这里讨论的系统的类型使用固相微萃取，浓缩乙醇在汽相中在密封的样品瓶中，通过使吸收的纤维放置在小瓶中的顶空。这提高了检测限相比，它通常允许50-100微升的蒸气注入直接顶空取样。顶空法的另一个主要优点是，用于分析的样品注入是非常干净，无盐。也可使用具有较高SENSIT液体微透析样品直接注射的ivity，但是这将需要更多的仪器停机时间，由于要经常进行保养清洁的注射盐。 故障排除和其他注意事项<ol><li>实验开始前，给自己足够的时间来检查你的微透析设置是否正常工作，并进行故障排除的任何问题。我们建议您有一个额外的设置，ACSF灌流，准备转出发生故障的设定，以节省自己的时间，作为最操作性的会议往往发生在特定的时间，每天。 </li><li>成为确保微透析探针插入超过约5分钟的一段时间，以尽量减少组织损伤所产生剪切的组织作为探针穿透大脑。 </li><li>当大鼠转移到操作性室，它是有帮助的第二个人辅助，使输送线（微透析的一部分设置）不成为纠结，等，操作性的程序，可以初始化iated时间。 </li><li>请确保您提供的自动进样器水浴足够的时间来达到合适的温度。我们的系统通常需要两个小时才能达到50°C。此外，在分析样品之前，确保您的样品完全汽化。我们通常目视确认，这已开始样品分析之前发生。 </li><li>标准良好的化学分析应遵循的做法。这些包括，但不限于，个人，设备，和程序验证的。总之，我们采用了以下指导原则：个人用户应表现出的能力产生重复的峰高值，为跨越多天的标准浓度和线性的标准曲线（R2≥0.99）。要验证GC-FID系统的正常工作，标准应为总，之前注入透析液样本进行分析。标准曲线应该有一个R2≥0.99。 </li><li>我们使用的设备购买瓦里安公司，安捷伦科技公司在2010年收购了。在这个时候，布鲁克科学仪器获得了瓦里安的实验室气相色谱仪业务。对于未来的购买，咨询任何一家公司。 </li><li>本协议的操作行为与影响，但观众应该知道，更多的破坏行为是可能发生的小的小鼠模型。需要注意的另一个问题是，放置的微透析探针插入大脑内侧前额叶皮质和伏隔核中可能会破坏在更大程度上比这里所显示的操作行为以外的其他地区。重要的是要仔细研究大量的行为所造成的损失由探头放置，以确定是否产生严重的行为变化的参数。 </li></ol>

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td> 95％乙醇</td><td> AAPER酒精和化工有限公司，谢尔比维尔，KY </td><td> E190，111000190 </td></tr><tr><td>超纯蔗糖</td><td> MP Biomedicals，有限责任公司，政治家，OH </td><td> 821721 </td></tr><tr><td>超高纯度的氦气</td><td>空气气体</td><td> HE UHP300 </td></tr><tr><td>空气</td><td>空气气体</td><td> AIZ300 </td></tr><tr><td>氢</td><td>空气气体</td><td> AIZ300 </td></tr><tr><td> GC 2毫升小瓶</td><td>安捷伦</td><td> ＃8010-0015 </td></tr><tr><td> GC样品瓶盖与PTFE /硅橡胶隔垫</td><td>安捷伦</td><td> ＃8010-0084 </td></tr></tbody></table> 表1中。具体试剂

microdialysis of ethanol during operant ethanol self-administration and ethanol determination by gas chromatography

操作性自我管理常用的方法有许多药物的滥用，包括乙醇的行为和药理作用的研究。然而，乙醇是典型的自我，而不是像许多其他药物的滥用静脉注射口服。口服药物的药代动力学静脉注射的药物要复杂得多。由于理解的药理和乙醇中的行为的影响之间的关系，需要知识乙醇到达大脑期间和饮酒后的时间过程中，我们在体内微透析和气相色谱法中使用，用氢火焰离子化检测监测脑透析液中的乙醇浓度随着时间的推移。 结合微透析行为实验涉及使用多种技术。在本文中，立体定位手术，行为训练和微透析，可适于测试大量的自我管理和神经包括rochemical中心的假设，只是为了说明他们是如何涉及到的样品采集和分析透析液中乙醇的后续阶段。透析液乙醇浓度通过气相色谱法分析，用火焰离子化检测器，这是特定的乙醇研究，进行详细说明。通过这些方法生产的数据揭示了图案乙醇期间自我管理程序到达大脑，神经化学相同的透析液样品的分析配对时，允许关于药理和乙醇中的行为的影响作出结论。

Watch this Scientific Journal Video about Microdialysis of Ethanol During Operant Ethanol Self-administration and Ethanol Determination by Gas Chromatography at JoVE.com

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

自我管理的方法，以确定在老鼠的大脑中乙醇浓度的时间过程，在操作性乙醇。用火焰离子化检测器的气相色谱分析是用来量化乙醇的透析样品中，因为它具有所需的所生成的小卷的灵敏度。

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.  ...

microdialysis-ethanol-during-operant-ethanol-self-administration

Research

JoVE Journal

Neuroscience

17.4K 次观看.  The University of Texas at Austin. 自我管理的方法，以确定在老鼠的大脑中乙醇浓度的时间过程，在操作性乙醇。用火焰离子化检测器的气相色谱分析是用来量化乙醇的透析样品中，因为它具有所需的所生成的小卷的灵敏度。

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

Assessment of Ultrasonic Vocalizations During Drug Self-administration in Rats

Drug self-administration procedures are commonly used to study behavioral and neurochemical changes associated with human drug abuse, addiction and relapse. Various types of behavioral activity are commonly utilized as measures of drug motivation in animals. However, a crucial component of drug abuse relapse in abstinent cocaine users is "drug craving", which is difficult to model in animals, as it often occurs in the absence of overt behaviors. Yet, it is possible that a class of ultrasonic vocalizations (USVs) in rats may be a useful marker for affective responses to drug administration, drug anticipation and even drug craving. Rats vocalize in ultrasonic frequencies that serve as a communicatory function and express subjective emotional states. Several studies have shown that different call frequency ranges are associated with negative and positive emotional states. For instance, high frequency calls ("50-kHz") are associated with positive affect, whereas low frequency calls ("22-kHz") represent a negative emotional state. This article describes a procedure to assess rat USVs associated with daily cocaine self-administration. For this procedure, we utilized standard single-lever operant chambers housed within sound-attenuating boxes for cocaine self-administration sessions and utilized ultrasonic microphones, multi-channel recording hardware and specialized software programs to detect and analyze USVs. USVs measurements reflect emotionality of rats before, during and after drug availability and can be correlated with commonly assessed drug self-administration behavioral data such lever responses, inter-response intervals and locomotor activity. Since USVs can be assessed during intervals prior to drug availability (e.g., anticipatory USVs) and during drug extinction trials, changes in affect associated with drug anticipation and drug abstinence can also be determined. In addition, determining USV changes over the course of short- and long-term drug exposure can provide a more detailed interpretation of drug exposure effects on affective functioning.

Drug self-administration and ultrasonic vocalizations (USV) are used as behavioral assessments in animal research, but rarely in combination. The purpose of this article is to describe the advantages of recording USVs during drug self-administration procedures to assess affective responses to drug experience.

在大鼠的药物自我管理的评估超声发声

Drug self-administration procedures are commonly used to study behavioral and neurochemical changes associated with human drug abuse, addiction and ...

科研

神经科学

Operant Sensation Seeking in the Mouse

Operant methods are powerful behavioral tools for the study of motivated behavior. These 'self-administration' methods have been used extensively in drug addiction research due to their high construct validity. Operant studies provide researchers a tool for preclinical investigation of several aspects of the addiction process. For example, mechanisms of acute reinforcement (both drug and non-drug) can be tested using pharmacological or genetic tools to determine the ability of a molecular target to influence self-administration behavior1-6. Additionally, drug or food seeking behaviors can be studied in the absence of the primary reinforcer, and the ability of pharmacological compounds to disrupt this process is a preclinical model for discovery of molecular targets and compounds that may be useful for the treatment of addiction3,7-9. One problem with performing intravenous drug self-administration studies in the mouse is the technical difficulty of maintaining catheter patency. Attrition rates in these experiments are high and can reach 40% or higher10-15. Another general problem with drug self-administration is discerning which pharmacologically-induced effects of the reinforcer produce specific behaviors. For example, measurement of the reinforcing and neurological effects of psychostimulants can be confounded by their psychomotor effects. Operant methods using food reinforcement can avoid these pitfalls, although their utility in studying drug addiction is limited by the fact that some manipulations that alter drug self-administration have a minimal impact on food self-administration. For example, mesolimbic dopamine lesion or knockout of the D1 dopamine receptor reduce cocaine self-administration without having a significant impact on food self-administration 12,16. 
Sensory stimuli have been described for their ability to support operant responding as primary reinforcers (i.e. not conditioned reinforcers)17-22. Auditory and visual stimuli are self-administered by several species18,21,23, although surprisingly little is known about the neural mechanisms underlying this reinforcement. The operant sensation seeking (OSS) model is a robust model for obtaining sensory self-administration in the mouse, allowing the study of neural mechanisms important in sensory reinforcement24. An additional advantage of OSS is the ability to screen mutant mice for differences in operant behavior that may be relevant to addiction. We have reported that dopamine D1 receptor knockout mice, previously shown to be deficient in psychostimulant self-administration, also fail to acquire OSS24. This is a unique finding in that these mice are capable of learning an operant task when food is used as a reinforcer. While operant studies using food reinforcement can be useful in the study of general motivated behavior and the mechanisms underlying food reinforcement, as mentioned above, these studies are limited in their application to studying molecular mechanisms of drug addiction. Thus, there may be similar neural substrates mediating sensory and psychostimulant reinforcement that are distinct from food reinforcement, which would make OSS a particularly attractive model for the study of drug addiction processes. The degree of overlap between other molecular targets of OSS and drug reinforcers is unclear, but is a topic that we are currently pursuing. While some aspects of addiction such as resistance to extinction may be observed with OSS, we have found that escalation 25 is not observed in this model24. Interestingly, escalation of intake and some other aspects of addiction are observed with self-administration of sucrose26. Thus, when non-drug operant procedures are desired to study addiction-related processes, food or sensory reinforcers can be chosen to best fit the particular question being asked. 
In conclusion, both food self-administration and OSS in the mouse have the advantage of not requiring an intravenous catheter, which allows a higher throughput means to study the effects of pharmacological or genetic manipulation of neural targets involved in motivation. While operant testing using food as a reinforcer is particularly useful in the study of the regulation of food intake, OSS is particularly apt for studying reinforcement mechanisms of sensory stimuli and may have broad applicability to novelty seeking and addiction.

In this protocol we describe a method of operant learning using sensory stimuli as a reinforcer in the mouse. It requires no prior training or food restriction, and it allows the study of motivated behavior without the use of a pharmacological or natural reinforcer such as food.

在鼠标的操作性感觉寻求

Operant methods are powerful behavioral tools for the study of motivated behavior.  These 'self-administration' methods have been used extensively in drug ...

A Simple Way to Measure Ethanol Sensitivity in Flies

Low doses of ethanol cause flies to become hyperactive, while high doses are sedating. The sensitivity to ethanol-induced sedation of a given fly strain is correlated with that strain s ethanol preference, and therefore sedation is a highly relevant measure to study the genetics of alcohol responses and drinking. We demonstrate a simple way to expose flies to ethanol and measure its intoxicating effects. The assay we describe can determine acute sensitivity, as well as ethanol tolerance induced by repeat exposure. It does not require a technically involved setup, and can therefore be applied in any laboratory with basic fly culture tools.

A simple assay to measure the sedating effects of ethanol on Drosophila flies, based on the loss of righting reflex, is described.

一个简单的方法来衡量在果蝇的乙醇灵敏度

Low doses of ethanol cause flies to become hyperactive, while high doses are sedating. The sensitivity to ethanol-induced sedation of a given fly strain ...

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 ...

Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings (GCMR) in the Insect Antennal Lobe

All organisms inhabit a world full of sensory stimuli that determine their behavioral and physiological response to their environment. Olfaction is especially important in insects, which use their olfactory systems to respond to, and discriminate amongst, complex odor stimuli. These odors elicit behaviors that mediate processes such as reproduction and habitat selection1-3. Additionally, chemical sensing by insects mediates behaviors that are highly significant for agriculture and human health, including pollination4-6, herbivory of food crops7, and transmission of disease8,9. Identification of olfactory signals and their role in insect behavior is thus important for understanding both ecological processes and human food resources and well-being.
	To date, the identification of volatiles that drive insect behavior has been difficult and often tedious. Current techniques include gas chromatography-coupled electroantennogram recording (GC-EAG), and gas chromatography-coupled single sensillum recordings (GC-SSR)10-12. These techniques proved to be vital in the identification of bioactive compounds. We have developed a method that uses gas chromatography coupled to multi-channel electrophysiological recordings (termed 'GCMR') from neurons in the antennal lobe (AL; the insect's primary olfactory center)13,14. This state-of-the-art technique allows us to probe how odor information is represented in the insect brain. Moreover, because neural responses to odors at this level of olfactory processing are highly sensitive owing to the degree of convergence of the antenna's receptor neurons into AL neurons, AL recordings will allow the detection of active constituents of natural odors efficiently and with high sensitivity. Here we describe GCMR and give an example of its use.
	Several general steps are involved in the detection of bioactive volatiles and insect response. Volatiles first need to be collected from sources of interest (in this example we use flowers from the genus Mimulus (Phyrmaceae)) and characterized as needed using standard GC-MS techniques14-16. Insects are prepared for study using minimal dissection, after which a recording electrode is inserted into the antennal lobe and multi-channel neural recording begins. Post-processing of the neural data then reveals which particular odorants cause significant neural responses by the insect nervous system.
	Although the example we present here is specific to pollination studies, GCMR can be expanded to a wide range of study organisms and volatile sources. For instance, this method can be used in the identification of odorants attracting or repelling vector insects and crop pests. Moreover, GCMR can also be used to identify attractants for beneficial insects, such as pollinators. The technique may be expanded to non-insect subjects as well.

Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings GCMR in the Insect Antennal Lobe

Olfactory cues mediate many different behaviors in insects, and are often complex mixtures comprised of tens to hundreds of volatile compounds. Using gas chromatography with multi-channel recording in the insect antennal lobe, we describe a method for the identification of bioactive compounds.

用气相色谱 - 多单元的录音（GCMR）昆虫触角叶嗅觉挥发物的鉴定

All  organisms inhabit a world full of sensory stimuli that determine their  behavioral and physiological response to their environment. Olfaction is ...

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance imaging. However, use of such methods in animal models requires anesthesia and hence both alters the brain state and prevents behavioral measures. An alternative method is the use of in vivo microdialysis to take continuous measurement of brain extracellular fluid concentrations of glucose, lactate, and related metabolites in awake, unrestrained animals. This technique is especially useful when combined with tasks designed to rely on specific brain regions and/or acute pharmacological manipulation; for example, hippocampal measurements during a spatial working memory task (spontaneous alternation) show a dip in extracellular glucose and rise in lactate that are suggestive of enhanced glycolysis1-3,4-5, and intrahippocampal insulin administration both improves memory and increases hippocampal glycolysis6. Substances such as insulin can be delivered to the hippocampus via the same microdialysis probe used to measure metabolites. The use of spontaneous alternation as a measure of hippocampal function is designed to avoid any confound from stressful motivators (e.g. footshock), restraint, or rewards (e.g. food), all of which can alter both task performance and metabolism; this task also provides a measure of motor activity that permits control for nonspecific effects of treatment. Combined, these methods permit direct measurement of the neurochemical and metabolic variables regulating behavior.

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Modulation of hippocampally-dependent spatial working memory by direct intrahippocampal microinjection, accompanied and followed by in vivo microdialysis for metabolites in conscious, behaving animals.

海马胰岛素显微注射和<em在体内</em&gt;微透析过程中空间记忆测试

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance ...

An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

Alcohol use disorder (AUD) is a serious health challenge. Despite a large hereditary component to AUD, few genes have been unambiguously implicated in their etiology. The fruit fly, Drosophila melanogaster, is a powerful model for exploring molecular-genetic mechanisms underlying alcohol-related behaviors and therefore holds great promise for identifying and understanding the function of genes that influence AUD. The use of the Drosophila model for these types of studies depends on the availability of assays that reliably measure behavioral responses to ethanol. This report describes an assay suitable for assessing ethanol sensitivity and rapid tolerance in flies. Ethanol sensitivity measured in this assay is influenced by the volume and concentration of ethanol used, a variety of previously reported genetic manipulations, and also the length of time the flies are housed without food immediately prior to testing. In contrast, ethanol sensitivity measured in this assay is not affected by the vigor of fly handling, sex of the flies, and supplementation of growth medium with antibiotics or live yeast. Three different methods for quantitating ethanol sensitivity are described, all leading to essentially indistinguishable ethanol sensitivity results. The scalable nature of this assay, combined with its overall simplicity to set-up and relatively low expense, make it suitable for small and large scale genetic analysis of ethanol sensitivity and rapid tolerance in Drosophila.

An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

Straightforward assays for measuring ethanol sensitivity and rapid tolerance in Drosophila facilitate the use of this model organism for investigating these important ethanol-related behaviors. Here, a relatively simple, scalable assay for measuring ethanol sensitivity and rapid tolerance in flies is described.

一种廉价的，可扩展的行为分析测量乙醇镇静灵敏度和快速耐受<em&gt;果蝇</em

Alcohol use disorder (AUD) is a serious health challenge. Despite a large hereditary component to AUD, few genes have been unambiguously implicated in ...

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the development of more effective treatments is the relative lack of understanding of the molecular underpinnings of the effects of ethanol on behavior. The nematode, Caenorhabditis elegans (C.&#160;elegans), provides a useful model in which to generate and test hypotheses about the molecular effects of ethanol. Here, we describe an assay that has been developed and used to examine the roles of particular genes and environmental factors in behavioral responses to ethanol, in which locomotion is the behavioral output. Ethanol dose-dependently causes an acute depression of crawling on an agar surface. The effects are dynamic; animals exposed to a high concentration demonstrate an initial strong depression of crawling, referred to here as initial sensitivity, and then partially recover locomotion speed despite the continued presence of the drug. This ethanol-induced behavioral plasticity is referred to here as the development of acute functional tolerance. This assay has been used to demonstrate that these two phenotypes are distinct and genetically separable. The straightforward locomotion assay described here is suitable for examining the effects of both genetic and environmental manipulations on these acute behavioral responses to ethanol in C.&#160;elegans.

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

C. elegans is a useful model for studying the effects of ethanol on behavior. We present a behavioral assay that quantifies the effects of ethanol on the locomotion speed of crawling worms; both initial sensitivity and the development of acute functional tolerance to ethanol can be measured with this assay.

测量乙醇的影响上的步态速度的测定<em&gt;秀丽隐杆线虫</em

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the ...

Custom-made Microdialysis Probe Design

Microdialysis is a commonly used technique in neuroscience research. Therefore commercial probes are in great demand to monitor physiological, pharmacological and pathological changes in cerebrospinal fluid. Unfortunately, commercial probes are expensive for research groups in public institutions. In this work, a probe assembly is explained in detail to build a reliable, concentric, custom-made microdialysis probe for less than $10. The microdialysis probe consists of a polysulfone membrane with a molecular cut-off of 30 kDa. Probe in vitro recoveries of substances with different molecular weight (in the range of 100-1,600 Da) and different physicochemical properties are compared. The probe yields an in vitro recovery of approximately 20% for the small compounds glucose, lactate, acetylcholine and ATP. In vitro recoveries for neuropeptides with a molecular weight between 1,000-1,600 Da amount to 2-6%. Thus, while the higher molecular weight of the neuropeptides lowered in vitro recovery values, dialysis of compounds in the lower range (up to 500 Da) of molecular weights has no great impact on the in vitro recovery rate. The present method allows utilization of a dialysis membrane with other cut-off value and membrane material. Therefore, this custom-made probe assembly has the advantage of sufficient flexibility to dialyze substances in a broad molecular weight range. Here, we introduce a microdialysis probe with an exchange length of 2 mm, which is applicable for microdialysis in mouse and rat brain regions. However, dimensions of the probe can easily be adapted for larger exchange lengths to be used in larger animals.

Microdialysis is a commonly used technique in neuroscience research. Therefore commercial probes are in great demand.In this work a probe assembly is explained in detail to build a reliable, concentric, custom-made microdialysis probe for less than $10.

定制的微透析探针设计

Microdialysis is a commonly used technique in neuroscience research. Therefore commercial probes are in great demand to monitor physiological, ...

Assessment of Dopaminergic Homeostasis in Mice by Use of High-performance Liquid Chromatography Analysis and Synaptosomal Dopamine Uptake

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) neurotransmission is strongly associated with several central nervous system-associated diseases such as Parkinson&#39;s disease, attention-deficit-hyperactivity disorder and drug addiction1,2,3,4. Delineating disease mechanisms involving DA imbalance is critically dependent on animal models to mimic aspects of the diseases, and thus protocols that assess specific parts of the DA homeostasis are important to provide novel insights and possible therapeutic targets for these diseases.
Here, we present two useful experimental protocols that when combined provide a functional read-out of the DAergic system in mice. Biochemical and functional parameters on DA homeostasis are obtained through assessment of DA levels and dopamine transporter (DAT) functionality5. When investigating the DA system, the ability to reliably measure endogenous levels of DA from adult brain is essential. Therefore, we present how to perform high-performance liquid chromatography (HPLC) on brain tissue from mice to determine levels of DA. We perform the experiment on tissue from dorsal striatum (dStr) and nucleus accumbens (NAc), but the method is also suitable for other DA-innervated brain areas.
DAT is essential for reuptake of DA into the presynaptic terminal, thereby controlling the temporal and spatial activity of released DA. Knowing the levels and functionality of DAT in the striatum is of major importance when assessing DA homeostasis. Here, we provide a protocol that allows to simultaneously deduce information on surface levels and function using a synaptosomal6 DA uptake assay.
Current methods combined with standard immunoblotting protocols provide the researcher with relevant tools to characterize the DAergic system.

Synaptosomal dopamine uptake and high-performance liquid chromatography analysis represent experimental tools to investigate dopamine homeostasis in mice by assessing the function of the dopamine transporter and levels of dopamine in striatal tissue, respectively. Here we present protocols to measure dopamine tissue content and assess the functionality of the dopamine transporter.

高效液相色谱分析和触多巴胺摄取对小鼠多巴胺能稳态的评价

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) ...