这项工作是由美国国立卫生研究院支持的补助CA163640，CA166590和AG041250到LC。

下面所描述的程序已经批准了美国俄亥俄州立大学的机构动物护理和使用委员会（IACUC）。 1.准备丰富环境之前，房屋小鼠<ol><li>清洁和维护仪器。 <ol><li>通过洗笼运行大型铀浓缩仓（120厘米×90厘米×76厘米）。一个典型的动物笼子设施清洗协议包括5至6次的洗涤，最后到达周期82ºC。整个清洗大约需要40分钟。空气干燥仓。 <ol><li>如果大富集bin和任何其他壳体仪器，无法通过笼洗涤被发送，然后喷雾和擦拭用按照制造商的说明，杀孢子消毒剂的bin的内部。擦去用自来水浸湿的纸巾或30％乙醇和用干纸巾的bin摆脱从以前的任何消毒剂气味。 </li></ol></li><li>使用PLAS抽动油管，用圆顶型飞碟车轮，而在其他EE各种塑料笼般的玩具。通过笼洗洗这些玩具。如果您没有访问笼清洗的后续步骤1.1.1.1。 </li><li>使用金​​属转轮在EE。通过笼洗洗车轮。如果在车轮的涂料开始褪色，使得车轮变得嘈杂当动物在其上运行，将少量凡士林到车轮和阿克塞尔连接。 </li><li>使用木原木在EE。通过消除手工任何被褥或其他杂物清理干净的日志。然后，通过高压灭菌它们在干燥循环或纺织品周期，这达到121°C持续15分钟的温度杀菌日志。 </li><li>使用的29厘米×18厘米×12.5厘米和30.5厘米×19厘米×14厘米维度之间笼成保持进料和饮用导线rack.Cut空穴至少5厘米，直径从笼的前端底部的老鼠从EE进出的。通过清洁洗笼喂养这些笼子。 </li></ol></li><li>设置环境<ol><li>倒入¼英寸的玉米棒实验动物的床上用品到大仓富集。覆盖仓的整个地板表面的2-2.5厘米的床上用品的深度。 </li><li>将两个或三个日志中的仓的一个角落。在边角处日志隔离约1225厘米2的面积。通过日志隔离的区域，放三冰屋在顶部飞碟车轮。确保车轮未接触的bin，日志，或彼此的壁，以便有足够的空间供车轮自由旋转。 </li><li>拼凑的塑料管，以便多个臂从森特拉尔连接件分支出来。把这个管子在bin中间。 </li><li>在仓的两个角，将每个填充有相同寝具作为箱柜1馈送笼，并在馈送笼前部的孔应朝向仓的中心。在这些笼子里喂一个干净的食物和水线之上应放置于保持食物和水。不要将任何盖子上笼。放的食物和水的笼子里的电线。 </li><li>将三个金属轮子的箱子，虽然不是所有的彼此相邻。三种不同类型的车轮是一体11.5厘米车轮一20.5厘米轮，以及一个28厘米车轮。然后将4-5更多的塑料笼般的玩具或隧道贯穿富集仓。 </li><li>覆盖微型隔离过滤纸仓。用长尾夹滤纸夹到bin。 </li><li>新颖性是重要的EE。使用该设置作为EE的框架和变化的放置上述笼，对象和玩具以创建一个新的环境，为动物。 </li></ol></li></ol> 2.房屋的动物<ol><li>动物繁殖在常规实验室的住房，而不是EE实验。或者，将动物EE年龄早21天断奶后。基因型和耳朵打孔/标签动物前EE住房。 </li><li>当放置进入动物从笼子标准富集箱，放置在其一侧仓笼，使得动物可以从标准笼富集仓自由移动。不允许嵌套的材料被转移到富集箱。 </li><li>众议院10-20动物一样的年龄，性别，并在EE基因菌株。 </li></ol> 3.维护环境和观察小鼠在研究<ol><li>日报<ol><li>静静地观察动物的疾病或痛苦的迹象。如果有必要，轻轻地处理动物进一步检查健康状况。记录异常的观察和咨询卫生技术人员或兽医人员如果需要去除的实验动物显示条件。 </li><li>提供三重抗生物药膏小伤口结痂或局部治疗。如果动物需要因慢性疾病或伤害被删除，从实验安乐死取出的动物。如果动物满足了监督动物研究护理委员会或兽医早期切除的标准，不回动物回EE。 </li></ol></li><li>一个星期两次<ol><li>检查饲料和饮水的水平。确保饲料和水将持续在未来3-4天。整顿运行车轮和富集玩具使它们展开并且可以使用小鼠。更换任何过于肮脏。 </li></ol></li><li>周刊<ol><li>更换水瓶用干净的矿泉水瓶。重新排列在下面的第1.2节设置的方式浓缩设备。然而从此前一周的设置不同。 注意:如果一个实验要求从富集的环境中取出为活性的动物（记录体重，抽血，行为试验等 ），最好是重新排列的环境，而动物是远离富集仓。 </li><li>手工清洁的EE饲养笼或通过洗笼发送详见第1.1.5。更换食物和水持有干净的线架以及导线架。 </li><li>按照第2.2节的指令。当放置在浓缩的动物回来。现货必要时检查大型铀浓缩bin和清洁的休息，由舀出肮脏的垃圾和推广新的下跌干净的被褥。 </li></ol></li><li>双周<ol><li>在执行每周饲养，手工清洗的玩具或者通过发送洗笼在1.1.2节详述。如果需要的话，将小鼠在标准的微隔离器笼清洁所需的时间。 </li></ol></li><li>每月一次<ol><li>清洁大多数垃圾出移动富集玩具和饲养笼和舀出的垃圾。更换新鲜垫料。确保有床上用品，在笼子里的各个领域之一英寸。如果需要的话，将小鼠在标准的微隔离器笼清洁所需的时间。 </li></ol></li><li>每三个月（或在实验结束） <ol><li>删除所有的床上用品和玩具，干净，如第1.1.1节所述消毒富集仓。如果实验正在进行中，更换新的床上用品，饲养笼，玩具琳琅满目，然后返回小鼠到bin。 </li></ol></li><li>如果需要的研究机构，其中EE坐落更频繁地进行消毒，整个富集比斌每三个月。如果这是必要的，另一家大型货柜充实可用，设置其他富集容器在1.2节详述。和动物转移到新的容器中，而前一个被清洗。 </li></ol> 4.控制住房情况<ol><li>房子的动物的对照组中尺寸30.5厘米×19厘米×14厘米的微隔离器笼中。包括用于盛装食物和水的瓶子，如果笼不挂接到一个自动洒水系统在笼子里的电线架。 </li><li>提供相同的玉米芯被褥在控制笼在EE。为4cm的×4厘米的nestlet的老鼠做窝出来。 </li><li>更换和/或清洁笼子，喂养一周一次的电线，食物和水。通过笼洗干净的笼子和铁丝衣架或第1.1.1.1尽可能详细。更加频繁地清洗鸡笼，如果动物小便比较频繁，或者如果饮用水泄漏到床上用品。 </li><li>监测控制住房日常动物的健康。保证动物有充足的食物和水。 </li><li>众议院3〜5只小鼠同性别的普通鼠笼的。不提供任何玩具，管，或在笼笼。 </li><li>机构实验动物护理设施的健康和实验室技术人员可以对这些控制老鼠提供饲养。 </li><li>饲养动物在12小时的光/暗周期作为对于小鼠中EE。 </li></ol>

<ol>
	<li>Cao, L., During, 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=What+is+the+brain-cancer+connection.">What is the brain-cancer connection.</a> Annu. Rev. Neurosci. 35, 331-345 (2012).</li><li>Nithianantharajah, J., Hannan, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enriched+environments,+experience-dependent+plasticity+and+disorders+of+the+nervous+system.">Enriched environments, experience-dependent plasticity and disorders of the nervous system.</a> Nat. Rev. Neurosci. 7, 697-709 (2006).</li><li>Milsum, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+model+of+the+eustress+system+for+health/illness.">A model of the eustress system for health/illness.</a> Behav. Sci. 30, 179-186 (1985).</li><li>Selye, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Stress Without Distress. , (1974).</li><li>Krech, D., Rosenzweig, M. R., Bennett, E. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+environmental+complexity+and+training+on+brain+chemistry.">Effects of environmental complexity and training on brain chemistry.</a> J. Comp. Physiol. Psychol. 53, 509-519 (1960).</li><li>Rosenzweig, M. R., Krech, D., Bennett, E. L., Diamond, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+environmental+complexity+and+training+on+brain+chemistry+and+anatomy:+A+replication+and+extension.">Effects of environmental complexity and training on brain chemistry and anatomy: A replication and extension.</a> J. Comp. Physiol. Psychol. 55, 429-437 (1962).</li><li>Frick, K. 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=Effects+of+Environmental+Enrichment+on+Spatial+Memory+and+Neurochemistry+in+Middle-Aged+Mice.">Effects of Environmental Enrichment on Spatial Memory and Neurochemistry in Middle-Aged Mice.</a> Learn. Mem. 10 (3), 187-198 (2003).</li><li>Dellen, A., Blakemore, C., Deacon, R., York, D., Hannan, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delaying+the+onset+of+Huntington&#8217;s+in+mice.">Delaying the onset of Huntington&#8217;s in mice.</a> Nature. 404, 721-722 (2000).</li><li>Spires, T. 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=Environmental+enrichment+rescues+protein+deficits+in+a+mouse+model+of+Huntington&#8217;s+disease,+indicating+a+possible+disease+mechanism.">Environmental enrichment rescues protein deficits in a mouse model of Huntington&#8217;s disease, indicating a possible disease mechanism.</a> J. Neurosci. 24 (9), 2270-2276 (2004).</li><li>Jankowsky, J. 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=Environmental+enrichment+mitigates+cognitive+deficits+in+a+mouse+model+of+Alzheimer&#8217;s+disease.">Environmental enrichment mitigates cognitive deficits in a mouse model of Alzheimer&#8217;s disease.</a> J. Neurosci. 25 (21), 5217-5224 (2005).</li><li>Westood, J. A., Darcy, P. K., Kershaw, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Environmental+Enrichment+does+not+impact+on+tumor+growth+in+mice.">Environmental Enrichment does not impact on tumor growth in mice.</a> F1000Research. 2, 140 (2013).</li><li>Cao, 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=VEGF+links+hippocampal+activity+with+neurogenesis,+learning.">VEGF links hippocampal activity with neurogenesis, learning.</a> Nat. Genet. 36 (8), 827-835 (2004).</li><li>Young, D., Lawlor, P. A., Leone, P., Dragunow, M., During, 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=Environmental+enrichment+inhibits+sponta-+neous+apoptosis,+prevents+seizures+and+is+neuroprotective.">Environmental enrichment inhibits sponta- neous apoptosis, prevents seizures and is neuroprotective.</a> Nat. Med. 5 (4), 448-453 (1999).</li><li>Cao, 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=Environmental+and+genetic+activation+of+a+brain-adipocyte+BDNF/leptin+axis+causes+cancer+remission+and+inhibition.">Environmental and genetic activation of a brain-adipocyte BDNF/leptin axis causes cancer remission and inhibition.</a> Cell. 142 (1), 52-64 (2010).</li><li>Cao, 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=White+to+brown+fat+phenotypic+switch+induced+by+genetic+and+environmental+activation+of+a+hypothalamic-adipocyte+axis.">White to brown fat phenotypic switch induced by genetic and environmental activation of a hypothalamic-adipocyte axis.</a> Cell Metab. 14 (3), 324-338 (2011).</li></ol>

作者宣称没有利益冲突。

EE对基因表达的健壮效果在大脑中，降低肥胖和癌症生长在小鼠中在很大程度上取决于研究者的能力，以保持新的环境和处理的动物中促进身体活动，社交和认知挑战的方式，但没有痛苦的动物。有两个最重要的程序性步骤，记住。首先，记住每星期更改放置玩具，物件，和笼子的。尽管玩具和物体可能与前一周相同，老鼠感知环境的小说。更改玩具和对象本身具有不同的人每隔一周。其次，记得要保持环境清洁。如果弄脏被褥没有得到消除，增加的氮含量可以是一个压力源给动物。 此外，还有许多技术来注意到本EE协议。利用不同的或独特的玩具和不同塑料管的迷宫设计的鼓励。环境可以是具有挑战性的，但应不构成对动物的健康构成威胁。此外，要注意玩具的功能失调的，因为不正常的玩具增加鼠标肢体的风险被抓住，或者消耗咀嚼它可能是有毒的动物。一恒定的EE是使用三个飞碟车轮上圆顶的顶部，由2-3日志螯合到bin的一角或一侧。运行日志后面的碟子这样的设置一直存在于我们所有的铀浓缩研究。在其他对象的使用和放置而言，布局的以上描述是EE设置的基本蓝图。变异的塑料管这个基础框架，车轮跑，跑碟，网箱和日志应提供足够的新的环境，以重现报告的结果设置的。 如果与患有糖尿病或其他代谢转基因动物工作疾病，狠抓食品和水的消耗，以确保动物不会跑出来的食物和水。如果动物有糖尿病，他们会小便较多，因此EE壳体的被褥可能需要现场检查和清洁更频繁。在EE房屋需要拆除的床上用品最常见的地方是死角。记得调整车轮等玩具，让他们可以使用该动物。动物是能够运行飞碟轮进被褥和运行敲了车轮的。如果动物是年轻的，3到5周龄，他们可能挖下在EE的笼中，并在那里隐藏起来，使研究人员无能，其中动物。当拆除弄脏被褥，玩具更换或清洗笼子，建议把动物在一个干净整洁的标准笼，以免他们强调了在调整他们的环境。这是比较容易得到所有的动物在一个笼子里，如果他们有较少的地方，在ENV隐藏ironment。因此，之前封存的动物关在笼子里取出玩具，使过程更容易。此外，不要将超过5-6动物在一个单一的标准笼子在等待清洗或调整自己的EE环境。雄性动物EE可以是积极的特别是当EE在成年期开始。因此，观察其环境的任何受伤或死亡的动物是很重要的。 此外，动物的典型数目，我们的房子在EE集装箱的这种大尺寸为10-20只。动物的数量其他研究者使用EE而变化，使用约10或更少的小鼠，和笼或仓的尺寸应确定的动物的数量，以房子在EE时予以考虑。有太多的动物的生活环境会导致侵略雄性小鼠。我们住在EE老鼠一直是C57BL / 6背景。其它菌株的小鼠可以是更多或更少的攻击性，或更容易出现健康问题这个EE设置。这应该被考虑在确定为小鼠和富集的环境的饲养计划。 四周EE壳体足以诱导减少肥胖，降瘦素水平，并抑制B16黑色素瘤的生长。再EE会导致新陈代谢和抑制癌症的作用更强。如果小鼠中或暴露于EE之前经受外科手术，可以将它们安置与缝线完好无损。是否动物反应良好的这个特定EE设置与外科U形钉或导管完好尚未研究。此外，我们目前正在调查这个特殊的EE如何影响老年动物的寿命或长期保健。初步数据表明，雌性C57BL / 6小鼠中，10个月的年龄，响应于类似EE到年轻小鼠。在EE壳体的一个限制是容纳这样一个大容器在动物设施的空间，并优先地在一个性别特定房间。因此，未来的研究EFFEE的规模较小学分，以及发现玩具易清洗，忍受高温清洁和安全的动物健康，将改善的EE中代谢和癌症研究中的应用。

丰富环境（EE）是用来探索的复杂和具有挑战性的环境中，不强调动物1效果的实验范式。 EE是一个增加的空间，体力活动，和社会交往，这反过来又增加感觉，认知，运动和社会刺激2组成动物壳体技术。冰屋，运行轮子，轮子飞碟，管迷宫，和其他对象的居住环境推动这一感官认知，社会和运动刺激，促进勘探和互动。在外壳的变化的对象的布局经常提出了一种新的环境中，向其中将小鼠被迫适应。这种策略提供了诱导有益的，良性的，健康的适应性反应，称为良性压力，而不是更厌恶，恶劣的环境或适应不良的困扰3,4-温和和短暂的挑战。通过促进良性压力和限制有害困扰，老鼠展示各种有利表型。 EE是一个可靠的实验模型具有广泛的应用，而具有不会产生传统上认为是压力1元素。已经研究了超过半个世纪EE对啮齿动物的影响。在20世纪60年代早期的研究表明，皮质胆碱酯酶水平下降和皮层下胆碱酯酶水平在被安置在环境与增加的生活空间，大鼠的数量增加，以及各种木制玩具，迷宫和平台5,6-老鼠增加。此外，EE与在不同年龄和遗传背景的小鼠中的脑，行为和代谢许多适应性变化相关联。因此，EE是进行的实验研究对小鼠的环境的影响的范围广阔的有用的范例。 EE效果可以在小鼠中一样迅速被观察为一个星期曝光，并且可以在动物中作为旧被视为18个月7。 EE是经常ü在sed的神经科学和神经行为实验，研究恢复功能，学习和记忆2。此外，EE对各种毒素诱导和人类神经系统疾病的转基因动物模型，包括亨廷顿氏病，阿尔茨海默氏病，帕金森氏症，癫痫症，中风的表型相当大的影响，并且更2。 然而，多年来一直存在矛盾与来自不同实验室的艰苦学习的充实设计进行比较。例如，每对相对于上面提到的人类神经系统疾病的EE效果的研究已经变参数的富集。亨廷顿氏病的研究收纳4-6小鼠的大笼（44×28×12.5厘米3）含有纸箱，隧道，和片材，以及木，塑料对象被改变每两天8,9。一对老年性痴呆所做的研究＃8217; S在包含2转轮，塑料管，纸箱被改变或重新安排每周10的排料较大的笼（1米3）装20只。 EE的许多设置有类似的主题，例如上运行或隐藏的物体，但每EE外壳，生活区的小鼠的数目，和改变环境的周期变化不大。 EE的不一致的设计导致了电子工程效果失败的复制。我们对癌症和新陈代谢数据一起表明，在下面的详细EE壳体协议导致显著抗癌和抗肥胖效果14,15提供刺激的结合。然而，通过不同的实验室可能影响EE的结果，如被认为是在一个EE设置的差值未能尝试重现上的植入小鼠11特定癌症肿瘤的生长EE影响。的不同的结果可能是由于多种原因的范围往复米的肿瘤细胞培养物，以用于清洁房屋设备11洗涤剂的类型的不同的增长率。为了复制看到在我们的实验的结果，浓缩和畜牧业的特定设计是必需的。下面的过程详细介绍了EE住房，将允许其他研究人员成功地复制我们的有效富集模式。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Large enrichment bin (120cm x 90cm x 76cm)</td>
			<td>Custom made.</td>
			<td>Custom made.</td>
			<td>Made of polypropylene. Edges of the bin are 1.5cm thick.</td>
		</tr>
		<tr>
			<td>Large enrichment bin alternative (120cm x 110cm x 76cm)</td>
			<td>Global Industrial</td>
			<td>155202</td>
			<td>If necessary, this bin works as an alternative to the one above.</td>
		</tr>
		<tr>
			<td>Habitrail Mini tubes</td>
			<td>Habitrail</td>
			<td>62720; 62705; 62725; 62715; 62054</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Plastic igloos with saucer wheels</td>
			<td>Bio Serv</td>
			<td>K3327; K3250</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Metal Running Wheels</td>
			<td>Super Pet</td>
			<td>61392; 61394; 61396</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Manzanita Wood Gnawing Sticks, 3&#34;</td>
			<td>Bio Serv</td>
			<td>W0016</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse Tunnels</td>
			<td>Bio Serv</td>
			<td>K3322</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Mouse cages that go in the large EE bin.</td>
			<td>Lab Products Inc</td>
			<td>10027 Mouse Cage</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Cage Klenz 180 Alkaline Cage Wash Detergent, Cage Klenz 250 Acid Detergent</td>
			<td>Steris Life Sciences</td>
			<td>1K1802; 1K2502</td>
			<td>These detergents are used in the cage wash for the toys, large enrichment bins, and cages.</td>
		</tr>
		<tr>
			<td>Sporklenz</td>
			<td>Steris Life Sciences</td>
			<td>6526Q8</td>
			<td>Alternative sporicide/disinfectant to washing toys, bins, and cages in cage wash.</td>
		</tr>
		<tr>
			<td>Bed-O-Cobs 1/4&#34; bedding</td>
			<td>The Andersons, Inc.</td>
			<td>4B</td>
			<td>Corn cob bedding is 1/4&#34; in diameter.</td>
		</tr>
		<tr>
			<td>Nestlets for control housed mice</td>
			<td>Ancare</td>
			<td></td>
			<td>Cotton squares</td>
		</tr>
	</tbody>
</table>

a protocol for housing mice in an enriched environment

Environmental enrichment (EE) is a housing environment for mice that boosts mental and physical health compared to standard laboratory housing. Our recent studies demonstrate that environmental enrichment decreases adiposity, increases energy expenditure, resists diet induced obesity, and causes cancer remission and inhibition in mice. EE typically consists of larger living space, a variety of &#8216;toys&#8217; to interact with, running wheels, and can include a number of other novel environmental changes. All of this fosters a more complex social engagement, cognitive and physical stimulations. Importantly, the toy location and type of toy is changed regularly, which encourages the mice to adapt to a frequently changing and novel environment. Many variables can be manipulated in EE to promote health effects in mice. Thus these approaches are difficult to control and must be properly managed to successfully replicate the associated phenotypes. Therefore, the goal of this video is to demonstrate how EE is properly set up and maintained to assure a complex, challenging, and controlled environment so that other researchers can easily reproduce the protective effects of EE against obesity and cancer.

EE影响生长因子的表达，增加了中枢神经系统（CNS）12,13内神经发生和细胞的存活。最近的研究已经表明，EE这种技术导致癌症生长的显着抑制在三个小鼠癌症模型包括B16黑色素瘤，MC38结肠癌和APC 分钟/ +小鼠包含种系突变和很容易受到自发腺瘤的形成肠子14。结果在图2中确认的B16黑色素瘤的肿瘤生长抑制植入暴露于EE 5周（〜减少肿瘤质量的80％）的小鼠。癌细胞生长抑制甚至在小鼠中观察到，当EE发起的肿瘤已经建立了14后。 EE造成了更大的抑制黑素瘤生长在高脂饮食诱导的肥胖小鼠（〜减少肿瘤质量的70％）相比，在正常体重（〜减少肿瘤质量的50％）14只小鼠。 EE IND<html> uced签名变化的血清生物标志物包括鲁棒降瘦素，减少的IGF-1，和脂联素水平14的上升。 此外，EE的这种技术减少肥胖，增加能量消耗，导致对饮食诱导的肥胖抗性，并诱导白色至棕色脂肪表型开关通过激活下丘脑- sympathoneural脂肪细胞轴线15。在肥胖的减少被认为是早期六天暴露于EE（ 图3A，B），并再次在动物暴露于EE四星期（ 图3C，D）的确认。同样重要的是要注意的是，一个，使用EE相关的抗癌和抗肥胖效果不能由体力活动记帐单，和，二，EE效果可以通过脑源性神经营养因子的过度表达被再现（BDNF ）下丘脑14,15。 <html> 2874fig1.jpg"/&gt; 图1:丰富环境仓建立的丰富环境有120厘米×90厘米面积包括两个修改笼，跑车轮，带轮子飞碟，塑料管材，木原木等塑料玩具富集冰屋。 <img alt="图2" src="/files/ftp_upload/52874/52874fig2.jpg" /> 图2:丰富环境降低了肿瘤的生长C57BL / 6雄性小鼠三周龄雄性小鼠的房子在电子工程或控制的笼子前，肿瘤接种5周。。 B16黑色素瘤肿瘤重量在每只动物皮下10 5个细胞的接种上述右胁17天后（每组n = 10组。学生t-检验 ，* P &lt;0.05）。 <img alt="图3a" src="/files/ftp_upload/52874/52874fig3a.jpg" /> <img alt="图3b" src="/files/ftp_upload/52874/52874fig3b.jpg" /> S / ftp_upload / 52874 / 52874fig3c.jpg"/&gt; <img alt="图3d" src="/files/ftp_upload/52874/52874fig3d.jpg" /> 图3:丰富环境减少肥胖的C57BL / 6雄性小鼠三周龄小鼠进行EE。一组6天，另四个星期。 （ 一 ）在EE六天减少脂肪垫的重量（N =每组5。学生的t检验 ，* P &lt;0.05）。 （ 二）在EE六天减少脂肪垫的重量相对于体重（N =每组5。学生的t检验 ，* P &lt;0.05）。 （C）四个星期下降EE脂肪垫的重量（N =每组5。学生的t检验 ，* P &lt;0.05）。 （D）四个星期下降EE脂肪垫的重量相对于体重（N =每组5。学生的t检验 ，* P &lt;0.05）。 BAT:棕色脂肪组织，IWAT:腹股沟白色脂肪组织，EWAT:附睾白色脂肪组织，RWAT:腹膜后脂肪组织。

Watch this Scientific Journal Video about A Protocol for Housing Mice in an Enriched Environment at JoVE.com

A Protocol for Housing Mice in an Enriched Environment

Environmental enrichment for mice requires a complex and challenging setup, as well as comprehensive husbandry and handling techniques to assure robust metabolic and anti-cancer effects in the mice. This protocol provides detailed procedures to reproduce the above mentioned effects in mice.

在丰富环境的协议房屋小鼠

Environmental enrichment (EE) is a housing environment for mice that boosts mental and physical health compared to standard laboratory housing. Our recent ...

a-protocol-for-housing-mice-in-an-enriched-environment

Research

JoVE Journal

Behavior

18.2K 次观看.  The Ohio State University. Environmental enrichment for mice requires a complex and challenging setup, as well as comprehensive husbandry and handling techniques to assure robust metabolic and anti-cancer effects in the mice. This protocol provides detailed procedures to reproduce the above mentioned effects in mice.

视频: A Protocol for Housing Mice in an Enriched Environment

Two-photon Calcium Imaging in Mice Navigating a Virtual Reality Environment

In recent years, two-photon imaging has become an invaluable tool in neuroscience, as it allows for chronic measurement of the activity of genetically identified cells during behavior1-6. Here we describe methods to perform two-photon imaging in mouse cortex while the animal navigates a virtual reality environment. We focus on the aspects of the experimental procedures that are key to imaging in a behaving animal in a brightly lit virtual environment. The key problems that arise in this experimental setup that we here address are: minimizing brain motion related artifacts, minimizing light leak from the virtual reality projection system, and minimizing laser induced tissue damage. We also provide sample software to control the virtual reality environment and to do pupil tracking. With these procedures and resources it should be possible to convert a conventional two-photon microscope for use in behaving mice.

Here we describe the experimental procedures involved in two-photon imaging of mouse cortex during behavior in a virtual reality environment.

在小鼠导航虚拟现实环境的双光子钙成像

In recent years, two-photon imaging has become an invaluable tool in neuroscience, as it allows for chronic measurement of the activity of genetically ...

科研

行为学

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

修改后的孔板 - 测量行为，认知和社会互动小鼠和大鼠

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

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, ...

Evaluation of a Smartphone-based Human Activity Recognition System in a Daily Living Environment

An evaluation method that includes continuous activities in a daily-living environment was developed for Wearable Mobility Monitoring Systems (WMMS) that attempt to recognize user activities. Participants performed a pre-determined set of daily living actions within a continuous test circuit that included mobility activities (walking, standing, sitting, lying, ascending/descending stairs), daily living tasks (combing hair, brushing teeth, preparing food, eating, washing dishes), and subtle environment changes (opening doors, using an elevator, walking on inclines, traversing staircase landings, walking outdoors). 
To evaluate WMMS performance on this circuit, fifteen able-bodied participants completed the tasks while wearing a smartphone at their right front pelvis. The WMMS application used smartphone accelerometer and gyroscope signals to classify activity states. A gold standard comparison data set was created by video-recording each trial and manually logging activity onset times. Gold standard and WMMS data were analyzed offline. Three classification sets were calculated for each circuit: (i) mobility or immobility, ii) sit, stand, lie, or walking, and (iii) sit, stand, lie, walking, climbing stairs, or small standing movement. Sensitivities, specificities, and F-Scores for activity categorization and changes-of-state were calculated.
The mobile versus immobile classification set had a sensitivity of 86.30% &#177; 7.2% and specificity of 98.96% &#177; 0.6%, while the second prediction set had a sensitivity of 88.35% &#177; 7.80% and specificity of 98.51% &#177; 0.62%. For the third classification set, sensitivity was 84.92% &#177; 6.38% and specificity was 98.17 &#177; 0.62. F1 scores for the first, second and third classification sets were 86.17 &#177; 6.3, 80.19 &#177; 6.36, and 78.42 &#177; 5.96, respectively. This demonstrates that WMMS performance depends on the evaluation protocol in addition to the algorithms. The demonstrated protocol can be used and tailored for evaluating human activity recognition systems in rehabilitation medicine where mobility monitoring may be beneficial in clinical decision-making.

A standardized evaluation method was developed for Wearable Mobility Monitoring Systems (WMMS) that includes continuous activities in a realistic daily living environment. Testing with a series of daily living activities can decrease activity recognition sensitivity; therefore, realistic testing circuits are encouraged for valid evaluation of WMMS performance.

在日常生活环境评价一个智能手机为基础的人类活动识别系统

An evaluation method that includes continuous activities in a daily-living environment was developed for Wearable Mobility Monitoring Systems (WMMS) that ...

A Method to Study Adaptation to Left-Right Reversed Audition

An unusual sensory space is one of the effective tools to uncover the mechanism of adaptability of humans to a novel environment. Although most of the previous studies have used special spectacles with prisms to achieve unusual spaces in the visual domain, a methodology for studying the adaptation to unusual auditory spaces has yet to be fully established. This study proposes a new protocol to set-up, validate, and use a left-right reversed stereophonic system using only wearable devices, and to study the adaptation to left-right reversed audition with the help of neuroimaging. Although individual acoustic characteristics are not yet implemented, and slight spillover of unreversed sounds is relatively uncontrollable, the constructed apparatus shows high performance in a 360&#176; sound source localization coupled with hearing characteristics with little delay. Moreover, it looks like a mobile music player and enables a participant to focus on daily life without arousing curiosity or drawing attention of other individuals. Since the effects of adaptation were successfully detected at the perceptual, behavioral, and neural levels, it is concluded that this protocol provides a promising methodology for studying adaptation to left-right reversed audition, and is an effective tool for uncovering the adaptability of humans to a novel environments in the auditory domain.

The present study proposes a protocol to investigate the adaptation to left-right reversed audition achieved only by wearable devices, using neuroimaging, which can be an effective tool for uncovering the adaptability of humans to a novel environment in the auditory domain.

一种研究左右反试镜的适应方法

An unusual sensory space is one of the effective tools to uncover the mechanism of adaptability of humans to a novel environment. Although most of the ...

The Unpredictable Chronic Mild Stress Protocol for Inducing Anhedonia in Mice

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by many patients prompts the need to develop new therapeutic alternatives and to better understand the etiology of the disorder. Pre-clinical models with translational merits are rudimentary for this task. Here we present a protocol for the unpredictable chronic mild stress (UCMS) method in mice. In this protocol, adolescent mice are chronically exposed to interchanging unpredictable mild stressors. Resembling the pathogenesis of depression in humans, stress exposure during the sensitive period of mice adolescence instigates a depressive-like phenotype evident in adulthood. UCMS can be used for screenings of antidepressants on the variety of depressive-like behaviors and neuromolecular indices. Among the more prominent tests to assess depressive-like behavior in rodents is the sucrose preference test (SPT), which reflects anhedonia (core symptom of depression). The SPT will also be presented in this protocol. The ability of UCMS to induce anhedonia, instigate long-term behavioral deficits and enable reversal of these deficits via chronic (but not acute) treatment with antidepressants strengthens the protocol&#39;s validity compared to other animal protocols for inducing depressive-like behaviors.

Here we present the unpredictable chronic mild stress protocol in mice. This protocol induces a long-term depressive-like phenotype and enables to assess the efficacy of putative antidepressants in reversing the behavioral and neuromolecular depressive-like deficits.

诱导小鼠快感缺失的不可预测慢性轻度应力协议

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by ...

Low-cost Protocol of Footprint Analysis and Hanging Box Test for Mice Applied the Chronic Restraint Stress

Gait disturbance is frequently observed in patients with movement disorders. In mouse models used for movement disorders, gait analysis is important behavioral test to determine whether the mice mimic the symptoms of patients. Motor deficits are often induced by stress when no spontaneous motor phenotype is observed in the mouse models. Therefore, gait analysis followed by stress loading would be a sensitive method for evaluating the motor phenotype in mouse models. However, researchers face the requirement of an expensive apparatus to obtain quantitative results automatically from gait analysis. For stress, stress loading by simple methods without expensive apparatuses required for electric shock and forced running is desirable. Therefore, we introduce a simple and low-cost protocol consisting of footprint analysis with paper and ink, hanging box test to evaluate motor function, and stress loading defined by restraint with a conical tube. The motor deficits of mice were successfully detected by this protocol.

The low-cost protocol consisting of footprint analysis and hanging box test after restraint stress is useful for evaluating the movement disorders of mouse model.

小鼠应用慢性约束应激的低成本足迹分析和吊箱试验协议

Gait disturbance is frequently observed in patients with movement disorders. In mouse models used for movement disorders, gait analysis is important ...

In Vivo Protocol of Controlled Subconcussive Head Impacts for the Validation of Field Study Data

Subconcussive hits pose a threat to neuronal health as they have shown to induce neuronal structural damage and functional impairment without causing outward symptomology and appear to be a key contributor to an irreversible neurodegenerative disease, chronic traumatic encephalopathy (CTE). In addition, athletes can incur more than 1,000 of these hits per season. The subconcussive soccer heading model (SSHM) is a relevant, reproducible, and leading method of isolating and examining the effects of these subconcussive head impacts. By controlling variables such as ball traveling speed, the frequency of impacts, interval, ball placement to the head, as well as by measuring head impact magnitude, the SSHM provides the scientific community with a superior avenue of investigating the acute subconcussive effects on neuronal health. In this paper, we demonstrate the utility of SSHM in studying a time-course expression of neurofilament-light polypeptide (NF-L) in plasma in a repeated measures fashion. NF-L is an axonal injury marker that has previously been shown to be elevated in boxers and football players following subconcussive head trauma. Thirty-four adult aged soccer players were recruited and randomly assigned to either a soccer heading (n = 18) or kicking (n = 16) group. The heading group executed 10 headers with soccer balls projected at a velocity of 25 mph over 10 min. The kicking group followed the same protocol with 10 kicks. Plasma samples were obtained before and at 0 h, 2 h, and 24 h after heading/kicking and assessed for NF-L expressions. The heading group showed a gradual increase in plasma NF-L expression and peaked at 24 h after the heading protocol, whereas the kicking group remained consistent across the time points. These results confirmed the NF-L data from clinical field studies, encouraging the use of SSHM to validate clinical subconcussion data.

The subconcussive soccer heading model is a safe and concise methodological approach to isolate and measure the effects of subconcussive head impacts.

控制下冲击头冲击的活体协议对现场研究数据的验证

Subconcussive hits pose a threat to neuronal health as they have shown to induce neuronal structural damage and functional impairment without causing ...

A Chronic Immobilization Stress Protocol for Inducing Depression-Like Behavior in Mice

Depression is not yet fully understood, but various causative factors have been reported. Recently, the prevalence of depression has increased. However, therapeutic treatments for depression or research on depression is scarce. Thus, in the present paper, we propose a mouse model of depression induced by movement restriction. Chronic mild stress (CMS) is a well-known technique to induce depressive-like behavior. However, it necessitates a complex procedure consisting of a combination of various mild stresses. In contrast, chronic immobilization stress (CIS) is a readily accessible chronic stress model, modified from a restraint model that induces depressive behavior by restricting movement using a restrainer for a certain period. To evaluate the depressive-like behaviors, the sucrose preference test (SPT), the tail suspension test (TST), and the ELISA assay to measure stress marker corticosterone levels are combined in the present experiment. The described protocols illustrate the induction of CIS and evaluation of the changes in behavior and physiological factors for the validation of depression.

This article provides a simplified and standardized protocol for induction of depressive-like behavior in chronically immobilized mice by using a restrainer. In addition, behavior and physiological techniques to verify induction of depression are explained.

诱导小鼠抑郁行为的慢性固定性压力协议

Depression is not yet fully understood, but various causative factors have been reported. Recently, the prevalence of depression has increased. However, ...

Nest Building Behavior as an Early Indicator of Behavioral Deficits in Mice

Nest building is an innate behavior in male and female rodents, even when raised in laboratory settings. As such, many researchers provide rodents synthetic and/or natural materials (such as twine, tissue, cotton, paper, and hay) as a gauge of their overall well-being and as an ancillary assessment to predict the possible decline in cognition. Typically, changes in nesting behaviors, such as failure to create a nest, indicate a change in health or welfare. In addition, nesting behavior is sensitive to many environmental and physiological challenges, as well as many genetic mutations underlying pathological disease states. The following protocol describes a nesting behavior paradigm that explores the usage of four types of nesting material. In addition, the protocol utilizes intraclass correlations to demonstrate that inter-rater reliability is higher when nests are constructed out of shredded paper compared to other common nesting materials such as cotton squares, paper twists, and soft cob bedding. The chosen methodology and statistical considerations (i.e., intraclass correlation) for this assay may be of interest for those conducting experiments assessing the quality of living of mice.&#160;

Here, we present a protocol to quantify nest building behavior in mice, which is known to be impaired in several neurological disorders and diseases. This protocol examines the utility of four materials and offers the opportunity to quantify the rater agreement in scoring, improving the validity and reliability of the assay.