威康信托基金会提供开放式访问牛津大学的资助。罗伯特执事牛津OXION组由威康信托基金赠款WT084655MA资助，是一个成员。

1。穴居 1。仪器各种容器设计的尝试（例如一个金属壶，可以很容易地清洗的，但老鼠都不愿意进入）。最终，它被发现的塑料溜子长度（房屋淌着连接）轻微升高的一端，以防止意外（非刻意）的食物颗粒的位移和密封在另一端，是一个高效的仪器。巴路士现在从塑料溜子，直径68毫米，切成20厘米长（ 图1）。一端是密封用12毫米的中密度纤维板（MDF）插头。 （MDF工作建议时，呼吸防护）。清洗的目的和防水，防水胶乳香化合物可应用于中密度纤维板的内外表面以及服务到穴居管胶水。机螺丝（5厘米长的）用于提升其他尾of的管离地面3厘米。两洞钻挖洞管的开口端的1厘米在一个90 °的螺丝插入的角度，拧紧4。 洞穴的直径可能是至关重要的;施密德，Holmes 等 19观察到，在野生小鼠“洞穴清理”主要是从洞穴大小相似，以人工的。 <img src="/files/ftp_upload/2607/2607fig1.jpg" alt="图1" /> 图1。鼠标在挖洞管。 2。程序<ol><li> 习惯 穴居通常是自发的，但有一定的学习元素。学习过程是提高社会便利。因此设立一个完整的洞穴到以后要测试一种治疗的小鼠之家笼过夜。每个洞穴是充满了200克的正常饮食供应的食物颗粒。 （这是指到T他密集压缩的饮食类型，如果“扩大”的饮食中使用的洞穴，将举行不少于200克）。实验者应考虑选择一个穴居基板等因素。 如果老鼠对食物配给（例如，激励他们运行并发迷宫学习任务），非食用物质可以使用​​，例如粘土球，用于线在室内盆栽的土壤表面;豌豆瓦（小石头〜1厘米直径）和沙子。豌豆碎石和沙子，食物颗粒较重的三倍，是有益的食物颗粒的替代品，如果老鼠挖洞很大力，避免“天花板效应”。 </li><li> 基线 挖洞测试，以同样的方式48小时外，将建议治疗后的测试运行（最好是两个）基准。下午4点左右，如果老鼠是一个0700-1900小时黑暗的光周期内测试是最佳开始。这是一个敏感的时刻进行测试，因为它们不是在这个时候通常清醒和穴居启动更慢，如果他们在黑暗阶段，因此非常活跃。两个小时后开始，是一个“快照”测量挖洞重量的食品;洞穴成皮重容器是空，颗粒替换到这是入笼的洞穴和权衡。如果鼠标是在洞穴，只需轻轻倒入称量容器和内容，并取代在笼子里的鼠标。然后在两个小时挖洞的重量是由减法计算，从原来的200克。最后的读数是第二天早晨。小鼠出尔反尔睡眠（经常在洞穴），不再积极挖洞，时间不是非常重要。使用2小时测量二审为基准性能的平衡，形成群体。穴居与实践17增加，基线也有助于小鼠，以达到更稳定的渐近渐近性能，从而揭示了治疗效果更好。 </li><li> 测试 测试，治疗后管理的老鼠挖洞测试在基线阶段完全一样。如果挖洞测试是长期使用，建议至少48小时单独的测试，以防止由于过度暴露于程序的性能下降。 </li><li> 请注意 不要替换进入洞穴笼地板上挖洞食品。即使鼠标没有任何挖洞，进行称重过程，你会如果挖洞发生了称重造成的干扰，往往刺激的行为，并已钻地基板的小鼠往往会开始挖洞后，重新焕发了生机称重过程。 </li></ol> 2。预期成果 C57〜BL / 6小鼠通常洞穴在首两小时，约70克，200克附近过夜。海马损毁老鼠经常洞穴少于5克11（ 图2）。朊病毒疾病抑制挖洞1,2，脂多糖管理20。穴居可以检测朊病毒疾病（痒病）在10-12周注射后病变脑组织匀浆，而只在22个星期13出现的临床症状。 （ 图3）。穴居已被证明是敏感的应变和钾离子通道亚基10的差异 15淘汰。 <img src="/files/ftp_upload/2607/2607fig2.jpg" alt="图2" /> 图2。钻地控制和海马损毁小鼠在两小时内的食物颗粒（中位数和四分位数范围）的重量。后者挖洞显著比对照组少（P = 0.0001）。 <img src="/files/ftp_upload/2607/2607fig3.jpg" alt="图3" /> 图3的重量。钻地通过控制和绵羊痒病感染的小鼠过夜的食物颗粒（手段）。穴居增加（含实践），在注射后都从每周7至10组，但在痒病组与组之间的差异12穴居下降，从一周变得统计学意义（超过平均值的两个标准误差）。 3。讨论我们已经测试在其他物种中的啮齿动物-鼠，沙鼠，仓鼠和埃及8刺小鼠挖洞。但后者可诱发大量地面基材，如碎石或沙子的洞穴，但挖洞的食物颗粒要少得多，即使与地面材料的丰富经验。这种从小鼠的差异的原因尚不清楚。已经做了这项工作后，它被发现埃及刺的小鼠，其实并不在野外洞穴，他们可能会占据洞穴沙鼠，还其自然公顷的表面bitat往往是坚硬的岩石。最近在内罗毕举行的观察表明，内属Acomys，威尔逊的多刺的鼠标（Acomys wilsoni）和波斯富街的黑刺鼠（Acomys percivali），在野外的洞穴，在实验室挖洞。 4。嵌套 1。仪器这个测试是在个别的笼子里，为穴居使用相同的是合适的。正常的被褥应包括地板深度为0.5厘米。 （深度的变化，非常深刻的被褥，可能影响鸟巢的建设）。每个网箱是提供一个“Nestlet”，按下棉絮（Ancare）5厘米见方的。 2。程序小鼠是单独放置到约一小时的嵌套笼前的黑暗阶段，并评估结果第二天早上。挖洞，出于同样的原因，时机是不是很关键。 3。评分巢是一个规模5点（ 图4-8），并评估untorn Nestlet量也重达5。 <ol><li> Nestlet是基本上不变（&gt; 90％完好）。 </li><li> Nestlet是部分撕裂（50-90％，保持不变）。 </li><li> Nestlet大多是切丝，但往往没有识别巢网站：&lt;50％的Nestlet保持不变，但&lt;90％是在笼楼面面积的四分之一，即棉花不聚集成巢，但周围的笼子里蔓延。注：材料有时会在广泛定义的巢区，但关键的定义是50-90％已被撕碎。 </li><li>识别，但平巢：&gt; 90％的Nestlet撕毁，物质聚集成一个巢内笼楼面面积的四分之一，但窝平坦，墙壁高于鼠标车身高度（蜷缩于第它的副作用）其周长小于50％。 </li><li> （附近）完美ÑEST：&gt; 90％的Nestlet被撕裂了，鸟巢是一个火山口与墙壁高于其周长超过50％的鼠标车身高度。 </li></ol>如果标准不同意分裂的区别。例如一个完美巢unshredded0.7克一块得分4.5。 <img src="/files/ftp_upload/2607/2607fig4.jpg" alt="图4" /> 图4。鸟巢得分1 <img src="/files/ftp_upload/2607/2607fig5.jpg" alt="图5" /> 图5。鸟巢得分2 <img src="/files/ftp_upload/2607/2607fig6.jpg" alt="图6" /> 图6。鸟巢3分<img src="/files/ftp_upload/2607/2607fig7.jpg" alt="图7" /> 图7。鸟巢得分4 <img src="/files/ftp_upload/2607/2607fig8.jpg" alt="图8" /> 图8。鸟巢SCORE 5 4。预期成果男性和女性的巢穴，他们的目的是体温调节，以及与生殖相关。 C57BL / 6小鼠，巢分数为男性和女性在同一范围内，虽然我们没有做一个控制在一个单一的实验比较。最C57BL / 6小鼠巢建设得分4-5，但是当海马损毁的中位数的得分将围绕1-2个; 3分是不可能被超越（ 图 9 ）。嵌套也被证明是大约10-12周1,2,16敏感的朊病 ​​毒疾病。 <img src="/files/ftp_upload/2607/2607fig9.jpg" alt="图9" /> 图9。鸟巢分数（中位数和四分位数范围）为对照组小鼠和小鼠的背，腹和完整的海马病变。只有完整的损毁小鼠抑制嵌套，所以选择性脊的影响和ventr人地区出现亚阈值，但添加剂。 5。囤积 1。仪器一个“基地”系列连接丝网制成的管子，在远端放置的食物颗粒，并暂时由一个木制的插头近端永久密封。这是用来防止老鼠进入管适应大本营前occurred7。 仪器（ 图10）可以以不同的方式，只要坚持的基本原则。我们一排8个木箱组成，每个30 × 13 × 15厘米的透明有机玻璃盖。每提供一瓶水，并在后面的洞囤积管推装。管是黑色塑料，长10厘米，直径40毫米制成，加入到一个丝网管，长45厘米，4厘米直径的，形成了总长度为50厘米。网格由13毫米的正方形，与MIS对齐网格在网格较小的孔和防止食物颗粒下降通过双辊。如果6平方毫米的网状，这是一个更好的材料一卷就足够了。管，放置的食物颗粒，末端是封闭的。近端是一个可移动的木制插头与密封。 <img src="/files/ftp_upload/2607/2607fig10.jpg" alt="图10" /> 在囤积管小鼠图10。 2。程序<ol><li>通常情况下，囤积是非常变量的第一次，有些小鼠可能不会囤积在所有。囤积居奇，喜欢挖洞，提高与实践。因此，实验设计允许，基线会议是最好的治疗前管理。显然，这是不实际的，如果自变量是天生的，作为一个（非条件）突变小鼠或测试时，不同的小鼠品系。 </li><li>鼠标是投入早在一天的每个方块habituate它。框提供了一个纸板隧道，其中有一批真正的家笼中至少一晚，也是一个慷慨的部分弄脏被褥，使它像“家”的感觉。访问囤积管是防止晚上，直到用木棍塞。 100克食物颗粒（囤积管末端放置的小型和大型的混合物）（浇在另一端举行垂直管）。为了确保测试开始前，鼠标轻度饥饿，家中没有提供食物。 </li><li>正如黑暗期开始前，取出木塞让鼠标进入囤积管。第二天早晨，（再次，时机是不是很关键的）收集和权衡所有的食物已囤积到大本营中的颗粒。 </li></ol> 6。预期成果使用雌性C57BL / 6小鼠，囤积的中位数金额将围绕50-70克。海马病变强烈抑制hoarding11，而前额叶皮层的病变只有微弱的影响12（ 图11）。 <img src="/files/ftp_upload/2607/2607fig11.jpg" alt="图11" /> 图11。囤积食物颗粒重量控制和内侧前额叶皮层损毁小鼠（中位数和四分位数范围）。虽然中位数差异很大，在控制高可变性意味着远，结果是从统计学意义（P = 0.3）。

<ol>
	<li>Cunningham, C., Deacon, R. M. J., Chan, K., Boche, D., Rawlins, J. N. P., Perry, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuropathologically+distinct+prion+strains+give+rise+to+similar+temporal+profiles+ofbehavioural+deficits.">Neuropathologically distinct prion strains give rise to similar temporal profiles ofbehavioural deficits.</a> Neurobiol. Dis. 18, 258-269 (2005).</li><li>Cunningham, C., Deacon, R., Wells, H., Boche, D., Waters, S., Picanco Diniz, C., Scott, H., Rawlins, J. N. P., Perry, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+changes+characterize+early+behavioural+signs+in+the+ME7+model+of+murine+prion+disease.">Synaptic changes characterize early behavioural signs in the ME7 model of murine prion disease.</a> Eur. J. Neurosci. 17, 2147-2215 (2003).</li><li>Damiani, R., Modesto, S., Yates, A., Neveling, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Earliest+evidence+of+cynodont+burrowing.">Earliest evidence of cynodont burrowing.</a> Proc. R. Soc. Lond. B. 270, 1747-1751 (2003).</li><li>Deacon, R. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burrowing+in+rodents:+a+sensitive+method+for+detecting+behavioral+dysfunction.">Burrowing in rodents: a sensitive method for detecting behavioral dysfunction.</a> Nat. Protocols. 1, 118-121 (2006).</li><li>Deacon, R. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+nest+building+in+mice.">Assessing nest building in mice.</a> Nat. Protocols. 1, 1117-1119 (2006).</li><li>Deacon, R. M. 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=Digging+in+Mice:+Marble+Burying,+Burrowing,+and+Direct+Observation+Reveal+Changes+in+Mouse+Behavior.">Digging in Mice: Marble Burying, Burrowing, and Direct Observation Reveal Changes in Mouse Behavior.</a> Mood and anxiety related phenotypes in mice. , (2009).</li><li>Deacon, R. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+hoarding+in+mice.">Assessing hoarding in mice.</a> Nat. Protocols. 1, 2828-2830 (2006).</li><li>Deacon, R. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burrowing:+A+sensitive+behavioural+assay,+tested+in+five+species+of+laboratory+rodents.">Burrowing: A sensitive behavioural assay, tested in five species of laboratory rodents.</a> Behav. Brain. Res. 200, 128-133 (2009).</li><li>Deacon, R. M. J., Bannerman, D. M., Kirby, B. P., Croucher, A., Rawlins, J. N. P. <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+cytotoxic+hippocampal+lesions+in+mice+on+a+cognitive+test+battery.">Effects of cytotoxic hippocampal lesions in mice on a cognitive test battery.</a> Behav. Brain. Res. 133, 57-68 (2002).</li><li>Deacon, R. M. J., Brook, R. C., Meyer, D., Haeckel, O., Ashcroft, F. M., Miki, T., Seino, S., Liss, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+phenotyping+of+mice+lacking+the+KATP+channel+subunit+Kir6.2.">Behavioral phenotyping of mice lacking the KATP channel subunit Kir6.2.</a> Physiol. Behav. 87, 723-733 (2006).</li><li>Deacon, R. M. J., Croucher, A., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+cytotoxic+lesion+effects+on+species-typical+behaviors+in+mice.">Hippocampal cytotoxic lesion effects on species-typical behaviors in mice.</a> Behav. Brain. Res. 132, 203-213 (2002).</li><li>Deacon, R. M. J., Penny, C., Rawlins, J. N. P. <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+medial+prefrontal+cortex+cytotoxic+lesions+in+mice.">Effects of medial prefrontal cortex cytotoxic lesions in mice.</a> Behav. Brain. Res. 139, 139-155 (2003).</li><li>Deacon, R. M. J., Raley, J. M., Perry, V. H., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burrowing+into+prion+disease.">Burrowing into prion disease.</a> Neuroreport. 12, 2053-2057 (2001).</li><li>Deacon, R. M. J., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+lesions,+species-typical+behaviours+and+anxiety+in+mice.">Hippocampal lesions, species-typical behaviours and anxiety in mice.</a> Behav. Brain. Res. 156, 241-249 (2005).</li><li>Deacon, R. M. J., Thomas, C. L., Rawlins, J. N. P., Morley, 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=A+comparison+of+the+behavior+of+C57BL/6+and+C57BL/10.">A comparison of the behavior of C57BL/6 and C57BL/10.</a> Behav. Brain. Res. 179, 239-247 (2007).</li><li>Guenther, K., Deacon, R. M. J., Perry, V. H., Rawlins, J. N. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+behavioural+changes+in+scrapie-affected+mice+and+the+influence+of+dapsone.">Early behavioural changes in scrapie-affected mice and the influence of dapsone.</a> Eur. J. Neurosci. 14, 401-409 (2001).</li><li>Mallucci, G. R., White, F. a. r. m. e. r., Dickinson, M., Khatun, A., Powell, H., Brandner, A. D., Jefferys, S., R, J. G., Collinge, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+Cellular+Prion+Protein+Reverses+Early+Cognitive+Deficits+and+Neurophysiological+Dysfunction+in+Prion-Infected+Mice.">Targeting Cellular Prion Protein Reverses Early Cognitive Deficits and Neurophysiological Dysfunction in Prion-Infected Mice.</a> Neuron. 53, 325-335 (2007).</li><li>Russell, W. M. S., Burch, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The principles of humane experimental technique. , (1959).</li><li>Schmid-Holmes, S., Drickamer, L. C., Robinson, A. S., Gillie, L. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burrows+and+burrow+cleaning+behaviour+of+house+mice.">Burrows and burrow cleaning behaviour of house mice.</a> Am. Mid. Nat. 146, 53-62 (2001).</li><li>Teeling, J. L., Felton, L. M., Deacon, R. M. J., Cunningham, C., Rawlins, J. N. P., Perry, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sub-pyrogenic+systemic+inflammation+impacts+on+brain+and+behavior,+independent+of+cytokines.">Sub-pyrogenic systemic inflammation impacts on brain and behavior, independent of cytokines.</a> Brain Behav. Immun. 21, 836-850 (2007).</li></ol>

没有利益冲突的声明。

<html>这些测试是非常简单的运行的，但效果大小是非常大，使他们极为敏感，处理，应变的差异，等他们符合罗素和伯奇的3 个 RS“18”精致“的方面。事实上，良好的性能测试，像穴居，似乎表明动物的健康情况大致良好。因此，这可能是一个很好的测试评估动物福利。 啮齿类实验动物的丰富环境的重要性是众所公认这些天。提供排料现在是标准的做法在许多动物的房子，这是很容易做到。的热情，使老鼠挖洞，挖洞一个潜在的优秀富集，但不幸的是一种“自我重载”的洞穴，这将是一个切实可行的办法提供这个尚未开发的。无论小鼠发现囤积丰富没有尚未确定。 _content“&gt;一个缺点是，这些试验产生的数据是一般非参数（高斯分布），因此适当的非参数统计，需要对它们进行分析，如采用Mann - Whitney U为两两比较测试，克鲁斯卡尔瓦利斯多组数据的方差分析，如果使用重复测量设计，因为没有合适的ANOVA检验非参数数据，结果可以转化，例如平方根或日志程序，使它们符合一个比较正常的分布。 仪器测量可略有不同，那些在文本中指定的（美国实验者有用），但应避免的巨大差异。然而，在某些情况下，其他实验者可能会发现不同的建设工作比这里描述的更好。 所有的行为测量，小鼠应测试，看看如果他们的活动水平是正常的的，不活动将明显有detrimental效果物种典型行为。当绵羊痒病感染的小鼠显示受损挖洞和筑巢，它们的活动水平是正常的（在一个开放的领域仪器测试）。事实上，一段时间多动症的发生，约2-4周后，这些物种典型行为是首先受损2。 由海马病变产生强劲的减值上挖洞，筑巢和囤积，所有这一切都是种典型的行为，在空间学习和memory9补充的损伤等行之有效的这些病变的影响。他们似乎被等同于公元14特有的日常生活活动障碍。 这些物种的典型测试可能被证明是用于治疗阿尔茨海默氏症和其他神经退行性疾病的14的临床前筛选的伟大。在许多人的大脑疾​​病的损失或减少能力执行正常的前夕ryday任务是其他疾病的功能，如认知功能障碍的重要。特别是，这种损失可能需要照顾者的参与和巨大的社会经济负担，这样往往规定。

assessing burrowing, nest construction, and hoarding in mice

在执行“日常生活活动能力（ADL）能力的恶化是阿尔茨海默氏病的早期征兆（公元）。可能的治疗AD的临床前行为筛查目前主要集中在认知测试，这往往需要昂贵的设备和大量的实验者时间。然而，人类的情节记忆（在AD方面受影响最严重的内存），是不同的啮齿动物的内存，这似乎在很大程度上非偶发。因此，目前AD在啮齿类动物中的新的AD治疗方法的筛选，本质上是不可能取得成功。临床前筛选的新方法将特征ADL小鼠。巧合的是，几个这样的实验最近已经开发在牛津大学，这里的三个最敏感和最良好的特点。 穴居是第一次在牛津 13 。它的发展需要制定一个鼠标囤积范式。大多数发布ED啮齿动物囤积范式需要一个遥远的食物来源，以挂钩连接通过家庭笼。这将涉及修改主页笼以及鼠标证明连接通道和食物来源。因此，它被认为是否有可能把笼子内的食物来源。结果发现，如果一个容器放置在地板上，第二天早上被掏空。食物颗粒，但是，只需存放在一个堆在货柜入口，而不是放置在一个独立的地方离的容器，可以预料，如果老鼠被真正囤积他们。仔细检查发现小鼠挖（“穴居”）的动作，没有携带自己的嘴巴颗粒到选定的地方，因为他们如果真正囤积。 粮食颗粒不挖洞的一个必不可少的基板;小鼠空管充满砂，砾石，甚至从自己的笼子弄脏被褥。更多以上，他们将空管，即使被放在一个空旁边，8 。 文献中存在的几个嵌套协议。本牛津简化了程序，有一个明确的巢质量 5的计分制。 后来发展的小鼠，而不是囤积回到真正的家笼，适应生活中的“大本营”的一个囤积仪器的一个囤积范式。这家基地被连接到一个管网，其中包含的食物来源的末端。这样的安排被证明取得了良好的囤地行为，只要小鼠适应生活中的“大本营”白天只允许在夜间进入囤积管。

Watch this Scientific Journal Video about Assessing Burrowing, Nest Construction, and Hoarding in Mice at JoVE.com

Assessing Burrowing, Nest Construction, and Hoarding in Mice

挖洞，筑巢和囤积是种典型的活动，在实验室小鼠容易执行。本文介绍了如何能轻松，廉价地评估。这些协议是极其敏感的的小鼠品系，脑损伤和疾病。此外，它们构成了“丰富环境”的老鼠，并体现“3个RS”18“精致”的方面。

评估挖洞，在小鼠的鸟巢建设和囤积

Deterioration in the ability to perform "Activities of daily living" (ADL) is an early sign of Alzheimer's disease (AD). Preclinical behavioural screening ...

assessing-burrowing-nest-construction-and-hoarding-in-mice

Research

JoVE Journal

Neuroscience

31.8K 次观看.  University of Oxford. 挖洞，筑巢和囤积是种典型的活动，在实验室小鼠容易执行。本文介绍了如何能轻松，廉价地评估。这些协议是极其敏感的的小鼠品系，脑损伤和疾病。此外，它们构成了“丰富环境”的老鼠，并体现“3个RS”18“精致”的方面。

视频: Assessing Burrowing, Nest Construction, and Hoarding in Mice

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 ...

科研

神经科学

Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors1,2 while the second involves a wholesale (global) activation of most or all such neurons3. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons.
The Drosophila larva is an established model system to study thermal nociception, a sensory response to potentially harmful temperatures that is evolutionarily conserved across species1,2. The advantages of Drosophila for such studies are the relative simplicity of its nervous system and the sophistication of the genetic techniques that can be used to dissect the molecular basis of the underlying biology4-6 In Drosophila, as in all metazoans, the response to noxious thermal stimuli generally involves a "nocifensive" aversive withdrawal to the presented stimulus7. Such stimuli are detected through free nerve endings or nociceptors and the amplitude of the organismal response depends on the number of nociceptors receiving the noxious stimulus8. In Drosophila, it is the class IV dendritic arborization sensory neurons that detect noxious thermal and mechanical stimuli9 in addition to their recently discovered role as photoreceptors10. These neurons, which have been very well studied at the developmental level, arborize over the barrier epidermal sheet and make contacts with nearly all epidermal cells11,12. The single axon of each class IV neuron projects into the ventral nerve cord of the central nervous system11 where they may connect to second-order neurons that project to the brain.
Under baseline conditions, nociceptive sensory neurons will not fire until a relatively high threshold is reached. The assays described here allow the investigator to quantify baseline behavioral responses or, presumably, the sensitization that ensues following tissue damage. Each assay provokes distinct but related locomotory behavioral responses to noxious thermal stimuli and permits the researcher to visualize and quantify various aspects of thermal nociception in Drosophila larvae. The assays can be applied to larvae of desired genotypes or to larvae raised under different environmental conditions that might impact nociception. Since thermal nociception is conserved across species, the findings gleaned from genetic dissection in Drosophila will likely inform our understanding of thermal nociception in other species, including vertebrates.

Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors1,2 while the second involves a wholesale (global) activation of most or all such neurons3. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons.

局部和全局的方法评估热痛觉<em&gt;果蝇</em&gt;幼虫

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Assessing Primary Neurogenesis in Xenopus Embryos Using Immunostaining

Primary neurogenesis is a dynamic and complex process during embryonic development that sets up the initial layout of the central nervous system. During this process, a portion of neural stem cells undergo differentiation and give rise to the first populations of differentiated primary neurons within the nascent central nervous system. Several vertebrate model organisms have been used to explore the mechanisms of neural cell fate specification, patterning, and differentiation. Among these is the African clawed frog, Xenopus, which provides a powerful system for investigating the molecular and cellular mechanisms responsible for primary neurogenesis due to its rapid and accessible development and ease of embryological and molecular manipulations. Here, we present a convenient and rapid method to observe the different populations of neuronal cells within Xenopus central nervous system. Using antibody staining and immunofluorescence on sections of Xenopus embryos, we are able to observe the locations of neural stem cells and differentiated primary neurons during primary neurogenesis.

Assessing Primary Neurogenesis in Xenopus Embryos Using Immunostaining

This article presents a convenient and rapid method for visualizing different neuronal cell populations in the central nervous system of Xenopus embryos using immunofluorescent staining on sections.

评估主要神经再生<em&gt;爪</em&gt;胚胎使用免疫染色

Primary neurogenesis is a dynamic and complex process during embryonic development that sets up the initial layout of the central nervous system. During ...

Assessing Spatial Learning and Memory in Small Squamate Reptiles

Clinical research has leveraged a variety of paradigms to assess cognitive decline, commonly targeting spatial learning and memory abilities. However, interest in the cognitive processes of nonmodel species, typically within an ecological context, has also become an emerging field of study. In particular, interest in the cognitive processes in reptiles is growing although experimental studies on reptilian cognition are sparse. The few reptilian studies that have experimentally tested for spatial learning and memory have used rodent paradigms modified for use in reptiles. However, ecologically important aspects of the physiology and behavior of this taxonomic group must be taken into account when testing for spatially based cognition. Here, we describe modifications of the dry land Barnes maze and associated testing protocol that can improve performance when probing for spatial learning and memory ability in small squamate reptiles. The described paradigm and procedures were successfully used with male side-blotched lizards (Uta stansburiana), demonstrating that spatial learning and memory can be assessed in this taxonomic group with an ecologically relevant apparatus and protocol.

This paper describes a modification of the Barnes maze, a standard rodent paradigm used to assess spatial memory and learning, for use in small squamate reptiles.

评估空间学习记忆小鳞类爬行动物

Clinical research has leveraged a variety of paradigms to assess cognitive decline, commonly targeting spatial learning and memory abilities. However, ...

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and parenting. Importantly, mice can be trained to associate novel odors with specific behavioral responses to provide insight into olfactory circuit function. This protocol details the procedure for training mice on a Go/No-Go operant learning task. In this approach, mice are trained on hundreds of automated trials daily for 2&#8211;4 weeks and can then be tested on novel Go/No-Go odor pairs to assess olfactory discrimination, or be used for studies on how odor learning alters the structure or function of the olfactory circuit. Additionally, the mouse olfactory bulb (OB) features ongoing integration of adult-born neurons. Interestingly, olfactory learning increases both the survival and synaptic connections of these adult-born neurons. Therefore, this protocol can be combined with other biochemical, electrophysiological, and imaging techniques to study learning and activity-dependent factors that mediate neuronal survival and plasticity.

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

小鼠嗅觉学习和鉴别的客观和可重复性试验

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and ...

Assessing Pupil-linked Changes in Locus Coeruleus-mediated Arousal Elicited by Trigeminal Stimulation

Current scientific literature provides evidence that trigeminal sensorimotor activity associated with chewing may affect arousal, attention, and cognitive performance. These effects may be due to widespread connections of the trigeminal system to the ascending reticular activating system (ARAS), to which noradrenergic neurons of the locus coeruleus (LC) belongs. LC neurons contain projections to the whole brain, and it is known that their discharge co-varies with pupil size. LC activation is necessary for eliciting task-related mydriasis. If chewing effects on cognitive performance are mediated by the LC, it is reasonable to expect that changes in cognitive performance are correlated to changes in task-related mydriasis. Two novel protocols are presented here to verify this hypothesis and document that chewing effects are not attributable to aspecific motor activation. In both protocols, performance and pupil size changes observed during specific tasks are recorded before, soon after, and half an hour following a 2 min period of either: a) no activity, b) rhythmic, bilateral handgrip, c) bilateral chewing of soft pellet, and d) bilateral chewing of hard pellet. The first protocol measures level of performance in spotting target numbers displayed within numeric matrices. Since pupil size recordings are recorded by an appropriate pupillometer that impedes vision to ensure constant illumination levels, task-related mydriasis is evaluated during a haptic task. Results from this protocol reveal that 1) chewing-induced changes in performance and task-related mydriasis are correlated and 2) neither performance nor mydriasis are enhanced by handgrip. In the second protocol, use of a wearable pupillometer allows measurement of pupil size changes and performance during the same task, thus allowing even stronger evidence to be obtained regarding LC involvement in the trigeminal effects on cognitive activity. Both protocols have been run in the historical office of Prof. Giuseppe Moruzzi, the discoverer of ARAS, at the University of Pisa.

To verify whether trigeminal effects on cognitive performance involve locus coeruleus activity, two protocols are presented that aim to evaluate possible correlations between the performance and task-related pupil size changes induced by chewing. These protocols may be applied to conditions in which locus coeruleus contribution is suspected.

评估由三角刺激引起的由Locus Coeruleus介导的觉醒中与学生相关的变化

Current scientific literature provides evidence that trigeminal sensorimotor activity associated with chewing may affect arousal, attention, and cognitive ...

Quantitative Measurement of Intrathecally Synthesized Proteins in Mice

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the CSF protein composition delivers crucial information in basic neuroscience research as well as neurological diseases. One caveat is that proteins measured in CSF may derive from both intrathecal synthesis and transudation from serum, and protein analysis of CSF can only determine the sum of these two components. To discriminate between protein transudation from the blood and intrathecally produced proteins in animal models as well as in humans, CSF protein profiling measurements using conventional protein analysis tools must include the calculation of the albumin CSF/serum quotient (Qalbumin), a marker of the integrity of the blood-brain interface (BBI), and the protein index (Qprotein/Qalbumin), an estimate of intrathecal protein synthesis. This protocol illustrates the entire procedure, from CSF and blood collection to quotients and indices calculations, for the quantitative measurement of intrathecal protein synthesis and BBI impairment in mouse models of neurological disorders.

Elevated spinal fluid protein levels can either be the result of diffusion of plasma protein across an altered blood-brain barrier or intrathecal synthesis. An optimized testing protocol is presented in this article that helps to discriminate both cases and provides quantitative measurements of intrathecally synthesized proteins.

小鼠体内合成蛋白的定量测量

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the ...

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings

Multichannel electrode arrays offer insight into the working brain and serve to elucidate neural processes at the single-cell and circuit levels. Development of these tools is crucial for understanding complex behaviors and cognition and for advancing clinical applications. However, it remains a challenge to densely record from cell populations stably and continuously over long time periods. Many popular electrodes, such as tetrodes and silicon arrays, feature large cross-diameters that produce damage upon insertion and elicit chronic reactive tissue responses associated with neuronal death, hindering the recording of stable, continuous neural activity. In addition, most wire bundles exhibit broad spacing between channels, precluding simultaneous recording from a large number of cells clustered in a small area. The carbon fiber microelectrode arrays described in this protocol offer an accessible solution to these concerns. The study provides a detailed method for fabricating carbon fiber microelectrode arrays that can be used for both acute and chronic recordings in vivo. The physical properties of these electrodes make them ideal for stable and continuous long-term recordings at high cell densities, enabling the researcher to make robust, unambiguous recordings from single units across months.

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings

This protocol describes a procedure for constructing carbon fiber microelectrode arrays for chronic and acute in vivo electrophysiological recordings in mouse (Mus musculus) and ferret (Mustela putorius furo) from multiple brain regions. Each step, following the purchase of raw carbon fibers to microelectrode array implantation, is described in detail, with emphasis on microelectrode array construction.

用于慢性和急性体内记录的碳纤维微电极阵列 的 构建和实现

Multichannel electrode arrays offer insight into the working brain and serve to elucidate neural processes at the single-cell and circuit levels. ...

用于记录幼年小鼠日常神经活动的 Micro-drive 植入物

A Lightweight Drive Implant for Chronic Tetrode Recordings in Juvenile Mice

In vivo electrophysiology provides unparalleled insight into the sub-second-level circuit dynamics of the intact brain and represents a method of particular importance for studying mouse models of human neuropsychiatric disorders. However, such methods often require large cranial implants, which cannot be used in mice at early developmental time points. As such, virtually no studies of in vivo physiology have been performed in freely behaving infant or juvenile mice, despite the fact that a better understanding of neurological development in this critical window would likely provide unique insights into age-dependent developmental disorders such as autism or schizophrenia. Here, a micro-drive design, surgical implantation procedure, and post-surgery recovery strategy are described that allow for chronic field and single-unit recordings from multiple brain regions simultaneously in mice as they age from postnatal day 20 (p20) to postnatal day 60 (p60) and beyond, a time window roughly corresponding to the human ages of 2 years old through to adulthood. The number of recording electrodes and final recording sites can be easily modified and expanded, thus allowing flexible experimental control of the in vivo monitoring of behavior- or disease-relevant brain regions across development.

作者聚焦：用于幼年小鼠慢性四极管记录的轻型驱动植入物  

Here, we describe a micro-drive design, surgical implantation procedure, and post-surgery recovery strategy that allow for chronic field and single-unit recordings from multiple brain regions simultaneously in juvenile and adolescent mice across a critical developmental window from postnatal day 20 (p20) to postnatal day 60 (p60) and beyond.

Methods Article

评估挖洞，在小鼠的鸟巢建设和囤积