这项工作是由贝勒大学的研究理事会，并授予从癫痫症基金会的研究经费支持。

在下面的实验中使用的小鼠产生和安置在贝勒大学在22℃的环境温度下，用14小时光照和10小时黑暗（20:00至下午六小时）昼夜周期。给予小鼠随意获取食物和水。所有程序的小鼠均符合卫生指引全国学院实验动物的护理和使用以及动物方案经贝勒大学动物护理和使用委员会。 概观跟踪空调恐惧任务是基于由Wiltgen及其同事9所述的程序。 1。设备的研制恐惧调节装置腔室（26厘米×22厘米x 18厘米）由双方是丙烯酸类，两侧是金属，和一格栅地板底部，用来提供一个轻微的足部电击的。测试室是房子D在一个声音衰减室。该室也轻紧，以防止外部光线影响运动检测软件。 <ol><li>校准休克水平，光照水平，以及声音强度水平的测试室中。测量测试腔室的背景水平。在这个房间的背景噪音为65分贝。用声级计测量这个水平。仪表应设置为70dB时，设置为C，并设置检测速度慢。 <ol><li>校准震撼级到0.5 mA。使用外部校准设备正确校准的冲击水平（见材料表）。冲击发生器的内部的测量是不准确的。电击发生器管理着一个加扰休克不能由一个标准安培计来精确测量。 </li><li>将一根导线对电网的酒吧之一，并将其​​他领先3或4条以上。使用冲击发生器来管理的震撼。震荡调整水平，直到正确的水平来实现。做到这一点的EACH怕调理室。 </li><li>测量室的光照水平时关闭大门室。校准光水平约为1.0。这是一个数字，具体到本实验中使用的FreezeFrame软件。外部测光表将读取此为2勒克斯。的光量可以通过移动的房子光的位置，或通过调节透镜的聚光器来调整。确保调整后收紧镜头调节螺丝。 </li><li>校正声音分贝为85分贝。使用测试笼内外部分贝仪校准分贝（见材料表）。呈现的声音将是一个2,700 Hz的音调。注意：如果使用超过6个月大的老鼠可能是更好的使用白噪声，因为年纪较大的老鼠能有听力缺陷。 </li></ol></li><li>之后该装置准备好采取受试者单独的夹持室。注意：不要房子的老鼠在同一个房间考场。 </li><li>标签日Ë尾巴会为了测试被测试的小鼠。最好能减少过多handling测试之前。或者，所述尾部可以被标记在实验的前一天，以减少处理压力。后的小鼠已被标记为允许它们适应于房间30分钟。有额外的清洁笼子住房小鼠测试完成后。 </li></ol> 2。跟踪调理1天<ol><li>从笼中取出每只小鼠，并将其放置在运输单独的笼子的恐惧制约室。使用干净的被褥每个笼子。广场上的转移笼子里的便条养老鼠测试正确的顺序。注意：如果老鼠被单独安置那么他们可以在他们的家笼运输。 </li><li>将鼠标放置在测试室并关上门。启动该软件程序。 </li><li>在训练日，让小鼠探索室3分钟。然后，该软件提供了一个20秒的音（85分贝2,700赫兹）笔Ø动物。经过20秒的扫描时段轻微的冲击（2秒，0.5毫安）施用给动物。 </li><li>记录拍摄对象的反应，确认他们收到的厌恶刺激通过查看视频。 A 200秒的试验间隔隔开5调理试验。每次试验包括一个20秒的音，然后是一个20秒的延迟，然后一个冲击。 </li><li>后测试完成允许从测试笼中取出之前，动物留在测试腔室，持续1分钟。 </li><li>将动物放回传输笼子里，它返回到其家笼。如果有额外的老鼠在他们的家笼然后分别容纳鼠标，直到所有小鼠完整的测试。这将减少应力，还没有被测试的其他小鼠。一种替代的解决方案是一个星期单独容纳所有的小鼠在测试前，减少连续地从一个笼子取出小鼠的影响。 </li><li>选管会在30％异丙醇清洁测试室ħ动物进行测试。 </li><li>重复步骤2.2-2.7在测试队列所有小鼠。 </li><li>返回所有小鼠，以他们的殖民地的房间，最后鼠标在队列已经过测试后</li></ol> 3。一丝恐惧条件第2天：跟踪存储器测试<ol><li>跟踪存储器测试将发生在2日。在这个协议中有3个音介绍。将小鼠进行跟踪调节测试一个新的上下文。 </li><li>准备了2分钟基准期，其后三年20秒语气演示运行程序的软件。还有就是每个音之间呈现SA 220秒ITI。 </li><li>对新的上下文状态，放置在腔室的地板上透明的丙烯酸嵌件，以改变形状，纹理和调节室的颜色。 </li><li>通过在地板下放置插入香草精在权衡船改变气味在室内。 </li><li>用70％乙醇代替30％异丙醇清洗室中。注意：这将有助于创建一个新的上下文。 </li><li>把小鼠的持房，必要时重新标记它们的尾巴进行测试。 </li><li>用碎纸​​更换床上用品准备新的上下文转移的笼子。注意：这将创建一个新的上下文帮助。 </li><li>把鼠标在测试室中，然后启动该程序。用70％乙醇清洗室测试完成之后。 </li><li>返回小鼠的笼子跟踪调节完成后。返回所有小鼠，以他们的殖民地客房时，所有小鼠进行了检验。 </li></ol> 4。一丝恐惧条件第3天：语境记忆检测<ol><li>在第三天情景条件下进行。准备运行一个程序来记录冻结行为8分钟的软件。 </li><li>在测试前和测试每个鼠标后用30％的异丙醇清洁室。上下文应该是相同的，1天的。转印笼子应该是相同的如1天。 </li><li&gt;把小鼠的持房并标记它们的尾巴进行测试。 </li><li>将每只小鼠在试验室中，然后启动该程序。用30％异丙醇清洗室测试完成之后。 </li><li>返回所有小鼠到他们的殖民地室完成时。 </li></ol>

<ol>
	<li>Powell, R. A., Honey, P. L., Symbaluk, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Introduction to learning and behavior. , .</li><li>Tsaltas, E., Preston, G. C., Gray, 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=The+effects+of+dorsal+bundle+lesions+on+serial+and+trace+conditioning.">The effects of dorsal bundle lesions on serial and trace conditioning.</a> Behav. Brain Res. 10, 361-374 (1983).</li><li>McAlonan, G. M., Dawson, G. R., Wilkinson, L. O., 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=The+effects+of+AMPA-induced+lesions+of+the+medial+septum+and+vertical+limb+nucleus+of+the+diagonal+band+of+Broca+on+spatial+delayed+non-matching+to+sample+and+spatial+learning+in+the+water+maze.">The effects of AMPA-induced lesions of the medial septum and vertical limb nucleus of the diagonal band of Broca on spatial delayed non-matching to sample and spatial learning in the water maze.</a> Eur. J. Neurosci. 7, 1034-1049 (1995).</li><li>Chowdhury, N., Quinn, J. J., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dorsal+hippocampus+involvement+in+trace+fear+conditioning+with+long,+but+not+short,+trace+intervals+in+mice.">Dorsal hippocampus involvement in trace fear conditioning with long, but not short, trace intervals in mice.</a> Behav. Neurosci. 119, 1396-1402 (2005).</li><li>Quinn, J. J., Oommen, S. S., Morrison, G. E., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post-training+excitotoxic+lesions+of+the+dorsal+hippocampus+attenuate+forward+trace,+backward+trace,+and+delay+fear+conditioning+in+a+temporally+specific+manner.">Post-training excitotoxic lesions of the dorsal hippocampus attenuate forward trace, backward trace, and delay fear conditioning in a temporally specific manner.</a> Hippocampus. 12, 495-504 (2002).</li><li>Misane, 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=Time-dependent+involvement+of+the+dorsal+hippocampus+in+trace+fear+conditioning+in+mice.">Time-dependent involvement of the dorsal hippocampus in trace fear conditioning in mice.</a> Hippocampus. 15, 418-426 (2005).</li><li>Quinn, J. J., Loya, F., Ma, Q. D., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dorsal+hippocampus+NMDA+receptors+differentially+mediate+trace+and+contextual+fear+conditioning.">Dorsal hippocampus NMDA receptors differentially mediate trace and contextual fear conditioning.</a> Hippocampus. 15, 665-674 (2005).</li><li>McEchron, M. D., Bouwmeester, H., Tseng, W., Weiss, C., Disterhoft, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampectomy+disrupts+auditory+trace+fear+conditioning+and+contextual+fear+conditioning+in+the+rat.">Hippocampectomy disrupts auditory trace fear conditioning and contextual fear conditioning in the rat.</a> Hippocampus. 8, 638-646 (1998).</li><li>Wiltgen, B. J., Sanders, M. J., Ferguson, C., Homanics, G. E., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+fear+conditioning+is+enhanced+in+mice+lacking+the+delta+subunit+of+the+GABAA+receptor.">Trace fear conditioning is enhanced in mice lacking the delta subunit of the GABAA receptor.</a> Learn. Mem. 12, 327-333 (2005).</li><li>Davis, R. R., 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=Genetic+basis+for+susceptibility+to+noise-induced+hearing+loss+in+mice.">Genetic basis for susceptibility to noise-induced hearing loss in mice.</a> Hear. Res. 155, 82-90 (2001).</li><li>Zheng, Q. Y., Johnson, K. R., Erway, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+hearing+in+80+inbred+strains+of+mice+by+ABR+threshold+analyses.">Assessment of hearing in 80 inbred strains of mice by ABR threshold analyses.</a> Hear. Res. 130, 94-107 (1999).</li><li>Moyer, J. R., Thompson, L. T., Disterhoft, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+eyeblink+conditioning+increases+CA1+excitability+in+a+transient+and+learning-specific+manner.">Trace eyeblink conditioning increases CA1 excitability in a transient and learning-specific manner.</a> 16, 5536-5546 (1996).</li><li>Leuner, B., Falduto, J., Shors, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Associative+memory+formation+increases+the+observation+of+dendritic+spines+in+the+hippocampus.">Associative memory formation increases the observation of dendritic spines in the hippocampus.</a> J. Neurosci. 23, 659-665 (2003).</li><li>McEchron, M. D., Disterhoft, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+encoding+of+non-spatial+trace+conditioning.">Hippocampal encoding of non-spatial trace conditioning.</a> Hippocampus. 9, 385-396 (1999).</li><li>McLaughlin, J., Skaggs, H., Churchwell, J., Powell, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Medial+prefrontal+cortex+and+pavlovian+conditioning:+trace+versus+delay+conditioning.">Medial prefrontal cortex and pavlovian conditioning: trace versus delay conditioning.</a> Behav. Neurosci. 116, 37-47 (2002).</li><li>Runyan, J. D., Moore, A. N., Dash, P. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+role+for+prefrontal+cortex+in+memory+storage+for+trace+fear+conditioning.">A role for prefrontal cortex in memory storage for trace fear conditioning.</a> J. Neurosci. 24, 1288-1295 (2004).</li><li>Gilmartin, M. R., McEchron, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+neurons+in+the+medial+prefrontal+cortex+of+the+rat+exhibit+tonic+and+phasic+coding+during+trace+fear+conditioning.">Single neurons in the medial prefrontal cortex of the rat exhibit tonic and phasic coding during trace fear conditioning.</a> Behav. Neurosci. 119, 1496-1510 (2005).</li><li>Crow, T., Xue-Bian, J. J., Siddiqi, V., Kang, Y., Neary, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phosphorylation+of+mitogen-activated+protein+kinase+by+one-trial+and+multi-trial+classical+conditioning.">Phosphorylation of mitogen-activated protein kinase by one-trial and multi-trial classical conditioning.</a> J. Neurosci. 18, 3480-3487 (1998).</li><li>Martin, K. C., 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=MAP+kinase+translocates+into+the+nucleus+of+the+presynaptic+cell+and+is+required+for+long-term+facilitation+in+Aplysia.">MAP kinase translocates into the nucleus of the presynaptic cell and is required for long-term facilitation in Aplysia.</a> Neuron. 18, 899-912 (1997).</li><li>Crestani, 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=Trace+fear+conditioning+involves+hippocampal+alpha5+GABA(A)+receptors.">Trace fear conditioning involves hippocampal alpha5 GABA(A) receptors.</a> Proc. Natl. Acad. Sci. U.S.A. 99, 8980-8985 (2002).</li><li>Crestani, 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=Decreased+GABAA-receptor+clustering+results+in+enhanced+anxiety+and+a+bias+for+threat+cues.">Decreased GABAA-receptor clustering results in enhanced anxiety and a bias for threat cues.</a> Nat. Neurosci. 2, 833-839 (1999).</li><li>Moore, M. 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=Trace+and+contextual+fear+conditioning+is+enhanced+in+mice+lacking+the+alpha4+subunit+of+the+GABA(A)+receptor.">Trace and contextual fear conditioning is enhanced in mice lacking the alpha4 subunit of the GABA(A) receptor.</a> Neurobiol. Learn. Mem. 93, 383-387 (2010).</li><li>Cushman, J. D., Moore, M. D., Jacobs, N. S., Olsen, R. W., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+pharmacogenetic+analysis+on+the+role+of+the+alpha4+GABA(A)+receptor+subunit+in+the+ethanol-mediated+impairment+of+hippocampus-dependent+contextual+learning.">Behavioral pharmacogenetic analysis on the role of the alpha4 GABA(A) receptor subunit in the ethanol-mediated impairment of hippocampus-dependent contextual learning.</a> Alcohol Clin. Exp. Res. 35, 1948-1959 (2011).</li><li>Raybuck, J. D., Lattal, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Double+dissociation+of+amygdala+and+hippocampal+contributions+to+trace+and+delay+fear+conditioning.">Double dissociation of amygdala and hippocampal contributions to trace and delay fear conditioning.</a> PLoS ONE. 6, (2011).</li><li>Kwapis, J. L., Jarome, T. J., Schiff, J. C., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+consolidation+in+both+trace+and+delay+fear+conditioning+is+disrupted+by+intra-amygdala+infusion+of+the+protein+synthesis+inhibitor+anisomycin.">Memory consolidation in both trace and delay fear conditioning is disrupted by intra-amygdala infusion of the protein synthesis inhibitor anisomycin.</a> Learn. Mem. 18, 728-732 (2011).</li><li>Gilmartin, M. R., Kwapis, J. L., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+and+contextual+fear+conditioning+are+impaired+following+unilateral+microinjection+of+muscimol+in+the+ventral+hippocampus+or+amygdala,+but+not+the+medial+prefrontal+cortex.">Trace and contextual fear conditioning are impaired following unilateral microinjection of muscimol in the ventral hippocampus or amygdala, but not the medial prefrontal cortex.</a> Neurobiol. Learn. Mem. 97, 452-464 (2012).</li><li>Baysinger, A. N., Kent, B. A., Brown, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscarinic+receptors+in+amygdala+control+trace+fear+conditioning.">Muscarinic receptors in amygdala control trace fear conditioning.</a> PLoS ONE. 7, (2012).</li><li>Wanisch, K., Tang, J., Mederer, A., Wotjak, C. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trace+fear+conditioning+depends+on+NMDA+receptor+activation+and+protein+synthesis+within+the+dorsal+hippocampus+of+mice.">Trace fear conditioning depends on NMDA receptor activation and protein synthesis within the dorsal hippocampus of mice.</a> Behav. Brain. 157, 63-69 (2005).</li><li>Smith, D. R., Gallagher, M., Stanton, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+background+differences+and+nonassociative+effects+in+mouse+trace+fear+conditioning.">Genetic background differences and nonassociative effects in mouse trace fear conditioning.</a> Learn. Mem. 14, 597-605 (2007).</li><li>Rudy, J. W., O'Reilly, 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=Contextual+fear+conditioning,+conjunctive+representations,+pattern+completion,+and+the+hippocampus.">Contextual fear conditioning, conjunctive representations, pattern completion, and the hippocampus.</a> Behav. Neurosci. 113, 867-880 (1999).</li><li>Wiltgen, B. J., Sanders, M. J., Anagnostaras, S. G., Sage, J. R., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Context+fear+learning+in+the+absence+of+the+hippocampus.">Context fear learning in the absence of the hippocampus.</a> J. Neurosci. 26, 5484-5491 (2006).</li><li>Reijmers, L. G., Perkins, B. L., Matsuo, N., Mayford, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localization+of+a+stable+neural+correlate+of+associative+memory.">Localization of a stable neural correlate of associative memory.</a> Science. 317, 1230-1233 (2007).</li><li>Huerta, P. T., Sun, L. D., Wilson, M. A., Tonegawa, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formation+of+temporal+memory+requires+NMDA+receptors+within+CA1+pyramidal+neurons.">Formation of temporal memory requires NMDA receptors within CA1 pyramidal neurons.</a> Neuron. 25, 473-480 (2000).</li><li>Jacobs, N. S., Cushman, J. D., Fanselow, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+accurate+measurement+of+fear+memory+in+Pavlovian+conditioning:+Resolving+the+baseline+issue.">The accurate measurement of fear memory in Pavlovian conditioning: Resolving the baseline issue.</a> J. Neurosci. Methods. 190, 235-239 (2010).</li></ol>

作者什么都没有透露。

已经有已经阐明了神经电路，跟踪的基础恐惧条件反射的几项研究。追溯恐惧条件，相信涉及的12-14海马CA1区。也有证据表明，内侧前额叶皮质（mPFC的）起着跟踪眼睛一眨不眨空调15大的作用，而内侧前额叶皮质已被发现参与了一丝恐惧条件。一项研究发现，内侧前额叶皮质的神经元在跟踪期间提供持续的活性，从 ​​而提供一个可以追踪间隔17期间维持内存的结构。 ...

在恐惧条件反射的中性刺激（NS）是搭配厌恶无条件刺激（美国）。的NS通常是一个音，并通过反复配对与美国成为条件刺激（CS）。然后将CS可以引起一个条件反应（CR），如冷冻，在没有厌恶美国。常用的恐惧制约的协议是延迟调节。在这个协议中的NS和美国的发病是相连或与在刺激呈现一些重叠。尽管延迟恐惧制约是最常用的类型时空关联空调之一，还有其他一些类型的关联调理时间安排：同步调理，调理落后，并跟踪调节1。在跟踪恐惧条件存在着造成了“跟踪”期间几秒钟的NS和美国之间的经济刺激，无间隔。 一些研究已经报道赤字在痕迹恐惧调理S当神经病变产生的结构，输入到海马区2-5或当药物制剂被用来阻止受体功能的海马。病变海马导致在痕迹条件和上下文调理缺陷，但不影响延迟的恐惧条件8。有几个好处使用痕迹恐惧条件。怕调理协议可以实现在一个为期三天的测试期，并允许海马依赖的记忆，是不是空间相关的。跟踪恐惧条件可以作为一个补充试验的Morris水迷宫，新物体识别测试中，或在调查的海马依赖性记忆的其他迷宫测试。

<table border="0" cellpadding="0" cellspacing="0" width="741">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">FreezeFrame</td>
			<td style="width:143px;">Coulbourn</td>
			<td style="width:119px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">30% Isopropanol</td>
			<td style="width:143px;"></td>
			<td style="width:119px;"></td>
			<td style="width:263px;">Purchase 90% isopropanol and dilute it down to 30%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">70% Ethanol</td>
			<td style="width:143px;"></td>
			<td style="width:119px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Amp-meter</td>
			<td style="width:143px;">Med-Associates</td>
			<td style="width:119px;">ENV-420</td>
			<td style="width:263px;">Windows XP, Vista, and 7 Compatible (32-bit only)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Digital Sound Level Meter</td>
			<td style="width:143px;"></td>
			<td style="width:119px;">33-2055</td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Vanilla Extract</td>
			<td style="width:143px;">McCormick Pure Vanilla Extract</td>
			<td style="width:119px;"></td>
			<td style="width:263px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sticky Notes</td>
			<td style="width:143px;">Post-it</td>
			<td style="width:119px;"></td>
			<td style="width:263px;">3X3 inch</td>
		</tr>
	</tbody>
</table>

trace fear conditioning in mice

在这个实验中，我们提出了一个方法来衡量学习和记忆。在这里介绍的一丝恐惧条件的协议，存在一个中性刺激和无条件刺激之间的五个配对。有分隔各个空调审判20秒跟踪期。翌日冻结呈现条件刺激（CS）和跟踪一段期间测量。在第三天有一个8分钟的试验来测量情景记忆。代表性的结果是从出席与厌恶无条件刺激（电击），相对于所接收的音调演示，而不无条件刺激小鼠的小鼠。一丝恐惧条件已被成功地用于检测细微的学习和记忆障碍和增强功能未发现与其他恐惧条件反射的方法小鼠。这种类型的恐惧条件的被认为是依赖于内侧前额叶皮质和海马之间的连接。一个电流争论是该方法是否被认为是杏仁核无关。因此，其他的恐惧条件测试是需要检查杏仁核依赖性学习和记忆的效果，如通过延迟恐惧条件反射。

对于有代表性的结果，我们提出从较收到的中性刺激，但没有收到非条件刺激（无震动状态）小鼠收到的中性刺激配对与非条件刺激（休克状态）的控制C57BL/6J成年小鼠的数据。它运行这个条件时，首先设置此行为测试，以确定是否该协议已经被正确执行是非常重要的。 图1中的数据代表了一丝恐惧条件测试C57BL/6J小鼠的训练日。它通常没有必要在训练天在所有条...

Watch this Scientific Journal Video about Trace Fear Conditioning in Mice at JoVE.com

Trace Fear Conditioning in Mice

在接下来的实验中，我们描述了一个协议，用于在小鼠体内微量恐惧条件。这种类型的联想记忆，包括分隔中性刺激和非条件刺激一丝时期。

跟踪恐惧条件小鼠

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings ...

Research

JoVE Journal

Behavior

32.5K 次观看.  Baylor University. 在接下来的实验中，我们描述了一个协议，用于在小鼠体内微量恐惧条件。这种类型的联想记忆，包括分隔中性刺激和非条件刺激一丝时期。 

视频: Trace Fear Conditioning in Mice

Contextual and Cued Fear Conditioning Test Using a Video Analyzing System in Mice

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association between environmental cues and aversive experiences. In this test, mice are placed into a conditioning chamber and are given parings of a conditioned stimulus (an auditory cue) and an aversive unconditioned stimulus (an electric footshock). After a delay time, the mice are exposed to the same conditioning chamber and a differently shaped chamber with presentation of the auditory cue. Freezing behavior during the test is measured as an index of fear memory. To analyze the behavior automatically, we have developed a video analyzing system using the ImageFZ application software program, which is available as a free download at http://www.mouse-phenotype.org/. Here, to show the details of our protocol, we demonstrate our procedure for the contextual and cued fear conditioning test in C57BL/6J mice using the ImageFZ system. In addition, we validated our protocol and the video analyzing system performance by comparing freezing time measured by the ImageFZ system or a photobeam-based computer measurement system with that scored by a human observer. As shown in our representative results, the data obtained by ImageFZ were similar to those analyzed by a human observer, indicating that the behavioral analysis using the ImageFZ system is highly reliable. The present movie article provides detailed information regarding the test procedures and will promote understanding of the experimental situation.

This article presents a protocol for a contextual and cued fear conditioning test using a video analyzing system to assess fear learning and memory in mice.

背景和线索恐惧制约试验小鼠中使用视频分析系统

The contextual and cued fear conditioning test is one of the behavioral tests that assesses the ability of mice to learn and remember an association ...

科研

行为学

Protocol for Studying Extinction of Conditioned Fear in Naturally Cycling Female Rats

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that neural mechanisms for certain behavioral tasks and response behaviors are different in females and males. Using females in research studies can represent a challenge because of the variation of gonadal hormones during their estrous cycle. This protocol describes well-established procedures that are useful in investigating the role of estrogen in fear extinction memory consolidation in female rats. Phase of the estrous cycle and exogenous estrogen administration prior to extinction training can influence extinction recall 24 hr later. The vaginal swabbing technique for estrous phase identification described here aids the examination and manipulation of naturally cycling gonadal hormones. The use of this basic rodent model may further delineate the mechanisms by which estrogen can modulate fear extinction memory in females.

Gonadal hormones such as estrogen modulate memory formation in a number of experimental paradigms including fear extinction memory. This protocol describes a set of methods for investigating the influence of gonadal hormones specifically during extinction in naturally cycling females, including estrous cycle monitoring and exogenous estrogen administration.

协议在骑自行车自然雌性大鼠学习条件性恐惧灭绝

Extinction of conditioned fear has been extensively studied in male rodents. Recently, there have been an increasing number of studies indicating that ...

Quantifying Social Motivation in Mice Using Operant Conditioning

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered shuttle boxes were equipped with two operant levers (left and right) and a food receptacle in one chamber, which was then divided from the second chamber by an automated guillotine door covered by a wire grid. Different stimulus mice, rotated across testing days, served as a social stimulus behind the wire grid, and were only visible following the opening of the guillotine door. Test mice were trained to lever press in order to open the door and gain access to the stimulus partner for 15 sec. The number of lever presses required to obtain the social reward progressively increased on a fixed schedule of 3. Testing sessions ended after test mice stopped lever pressing for 5 consecutive minutes. The last reinforced ratio or breakpoint can be used as a quantitative measure of social motivation. For the second paradigm, test mice were trained to discriminate between left and right lever presses in order to obtain either a food reward or the social reward. Mice were rewarded for every 3 presses of each respective lever. The number of food and social rewards can be compared as a measurement of the value placed upon each reward. The ratio of each reward type can also be compared between mouse strains and the change in this ratio can be monitored within testing sessions to measure satiation with a given reward type. Both of these operant conditioning paradigms are highly useful for the quantification of social motivation in mouse models of autism and other disorders of social behavior.

This article describes a new protocol for the quantitative measurement of social motivation in mice using operant conditioning for a social reward. The protocol is useful for the measurement of social motivation in mouse models of autism and other disorders of social behavior.

量化的社会动机小鼠使用操作条件反射

In this protocol, social motivation is measured in mice through a pair of operant conditioning paradigms. To conduct the experiments, two-chambered ...

Whisker-signaled Eyeblink Classical Conditioning in Head-fixed Mice

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive repertoire of scientific techniques available to study learning and memory at the cellular level, it is ideal to have a stable cranial platform. Because mice do not readily tolerate restraint, they are usually trained while moving about freely in a chamber. Conditioned stimulus (CS) and unconditioned stimulus (US) information are delivered and eyeblink responses recorded via a tether connected to the mouse's head. In the head-fixed apparatus presented here, mice are allowed to run as they desire while their heads are secured to facilitate experimentation. Reliable conditioning of the eyeblink response is obtained with this training apparatus, which allows for the delivery of whisker stimulation as the CS, a periorbital electrical shock as the US, and analysis of electromyographic (EMG) activity from the eyelid to detect blink responses.

The preparation presented here for whisker-signaled eyeblink conditioning in head-fixed mice precisely stimulates specific whiskers while allowing mice to ambulate on a cylindrical treadmill. A whisker stimulation conditioned stimulus (CS) paired with a periorbital shock unconditioned stimulus (US) results in reliable associative learning on this apparatus.

在头固定小鼠胡须信号眨眼经典条件反射

Eyeblink conditioning is a common paradigm for investigating the neural mechanisms underlying learning and memory. To better utilize the extensive ...

Reducing State Anxiety Using Working Memory Maintenance

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working Memory (WM) and the threat of shock paradigms. In the Sternberg WM paradigm, subjects are required to maintain a series of letters in the WM for a brief interval and respond by identifying whether the position of a given letter in the series matches a numerical prompt. In the threat of shock paradigm, subjects are exposed to alternating blocks where they are either at risk of receiving unpredictable presentations of a mild electric shock or are safe from the shock. Anxiety is probed throughout the safe and threat blocks using the acoustic startle reflex, which is potentiated under threat (Anxiety-Potentiated Startle (APS)). By conducting the Sternberg WM paradigm during the threat of shock and probing the startle response during either the WM maintenance interval or the intertrial interval, it is possible to determine the effect of WM maintenance on APS.

This protocol demonstrates how to measure anxiety-potentiated startle during the Sternberg Working Memory paradigm.

使用工作记忆维护降低状态焦虑

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working ...

Classical Short-Delay Eyeblink Conditioning in One-Year-Old Children

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a puff of air to the cornea). Eyeblink conditioning is often used experimentally to detect abnormalities in cerebellar-dependent learning and memory that underlies this type of associative learning. While experiments in adults and older children are relatively simple to administer using commercial equipment, eyeblink conditioning in infants is more challenging due to their poor compliance, which makes correct positioning of the equipment difficult. To achieve conditioning in one-year-old infants, a custom-made or an adapted commercial system can be used to deliver the air puff to the infant's cornea. The main challenge lies in successfully detecting and classifying the behavioral responses. We report that automated blink detection methods are unreliable in this population, and that conditioning experiments should be analyzed using frame-by-frame analysis of supplementary video camera recordings. This method can be applied to study developmental changes in eyeblink conditioning and to examine whether this paradigm can detect children with neurological disorders.

This protocol describes an eyeblink conditioning paradigm appropriate for experiments with one-year-old infants. Commercial or custom-made equipment can be used to deliver the stimuli, and data collection and analysis should be performed on the video recordings.

经典短时滞上只是一眨眼在一岁儿童中的调节作用

Classical eyeblink conditioning (EBC) refers to the learned association between a conditioned stimulus (an auditory tone) and an unconditioned stimulus (a ...

Stress-Enhanced Fear Learning, a Robust Rodent Model of Post-Traumatic Stress Disorder

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders such as post-traumatic stress disorder (PTSD). To understand the biological mechanisms of fear dysregulation in PTSD, it is important to start with a valid animal model of the disorder. This protocol describes the methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of PTSD, in both rats and mice. SEFL was developed to recapitulate critical aspects of PTSD, including long-term sensitization of fear learning caused by an acute stressor. SEFL uses aspects of Pavlovian fear conditioning but produces a distinct and robust sensitized fear response far greater than normal conditional fear responses. The trauma procedure involves placing a rodent in a conditioning chamber and administering 15 unsignaled shocks randomly distributed over 90 minutes (for rat experiments; for mouse experiments, 10 unsignaled shocks randomly distributed over 60 minutes are used). On day 2, rodents are placed in a novel conditioning context where they receive a single shock; then, on day 3 they are placed back in the same context as on day 2 and tested for changes in freezing levels. Rodents that previously received the trauma display enhanced levels of freezing on the test day compared to those that received no shocks on the first day. Thus, with this model, a single highly stressful experience (the trauma) produces extreme fear of the stimuli associated with the traumatic event.

Here we describe the detailed methodology required to conduct stress-enhanced fear learning (SEFL) experiments, a preclinical model of post-traumatic stress disorder, in both rats and mice. The model utilizes aspects of Pavlovian fear conditioning and freezing as an index of enhanced fear in rodents.

压力增强恐惧学习--创伤后应激障碍的健壮啮齿动物模型

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders ...

Visual Classical Conditioning in Wood Ants

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, studies implementing classical conditioning paradigms with harnessed insects have been important for investigating the exact cues that individuals learn and the neural mechanisms underlying learning and memory formation. Here we present a protocol for evoking visual associative learning in wood ants through classical conditioning. In this paradigm, ants are harnessed and presented with a visual cue (a blue cardboard), the conditional stimulus (CS), paired with an appetitive sugar reward, the unconditional stimulus (US). Ants perform a Maxilla-Labium Extension Reflex (MaLER), the unconditional response (UR), which can be used as a readout for learning. Training consists of 10 trials, separated by a 5-minute intertrial interval (ITI). Ants are also tested for memory retention 10 minutes or 1 hour after training. This protocol has the potential to allow researchers to analyze, in a precise and controlled manner, the details of visual memory formation and the neural basis of learning and memory formation in wood ants.

We present a protocol for the classical conditioning of harnessed ants that permits researchers to study visual learning and memory formation at a level of analysis not possible with freely moving individuals.

木蚁视觉经典调理

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, ...

Fear Incubation Using an Extended Fear-Conditioning Protocol for Rats

Emotional memory has been primarily studied with fear-conditioning paradigms. Fear conditioning is a form of learning through which individuals learn the relationships between aversive events and otherwise neutral stimuli. The most-widely utilized procedures for studying emotional memories entail fear conditioning in rats. In these tasks, the unconditioned stimulus (US) is a footshock presented once or several times across single or several sessions, and the conditioned response (CR) is freezing. In a version of these procedures, called cued fear conditioning, a tone (conditioned stimulus, CS) is paired with footshocks (US) during the training phase. During the first test, animals are exposed to the same context in which training took place, and freezing responses are tested in the absence of footshocks and tones (i.e., a context test). During the second test, freezing is measured when the context is changed (e.g., by manipulating the smell and walls of the experimental chamber) and the tone is presented in the absence of footshocks (i.e., a cue test). Most cued fear conditioning procedures entail few tone-shock pairings (e.g., 1-3 trials in a single session). There is a growing interest in less common versions involving an extensive number of pairings (i.e., overtraining) related to the long-lasting effect called fear incubation (i.e., fear responses increase over time without further exposure to aversive events or conditioned stimuli). Extended fear-conditioning tasks have been key to the understanding of fear incubation&#8217;s behavioral and neurobiological aspects, including its relationship with other psychological phenomena (e.g., post-traumatic stress disorder). Here, we describe an extended fear-conditioning protocol that produces overtraining and fear incubation in rats. This protocol entails a single training session with 25 tone-shock pairings (i.e., overtraining) and a comparison of conditioned freezing responses during context and cue tests 48 h (short-term) and 6 weeks (long-term) after training.

We describe an extended fear-conditioning protocol that produces overtraining and fear incubation in rats. This protocol entails a single training session with 25 tone-shock pairings (i.e., overtraining) and a comparison of conditioned freezing responses during context and cue tests 48 h (short-term) and 6 weeks (long-term) after training.

对大鼠使用扩展的恐惧调节协议进行恐惧孵化

Emotional memory has been primarily studied with fear-conditioning paradigms. Fear conditioning is a form of learning through which individuals learn the ...

Operant Conditioning Task to Measure Song Preference in Zebra Finches

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

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

操作调节任务，以测量斑马芬奇中的歌曲首选项

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