该研究得到了翻译健康科学研究所试点项目授予机会（UL1TR000423），华盛顿人类发展与残疾中心大学和华盛顿大学动物行为核心与脑成像核心的支持。我们要感谢Warren Ladiges博士在制定和传播这里提出的原始径向水面踏步迷宫设计和协议方面的作用。我们也感谢Toby Cole对这个项目的协助。

所有程序和动物处理均按照国家卫生研究院和华盛顿大学动物护理与使用委员会颁布的动物护理指南进行。 手术<ol><li>在诱导盒中麻醉5％异氟烷的老鼠，直到无意识。通过降低呼吸频率和在趾部夹紧后不存在撤回反射来确认麻醉。 </li><li>在手术过程中，通过鼻锥保持2-2.5％的麻醉。在整个手术过程中监测呼吸频率，以确保鼠标无意识。 </li><li>将鼠标放在加热垫上，并使用耳杆将鼠标放置在立体定向装置中，确保头部牢固且平整。 </li><li>使用脱毛霜从头皮上去除头发。用盐水彻底冲洗头皮。 </li><li>用交替碘和70％乙醇洗涤清洁手术部位。 </li><li> Administe在头皮皮下注射利多卡因和布比卡因（1mg / kg）。 </li><li>用手术剪刀做纵向中线切口，并收回皮肤以露出头骨。 </li><li>使用5毫米环钻预制件在左前额叶皮质上进行开颅手术，中心点在正前方2.5 mm处，中线左侧2.5 mm。小心地取出骨头露出大脑。 </li><li>将冲击器设备设置为6 m / s和200 ms驻留时间的速度。 </li><li>定位冲击器装置，直到3毫米凸出的撞击尖端轻轻地接触大脑后方2.5英寸后2.55毫米的中线。将冲击头缩回，并降低1 mm（冲击深度）。准备好后，启动设备，产生所需的影响。 </li><li>用无菌聚丙烯盘覆盖开颅手术，用组织粘合剂胶合到皮肤，并缝合切口。 </li><li>从麻醉中取出鼠标，并给予Bupr的IP注射（0.5mg / kg）。 </li><li>让鼠标在一个干净的笼子里恢复，加热垫加热。在接下来的24小时内，应该监测老鼠的疼痛或痛苦的迹象。 注意:假手术控制应该得到与上述相同的治疗，省略步骤1.8-1.9。 </li></ol>径向水踏面迷宫建筑<ol><li> Bore 9个出口孔，足够大以容纳1-1 / 2英寸ABS DWV SPG x SJ陷阱适配器，围绕32英寸直径镀锌钢桶的圆周等间隔。将这些出口孔围在浴缸地板的大约2-1 / 4英寸处。 </li><li>将1-1 / 2英寸ABS DWV SPG x SJ陷阱适配器安装到每个出口孔中，并使用附带的环形螺母固定。 </li><li>使用橡胶塞子，九口八口从设备外部出口。最后的未封顶出口将作为逃生路线。将退出指定为逃生路线无关紧要。 </li><li>安装90°ABS肘至剩余出口的外端。 90°弯曲用于防止测试对象从迷宫内部目视确定正确的逃生路线。 </li><li>从任何能够消毒的不透明盒子构成逃生箱，大小约为30厘米x 15厘米x 15厘米。在地板的正上方，在盒子的侧面切开一个孔，足够大以容纳90°的ABS弯头。 </li><li>将逃生箱安装在90°ABS弯头的末端。 </li><li>略微提升地板表面上方的逃生箱（小于一英寸）。这允许将电加热垫或其他加热源放置在逃逸箱下面的充足空间。 </li><li>打印和层压至少5个独特的视觉线索。使用简单，高对比度的图像，可以很容易地从设备中辨别出来。建议使用黑白剪贴画形状（三角形，正方形，圆形）。 </li><li>使用磁铁，将视觉线索粘贴到内壁仪器。围绕设备周围的距离应大致相等。 </li></ol>径向水踏面迷宫方案注意:水迷宫测试应仅在手术部位愈合后（术后约1周）开始。 <ol><li> 准备测试 <ol><li>允许小鼠在开始测试前至少30分钟适应测试室。 </li><li>使用70％乙醇喷雾消毒设备。 </li><li>用大约1英寸的冷（12-14℃）水填充设备。 </li><li>将电加热垫或其他加热源直接放在逃生箱下面。在整个测试期间保持逃生箱的黑暗和温暖。 </li><li>在设备上放置一个明亮的光源。 注意:如果使用从设备本身研究动物可能可见的灯，请注意确保灯泡处于同一波段每天都有一个意见灯本身可能代表了老鼠用于定位逃生箱的另一种视觉提示，并且每天都会大幅度地移动它可能会使结果复杂化。 </li></ol></li><li> 测试协议 <ol><li>将尾巴轻轻地从鼠笼中取出鼠标，放在设备的中央。 </li><li>一旦动物进入装置，开始计时。 </li><li>一旦动物找到正确的出口，并且已经找到/进入逃生箱，停止计时并记录找到正确路径所需的秒数。 </li><li>如果动物试图爬上一个终止孔，10秒后不会自动重入迷宫，用手将动物引回到迷宫的中心。 </li><li>如果动物未能在3分钟（180秒）内找到逃生箱的正确路径，则将该试验评定为失败记录为180秒。用手小心地将动物引导到正确的路径。 </li><li>允许小鼠留在逃生箱中1分钟，试用休息。 </li><li>一分钟的休息已经过去了，从逃生箱中取出动物并返回家中笼子。 </li><li>彻底消毒逃生箱，并用70％乙醇喷雾出口，以防止鼠标使用嗅觉线索找到正确的逃生路线。此步骤不应超过几秒钟。 </li><li>将鼠标放回迷宫进行下一次试验。 </li><li>重复步骤3.2.1-3.2.9，直到鼠标完成了三次试验，并已经定位或被引导到逃生箱三次。 </li><li>最后1分钟休息后，将鼠标移回家中的笼子。 </li><li>在动物之间排放并更换设备中的水，以确保整个测试过程中温度一致。 </li><li>对要测试的每只鼠标重复步骤3.2.1-3.2.12。 </li><li>第二天重复准备步骤3.2.1-3.2.5。特别注意确保视觉线索保留在测试日之间保持一致的位置。 </li><li>使用上述日常测试方案测试动物，每天进行三次试验，持续四天（训练期），第五天进行最后三次记忆保留试验。第十二个三年试验（长期记忆保留）可以在第十二天给予。 </li><li>在第五天和第十二天之间不要进行任何测试。 </li></ol></li><li> 分析 <ol><li>如果在任何两天的时间内，如果一只老鼠在180秒以内无法完成迷宫（ 即连续6次测试全部得分为180秒），请考虑由测试条件导致的鼠标不足，并从分析中删除。 </li><li>计算每个受试者完成迷宫的平均延迟时间，通过对其当天的三次日常试验进行平均化来测试日期。 </li><li>使用标准t检验在第五天测试和长期记忆测试日期比较组获得记忆保留的组差异。如果m比较两组比较，应采用单因素方差分析，然后进行适当的事后分析（如Tukey检验）跟踪所获得的任何显着性。 </li><li> 通过反复测量方差分析获得收购期（第1-4天）的组差异。 </li></ol></li></ol>

<ol>
	<li>Levin, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurobehavioral+outcome+of+closed+head+injury:+Implications+for+clinical+trials.">Neurobehavioral outcome of closed head injury: Implications for clinical trials.</a> J. Neurotrauma. 12 (4), 601-610 (1995).</li><li>Schretlen, D., Shapiro, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+quantitative+review+of+the+effects+of+traumatic+brain+injury+on+cognitive+functioning.">A quantitative review of the effects of traumatic brain injury on cognitive functioning.</a> Int Rev Psychiatry. 15 (4), 341-349 (2003).</li><li>Cline, M. M., 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=Novel+application+of+a+radial+water+tread+maze+can+distinguish+cognitive+deficits+in+mice+with+traumatic+brain+injury.">Novel application of a radial water tread maze can distinguish cognitive deficits in mice with traumatic brain injury.</a> Brain Res. 1657, 140-147 (2017).</li><li>Ellenbroek, B., Youn, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+in+neuroscience+research:+Is+it+a+rat+race?.">Rodent models in neuroscience research: Is it a rat race?.</a> Dis. Model. Mech. 9 (10), 1079-1087 (2016).</li><li>Morris, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developments+of+a+water-maze+procedure+for+studying+spatial+learning+in+the+rat.">Developments of a water-maze procedure for studying spatial learning in the rat.</a> J. Neurosci Methods. 11 (1), 47-60 (1984).</li><li>Barnes, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+deficits+associated+with+senescence:+A+neurophysiological+and+behavioral+study+in+the+rat.">Memory deficits associated with senescence: A neurophysiological and behavioral study in the rat.</a> J. Comp. Physiol. Psych. 93 (1), 74-104 (1979).</li><li>Frick, K., Stillner, E., Berger-Sweeney, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mice+are+not+little+rats:+Species+differences+in+a+one-day+water+maze+task.">Mice are not little rats: Species differences in a one-day water maze task.</a> Neuroreport. 11 (16), 3461-3465 (2000).</li><li>Whishaw, I., Tomie, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Of+Mice+and+Mazes:+Similarities+Between+Mice+and+Rats+on+Dry+Land+But+Not+Water+Mazes.">Of Mice and Mazes: Similarities Between Mice and Rats on Dry Land But Not Water Mazes.</a> Physiol Behav. 60 (5), 1191-1197 (1995).</li><li>Francis, D., Zaharia, M., Shanks, N., Anisman, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress-induced+disturbances+in+Morris+water-maze+performance:+Interstrain+variability.">Stress-induced disturbances in Morris water-maze performance: Interstrain variability.</a> Physiol Behav. 58 (1), 57-65 (1995).</li><li>Wahlsten, D., Rustay, N., Metten, P., Crabbe, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+search+of+a+better+mouse+test.">In search of a better mouse test.</a> Trends Neurosci. 26 (3), 132-136 (2003).</li><li>Crawley, , 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=Behavioral+phenotypes+of+inbred+mouse+strains:+implications+and+recommendations+for+molecular+studies.">Behavioral phenotypes of inbred mouse strains: implications and recommendations for molecular studies.</a> Psychopharmacology. (Berl). 132 (2), 107-124 (1997).</li><li>Wahlsten, 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=Different+data+from+different+labs:+lessons+from+studies+of+gene-environment+interaction.">Different data from different labs: lessons from studies of gene-environment interaction.</a> J. Neurobiol. 54 (1), 283-311 (2002).</li><li>Rogers, D. 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=Use+of+SHIRPA+and+discriminant+analysis+to+characterise+marked+differences+in+the+behavioural+phenotype+of+six+inbred+mouse+strains.">Use of SHIRPA and discriminant analysis to characterise marked differences in the behavioural phenotype of six inbred mouse strains.</a> Behav Brain Res. 105 (2), 207-217 (1999).</li><li>Wolfer, D. P., Stagljar-Bozicevic, M., Errington, M. L., Lipp, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+Memory+and+Learning+in+Transgenic+Mice:+Fact+or+Artifact?.">Spatial Memory and Learning in Transgenic Mice: Fact or Artifact?.</a> Physiology. 13 (3), 118-123 (1998).</li><li>Koopmans, G., Blokland, A., Vannieuwenhuijzen, P., Prickaerts, J. <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+spatial+learning+abilities+of+mice+in+a+new+circular+maze.">Assessment of spatial learning abilities of mice in a new circular maze.</a> Physiol Behav. 79 (4-5), 683-693 (2003).</li><li>Deacon, R., Rawlins, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Learning+impairments+of+hippocampal-lesioned+mice+in+a+paddling+pool.">Learning impairments of hippocampal-lesioned mice in a paddling pool.</a> Behav Neurosci. 116 (3), 472-478 (2002).</li><li>Pettan-Brewer, 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=A+novel+radial+water+tread+maze+tracks+age-related+cognitive+decline+in+mice.">A novel radial water tread maze tracks age-related cognitive decline in mice.</a> Pathobiol Aging Age Relat Dis. 3, 1-4 (2013).</li><li>Wiley, J., Pettan-Brewer, C., Ladiges, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenylbutyric+acid+reduces+amyloid+plaques+and+rescues+cognitive+behavior+in+AD+transgenic+mice.">Phenylbutyric acid reduces amyloid plaques and rescues cognitive behavior in AD transgenic mice.</a> Aging Cell. 10 (3), 418-428 (2011).</li><li>Enns, L., 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=Disruption+of+Protein+Kinase+A+in+Mice+Enhances+Healthy+Aging.">Disruption of Protein Kinase A in Mice Enhances Healthy Aging.</a> PLoS ONE. 4 (6), (2009).</li><li>Ivonen, H., Nurminen, L., Harri, M., Tanila, H., Puolivali, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothermia+in+mice+tested+in+Morris+water+maze.">Hypothermia in mice tested in Morris water maze.</a> Behav Brain Res. 141 (2), 207-213 (2003).</li><li>Shultz, S. 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=Granulocyte-macrophage+colony-stimulating+factor+is+neuroprotective+in+experimental+traumatic+brain+injury.">Granulocyte-macrophage colony-stimulating factor is neuroprotective in experimental traumatic brain injury.</a> J Neurotrauma. 31 (10), 976-983 (2014).</li></ol>

作者没有什么可以披露的。

这里介绍的RWT迷宫协议成功地区分了CCI诱导的TBI小鼠和假手术对照，代表了可行的，以鼠标为中心的替代MWM和Barnes圆形迷宫。而在这里报道的结果只会讲的使用在TBI小鼠模型中RWT迷宫，该设备已成功应用于老年人和转基因模型，其中应力诱导基于游泳测试导致违规使用MWM不切实际15,17国产，18，19。不?...

记忆障碍是由创伤性脑损伤（TBI）1，2名患者报告的最常见的症状之一。因此，准确识别和评估TBI动物模型中的类似记忆缺陷对于我们对这种状况及其管理的理解至关重要。在这里，我们提出了一个使用径向水胎（RWT）迷宫测试TBI鼠标模型中的空间记忆的协议。以前显示该装置可以评估受控皮层影响（CCI）诱导的TBI 3的小鼠模型中的认知缺陷，并且代表了大鼠验证的，基于游泳的认知测试的潜在替代方案。 转基因小鼠模型的日益多样化和可用性已经导致科研4使用小鼠大鼠上最近有所增加。尽管有这种转变，研究人员继续依靠行为最初为老鼠使用设计和验证的认知任务。目前用于评估在小鼠中的认知中最常见的测试，Morris水迷宫（MWM）和巴恩斯圆形迷宫，是专门设计，以利用大鼠5,6发现本能的行为。考虑到这两个物种4之间存在的遗传，神经行为和认知的差异，这是不足为奇的小鼠上这些任务7,8始终表现不佳。 测试能力方面的物种依赖性差异特别涉及基于游泳的认知测试，如MWM。虽然大鼠和小鼠是基于游泳认知任务9,10显着表现不佳精通游泳，研究人员已经确定了几个小鼠品系， </SUP&gt; 11，12，13。即使是在野生型动物，老鼠通常优于小鼠7,8。虽然这可以解释为空间记忆中的物种特异性差异，但使用干地迷宫的类似随访测试显示认知表现没有物种依赖性差异8 。与认知无关的一些因素可以解释这一发现，包括游泳能力或搜索策略中的物种依赖性差异。事实上，MWM中小鼠特异性搜索策略的因素分析表明，非空间因素（特别是Thigomotaxis和被动[ 即浮动]）可能在MWM表现中起到比空间学习更重要的作用14 。 在这里，我们展示了使用认知测试来设计来利用它小鼠的直觉行为，不需要游泳，测量CCI诱导的TBI小鼠模型中的空间记忆障碍。虽然RWT迷宫（ 图1 的A-B）被设想为在MWM和Barnes圆形迷宫的新颖混合型，它是专为采取的行为thigmotactic本能给小鼠15,16优势。该设备包括一个32英寸直径的镀锌钢桶，其中九个均匀分布的出口孔已经钻孔。孔位于浴缸底部的2-1 / 4英寸处，并且尺寸适合普通可用的1-1 / 2英寸ABS DWV SPG x SJ陷阱适配器。八个出口被外部盖住，并且用橡皮塞挡住1英寸的深度。第九个通过90°丙烯腈丁二烯苯乙烯（ABS）弯头连接到不透明的塑料盒上，测试后可以从中轻松取出鼠标。在一个过程中短暂采集期间，老鼠训练使用迷宫内的独特视觉线索找到这个逃生箱。在测试过程中，迷宫充满了一英寸冷水（12-14°C），足够的足够强大的逃逸能力，但不足以使鼠标游泳。 在RWT迷宫表示一种低成本，低维护的替代MWM，并已在老年和转基因小鼠15，17，18，19成功地使用，和CCI诱导的TBI 3的小鼠模型。这里概述的方案代表了一种测量空间记忆障碍的简单有效的方法，不需要进行伤前训练，可以轻松修改以适应研究实验室的特殊需要。

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			<td height="40" style="height:40px;width:304px;">35 Gal. Hot Dipped Steel Round Tub</td>
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			<td height="40" style="height:40px;">Vernier Stereotaxic w/ Manual Fine Drive Stereotaxic Instrument for Small Animals</td>
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assessing spatial memory impairment in a mouse model of traumatic brain injury using a radial water tread maze

尽管近来在科学研究中使用小鼠模型的增加，研究人员继续使用最初设计和验证大鼠使用的认知任务。以前已经显示了空间记忆的径向水胎（RWT）迷宫测试（专为小鼠而设计，不需要游泳），以成功区分受控的皮层撞击诱导的TBI小鼠和假手术对照。在这里，介绍了此任务的详细协议。 RWT迷宫利用小鼠自然倾向避免开放区域，有利于拥抱设备的两侧（thigmotaxis）。迷宫的墙壁上装有九个放置在设备地板上的逃生孔，老鼠训练使用视觉线索来定位引出迷宫的逃生孔。迷宫充满了一英寸的冷水，足以激发逃脱，但不够深，要求鼠标游泳。收购期仅需四次天，在第5天进行记忆保留测试，并在第12天进行长期记忆测试。这里报告的结果表明，RWT迷宫是评估空间中大鼠验证的，基于游泳的认知测试的可行替代方案TBI小鼠模型的记忆缺陷。

RWT迷宫（ 图1 ）用于研究随机分配接受受控皮质冲击诱导的TBI或假手术的小鼠中损伤依赖性空间记忆缺陷。使用电磁驱动的皮质冲击产生损伤，其具有3mm的凸尖，并且以下损伤参数:6m / s的冲击速度，1mm的穿透深度和200ms的接触时间。小鼠从手术后35天开始接受认知测试，并进行了四天的培训（收购期），然后在第5天进行记忆保留测试，并在第12天进?...

Watch this Scientific Journal Video about Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze at JoVE.com

Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

在这里，我们提出了一个不需要游泳的认知的小鼠特异性测试方案。该测试可用于成功地区分受控皮质冲击引起的创伤性脑损伤小鼠与假手术对照。

使用径向水痕迷宫评估外伤性脑损伤小鼠模型中的空间记忆障碍

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and ...

assessing-spatial-memory-impairment-mouse-model-traumatic-brain

Universidad de Cartagena UDEC

Research

JoVE Journal

Behavior

7.0K 次观看.  University of Washington. 在这里，我们提出了一个不需要游泳的认知的小鼠特异性测试方案。该测试可用于成功地区分受控皮质冲击引起的创伤性脑损伤小鼠与假手术对照。 

视频: Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

Shallow Water (Paddling) Variants of Water Maze Tests in Mice

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to the mouse becoming increasingly important. But researchers found that some strains of mutant mice were prone to problems like passively floating or diving when they were tested in the Morris water maze11. This was unsurprising considering their natural habitat; rats swim naturally (classically, the "sewer rat"), whereas mice evolved in the dry areas of central Asia.
To overcome these problems, it was considered whether shallow water would be a sufficient stimulus to provide escape motivation for mice. This would also avoid the problems of drying the small creatures with a towel and then putting them in a heated recovery chamber to avoid hypothermia, which is a much more serious problem than with rats; the large ratio of surface area to volume of a mouse makes it particularly vulnerable to rapid heat loss.
Another consideration was whether a more natural escape strategy could be used, to facilitate learning. Since animals that fall into water and swim away from the safety of the shore are unlikely to pass on their genes, animals have evolved a natural tendency to swim to the edge of a body of water. The Morris water maze, however, requires them to swim to a hidden platform towards the center of the maze - exactly opposite to their evolved behavior. Therefore the paddling maze should incorporate escape to the edge of the apparatus. This feature, coupled with the use of relatively non-aversive shallow water, embodies the "Refinement" aspect of the "3 Rs" of Russell and Burch8.
Various types of maze design were tried; the common feature was that the water was always shallow (2 cm deep) and escape was via a tube piercing the transparent wall of the apparatus. Other tubes ("false exits") were also placed around the walls but these were blocked off. From the inside of the maze all false exits and the single true exit looked the same. Currently a dodecagonal (12-sided) maze is in use in Oxford, with 12 true/false exits set in the corners. In a recent development a transparent paddling Y-maze has been tested successfully.

Shallow Water Paddling Variants of Water Maze Tests in Mice

Mice can swim, but many strains appear to find this activity stressful. To overcome this problem mazes have been devised where escape from shallow water is used to motivate behaviour. These have been demonstrated to support learning at least as good as the traditional and widely used Morris water maze.

浅水（杖刑）的变体在小鼠体内的水迷宫测试

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to ...

科研

行为学

Use of an Eight-arm Radial Water Maze to Assess Working and Reference Memory Following Neonatal Brain Injury

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food deprivation, and may introduce scent cue confounds. The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs. Specifically, subjects are required to avoid the arms previously used for escape during each testing day (working memory) as well as avoid the fixed arms, which never contain escape platforms (reference memory). Re-entries into arms that have already been used for escape during a testing session (and thus the escape platform has been removed) and re-entries into reference memory arms are indicative of working memory deficits. Alternatively, first entries into reference memory arms are indicative of reference memory deficits. We used this maze to compare performance of rats with neonatal brain injury and sham controls following induction of hypoxia-ischemia and show significant deficits in both working and reference memory after eleven days of testing. This protocol could be easily modified to examine many other models of learning impairment.

The eight-arm radial water maze is designed to evaluate reference and working memory performance simultaneously by requiring subjects to use extra-maze cues to locate escape platforms and remedies the limitations observed in land based radial arm maze designs.

Working and reference memory are commonly assessed using the land based radial arm maze.&#160;However, this paradigm requires pretraining, food ...

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their behavioral flexibility and required memory duration, the number of goals and pathways, and also the overall task complexity. A confounding feature in many of these tasks is the lack of control over the strategy employed by the rodents to reach the goal, e.g., allocentric (declarative-like) or egocentric (procedural) based strategies. The double-H maze is a novel water-escape memory task that addresses this issue, by allowing the experimenter to direct the type of strategy learned during the training period. The double-H maze is a transparent device, which consists of a central alleyway with three arms protruding on both sides, along with an escape platform submerged at the extremity of one of these arms.
Rats can be trained using an allocentric strategy by alternating the start position in the maze in an unpredictable manner (see protocol 1; &#167;4.7), thus requiring them to learn the location of the platform based on the available allothetic cues. Alternatively, an egocentric learning strategy (protocol 2; &#167;4.8) can be employed by releasing the rats from the same position during each trial, until they learn the procedural pattern required to reach the goal. This task has been proven to allow for the formation of stable memory traces.
Memory can be probed following the training period in a misleading probe trial, in which the starting position for the rats alternates. Following an egocentric learning paradigm, rats typically resort to an allocentric-based strategy, but only when their initial view on the extra-maze cues differs markedly from their original position. This task is ideally suited to explore the effects of drugs/perturbations on allocentric/egocentric memory performance, as well as the interactions between these two memory systems.

The goal of this protocol is to investigate spatial cognition in rodents. The double-H water maze is a novel test, which is particularly useful to elucidate the different components of learning, consolidation and memory, as well as the interplay of memory systems.

双-H迷宫:一个强大的行为测试学习和记忆的鼠害

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their ...

Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment

The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, including neurocognitive disorders such as Alzheimer&#8217;s disease. However, the background strain of the mouse model used can have a substantial effect on the observed behavioral phenotype, with some strains exhibiting superior learning ability relative to others. To ensure differences between transgene negative and transgene positive mice can be detected, identification of a training procedure sensitive to the background strain is essential. Failure to tailor the MWM protocol to the background strain of the mouse model may lead to under- or over- training, thereby masking group differences in probe trials. Here, a MWM protocol tailored for use with the F1 FVB/N x 129S6 background is described. This is a frequently used background strain to study the age-dependent effects of mutant P301L tau (rTg(TauP301L)4510 mice) on the memory deficits associated with Alzheimer&#8217;s disease. Also described is a strategy to re-optimize, as dictated by the particular testing environment utilized.

This manuscript describes a Morris water maze (MWM) protocol tailored for use with a commonly used mouse model of Alzheimer's disease. The MWM is widely used in transgenic mouse models. Implementation of a procedure sensitive to the background strain of the mouse model is essential for detecting group differences.

Morris水迷宫实验:优化小鼠品系和测试环境

 The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

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

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

颅脑损伤后大鼠行为缺陷的检测

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

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory alterations of the spinal cord. In this article, our goal was to use both Laser Doppler imaging and monitoring to analyze the change of BF after spinal cord injury. Both the laser Doppler image scanner and the probe/monitor were being employed to obtain each readout. The data of LDPI provided a local distribution of BF, which gave an overview of perfusion around the injury site and made it accessible for comparative analysis of BF among different locations. By intensely measuring the probing area over a period of time, a combined probe was used to simultaneously measure the BF and oxygen saturation of the spinal cord, showing overall spinal cord perfusion and oxygen supply. LDF itself has a few limitations, such as relative flux, sensitivity to movement, and biological zero signal. However, the technology has been applied in clinical and experimental study due to its simple setup and rapid measurement of BF.

Here we present a combination of laser Doppler perfusion imaging (LDPI) and laser Doppler perfusion monitoring (LDPM) to measure spinal cord local blood flows and oxygen saturation (SO2), as well as a standardized procedure for introducing spinal cord trauma on rat.

激光多普勒成像与监测在大鼠脊髓微循环分析中的应用

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory ...

A Behavioral Assay for Investigating the Role of Spatial Memory During Instinctive Defense in Mice

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often stereotyped actions elicited in response to innately aversive sensory stimuli, but their success requires enough flexibility for adapting to different spatial environments, which can change rapidly. Here, we describe a behavioral assay to evaluate the influence of learned spatial knowledge on defensive behaviors in mice. We have adapted the widely used Barnes maze spatial memory assay to investigate how mice navigate to a shelter during escape responses to innately aversive sensory stimuli in a novel environment, and how they adapt to acute changes in the environment. This new assay is an ethological paradigm that does not require training and exploits the natural exploration patterns and navigation strategies in mice. We propose that the set of protocols described here are a powerful means of studying goal-directed behaviors and stimulus-triggered navigation, which should be of interest to both the fields of instinctive behaviors and spatial memory.

This protocol describes a behavioral assay, based on the Barnes maze test, to study how instinctive defensive actions are modified by the knowledge of spatial environment.

一种研究空间记忆在小鼠本能防御中作用的行为分析法

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often ...

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 ...

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various physiological or pathological changes. The object location task (OLT) and the novel object recognition task (NORT) are two effective behavioral tasks commonly used to reveal the function and relative health of specific brain regions involved in memory. While both of these tests exploit the inherent preference of mice for the novelty to reveal memory for previously encountered objects, the OLT primarily evaluates spatial learning, which relies heavily on hippocampal activity. The NORT, in contrast, evaluates non-spatial learning of object identity, which relies on multiple brain regions. Both tasks require an open-field-testing arena, objects with equivalent intrinsic value to mice, appropriate environmental cues, and video recording equipment and the software. Commercially available systems, while convenient, can be costly. This manuscript details a simple, cost-effective method for building the arenas and setting up the equipment necessary to perform the OLT and NORT. Furthermore, the manuscript describes an efficient testing protocol that incorporates both OLT and NORT and provides typical methods for data acquisition and analysis, as well as representative results. Successful completion of these tests can provide valuable insight into the memory function of various mouse model systems and appraise the underlying neural regions that support these functions.

Here we provide a protocol which includes comprehensive instructions for the economical establishment of murine object location and novel object recognition behavioral testing, including the design, cost, and construction of required equipment as well as execution of behavioral testing, data collection, and analysis.

一种新的小鼠目标识别与目标定位行为测试

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various ...

Analysis of Learning and Memory Ability in an Alzheimer's Disease Mouse Model using the Morris Water Maze

A Morris water maze (MWM) experiment forces experimental animals to swim and learn to find a platform hidden in the water. It is widely used in scientific research to assess the learning and memory of animals. Due to the extensive use of the MWM test, visual experimental protocols are essential for researchers. This manuscript uses the latest studies to introduce the protocol of the MWM test. Alzheimer&#8217; Disease (AD) is characterized by a progressive loss of memory and cognitive function. An alternative and complementary treatment used for AD is Manual Acupuncture (MA). To assess the learning and memory ability of AD model mice, the MWM test was conducted. The visible platform trial, hidden platform trial, probe trial, and reversal trial of MWM were used to evaluate spatial learning and memory ability. In the visible platform trial, the swimming speed and escape latency of mice in different groups was not significantly different. In the hidden platform and reversal trials, the AD group showed a long escape latency. The escape latency decreased significantly after the MA treatment. Low platform crossover number and the proportion of time in the SW quadrant in the probe trial increased after the MA treatment (p &#60; 0.05 or p &#60; 0.01). The results of the MWM tests suggest that MA can effectively improve the spatial learning and memory abilities of AD model mice. Rigorous experimental operations provided assurance of the reliability of the results.

Herein, a protocol to conduct the Morris water maze tests to evaluate the ability of learning and memory of Alzheimer&#8217;s Disease model mice and to assess the effect of manual acupuncture for treating them is described.

使用莫里斯水迷宫对阿尔茨海默氏病小鼠模型学习和记忆能力的分析

A Morris water maze (MWM) experiment forces experimental animals to swim and learn to find a platform hidden in the water. It is widely used in scientific ...