这项研究已被授予NIHM MH091816 R01和R21的MH084043到TDG支持。

1。材料与方法 1.1。水箱 （30厘米高x 20厘米直径）的圆柱型坦克所需的小鼠强迫游泳实验（FST）的在我们的实验室，是透明的有机玻璃建造，这种材料能够承受的坦克和事故比玻璃更好的频繁流动。从底部水位15厘米，并应标明坦克，以确保水的体积是在小鼠一致。理想情况下，坦克的数量应该至少两次一次测试小鼠的数量很多，而第一套使用的是第二个水箱设置，使可充满。应选择的方式，老鼠将无法触摸到水箱底部，要么用自己的脚或它们的尾巴游泳测试期间，坦克的尺寸。坦克的高度应足够高，以防止从逃避坦克的老鼠。请注意:罐直径和水深的重要参数都可以调整，以改变对小鼠的行为（对这些问题的详细分析见1-3 ）。 1.2。温度计一个防水红外测温仪是可取的，因为温度的快速测量，减少了所需的时间进行测试。然而，玻璃水银温度计，也将这个任务足够。 1.3。定时器 1.4。录像设备我们使用一台摄像机三脚架支持。由于本次测试通常涉及多个动物在同一时间测试，现场打分将是非常困难的，是不可取的。摄像头应该记录在足够高的分辨率，呈现高质量的画面，行为得分后，将用于。务必确保有足够的记录存储在摄像机前starting的测试。我们使用一台摄像机，记录数字化无机械媒体的使用（如录像带），使数字视频传输。如果有过多的坦克，这可能发生在实验室环境与架空荧光灯照明反射，您可能要使用一个偏光镜片的过滤器与您的相机。 1.5。分频器在我们的实验室中，我们有两个分频器（高35厘米× 22厘米宽× 22厘米深度）。这些都是长方形三面墙壁和背景和坦克之间的分隔，以防止在测试过程中看到对方，并有可能改变他们的行为的小鼠。一组可以是黑色的白化和浅色的动物;另一套可轻深色动物着色，以呈现高对比度。实验者应确保分频器表面不会过度反射，使他们改变摄像机的图像，或使主要ðifferences之间的照度水平。 1.6。白噪声发生器这是需要在实验室环境中，突然大声喧哗，可以听到，可能不惊人死不休动物。噪声发生器会掩盖这种间歇性的令人不安的声音。应选择白噪声发生器的音量水平要高于其他环境和突发噪音。在我们的试验室的环境噪声水平（不带白噪声发生器激活）为60分贝。在坦克被放置的位置激活与白噪声发生器总的噪音水平是70-72分贝。但是，应该指出，这些数字只为例，每个实验室应根据其独特的环境和情况，选择合适的噪音水平。 1.7。干燥的纸张和热灯动物返回自己的家乡笼之前，重要的是要干他们轻轻地用纸巾是很有帮助的使用加热灯（某些暴露温度不超过32℃），以防止体温过低。 2。行为程序<ol><li>整体实验设计应反映具体到您的实验变量之间的适当制衡。例如，在我们的实验中，我们尝试以同样在每个FST会议（即，如果有四个治疗组，每个将在每届会议的代表）代表各组。此外，小鼠的旋转，这样，每个治疗组小鼠被放置在不同的坦克，在每届会议。 </li><li>将摄像头和位置的分隔。相机应尽可能靠近，以获得最高的小鼠可能的解决方案。 </li><li>坦克应填写在室温（23-25​​℃）确定的水平，这是对罐壁标记的自来水。如果您的设备没有恒定的热/冷WA之三，你可能要准备热水和/或冰，从而迅速地把水到合适的温度。检查水的温度，用红外测温仪。另外，如果冷热水的温度恒定在您的设施，你可以借鉴的坦克两个标记 - 一个水平热水和冷水此外第二个标记 - 亲近正确决赛水温迅速。 </li><li>开始白噪声发生器，如果正在使用，小鼠之前推出的试验室。白噪声水平只应足以屏蔽外部噪声。避免以高音量，并确保所有动物相同，白噪声水平。 </li><li>动物带入考场。如果殖民地的动物居住的房间里和试验室相邻或非常接近对方，环境条件相似，在笼的移动的干扰是最小的，然后没有驯化近郊OD将是必要的的。否则，放置在试验室动物的驯化期（一般至少一小时）。如果一个驯化期是必要的，确保驯化动物不会受到影响测试在同一时间在同一房间内的小鼠。请注意，嗅觉和超声波线索可以被放置在同一个房间里的其他动物感觉到。 </li><li>视频录制开始前放入水箱动物。 </li><li>拿住它的尾巴的动物，轻轻地，慢慢地放置在水中。一旦小鼠在水中，缓慢地释放尾部。通常情况下，使用此程序将防止从动物的头部被水淹没。 </li><li>录制的阻挠将减少的顺序放置在坦克的老鼠。当然，这样的顺序，应结合团体和其他要求具体到你的实验设计的制衡决定。 </li><li>一旦所有的小鼠在坦克开始倒计时秒表。对小鼠的一般测试长度是6分钟，在FST。 </li><li>在测试过程中，你是在一个合理的距离从动物，不作任何变动或动物可能注意到的噪声。老鼠可以很容易地浮在水中，但是，如果新菌株的小鼠或一种新的化合物，其效果没有以前的知识测试游泳行为，实验者应监测动物进行更密切的合作。在大鼠，小鼠通常不潜水在FST的，但是在潜水时，鼠标应该从油箱中删除。如果实验者离开小鼠应的事件，鼠标不能保持游泳和漂浮行为，并停止测试，如果有必要通过视频监控室。 </li><li>在6分钟的测试期间结束，停止录音。在我们的实验室，我们在前面的摄像头，标识的动物，在最后一个注意每个记录。使用这种方法时，随后得分记录的个人将不知道动物的身份，因为身份只有在录音结束。这可以防止任何可能出现的问题后有关录音的识别和记录保存。无论正在使用的记录保存策略，它应该清楚地识别动物和组分配知识的测试后得分，还可以防止个别。 </li><li>在同一顺序的尾巴的动物从水中取出，你把它们轻轻擦干干燥纸张和他们回他们homecage。 </li></ol> 3。行为分析<ol><li>福斯特鼠标版本是通常的，从开始到结束，6分钟之久。然而，一般只在最后四分钟的测试分析。这是由于的事实，大多数小鼠在FST的开始非常活跃，而P治疗的otential影响可以被遮蔽在头两分钟。 </li><li>在我们的实验室中，我们上传的视频文件直接从相机到PC，并在PC上做分析。 </li><li>在行为分析，测量每个小鼠花费的时间，移动。总额减去流动性的时间从240秒的测试时间，然后不动时间表示。虽然可以直接测量不动时间，在我们的实验室中，我们发现更容易发现和措施，而不是缺乏这种流动活跃运动。 </li><li>行为分析和观察员之间的可变性最大的来源通常在FST中最重要的是，作为真正的流动计算运动的正确识别。我们在FST流动性的业务定义是必要以外的任何动作，以平衡身体和头部保持在水面 之上 4。小鼠一般漂浮在水中很容易，但是他们仍然体现小的动作，以平衡自己的身体和头保持在水面之上。这些行为是不是企图逃跑，不应被视为取得了流动性。此外，单回合后的流动性，即使基本上不动，小鼠仍然可以漂在水中的势头。这些运动不应该拿下的流动性。 </li><li>在我们的实验室中，我们使用屏幕上的秒表时间测量软件（Xnote秒表，dnSoft研究小组）。在屏幕上使用两个独立的秒表。第一个秒表倒计时240秒和警报行为分析期结束时的观察员。第二个秒表措施所花费的时间移动。有些秒表软件已分配键启动和停止功能，使屏幕上的秒表，可以通过键盘控制的能力。在我们的实验室，而不是一个普通的键盘，我们使用的输入设备，俗称一个"手柄"控制秒表。 </li><li>当使用一台PC量化不动，如果有多个鼠标经过测试，并在屏幕上，它是一个好主意，以弥补其他动物（您可以使用另一个程序窗口或物理覆盖与纸屏幕），使他们的动作不会分散的观察员。 </li><li>如果使用屏幕上的秒表是，一定涵盖所有问题，但秒表毫秒小数。这样做的原因是为了防止在观察员的偏见，同时仍然允许的能力，以确定是否手表运行或者没有运行。由于观察员，而盲目的动物组分配，将有一个在小鼠体内的流动性水平的总体思路，有可能会发生一些偏差，如果她是允许流动时间的总金额为特定的鼠标经过分析会议之前完成。覆盖秒表，她只知道秒表是否打开或关闭在任何时候，但西港岛线升不知道总的时间，因此不能带任何偏见的影响。 </li><li>应当进行跨观察员可靠性测试才开始收集数据从实验动物为每一个新的观察员。在我们的实验室，每一个新的观察员第一只手表一位经验丰富的观察员得分。新的观察员收益的信心，以区别于万马齐喑流动后，他们再得分，与经验丰富的观察员观看，并指出任何错误。一旦成功完成这一阶段，新的观察员将分析具体的培训目的，我们在我们的实验室保持福斯特的视频设置。只有在一个高层次的跨观察员相关与经验丰富的观察员获得的调查员开始，在实际的实验分析FST的影片。我们从这些培训分析的数据进行归档，以构成一个内部标准，以供将来使用的实验室。我们观察的方式在菌株之间的差异，他们表达MOBIlity（不动）的行为，男女之间的平均不动时间。一个新的应变，性别，或转基因小鼠模型在实验室测试时，有必要再次进行这种类型的可靠性分析。 </li></ol> 4。代表性的成果基因不同的近交系和远交小鼠品系之间有显着差异，在其基准万马齐喑和响应一个特定的药物5-11方面。例如，我们发现差的抗抑郁反应锂，在小鼠品系的面板（ 图 1 ）5。本实验的实验的详细资料载于能等，2011年5。 <img src="/files/ftp_upload/3638/3638fig1.jpg" alt="图1" /> 图1。强迫游泳测试的不动时间（秒），5小时后，在不同的自交系和OU一个单一的IP注射生理盐水，200，300或400毫克/公斤tbred小鼠品系。 *:P &lt;0.05，**:P &lt;0.01，***:P &lt;0.001表示一个显著的差异相比，生理盐水组，Dunnett 的事后检验。数据表示为平均值± SEM。每一株每组动物的数量是6-8（复制图 5） 。 并非所有的小鼠品系适合在FST。一些菌株，如黑瑞士，瑞士，NIH和FVB / NJ显示控制的情况下很少或几乎没有不动，因此代表了地板的效果（图1）5。缺乏基准不动，有效地防止了检测实验操作的抗抑郁效果。虽然非常罕见的，这也是可能的，一些小鼠品系的行为可能aberrantly和潜水坦克在测试过程中，即使他们可以浮动。这样一个应变DBA/1OlaHsd（本实验室未发表的观察）。这种菌株不适合在FST。由于这种潜水的风险，但SMaL公司L，在FST或窝藏一种新型的基因操纵鼠标，过去一直没有被测试的一个新的应变测试时，必须仔细观察抢救，如果他们在从事可能有害行为的小鼠的初步试验。 在这里描述的实验设计，多种动物（最多五个）在同一时间测试。虽然我们所使用的分隔看到对方在测试过程中防止老鼠，白噪声发生器抑制发声的发声，我们的设置不防止被传输的所有超声波或嗅觉线索。虽然不太可能，因为测试的性质，这些可能会影响小鼠的行为。这个问题的一个解决方案，将单独进行测试的动物。但是，这种方法有其自身的问题。例如，常见的是，在每个会话中测试的动物来自同一homecage。这让实验变量的随机和制衡。测试小鼠个人LY意味着在从homecage时间删除一个鼠标。这将导致反复强调，在笼子之间留下的其他扰乱社会等级。与测试的另一个问题单的时间限制。一次测试一个鼠标，将扩展到许多小时的实验，在小鼠中，昼夜周期的不同时间测试的情况。这一天的效果可能会造成混淆时间。研究人员应该牢记这些问题，而他们的实验设计。

<ol>
	<li>Hasco&#233;t, M., Bourin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> In Mood and Anxiety Related Phenotypes in Mice. 42, 85-118 (2009).</li><li>Sunal, R., G&#252;m&#252;&#231;el, a. B., Kayaalp, S. O. <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+changes+in+swimming+area+on+results+of+"behavioral+despair+test".">Effect of changes in swimming area on results of "behavioral despair test".</a> Pharmacology Biochemistry and Behavior. 49, 891-896 (1994).</li><li>Petit-Demouliere, B., Chenu, F., Bourin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Forced+swimming+test+in+mice:+a+review+of+antidepressant+activity.">Forced swimming test in mice: a review of antidepressant activity.</a> Psychopharmacology. (Berl). 177, 245-255 (2005).</li><li>Cryan, J. F., Markou, A., Lucki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+antidepressant+activity+in+rodents:+recent+developments+and+future+needs.">Assessing antidepressant activity in rodents: recent developments and future needs.</a> Trends in Pharmacological Sciences. 23, 238-245 (2002).</li><li>Can, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antidepressant-like+responses+to+lithium+in+genetically+diverse+mouse+strains.">Antidepressant-like responses to lithium in genetically diverse mouse strains.</a> Genes, Brain and Behavior. 10, 434-443 (2011).</li><li>Lucki, I., Dalvi, A., Mayorga, 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=Sensitivity+to+the+effects+of+pharmacologically+selective+antidepressants+in+different+strains+of+mice.">Sensitivity to the effects of pharmacologically selective antidepressants in different strains of mice.</a> Psychopharmacology. (Berl). 155, 315-322 (2001).</li><li>David, D. J., Renard, C. E., Jolliet, P., Hascoet, M., Bourin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antidepressant-like+effects+in+various+mice+strains+in+the+forced+swimming+test.">Antidepressant-like effects in various mice strains in the forced swimming test.</a> Psychopharmacology (Berl). 166, 373-382 (2003).</li><li>Bai, F., Li, X., Clay, M., Lindstrom, T., Skolnick, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-+and+interstrain+differences+in+models+of+"behavioral+despair".">Intra- and interstrain differences in models of "behavioral despair".</a> Pharmacol. Biochem. Behav. 70, 187-192 (2001).</li><li>Guzzetti, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strain+differences+in+paroxetine-induced+reduction+of+immobility+time+in+the+forced+swimming+test+in+mice:+Role+of+serotonin.">Strain differences in paroxetine-induced reduction of immobility time in the forced swimming test in mice: Role of serotonin.</a> European Journal of Pharmacology. 594, 117-124 (2008).</li><li>Cervo, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genotype-dependent+activity+of+tryptophan+hydroxylase-2+determines+the+response+to+citalopram+in+a+mouse+model+of+depression.">Genotype-dependent activity of tryptophan hydroxylase-2 determines the response to citalopram in a mouse model of depression.</a> J. Neurosci. 25, 8165-8172 (2005).</li><li>Jiao, J., Nitzke, A., Doukas, D., Seiglie, M., Dulawa, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antidepressant+response+to+chronic+citalopram+treatment+in+eight+inbred+mouse+strains.">Antidepressant response to chronic citalopram treatment in eight inbred mouse strains.</a> Psychopharmacology. 213, 509-520 (2011).</li><li>Porsolt, R. D., Pichon, M. L. e., Jalfre, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Depression:+a+new+animal+model+sensitive+to+antidepressant+treatments.">Depression: a new animal model sensitive to antidepressant treatments.</a> Nature. 266, 730-732 (1977).</li><li>Porsolt, R. D., Bertin, A., Jalfre, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+despair+in+mice:+a+primary+screening+test+for+antidepressants.">Behavioral despair in mice: a primary screening test for antidepressants.</a> Arch. Int. Pharmacodyn. Ther. 229, 327-336 (1977).</li><li>Porsolt, R. D., Bertin, A., Jalfre, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term="Behavioural+despair"+in+rats+and+mice:+strain+differences+and+the+effects+of+imipramine.">"Behavioural despair" in rats and mice: strain differences and the effects of imipramine.</a> Eur. J. Pharmacol. 51, 291-294 (1978).</li><li>Mineur, Y. S., Belzung, C., Crusio, W. E. <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+unpredictable+chronic+mild+stress+on+anxiety+and+depression-like+behavior+in+mice.">Effects of unpredictable chronic mild stress on anxiety and depression-like behavior in mice.</a> Behav. Brain. Res. 175, 43-50 (2006).</li><li>Millstein, R. A., Holmes, A. <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+repeated+maternal+separation+on+anxiety-+and+depression-related+phenotypes+in+different+mouse+strains.">Effects of repeated maternal separation on anxiety- and depression-related phenotypes in different mouse strains.</a> Neuroscience &#38; Biobehavioral Reviews. 31, 3-17 (2007).</li><li>Cryan, J. F., Valentino, R. J., Lucki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+substrates+underlying+the+behavioral+effects+of+antidepressants+using+the+modified+rat+forced+swimming+test.">Assessing substrates underlying the behavioral effects of antidepressants using the modified rat forced swimming test.</a> Neuroscience &#38; Biobehavioral Reviews. 29, 547-569 (2005).</li><li>Crowley, J. J., Jones, O. '. L. e. a. r. y., F, O., Lucki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+tests+for+measuring+the+effects+of+antidepressants+in+mice.">Automated tests for measuring the effects of antidepressants in mice.</a> Pharmacology Biochemistry and Behavior. 78, 269-274 (2004).</li><li>Kurtuncu, M., Luka, L. J., Dimitrijevic, N., Uz, T., Manev, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reliability+assessment+of+an+automated+forced+swim+test+device+using+two+mouse+strains.">Reliability assessment of an automated forced swim test device using two mouse strains.</a> Journal of Neuroscience Methods. 149, 26-30 (2005).</li><li>Hayashi, E., Shimamura, M., Kuratani, K., Kinoshita, M., Hara, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+experimental+system+capturing+three+behavioral+components+during+murine+forced+swim+test.">Automated experimental system capturing three behavioral components during murine forced swim test.</a> Life Sciences. 88, 411-417 (2011).</li><li>Cryan, J., Page, M., Lucki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+behavioral+effects+of+the+antidepressants+reboxetine,+fluoxetine,+and+moclobemide+in+a+modified+forced+swim+test+following+chronic+treatment.">Differential behavioral effects of the antidepressants reboxetine, fluoxetine, and moclobemide in a modified forced swim test following chronic treatment.</a> Psychopharmacology. 182, 335-344 (2005).</li><li>Gould, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beta-catenin+overexpression+in+the+mouse+brain+phenocopies+lithium-sensitive+behaviors.">Beta-catenin overexpression in the mouse brain phenocopies lithium-sensitive behaviors.</a> Neuropsychopharmacology. 32, 2173-2183 (2007).</li><li>Can, A., Grahame, N. J., Gould, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Affect-related+related+behaviors+in+mice+selectively+bred+for+high+and+low+voluntary+alcohol+consumption.">Affect-related related behaviors in mice selectively bred for high and low voluntary alcohol consumption.</a> Behav. Genet. , (2011).</li><li>McKinney, W. T., Bunney, W. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Model+of+Depression:+I+Review+of+Evidence:+Implications+for+Research..">Animal Model of Depression: I Review of Evidence: Implications for Research..</a> Arch. Gen. Psychiatry. 21, 240-248 (1969).</li><li>Willner, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+validity+of+animal+models+of+depression.">The validity of animal models of depression.</a> Psychopharmacology.(Berl). 83, 1-16 (1984).</li><li>Detke, M. J., Lucki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+serotonergic+and+noradrenergic+antidepressants+in+the+rat+forced+swimming+test:+the+effects+of+water+depth.">Detection of serotonergic and noradrenergic antidepressants in the rat forced swimming test: the effects of water depth.</a> Behav. Brain Res. 73, 43-46 (1995).</li><li>Gould, T. D., Dao, D. T., Kovacsics, C. E., Gould, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> In Mood and Anxiety Related Phenotypes in mice: characterization using behavioral tests. 42, (2009).</li><li>Bourin, M., Fiocco, A. J., Clenet, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+valuable+are+animal+models+in+defining+antidepressant+activity.">How valuable are animal models in defining antidepressant activity.</a> Human Psychopharmacology: Clinical and Experimental. 16, 9-21 (2001).</li></ol>

作者宣称没有利益冲突。

FST的（有时也被称为Porsolt游泳测试）是开发的第一对大鼠，小鼠Porsolt和同事 12,13，然后修改。除了 ​​上述的协议，在我们的实验室中成功，主要是微妙的测试修改已公布（见Hascoët和Bourin 为一个完整的审查1）。它是一种常见的测试，使用的抗抑郁药物和各种行为和神经生物学基础和临床研究3,14-16操作的影响疗效评价。它被形容为渲染"行为绝望"是诱导情况，即动物?...

<ul><li>水箱</li><li>温度计</li><li>定时器</li><li>摄像机</li><li>白噪声发生器（可选） </li><li>干燥纸</li></ul>

the mouse forced swim test

强迫游泳试验是一种啮齿动物行为学测试，用于评价抗抑郁药物，抗抑郁疗效的新化合物，实验操作，目的是在渲染或防止国家像抑郁。小鼠被放置在透明水箱，装满水，并测量其逃避有关流动性的行为是负有不可推卸的。强迫游泳测试是直接进行可靠的，它需要最少的专用设备。强迫游泳试验的成功实施需要遵守一定的程序细节，并尽量减少不必要的压力的老鼠。在协议的描述和影随行中，我们解释了如何进行鼠标本次测试的版本，强调对潜在的隐患，可能危害结果的解释，以及如何避免它们。此外，我们解释了如何在测试中表现出来的行为评估。

Watch this Scientific Journal Video about The Mouse Forced Swim Test at JoVE.com

The Mouse Forced Swim Test

强迫游泳测试是在啮齿类动物实验的方法来评估潜在的抗抑郁疗效验证。实验动物被放置在一缸水和逃避相关的流动性行为，量化。共同的这个测试鼠标版的程序进行了阐述。

小鼠强迫游泳实验

The forced swim test is a rodent behavioral test used for evaluation of antidepressant drugs, antidepressant efficacy of new compounds, and experimental ...

Research

JoVE Journal

Neuroscience

104.6K 次观看.  University of Maryland School of Medicine. 强迫游泳测试是在啮齿类动物实验的方法来评估潜在的抗抑郁疗效验证。实验动物被放置在一缸水和逃避相关的流动性行为，量化。共同的这个测试鼠标版的程序进行了阐述。

视频: The Mouse Forced Swim Test

Multiple-mouse Neuroanatomical Magnetic Resonance Imaging

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and evaluating mouse models of human disease. MRI is an excellent modality for investigating genetically altered animals. It is capable of whole brain coverage, can be used in vivo, and provides multiple contrast mechanisms for investigating different aspects of neuranatomy and physiology. The advent of high-field scanners along with the ability to scan multiple mice simultaneously allows for rapid phenotyping of novel mutations.
Effective mouse MRI studies require attention to many aspects of experiment design. In this article, we will describe general methods to acquire quality images for mouse phenotyping using a system that images mice concurrently in shielded transmit/receive radio frequency (RF) coils in a common magnet (Bock et al., 2003). We focus particularly on anatomical phenotyping, an important and accessible application that has shown a high potential for impact in many mouse models at our imaging centre. Before we can provide the detailed steps to acquire such images, there are important practical considerations for both in vivo brain imaging (Dazai et al., 2004) and ex vivo brain imaging (Spring et al., 2007) that should be noted. These are discussed below.

Magnetic resonance imaging (MRI) has become an increasingly popular tool for examining the phenotype of genetically altered mice. This article illustrates the methods necessary to achieve high-throughput phenotyping of genetically altered mice using multiple-mouse MRI.

多鼠神经解剖学磁共振成像

The field of mouse phenotyping with magnetic resonance imaging (MRI) is rapidly growing, motivated by the need for improved tools for characterizing and ...

科研

神经科学

Detection of Neuritic Plaques in Alzheimer's Disease Mouse Model

Alzheimer's disease (AD) is the most common neurodegenerative disorder leading to dementia. Neuritic plaque formation is one of the pathological hallmarks of Alzheimer's disease. The central component of neuritic plaques is a small filamentous protein called amyloid &beta; protein (A&beta;)1, which is derived from sequential proteolytic cleavage of the beta-amyloid precursor protein (APP) by &beta;-secretase and &gamma;-secretase. The amyloid hypothesis entails that A&gamma;-containing plaques as the underlying toxic mechanism in AD pathology2. The postmortem analysis of the presence of neuritic plaque confirms the diagnosis of AD. To further our understanding of A&gamma; neurobiology in AD pathogenesis, various mouse strains expressing AD-related mutations in the human APP genes were generated. Depending on the severity of the disease, these mice will develop neuritic plaques at different ages. These mice serve as invaluable tools for studying the pathogenesis and drug development that could affect the APP processing pathway and neuritic plaque formation. In this protocol, we employ an immunohistochemical method for specific detection of neuritic plaques in AD model mice. We will specifically discuss the preparation from extracting the half brain, paraformaldehyde fixation, cryosectioning, and two methods to detect neurotic plaques in AD transgenic mice: immunohistochemical detection using the ABC and DAB method and fluorescent detection using thiofalvin S staining method.

One of the pathological characteristics of AD is the formation of Amyloid &#x03B2; protein positive neuritic plaques. In this protocol we describe two methods to detect neuritic plaques in transgenic AD model mice: immunohistochemical detection using the ABC and DAB method and fluorescent detection using thioflavin S staining method.

在阿尔茨海默氏症小鼠模型神经炎斑检测

Alzheimer's disease (AD) is the most common neurodegenerative disorder leading to dementia. Neuritic plaque formation is one of the pathological hallmarks ...

T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in rodents. In our laboratory, we evaluated the two types of memory in more than 30 strains of genetically engineered mice using the automated version of this apparatus. Here, we present the modified T-maze apparatus operated by a computer with a video-tracking system and our protocols in a movie format. The T-maze apparatus consists of runways partitioned off by sliding doors that can automatically open downward, each with a start box, a T-shaped alley, two boxes with automatic pellet dispensers at one side of the box, and two L-shaped alleys. Each L-shaped alley is connected to the start box so that mice can return to the start box, which excludes the effects of experimenter handling on mouse behavior. This apparatus also has an advantage that in vivo microdialysis, in vivo electrophysiology, and optogenetics techniques can be performed during T-maze performance because the doors are designed to go down into the floor. In this movie article, we describe T-maze tasks using the automated apparatus and the T-maze performance of &alpha;-CaMKII+/- mice, which are reported to show working memory deficits in the eight-arm radial maze task. Our data indicated that &alpha;-CaMKII+/- mice showed a working memory deficit, but no impairment of reference memory, and are consistent with previous findings using the eight-arm radial maze task, which supports the validity of our protocol. In addition, our data indicate that mutants tended to exhibit reversal learning deficits, suggesting that &alpha;-CaMKII deficiency causes reduced behavioral flexibility. Thus, the T-maze test using the modified automatic apparatus is useful for assessing working and reference memory and behavioral flexibility in mice.

This article presents the protocol of T-maze tests using a modified automated apparatus for assessing the learning and memory functions in mice.

T-迷宫强迫交替和评估工作，并参考在小鼠记忆的左，右的歧视任务

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in ...

Tissue Preparation and Immunostaining of Mouse Sensory Nerve Fibers Innervating Skin and Limb Bones

Detection and primary processing of physical, chemical and thermal sensory stimuli by peripheral sensory nerve fibers is key to sensory perception in animals and humans. These peripheral sensory nerve fibers express a plethora of receptors and ion channel proteins which detect and initiate specific sensory stimuli. Methods are available to characterize the electrical properties of peripheral sensory nerve fibers innervating the skin, which can also be utilized to identify the functional expression of specific ion channel proteins in these fibers. However, similar electrophysiological methods are not available (and are also difficult to develop) for the detection of the functional expression of receptors and ion channel proteins in peripheral sensory nerve fibers innervating other visceral organs, including the most challenging tissues such as bone. Moreover, such electrophysiological methods cannot be utilized to determine the expression of non-excitable proteins in peripheral sensory nerve fibers. Therefore, immunostaining of peripheral/visceral tissue samples for sensory nerve fivers provides the best possible way to determine the expression of specific proteins of interest in these nerve fibers. So far, most of the protein expression studies in sensory neurons have utilized immunostaining procedures in sensory ganglia, where the information is limited to the expression of specific proteins in the cell body of specific types or subsets of sensory neurons. Here we report detailed methods/protocols for the preparation of peripheral/visceral tissue samples for immunostaining of peripheral sensory nerve fibers. We specifically detail methods for the preparation of skin or plantar punch biopsy and bone (femur) sections from mice for immunostaining of peripheral sensory nerve fibers. These methods are not only key to the qualitative determination of protein expression in peripheral sensory neurons, but also provide a quantitative assay method for determining changes in protein expression levels in specific types or subsets of sensory fibers, as well as for determining the morphological and/or anatomical changes in the number and density of sensory fibers during various pathological states. Further, these methods are not confined to the staining of only sensory nerve fibers, but can also be used for staining any types of nerve fibers in the skin, bones and other visceral tissue.

Immunocytochemical identification of peripheral sensory nerve fiber subtypes (and detection of protein expression therein) are key to the understanding of molecular mechanisms underlying peripheral sensation. Here we describe methods for preparation of peripheral/visceral tissue samples, such as skin and limb bones, for specific immunostaining of peripheral sensory nerve fibers.

鼠标感觉神经支配的皮肤和四肢骨骼纤维组织的制备及免疫染色

Detection and primary processing of physical, chemical and thermal sensory stimuli by peripheral sensory nerve fibers is key to sensory perception in ...

The Tail Suspension Test

The tail-suspension test is a mouse behavioral test useful in the screening of potential antidepressant drugs, and assessing of other manipulations that are expected to affect depression related behaviors. Mice are suspended by their tails with tape, in such a position that it cannot escape or hold on to nearby surfaces. During this test, typically six minutes in duration, the resulting escape oriented behaviors are quantified. The tail-suspension test is a valuable tool in drug discovery for high-throughput screening of prospective antidepressant compounds. Here, we describe the details required for implementation of this test with additional emphasis on potential problems that may occur and how to avoid them. We also offer a solution to the tail climbing behavior, a common problem that renders this test useless in some mouse strains, such as the widely used C57BL/6. Specifically, we prevent tail climbing behaviors by passing mouse tails through a small plastic cylinder prior to suspension. Finally, we detail how to manually score the behaviors that are manifested in this test.

The tail-suspension test is validated as an experimental procedure to assess antidepressant efficacy of drug treatments in mice. Mice are suspended by their tails for six minutes and escape-related behaviors are assessed. We describe procedures used in conducting the tail suspension test.

悬尾实验

The tail-suspension test is a mouse behavioral test useful in the screening of potential antidepressant drugs, and assessing of other manipulations that ...

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol (TRAP) Assay

Telomerase, a ribonucleoprotein, is responsible for maintaining the telomere length and therefore promoting genomic integrity, proliferation, and lifespan. In addition, telomerase protects the mitochondria from oxidative stress and confers resistance to apoptosis, suggesting its possible importance for the surviving of non-mitotic, highly active cells such as neurons. We previously demonstrated the ability of novel telomerase activators to increase telomerase activity and expression in the various mouse brain regions and to protect motor neurons cells from oxidative stress. These results strengthen the notion that telomerase is involved in the protection of neurons from various lesions. To underline the role of telomerase in the brain, we here compare the activity of telomerase in male and female mouse brain and its dependence on age. TRAP assay is a standard method for detecting telomerase activity in various tissues or cell lines. Here we demonstrate the analysis of telomerase activity in three regions of the mouse brain by non-denaturing protein extraction using CHAPS lysis buffer followed by modification of the standard TRAP assay.
In this 2-step assay, endogenous telomerase elongates a specific telomerase substrate (TS primer) by adding TTAGGG 6 bp repeats (telomerase reaction). The telomerase reaction products are amplified by PCR reaction creating a DNA ladder of 6 bp increments. The analysis of the DNA ladder is made by 4.5% high resolution agarose gel electrophoresis followed by staining with highly sensitive nucleic acid stain.
Compared to the traditional TRAP assay that utilize 32P labeled radioactive dCTP&#39;s for DNA detection and polyacrylamide gel electrophoresis for resolving the DNA ladder, this protocol offers a non-toxic time saving TRAP assay for evaluating telomerase activity in the mouse brain, demonstrating the ability to detect differences in telomerase activity in the various female and male mouse brain region.

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol TRAP Assay

Telomerase is expressed in the neonatal brain and also in distinct regions of the adult brain. We present a non-toxic time saving TRAP assay for the analysis of telomerase activity in various regions of the mouse brain and detection of differences in telomerase activity between male and female mouse brains.

在小鼠大脑的各个区域端粒酶活性:非放射性端粒酶重复序列扩增法（TRAP）分析

Telomerase, a ribonucleoprotein, is responsible for maintaining the telomere length and therefore promoting genomic integrity, proliferation, and ...

Stereotaxic Infusion of Oligomeric Amyloid-beta into the Mouse Hippocampus

Alzheimer&#8217;s disease is a neurodegenerative disease affecting the aging population. A key neuropathological feature of the disease is the over-production of amyloid-beta and the deposition of amyloid-beta plaques in brain regions of the afflicted individuals. Throughout the years scientists have generated numerous Alzheimer&#8217;s disease mouse models that attempt to replicate the amyloid-beta pathology. Unfortunately, the mouse models only selectively mimic the disease features. Neuronal death, a prominent effect in the brains of Alzheimer&#8217;s disease patients, is noticeably lacking in these mice. Hence, we and others have employed a method of directly infusing soluble oligomeric species of amyloid-beta - forms of amyloid-beta that have been proven to be most toxic to neurons - stereotaxically into the brain. In this report we utilize male C57BL/6J mice to document this surgical technique of increasing amyloid-beta levels in a select brain region. The infusion target is the dentate gyrus of the hippocampus because this brain structure, along with the basal forebrain that is connected by the cholinergic circuit, represents one of the areas of degeneration in the disease. The results of elevating amyloid-beta in the dentate gyrus via stereotaxic infusion reveal increases in neuron loss in the dentate gyrus within 1 week, while there is a concomitant increase in cell death and cholinergic neuron loss in the vertical limb of the diagonal band of Broca of the basal forebrain. These effects are observed up to 2 weeks. Our data suggests that the current amyloid-beta infusion model provides an alternative mouse model to address region specific neuron death in a short-term basis. The advantage of this model is that amyloid-beta can be elevated in a spatial and temporal manner.

Here, we present a protocol for direct stereotaxic brain infusion of amyloid-beta. This methodology provides an alternative in vivo mouse model to address the short-term effects of amyloid-beta on brain neurons.

低聚淀粉样蛋白-β的立体灌注到小鼠海马

Alzheimer&#8217;s disease is a neurodegenerative disease affecting the aging population. A key neuropathological feature of the disease is the ...

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has proven to be an invaluable model organism in helping to tackle this challenge, from inspiring technological approaches,&#160;building the human brain connectome, to actually shedding light on the specific molecular drivers of basic functional patterns. The bulk of the behavioral studies in C. elegans have been performed on solid substrates. In liquid, animals exhibit behavioral patterns that include movement at a range of speeds in 3D, as well as partial body movements, such as a posterior curl without anterior shape change, which introduce new challenges for quantitation. The steps of a simple procedure, and use of a software that enables high-resolution analysis of C. elegans swim behavior, are presented here. The software, named CeleST, uses a specialized computer program that tracks multiple animals simultaneously and provides novel measures of C. elegans locomotion in liquid (swimming). The measures are mostly grounded in animal posture and based on mathematics used in computer vision and pattern recognition, without computational requirements for threshold cut-offs. The software tool can be used to both assess overall swimming prowess in hundreds of animals from combined small batch trials and to reveal novel phenotypes even in well-characterized genetic mutants. The preparation of specimens for analysis with CeleST is simple and low-tech, enabling wide adaptation by the scientific community. Use of the computational approach described here should therefore contribute to the greater understanding of behavior and behavioral circuits in the C. elegans model.

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

An efficient and simple methodology for computer-based analysis of nematode swimming behavior in liquid is described. The method requires little to no investment for C. elegans laboratories. The hardware used is standard, and the computer software for the behavioral analysis (CeleST) is an open source one.

的自动分析<em&gt;℃。线虫</em&gt;游泳行为使用天书软件

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has ...

Diffuse Reflectance Spectroscopy: Getting the Capillary Refill Test Under One's Thumb

The capillary refill test was introduced in 1947 to help estimate circulatory status in critically ill patients. Guidelines commonly state that refill should occur within 2 s after releasing 5 s of firm pressure (e.g., by the physician&#39;s finger) in the normal healthy supine patient. A slower refill time indicates poor skin perfusion, which can be caused by conditions including sepsis, blood loss, hypoperfusion, and hypothermia. Since its introduction, the clinical usefulness of the test has been debated. Advocates point out its feasibility and simplicity and claim that it can indicate changes in vascular status earlier than changes in vital signs such as heart rate. Critics, on the other hand, stress that the lack of standardization in how the test is performed and the highly subjective nature of the naked eye assessment, as well as the test&#39;s susceptibility to ambient factors, markedly lowers the clinical value. The aim of the present work is to describe in detail the course of the refill event and to suggest potentially more objective and exact endpoint values for the capillary refill test using diffuse polarization spectroscopy.

This protocol describes how the use of diffuse polarization spectroscopy can improve the clinical usefulness of the capillary refill test. We suggest a more detailed analysis of the course of the capillary refill in healthy volunteers using diffuse reflectance spectroscopy videos and new informatic endpoints.

漫反射光谱分析: 在拇指下进行毛细管填充试验

The capillary refill test was introduced in 1947 to help estimate circulatory status in critically ill patients. Guidelines commonly state that refill ...

Localization of the Locus Coeruleus in the Mouse Brain

The locus coeruleus (LC) is a major hub of norepinephrine producing neurons that modulate a number of physiological functions. Structural or functional abnormalities of LC impact several brain regions including cortex, hippocampus, and cerebellum and may contribute to depression, bipolar disorder, anxiety, as well as Parkinson disease and Alzheimer disease. These disorders are often associated with metal misbalance, but the role of metals in LC is only partially understood. Morphologic and functional studies of LC are needed to better understand the human pathologies and contribution of metals. Mice are a widely used experimental model, but the mouse LC is small (~0.3 mm diameter) and hard to identify for a non-expert. Here, we describe a step-by-step immunohistochemistry-based protocol to localize the LC in the mouse brain. Dopamine-&#946;-hydroxylase (DBH), and alternatively, tyrosine hydroxylase (TH), both enzymes highly expressed in the LC, are used as immunohistochemical markers in brain slices. Sections adjacent to LC-containing sections can be used for further analysis, including histology for morphological studies, metabolic testing, as well as metal imaging by X-ray fluorescence microscopy (XFM).

The locus coeruleus is a small cluster of neurons involved in a variety of physiological processes. Here, we describe a protocol to prepare mouse brain sections for studies of proteins and metals in this nucleus.

小鼠大脑中耳蜗的定位

The locus coeruleus (LC) is a major hub of norepinephrine producing neurons that modulate a number of physiological functions. Structural or functional ...