This work has been funded by the Israel Science Foundation Grant (ISF # 393/12), Israel Centers of Research Excellence Grant (I-CORE 1796/12), German-Israel Foundation Grant (GIF # 2299-2291.1/2011) and the Marie Curie Career Integration Grant (FP7-PEOPLE-2013-CIG #618201). Initial steps in the project were funded by an AXA postdoctoral fellowship to AC. We acknowledge the generous startup funds provided by the Edmond and Lily Safra Center for Brain Sciences.
Critical reading by members of the Citri lab is greatly appreciated.

注：该协议遵循耶路撒冷希伯来大学的动物护理指引。 1，准备学联解决方案<ol><li>如表1中所述制备ACSF溶液。车型1升在DDH 2 O（&gt; 18兆欧的纯度），使渗透压〜300毫渗透摩尔/升以适当添加水或氯化钠。 </li></ol> 2，设备及室内设置该设备用于监控的可卡因诱导的运动行为由行为室（50×45cm）上连接有内光源，风扇和相机（或红外传感器），并配有在其内的有机玻璃箱（30×30厘米）该小鼠可以自由移动，同时其运动受到监视。在行为测试应1小时之间灯之后进行到1小时灯之前关闭，在常规的光/暗循环。动物要坚持在每天同一时间进行培训。 <ol><li&gt;连接​​摄像机或红外线系统到一个适当的系统监测和记录小鼠的自发行为。 </li><li>清洗每次试验用小鼠设施和/或除臭剂提供消毒后的腔室，以防止嗅觉提示偏压，并确保适当的消毒。 </li></ol>重要提示：当处理的小鼠安静，冷静和谨慎。 3，动物准备<ol><li>众议院5 C57BL6雄性小鼠（6-12周龄〜25〜35克的重量）每笼，在一个配有12小时的光/暗周期和免费获得食物和水，按照标准和当地的动物的要求护理委员会的指导和协议。在启动实验，权衡小鼠和日志数据。可卡因是根据小鼠的体重后施用。 </li><li>在开始第一次庭审前将全部含有老鼠进入室内行为30分钟的笼子里。 </li><li> ，观察者所有的在实验中使用的小鼠对实验者处理剂，注射剂，并在行为设置监测。这是通过加入250μl盐水连续3次每日腹膜内注射，如下所述来实现。 </li><li>辖腹膜内注射250μl的生理盐水。紧接在注射，将鼠标在一个行为室来监测运动活性15分钟。每天重复这个动作，连续3天，在相同的时间。 </li><li>在第四天，分化的小鼠分成两组。制备可卡因的原液在2毫克/毫升的盐水中。管理一个单一的注射可卡因溶液（最后20毫克/千克）的实验组根据鼠标的重量（ 如 25 G滑鼠-注入250微升，而30克鼠标将获得300微升）。施用生理盐水的对照组。紧接射出，将鼠标20分钟的行为室监测自发活动（炎热的典型盟友，前15分钟进行监控）。 </li><li>可卡因注射后，牺牲小鼠在不同时间点（0，1，2，4，8和24小时以下的注射）。重要的是，确保参考老鼠（0时刻）将不会收到任何注射的这一天。由于其重要性，就必须有参​​照组的复制。 </li></ol> 4，解剖的伏隔核<ol><li>麻醉的小鼠用异氟烷在可卡因/盐水注射后适当的时间。异氟醚必须直接排放走出了房间。因此，该程序应在通风橱中进行。 </li><li>通过测试后足部反射监测麻醉深度。捏爪子引发的动物提款回应。动物，显示了反射是不是麻醉的手术水平，而不是在国家安乐死。 </li><li>提取的大脑： <ol><li>断头安乐死鼠标。 </li> &lt;LI&gt;删除上面的头骨中间的皮，切用锋利的剪刀头骨。 </li><li>将剪刀在那里的脊髓进入脑中（枕骨大孔）点，使两侧切口。 </li><li>从同一点，使沿矢状缝长矢状切。在到达嗅球，夹在左侧和右侧。 </li><li>有镊子，小心地取出颅骨，抓颅骨各占一半牢固，拉在一旁，小心不要按到或损伤大脑。 </li><li>取出大脑，而通过使用一个倒置的微勺铲切断脑神经。 </li></ol></li><li>将大脑中的冰冷ACSF溶液1-2分钟来冷却和硬化。 </li><li>从学联取出大脑，灯芯多余的液体用纸巾，创造小脑和大脑的其余部分之间的冠状切口和胶水的脑，有喙的部分朝上，上vibratome阶段。 </li><li>切割的伏隔核： <ol&gt; <li>维持学联的解决方案，在冰冷的气温在vibratome的切片室。在200微米到伏隔核（NAc的）部分被清楚地识别根据Paxinos和Franklin小鼠脑图谱（约1.8毫米前到前囟门;注意胼胝体，前连合的位置和形状的形状和心室，以及在脑切片的整体的形态）。在这一点上，创建两个400微米的部分，从该伏隔将解剖。 </li><li>根据立体镜，用细刀片从解剖切片的伏隔核区域了。根据Paxinos和Franklin小鼠脑图谱的伏隔核的定义，并可以方便地通过在组织上的刀片4快速传递解剖中相应的部分。 （ 图2）。 </li><li>将立刻伏隔段在离心管和存储900微升的Trizol裂解试剂在-80°C，直到RNA的抽行动。 </li></ol></li></ol> 5，RNA提取和cDNA反转录<ol><li>解冻含核管伏在Trizol试剂裂解试剂部分在室温下和均化用1ml注射器连接至25g针溶解物直至组织完全均化（至少15-20倍）。第一几轮是困难的，并且需要推压管的壁的组织，以将其分解到小块可能进入针的尖端。 </li><li>以确保均质，吸管溶解物到QIAshredder微型旋转柱和离心机以12,000 xg离心1分钟。 </li><li>根据制造商的说明吸管溶解物到新的离心管中，并提取RNA。存储所述RNA在-80℃直到使用。 </li><li>测量RNA浓度，使用生物分析仪或等同设备分析的完整性和质量。使用100-500纳克的RNA每一轮的cDNA的制备有Random六聚体引物。 </li></ol> 6，引物设计和检测引物<ol><li>选择良好的引物：为获得最佳引物对mRNA转录的定量PCR，请按照下列要求（ 图3）： <ol><li>设计引物含有不超过17-28个碱基。避免核苷酸重复，因为他们推动错配。 </li><li>设计引物之间56-68℃（最佳60-64℃）熔化温度（Tm值）。该引物对中的Tm值应该是在彼此的1℃。 </li><li>请注意，该产品的尺寸最好应在80和150个基点之间。 </li><li>确保引物中的至少一个包括一个外显子 - 外显子交界处，或扩增子中含有大量的内含子（内含子跨越），以避免基因组DNA污染的扩增。 </li><li>为了提高引物的设计采用基于引物3（对于实施例参照表2）的可靠的软件。 </li></ol></li><li>测试PRimers：为了确保引物的专一性和效率，控制定量PCR反应应进行： <ol><li>用积极的样品（含cDNA模板为所有相关的引物对）和滴定的基因（ 如 8从〜10毫微克的初始浓度3倍稀释）一式两份平行反应。包括无模板对照，这对于用于反应的溶液中的污染控制。 </li><li> （ - 1 /斜率）-1 10（）}×100，使得它们的最佳斜率的曲线是- 3.333（ 即 10（ - 1 / 3.333）= 2）（ 图4）：使用下面的公式计算出的引物效率。 </li><li>确定扩增的特异性。特异性扩增证明由共同的扩增产物所有的熔解曲线的单一主峰，而脱靶扩增会出现一个明显的偏差FROM单大峰。 </li></ol></li></ol> 7，定量PCR分析<ol><li>前置放大： <ol><li>综合定量PCR分析是基于限制的RNA的量与大量的引物对的并行查询。因此，预扩增步骤是必不可少的。在此步骤中，cDNA经受预扩增14-18个循环的所有将被用于查询该样品在未来（最多800个引物对）中的引物的混合物。 </li><li>稀释于TE（低EDTA）的所有引物，以50为针对特定目标扩增（STA）。 </li><li>池一起所有的50μM引物的1微升等分设置要包括在该STA反应，高达800测定。 </li><li>制备预混物和样品的STA，如表3所述。通过制备混合为需要被放大的样本数的110％允许误差。 </li><li>等分3.75微升STA预混每个样品。添加1.25微升的cDNA的每一个。 </li><li>涡流以混合反应并离心10秒。 </li><li>将STA的反应在热循环和周期，在表4所示。 </li></ol></li><li>核酸外切酶I（ExoI）治疗： <ol><li>稀释的外切余如表5所示。 </li><li>加入2微升稀释ExoI（4 U /μL），以每5微升的STA反应，漩涡，降速（&gt; 6000 XG），并将其放置在热循环仪，37 ℃下进行30分钟，然后在80℃下进行15分钟。 </li><li>稀释的最终产品，以适当的浓度进行测试。用TE缓冲液（加43μLTE缓冲液到7微升TSA样品）。 </li><li>存储稀释STA产品在-20°C或立即使用。 </li></ol></li><li>底漆和样品安装在定量PCR： <ol><li>制备的样品，如表6。根据选择的实验中，芯片准备混合，并添加样品适当。 </li><li>敏捷射孔样品混合解决方案，最少20秒，而离心机（&gt; 6000 XG）至少30秒。制备的反应可以被存储为短的持续时间，在4℃，直至该芯片已准备好装载。 </li><li>如表7中所述制备的测定。 </li><li>搅动测定混合物最少20秒，离心机（&gt; 6000 XG）的至少30秒降速所有组件。 重要提示：涡彻底离心吸取之前的所有样品制备和分析解决方案集成到芯片的入口。如果不这样做，可能会导致数据质量的下降。 注意：每个引物的终浓度为5μM的在入口和500nM的最终反应。 </li></ol></li><li>底漆和加载动态数组国际金融公司（集成流体通路）芯片。在IFC芯片具有入口的样品和测定法，以及指定的入口（累加器）为用于加压的微流体腔室的控制管道流体（矿物油）（Figu重新4A）。 <ol><li>注入控制线流体到芯片上的每个累加器。 </li><li>移除并从芯片的底部丢弃蓝色保护膜。 </li><li>将芯片插入相应的国际金融公司（IFC）控制器（控制器MX的48.48动态阵列国际金融公司（IFC）和国际金融公司（IFC）控制器HX的96.96动态阵列国际金融公司），然后运行首相（113X）脚本的48.48动态阵列国际金融公司（IFC）和总理（136X）脚本的96.96动态阵列国际金融公司（IFC），以素片。 </li><li>当剧本完成后，请从国际金融公司控制器底漆芯片。 </li><li>移液管将5μl各测定混合物和5μl各样品混合到其入口。 </li><li>返回芯片到IFC控制器。 </li><li>利用国际金融公司控制器软件，运行负载混合（113X）脚本的48.48动态阵列国际金融公司）或混合装载（136X）脚本的96.96动态阵列国际金融公司装载样品和检测到芯片中。当负载混合脚本已经完成，除去负荷版芯片从国​​际金融公司控制器。 </li><li>装载在热循环仪的芯片，创建一个新的芯片运行（根据制造商的指导方针），包括熔解曲线分析。使用热循环仪软件，确保该反应室被正确识别，并允许该反应进行至完成。 </li></ol></li></ol> 8，数据分析<ol><li>质量保证：为了确保数据的质量，通过寻址稀释度曲线验证引物的质量（如上述）。通过观察扩增子的熔化曲线监测扩增的特异性，其中峰值应最佳地紧密排列29。 </li><li>可视化：对于大批量的高通量定量PCR获得的数据可视化，使用软件生成的热图，其中表现为颜色编码的强度。此外，显示单个基因作为内衬散点图的转录动力学。 </li><li>出版：出版基因表达的结果，建议遵循MIQE准则发布实时定量PCR实验，详细说明了必须报告的定量PCR实验，以确保其相关性，准确性，正确的解释，和可重复性的最少信息。 </li></ol>

<ol>
	<li>Amit, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+module+of+negative+feedback+regulators+defines+growth+factor+signaling.">A module of negative feedback regulators defines growth factor signaling.</a> Nature genetics. 39, 503-512 (2007).</li><li>Citri, A., Yarden, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EGF-ERBB+signalling:+towards+the+systems+level.">EGF-ERBB signalling: towards the systems level.</a> Nature reviews. Molecular cell biology. 7, 505-516 (2006).</li><li>Holtmaat, A., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experience-dependent+structural+synaptic+plasticity+in+the+mammalian+brain.">Experience-dependent structural synaptic plasticity in the mammalian brain.</a> Nature reviews. Neuroscience. 10, 647-658 (2009).</li><li>Kleim, J. A., Jones, 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=Principles+of+experience-dependent+neural+plasticity:+implications+for+rehabilitation+after+brain+damage.">Principles of experience-dependent neural plasticity: implications for rehabilitation after brain damage.</a> Journal of speech, language, and hearing research. 51, S225-S239 (2008).</li><li>Kauer, J. A., Malenka, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+plasticity+and+addiction.">Synaptic plasticity and addiction.</a> Nature reviews. Neuroscience. 8, 844-858 (2007).</li><li>Grueter, B. A., Rothwell, P. E., Malenka, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrating+synaptic+plasticity+and+striatal+circuit+function+in+addiction.">Integrating synaptic plasticity and striatal circuit function in addiction.</a> Current opinion in neurobiology. 22, 545-551 (2012).</li><li>Robinson, T. E., Kolb, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+plasticity+associated+with+exposure+to+drugs+of+abuse.">Structural plasticity associated with exposure to drugs of abuse.</a> Neuropharmacology. 47, 33-46 (2004).</li><li>Koob, G. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurobiological+mechanisms+in+the+transition+from+drug+use+to+drug+dependence.">Neurobiological mechanisms in the transition from drug use to drug dependence.</a> Neuroscience and biobehavioral reviews. 27, 739-749 (2004).</li><li>Hyman, S. E., Malenka, R. C., Nestler, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+mechanisms+of+addiction:+the+role+of+reward-related+learning+and+memory.">Neural mechanisms of addiction: the role of reward-related learning and memory.</a> Annual review of neuroscience. 29, 565-598 (2006).</li><li>Beurrier, C., Malenka, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhanced+inhibition+of+synaptic+transmission+by+dopamine+in+the+nucleus+accumbens+during+behavioral+sensitization+to+cocaine.">Enhanced inhibition of synaptic transmission by dopamine in the nucleus accumbens during behavioral sensitization to cocaine.</a> The Journal of neuroscience. 22, 5817-5822 (2002).</li><li>Robinson, T. E., Berridge, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+psychology+and+neurobiology+of+addiction:+an+incentive-sensitization+view.">The psychology and neurobiology of addiction: an incentive-sensitization view.</a> Addiction. 95, S91-S117 (2000).</li><li>Boening, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurobiology+of+an+addiction+memory.">Neurobiology of an addiction memory.</a> Journal of neural transmission. 108, 755-765 (2001).</li><li>Everitt, B. J., Robbins, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+systems+of+reinforcement+for+drug+addiction:+from+actions+to+habits+to+compulsion.">Neural systems of reinforcement for drug addiction: from actions to habits to compulsion.</a> Nature neuroscience. 8, 1481-1489 (2005).</li><li>Volkow, N. D., Fowler, J. S., Wang, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+addicted+human+brain:+insights+from+imaging+studies.">The addicted human brain: insights from imaging studies.</a> The Journal of clinical investigation. 111, 1444-1451 (2003).</li><li>Carlezon, W. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+cocaine+reward+by+CREB.">Regulation of cocaine reward by CREB.</a> Science. 282, 2272-2275 (1998).</li><li>Hope, B., Kosofsky, B., Hyman, S. E., Nestler, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+immediate+early+gene+expression+and+AP-1+binding+in+the+rat+nucleus+accumbens+by+chronic+cocaine.">Regulation of immediate early gene expression and AP-1 binding in the rat nucleus accumbens by chronic cocaine.</a> Proceedings of the National Academy of Sciences of the United States of America. 89, 5764-5768 (1992).</li><li>Hope, B. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+a+long-lasting+AP-1+complex+composed+of+altered+Fos-like+proteins+in+brain+by+chronic+cocaine+and+other+chronic+treatments.">Induction of a long-lasting AP-1 complex composed of altered Fos-like proteins in brain by chronic cocaine and other chronic treatments.</a> Neuron. 13, 1235-1244 (1994).</li><li>Pulipparacharuvil, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cocaine+regulates+MEF2+to+control+synaptic+and+behavioral+plasticity.">Cocaine regulates MEF2 to control synaptic and behavioral plasticity.</a> Neuron. 59, 621-633 (2008).</li><li>Robison, A. J., Nestler, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transcriptional+and+epigenetic+mechanisms+of+addiction.">Transcriptional and epigenetic mechanisms of addiction.</a> Nature reviews. Neuroscience. 12, 623-637 (2011).</li><li>Hyman, S. E., Malenka, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Addiction+and+the+brain:+the+neurobiology+of+compulsion+and+its+persistence.">Addiction and the brain: the neurobiology of compulsion and its persistence.</a> Nature reviews. Neuroscience. 2, 695-703 (2001).</li><li>Nestler, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neurobiology+of+cocaine+addiction.+Science+&amp;+practice+perspectives+/+a+publication+of+the.">The neurobiology of cocaine addiction. Science &amp; practice perspectives / a publication of the.</a> National Institute on Drug Abuse, National Institutes of Health. 3, 4-10 (2005).</li><li>Robbins, T. W., Everitt, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurobehavioural+mechanisms+of+reward+and+motivation.">Neurobehavioural mechanisms of reward and motivation.</a> Current opinion in neurobiology. 6, 228-236 (1996).</li><li>Kaplan, G. B., Moore, K. 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+use+of+cognitive+enhancers+in+animal+models+of+fear+extinction.">The use of cognitive enhancers in animal models of fear extinction.</a> Pharmacology, biochemistry, and behavior. 99, 217-228 (2011).</li><li>Chauvet, C., Goldberg, S. R., Jaber, M., Solinas, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+environmental+enrichment+on+the+incubation+of+cocaine+craving.">Effects of environmental enrichment on the incubation of cocaine craving.</a> Neuropharmacology. 63, 635-641 (2012).</li><li>Nithianantharajah, J., Hannan, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enriched+environments,+experience-dependent+plasticity+and+disorders+of+the+nervous+system.">Enriched environments, experience-dependent plasticity and disorders of the nervous system.</a> Nature reviews. Neuroscience. 7, 697-709 (2006).</li><li>Silingardi, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ERK+pathway+activation+bidirectionally+affects+visual+recognition+memory+and+synaptic+plasticity+in+the+perirhinal+cortex.">ERK pathway activation bidirectionally affects visual recognition memory and synaptic plasticity in the perirhinal cortex.</a> Frontiers in behavioral neuroscience. 5, 84 (2011).</li><li>Tropea, D., Majewska, A. K., Garcia, R., Sur, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+dynamics+of+synapses+in+vivo+correlate+with+functional+changes+during+experience-dependent+plasticity+in+visual+cortex.">Structural dynamics of synapses in vivo correlate with functional changes during experience-dependent plasticity in visual cortex.</a> The Journal of neuroscience. 30, 11086-11095 (2010).</li><li>Steketee, J. D., Kalivas, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug+wanting:+behavioral+sensitization+and+relapse+to+drug-seeking+behavior.">Drug wanting: behavioral sensitization and relapse to drug-seeking behavior.</a> Pharmacological reviews. 63, 348-365 (2011).</li><li>Citri, A., Pang, Z. P., Sudhof, T. C., Wernig, M., Malenka, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+qPCR+profiling+of+gene+expression+in+single+neuronal+cells.">Comprehensive qPCR profiling of gene expression in single neuronal cells.</a> Nature protocols. 7, 118-127 (2012).</li></ol>

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从脑组织以下行为范式成功的表征基因的表达依赖于：行为范式中1）认真小鼠的处理; 2）快速，精确的利益组织的解剖; 3）RNA的保护措施以确保RNA的完整性; 4）引物和实验布局精心规划，以及精度和对细节的关注，准备的qPCR分析。 描述的步骤的目的是表征诱发一种行为体验的转录活动动态;因此，认真注意是必要的，以确保该行为范式所经历的小鼠确实表示打算由研究者的?...

大脑的充满活力的组织能够认知和行为的灵活性。经验是通过神经元之间的连接的结构和强度的变型在大脑3编码。这个“经验依赖性可塑性”是诱导特异性基因表达，可提供必要的蛋白质的突触结构和强度4的变形例的模式的结果。基因调控网络的识别中介长期记忆的形成是分子神经科学的核心原则，并期望识别转录程序的主导因素将提供洞察调节记忆的形成，以及对目标的基本原则治疗神经变性和神经精神障碍。转录程序展开在时间上定义的波，其中每一个编码的基因不同的字符，这是当d重要ifferent阶段的信号事件1,2的结果的执行情况。因此重要的是要解决的转录动力学进行了详细的时间时间表，以便确定诱导的基因的全面补充，并洞察他们的潜在功能，根据自己归纳的动态。 吸毒是造成药物滥用的神经回路在大脑5,6长期影响的经验依赖可塑性强大的表单。最初，急性暴露于药物可能导致成瘾的发展和过渡到长期使用。上下文信息是网瘾发展的关键因素。药物相关环境线索在药物滥用者的头脑中分配显著重要性。上下文信息，提醒过往用药经验滥用药物可诱发复吸毒品的渴求，甚至禁欲以下长时间的药物暴露7,8。因此，重要的临床挑战成瘾-成瘾的倾向，停药后症状已经消退9复发甚至长。 行为敏可卡因是可卡因经验的毒瘾机制的研究很有用一个简单的模型。在本作中持久过敏引起的慢性接触到滥用药物被广泛研究的模式，鼠类首先习惯于注射生理食盐水（腹腔，IP）的一个新的环境中（一个开放的领域腔中的自发活动监测） ;然后，他们收到可卡因每天注射，在旷场箱，而它们的活性被监测10（ 图1）。这种行为模式通常导致的自发行为（8-12倍以上的基线活动）11，这是保持以下停止注射可卡因数月的强劲敏，显示了PERV的形成的用药经验asive记忆痕迹。 奖励的神经回路，自然参与加强行为的一个物种的成功（ 例如喂食，性别）必不可少的，是由药物滥用，加强药物相关的行为12,13的利用。由药物滥用的经验增强的分子和细胞机制似乎是类似的底层声明或语义记忆在其他大脑结构14的形成机制。因此，该行为敏化模型的稳健性，使其成为有吸引力的模型系统来研究的经验依赖性可塑性机制。 伏隔核（NAC）是大脑奖励环路的中央集成商，并已被广泛使用成瘾5,6的发展有关。瘾的形成取决于在伏隔核的新的蛋白质的转录，并在鲁棒的结构清晰的转录程序duction是在伏隔核以下可卡因经验15-19 观察到可卡因曝光的急性转录反应是可能的，多层次的，以适应于强诱导刺激，并直接生产新的蛋白质的功能即负责通过照射诱发的药物6,19-22的结构和电学的改变。 为了促进的经验依赖性可塑性在大脑中的分子机制的研究中，一个协议被描述为转录动力学的脑组织样品中的下列行为操作的全面分析。该协议说明的行为经验研究的生活更轻松实验室环境 - 行为敏可卡因，利用微流体动态数组进行转录分析。中描述的协议显然不限于学习吨他细胞核中行为敏化的背景下伏，而是可以应用到大量的行为范例和大脑区域。事实上，这个协议可以被应用到身体组织中的外脑，和各种经历或生物体的操作的影响。 该协议被粗略地分为四个步骤。在第一步骤中，将动物进行的行为模式;在第二步骤中，组织是显微切割的;在第三步骤 - 的mRNA的纯化，逆转录和探测，并在最后的步骤中的数据进行了分析。 在研究转录动力学的情况下，精确定时的经验和定义可能是最重要的实验参数来控制。出于这个原因，我们的行为选择的模式是，行为敏可卡因，一种系统，其允许实验者控制的高含量超过experien的参数CE。提供额外的行为范式，使精确定时和解决不同型号的经验依赖可塑性和记忆的形成。这些模型包括恐惧条件23，急性丰富环境24,25，新物体探索26项黑暗饲养27的视觉体验。然而，行为敏可卡因是一贯稳健的操纵行为，创造一个高度普遍的记忆痕迹而持续数月以下的可卡因经验28。 大脑切片，随后是伏隔核的手工显微​​切割。一直以来，我们的经验，从快速制备脑切片手工显微切割提供提取相关的行为范例组织的最可靠和快速的方法，并且有经验的，组织的边界变得明显和容易地识别。另外，精细的切片可以prepar编，接着通过激光捕获显微切割。虽然这种方法能够高度限定的感兴趣区域的分界，它是慢（因而冒不稳定的mRNA的丢失），繁琐，并且需要昂贵的专用设备（配有激光捕获设置的显微镜）。本文所定义的协议，也可以适用于单细胞转录分析，通过使用膜片移液管29在视觉上识别细胞的细胞质中的手动抽吸。要注意的是所描述的协议提供了人口平均，而这是极可能的是，在大多数情况下，在组织内的细胞中，只有亚群实际参与反应的经验是很重要的。这是感兴趣的，从特定的细胞群响应经验内轮廓的转录以选择性方式，但这些方法的讨论已超出当前范围。 对于mRNA的纯化，反转录和定量PCR的查询中，组织是通过细针，其次是市售试剂盒的使用传递给它打乱了（有关详细信息，请参阅表8）。的选择是通过用这些方法，这确保了高品质的RNA和从下游应用健壮的结果可靠提取经验得知。 而该协议描述了用于高通量定量PCR使用动态阵列，样品可以用终点PCR，低通量定量PCR，基因表达的微阵列或深度测序被探测的基因表达。偏爱高通量定量PCR使用动态阵列是由于mRNA的脑细胞得到下列行为范例是常常限制量的事实。动态数组提供了一个平台，使成绩单高效综合分析从一个单一的实验大量平行样品。最初的收购微流体系统（通常是机构PU后rchase），实验是相对便宜的运行。根据这一分析，可以用更昂贵的平台，以寻找新的成绩单（由微阵列或RNAseq）与动态数组提供质量保证全面的参考来进行样品的进一步查询。最后，对数据进行分析，标准的方法被利用​​。对于可能出现的问题的具体指针将在协议的文本进行讨论。 该协议是最适合感兴趣的是他们的利益制度进行彻底的调查调查，研究多个条件和重复。该协议也是谁已经磨练中（通过微阵列或RNAseq实验）在50-500基因的兴趣，他们有兴趣在反复询问一个子集，研究者最合适的。

<table border="0" cellpadding="0" cellspacing="0" width="453">
	<tbody>
		<tr>
			<td>Virusol</td>
			<td>Oriek Medical</td>
			<td>J29D</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane, USP 100%</td>
			<td>MINRAD INC</td>
			<td>NDC 60307-110-25</td>
			<td></td>
		</tr>
		<tr>
			<td>RNeasy plus Universal Mini Kit</td>
			<td>QIAGENE</td>
			<td>73404</td>
			<td></td>
		</tr>
		<tr>
			<td>QIAshredder</td>
			<td>QIAGENE</td>
			<td>79654</td>
			<td></td>
		</tr>
		<tr>
			<td>High Capacity cDNA Reverse Transcription kit</td>
			<td>Invitrogene</td>
			<td>AB-4368814</td>
			<td></td>
		</tr>
		<tr>
			<td>TE Buffer</td>
			<td>Invitrogene</td>
			<td>1355656</td>
			<td></td>
		</tr>
		<tr>
			<td>Behaviour Chamber (MDF; 50X45cm)</td>
			<td>Self assembled</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Inner Perspex box (30X30cm)</td>
			<td>Self assembled</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>camera and video recorder</td>
			<td>Campden Inst</td>
			<td>CMD-80051</td>
			<td></td>
		</tr>
		<tr>
			<td>Media Recorder software</td>
			<td>Noldus</td>
			<td>NDS-NMR3-00M</td>
			<td></td>
		</tr>
		<tr>
			<td>Iris Scissors</td>
			<td>FST</td>
			<td>FST-14062-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Sagital Brain slicer with a 0.5mm section</td>
			<td>Brain Tree Scientific</td>
			<td>BS-AL-505S</td>
			<td></td>
		</tr>
		<tr>
			<td>Bioanalyzer</td>
			<td>Agilent Technologies</td>
			<td>The Agilent 2100 Bioanalyzer</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermal cycler</td>
			<td>Bio-Rad</td>
			<td>1852048</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted microspun spatula</td>
			<td>Bochem Instrument GmbH</td>
			<td>3213</td>
			<td></td>
		</tr>
		<tr>
			<td>Biomark HD Reader</td>
			<td>Fluidigm</td>
			<td>BMHD-BMKHD</td>
			<td></td>
		</tr>
		<tr>
			<td>Dynamic array Chip for 96.96gene expression</td>
			<td>Fluidigm</td>
			<td>BMK-M-96.96</td>
			<td></td>
		</tr>
	</tbody>
</table>

comprehensive analysis of transcription dynamics from brain samples following behavioral experience

在大脑和长期记忆巩固经验的编码依赖于基因转录。识别编码经验的特定基因的功能的分子神经科学的主要目标之一。此外，确定基因与特定行为的功能联系对于理解神经精神疾病的基础意义。感应的鲁棒转录方案已在下列各种行为操作小鼠的大脑中观察到。而一些遗传元件被用于循环地以下不同行为的操作和在不同脑核，转录方案是整体独特的诱导刺激物和它们所研究1,2的结构。 在本出版物，协议是针对操纵行为的描述，从小鼠脑核强大和全面的转录谱。该协议中演示了在细胞核中基因表达的动力学分析的情况下伏隔急性可卡因经验。随后到体内的经验定义，目标神经组织被解剖;其次是RNA纯化，反转录和利用微阵列为多个目标基因的综合定量PCR分析。该协议是着眼于全面的分析（处理50-500个基因）限制原料的数量，如小的大脑标本，甚至单个细胞。 该协议是对多个样品的平行分析（ 如单细胞，动态分析下列药物，病毒或行为紊乱）最有利的。然而，该协议也可以事先对全基因组研究中起到了样品的特性和质量保证通过微阵列或RNAseq，以及从全基因组研究中获得的数据的验证。

通过施加这个协议中得到的结果的质量在很大程度上取决于于许多参数。适当的试验计划将造成最小干扰的实验小鼠，如所测试的经验（在本例中，即暴露于可卡因）将在其最近历史上最占优势的经验，因此，将导致一个强大的和特异性的转录程序。 图1描述了实验计划行为敏可卡因，限定的时间点以下可卡因经验在其中的伏隔核是从小鼠解剖和分析。下一个关键点是，限定了精确的...

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Comprehensive Analysis of Transcription Dynamics from Brain Samples Following Behavioral Experience

This manuscript describes a protocol that applies comprehensive profiling for analysis of transcriptional programs induced in specific brain nuclei of rodents following behavioral paradigms. Herein, this approach is illustrated in the context of profiling genes induced in the nucleus accumbens (NAc) of mice following acute cocaine exposure, utilizing microfluidic qPCR arrays.

转录动力学的脑组织样本在行为经验综合分析

The encoding of experiences in the brain and the consolidation of long-term memories depend on gene transcription. Identifying the function of specific ...

comprehensive-analysis-transcription-dynamics-from-brain-samples

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11.5K 次观看.  The Hebrew University of Jerusalem. This manuscript describes a protocol that applies comprehensive profiling for analysis of transcriptional programs induced in specific brain nuclei of rodents following behavioral paradigms. Herein, this approach is illustrated in the context of profiling genes induced in the nucleus accumbens (NAc) of mice following acute cocaine exposure, utilizing microfluidic qPCR arrays. 

视频: Comprehensive Analysis of Transcription Dynamics from Brain Samples Following Behavioral Experience

Stereotactic Injection of MicroRNA-expressing Lentiviruses to the Mouse Hippocampus CA1 Region and Assessment of the Behavioral Outcome

MicroRNAs (miRNAs) are small regulatory single-stranded RNA molecules around 22 nucleotides long that may each target numerous mRNA transcripts and dim an entire gene expression pathway by inducing destruction and/or inhibiting translation of these targets. Several miRNAs play key roles in maintaining neuronal structure and function and in higher-level brain functions, and methods are sought for manipulating their levels for exploring these functions. Here, we present a direct in vivo method for examining the cognitive consequences of enforced miRNAs excess in mice by stereotactic injection of miRNA-encoding virus particles. Specifically, the current protocol involves injection into the hippocampal CA1 region, which contributes to mammalian memory consolidation, learning, and stress responses, and offers a convenient injection site. The coordinates are measured according to the mouse bregma and virus perfusion is digitally controlled and kept very slow. After injection, the surgery wound is sealed and the animals recover. Lentiviruses encoding silencers of the corresponding mRNA targets serve to implicate the specific miRNA/target interaction responsible for the observed effect, with na&iuml;ve mice, mice injected with saline and mice injected with "empty" lentivirus vectors as controls. One month post-injection, the animals are examined in the Morris Water Maze (MWM) for assessing their navigation learning and memory abilities. The MWM is a round tank filled with colored water with a small platform submerged 1 cm below the water surface. Steady visual cues around the tank allow for spatial navigation (sound and the earth's magnetic field may also assist the animals in navigating). Video camera monitoring enables measuring the route of swim and the time to find and amount the platform. The mouse is first taught that mounting the hidden platform offers an escape from the enforced swimming; it is then tested for using this escape and finally, the platform is removed and probe tests examine if the mouse remembers its previous location. Repeated tests over several consecutive days highlight improved performance of tested mice at shorter latencies to find and mount the platform, and as more direct routes to reach the platform or its location. Failure to show such improvement represents impaired learning and memory and/or anxiety, which may then be tested specifically (e.g. in the elevated plus maze). This approach enables validation of specific miRNAs and target transcripts in the studied cognitive and/or stress-related processes.

MicroRNAs have significant roles in brain structure and function. Here we describe a method to enforce hippocampal miRNA over-expression using stereotactic injection of an engineered miRNA-expressing lentivirus. This approach can serve as a relatively rapid way to assess the in vivo effects of over-expressed miRNAs in specific brain regions.

立体定向注射的microRNA表达慢病毒的小鼠海马CA1区和行为结果的评估

MicroRNAs (miRNAs) are small regulatory single-stranded RNA molecules around 22 nucleotides long that may each target numerous mRNA transcripts and dim an ...

科研

行为学

Compensatory Limb Use and Behavioral Assessment of Motor Skill Learning Following Sensorimotor Cortex Injury in a Mouse Model of Ischemic Stroke

Mouse models have become increasingly popular in the field of behavioral neuroscience, and specifically in studies of experimental stroke. As models advance, it is important to develop sensitive behavioral measures specific to the mouse. The present protocol describes a skilled motor task for use in mouse models of stroke. The Pasta Matrix Reaching Task functions as a versatile and sensitive behavioral assay that permits experimenters to collect accurate outcome data and manipulate limb use to mimic human clinical phenomena including compensatory strategies (i.e., learned non-use) and focused rehabilitative training. When combined with neuroanatomical tools, this task also permits researchers to explore the mechanisms that support behavioral recovery of function (or lack thereof) following stroke. The task is both simple and affordable to set up and conduct, offering a variety of training and testing options for numerous research questions concerning functional outcome following injury. Though the task has been applied to mouse models of stroke, it may also be beneficial in studies of functional outcome in other upper extremity injury models.

Mouse models have become increasingly popular in studies of behavioral neuroscience. As models advance, it is important to develop sensitive behavioral measures specific to the mouse. This protocol describes the Pasta Matrix Reaching Task, which is a skilled motor task for use in mouse models of stroke.

补偿肢使用和运动技能学习行为评估以下感觉运动皮层损伤缺血性脑卒中的小鼠模型

Mouse models have become increasingly popular in the field of behavioral neuroscience, and specifically in studies of experimental stroke. As models ...

Transcranial Magnetic Stimulation for Investigating Causal Brain-behavioral Relationships and their Time Course

Transcranial magnetic stimulation (TMS) is a safe, non-invasive brain stimulation technique that uses a strong electromagnet in order to temporarily disrupt information processing in a brain region, generating a short-lived &#8220;virtual lesion.&#8221; Stimulation that interferes with task performance indicates that the affected brain region is necessary to perform the task normally. In other words, unlike neuroimaging methods such as functional magnetic resonance imaging (fMRI) that indicate correlations between brain and behavior, TMS can be used to demonstrate causal brain-behavior relations. Furthermore, by varying the duration and onset of the virtual lesion, TMS can also reveal the time course of normal processing. As a result, TMS has become an important tool in cognitive neuroscience. Advantages of the technique over lesion-deficit studies include better spatial-temporal precision of the disruption effect, the ability to use participants as their own control subjects, and the accessibility of participants. Limitations include concurrent auditory and somatosensory stimulation that may influence task performance, limited access to structures more than a few centimeters from the surface of the scalp, and the relatively large space of free parameters that need to be optimized in order for the experiment to work. Experimental designs that give careful consideration to appropriate control conditions help to address these concerns. This article illustrates these issues with TMS results that investigate the spatial and temporal contributions of the left supramarginal gyrus (SMG) to reading.

Transcranial magnetic stimulation (TMS) is a technique for non-invasively disrupting neural information processing and measuring its effect on behavior. When TMS interferes with a task, it indicates that the stimulated brain region is necessary for normal task performance, allowing one to systematically relate brain regions to cognitive functions.

经颅磁刺激对调查因果脑 - 行为关系及其时间进程

Transcranial magnetic stimulation (TMS) is a safe, non-invasive brain stimulation technique that uses a strong electromagnet in order to temporarily ...

Combined Shuttle-Box Training with Electrophysiological Cortex Recording and Stimulation as a Tool to Study Perception and Learning

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the necessary equipment is now available by several commercial companies. This protocol provides a detailed description of a two-way shuttle-box avoidance learning paradigm in rodents (here Mongolian gerbils; Meriones unguiculatus) in combination with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical electrophysiological in vivo recordings. The detailed protocol is applicable to study multiple aspects of learning behavior and perception in different rodent species.
Site-specific ICMS of auditory cortical circuits as conditioned stimuli here is used as a tool to test the perceptual relevance of specific afferent, efferent and intracortical connections. Distinct activation patterns can be evoked by using different stimulation electrode arrays for local, layer-dependent ICMS or distant ICMS sites. Utilizing behavioral signal detection analysis it can be determined which stimulation strategy is most effective for eliciting a behaviorally detectable and salient signal. Further, parallel multichannel-recordings using different electrode designs (surface electrodes, depth electrodes, etc.) allow for investigating neuronal observables over the time course of such learning processes. It will be discussed how changes of the behavioral design can increase the cognitive complexity (e.g. detection, discrimination, reversal learning).

Shuttle-box avoidance learning is well-established in behavioral neuroscience. This protocol describes how shuttle-box learning in rodents can be combined with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical in vivo recordings as a tool to study multiple aspects of learning and perception.

联合穿梭盒子训练与电的Cortex记录和刺激作为一种工具来研究感知和学习

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the ...

Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as appropriate models, partly because the response to behaviorally relevant stimuli and the circuitry parsing stimulus-action outcomes is astonishingly similar between humans and rodents. In studying molecular substrates of complex behaviors, the microinjection of reagents that modify, augment, or silence specific systems is an invaluable technique. However, it is crucial that the microinjection site is precisely targeted in order to aid interpretation of the results. We present a method for the manufacture of surgical implements and microinjection needles that enables accurate microinjection and unlimited customizability with minimal cost. Importantly, this technique can be successfully completed in awake rodents if conducted in conjunction with other JoVE articles that covered requisite surgical procedures. Additionally, there are many behavioral paradigms that are well suited for measuring motivation. The progressive ratio is a commonly used method that quantifies the efficacy of a reinforcer to maintain responding despite an (often exponentially) increasing work requirement. This assay is sensitive to reinforcer magnitude and pharmacological manipulations, which allows reinforcing efficacy and/ or motivation to be determined. We also present a straightforward approach to program operant software to accommodate a progressive ratio reinforcement schedule.

Rodents are an appropriate model to investigate the molecular substrates of behavior and complex psychiatric disorders. Brain microinjection in awake rodents can be used to elucidate disease substrates. An efficient and customizable brain microinjection method as well as the execution of an operant paradigm that quantifies motivation is presented.

啮齿动物脑显微注射学习动机行为的分子基板

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as ...

Vagus Nerve Stimulation as a Tool to Induce Plasticity in Pathways Relevant for Extinction Learning

Extinction describes the process of attenuating behavioral responses to neutral stimuli when they no longer provide the reinforcement that has been maintaining the behavior. There is close correspondence between fear and human anxiety, and therefore studies of extinction learning might provide insight into the biological nature of anxiety-related disorders such as post-traumatic stress disorder, and they might help to develop strategies to treat them. Preclinical research aims to aid extinction learning and to induce targeted plasticity in extinction circuits to consolidate the newly formed memory. Vagus nerve stimulation (VNS) is a powerful approach that provides tight temporal and circuit-specific release of neurotransmitters, resulting in modulation of neuronal networks engaged in an ongoing task. VNS enhances memory consolidation in both rats and humans, and pairing VNS with exposure to conditioned cues enhances the consolidation of extinction learning in rats. Here, we provide a detailed protocol for the preparation of custom-made parts and the surgical procedures required for VNS in rats. Using this protocol we show how VNS can facilitate the extinction of conditioned fear responses in an auditory fear conditioning task. In addition, we provide evidence that VNS modulates synaptic plasticity in the pathway between the infralimbic (IL) medial prefrontal cortex and the basolateral complex of the amygdala (BLA), which is involved in the expression and modulation of extinction memory.

Vagus nerve stimulation (VNS) has emerged as a tool to induce targeted synaptic plasticity in the forebrain to modify a range of behaviors. This protocol describes how to implement VNS to facilitate the consolidation of fear extinction memory.

迷走神经刺激作为一种工具来诱导可塑性相关途径灭绝学习

Extinction describes the process of attenuating behavioral responses to neutral stimuli when they no longer provide the reinforcement that has been ...

Assessing Working Memory in Children: The Comprehensive Assessment Battery for Children &#8211; Working Memory (CABC-WM)

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of working memory in young school-age children. Working memory deficits have been identified in children with language-based learning disabilities, including dyslexia1,2 and language impairment3,4, but it is not clear whether these children exhibit deficits in subcomponents of working memory, such as visuospatial or phonological working memory. The CABC-WM is administered on a desktop computer with a touchscreen interface and was specifically developed to be engaging and motivating for children. Although the long-term goal of the CABC-WM is to provide individualized working memory profiles in children, the present study focuses on the initial success and utility of the CABC-WM for measuring central executive, visuospatial, phonological loop, and binding constructs in children with typical development. Immediate next steps are to administer the CABC-WM to children with specific language impairment, dyslexia, and comorbid specific language impairment and dyslexia.

Assessing Working Memory in Children: The Comprehensive Assessment Battery for Children &amp;#8211; Working Memory (CABC-WM)

Working memory predicts a significant amount of variance for a variety of cognitive tasks, including speaking, reading, and writing. However, few tools are available to assess working memory in children. We present an innovative, computer-based battery that comprehensively assesses different components of working memory in school-age children.

评估儿童工作记忆：儿童综合评估电池 - 工作记忆（CABC-WM）

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, few studies have investigated the long-term effects of injury in rodent models of TBI. Shown here are behavioral measures that are well-established in TBI research for times early after injury, such as two weeks, until two months. Some of these methods have previously been used at later times after injury, up to one year, but by very few laboratories. The methods demonstrated here are a short neurological assessment to test reflexes, a Beam-Balance to test balance, a Beam-Walk to test balance and motor coordination, and a working memory version of the Morris water maze that can be sensitive to deficits in reference memory. Male rats were handled and pre-trained to neurological, balance, and motor coordination tests prior to receiving parasagittal fluid percussion injury (FPI) or sham injury. Rats can be tested on the short neurological assessment (neuroscore), the beam-balance, and the Beam-Walk multiple times, while testing on the water maze can only be done once. This difference is because rats can remember the task, thus confounding the results if repeated testing is attempted in the same animal. When testing from one to three days after injury, significant differences are detected in all three non-cognitive tasks. However, differences in the Beam-Walk task were not detectable at later time points (after 3 months). Deficits were detected at 3 months in the Beam-Balance and at 6 months in the neuroscore. Deficits in working memory were detected out to 12 months after injury, and a deficit in a reference memory first appeared at 12 months. Thus, standard behavioral tests can be useful measures of persistent behavioral deficits after FPI.

The goal of the behavioral tests presented here is to detect functional deficits in rats after traumatic brain injury. Four specific tests are presented that detect deficits in behaviors to reflect the damage to specific brain areas at times extending to one year after injury.

颅脑损伤后大鼠行为缺陷的检测

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, ...

An Automated T-maze Based Apparatus and Protocol for Analyzing Delay- and Effort-based Decision Making in  Free Moving Rodents

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper understanding of the neurobiological causes underlying the decision-making problems. A cost-benefit based T-maze task is used for measuring decision making in which rodents choose between a high reward arm (HRA) and a low reward arm (LRA). There are two paradigms of the T-maze decision-making task, one in which the cost is a time delay and the other in which it is physical effort. Both paradigms require a tedious and labor-intensive management of experimental animals, multiple doors, pellet reward, and arm choice recordings. In the current work, we invented an apparatus based on traditional T-maze with full automation for pellet delivery, door management and choice recordings. This automated setup can be used for the evaluation of both delay- and effort-based decision making in rodents. With the protocol described here, our lab investigated the decision-making phenotypes of multiple genetically modified mice. In the representative data, we showed that the mice with ablated medial habenular showed aversions of both delay and effort and tended to choose the immediate and effortless reward. This protocol helps to decrease the variability caused by experimenter intervention and to enhance experiment efficiency. In addition, chronic silicon probe or microelectrode recording, fiber-optic imaging and/or manipulation of neural activity can be easily applied during the decision-making task using the setup described here.

This article introduces an automated T-maze apparatus that we invented, and a protocol based on this apparatus for analyzing delay-based decision making and effort-based decision making in free moving rodents.

基于自动 T 迷宫的自由移动啮齿动物延迟和努力决策分析装置及协议

Many neurological and psychiatric patients demonstrate difficulties and/or deficits in decision making. Rodent models are helpful to produce a deeper ...

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive decline, such as Alzheimer&#39;s disease (AD), have become a promising tool to elucidate the underlying mechanisms and promote the therapeutic advances. An important step is the validation and characterization of expected behavioral abnormality in the models, in the case of AD, cognitive decline. The long-term behavioral investigations of laboratory animals to study the effect of aging demand substantial efforts from researchers. The IntelliCage system is a high-throughput and cost-effective test battery for mice that eliminates the need for daily human handling. Here, we describe how the system is utilized in the long-term phenotyping of a genetic Alzheimer&#39;s disease model, specifically focusing on the cognitive functions. The experiment employs repeated battery of tests that assess spatial learning and executive functions. This cost-effective age-dependent phenotyping allows us to identify the transient and/or permanent effects of genes on various cognitive aspects.

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer&#039;s Disease, Using IntelliCage

This paper describes a protocol for cognitive assessments for genetic models of the Alzheimer&#39;s disease using the IntelliCage system, which is a high throughput automated behavioral monitoring system with operant conditioning.

基于 IntelliCage 的阿尔茨海默病多基因模型认知功能的自动、长期行为检测

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive ...