Name: assessing activity-based anorexia in mice
Uploaded: 2018-05-14T14:00:00.000+00:00
Duration: 8 min 26 s
Description: Watch this Scientific Journal Video about Assessing Activity-based Anorexia in Mice at JoVE.com

这项工作由一个 NARSAD 独立调查员奖和一个 IMHRO 上升的星抑郁症研究奖的资助为记忆乔治 Largay 到 SCD。

这里描述的所有方法都已被加州大学圣地亚哥分校的机构动物护理和使用委员会 (IACUC) 批准。
1. 老鼠
<ol><li>为研究选择合适的鼠标应变。</li>
	<li>从供应商处购买鼠标或放大一条线以获得适当的实验数据。</li>
	<li>小组房子购买了动物设施的老鼠至少1周前开始研究, 以允许一个充分的驯化期。</li>
	<li>在老鼠8周大的时候开始实验性的驯化阶段。</li>
</ol>2. 住房
<ol><li>选择足够大的笼子来容纳不受阻碍的跑步轮、食物贮器和水瓶。</li>
	<li>调节室温和湿度, 以补充所测试的假说。 注: 较高的温度会降低 ABA28的开发。</li>
</ol>3. 跑步轮
<ol><li>选择带有无线通信的车轮, 以避免在笼子内发生潜在的干扰和纠缠, 请参阅材料目录以了解示例。</li>
</ol>4. 水瓶
<ol><li>选择一个水瓶, 将不会与运行轮或在家庭笼中的食物插座争夺空间。</li>
	<li>如果在饮用水中提供药物治疗, 则用胶带将瓶子装在瓶中, 创造出光敏感的环境。</li>
</ol>5. 食品
<ol><li>选择适合于正在测试的假设的低脂周 (例如, Envigo 的啮齿动物饮食 8604)。 注意: 高脂肪和糖的饮食可以减少 ABA29的发展。</li>
	<li>使用一个小玻璃瓶提供周, 大约65厘米直径 x 50 厘米的高度。 注: 可使用自动进纸器代替玻璃罐, 通过限制调查人员的干扰, 可以减轻压力。这可能是最有利的, 如果进行 ABA 研究与药物操纵。</li>
</ol>6. 实验前驯化阶段
<ol><li>通过选择靠近笔记本电脑和轮毂的笼子架来设置实验区。这将限制数据传输的问题。</li>
	<li>打开笔记本电脑和无线集线器。</li>
	<li>打开笔记本电脑上的轮子管理软件。<ol>
<li>查看软件以确保两个集线器都处于活动状态。</li>
		<li>打开 "工具" 并选择 "删除车轮"。重复此步骤5次。</li>
		<li>关闭软件, 然后重新打开它。</li>
	</ol>
</li>
	<li>将3个 AAA 电池放入车轮底座的电池组中。<ol>
<li>将电池组线连接到车轮底座的硬件上。这将在使用之前断开连接。将电池组关闭到车轮底座。</li>
	</ol>
</li>
	<li>检查软件以确保车轮底座在相应的无线集线器下列出, ID 为 "1"。如果基未列出为 ID "1", 则松开电池包, 然后在步骤6.2 中重新启动。</li>
	<li>将车轮 id 从 "1" 更改为 "名称" 列下的鼠标 id。</li>
	<li>坚持运行车轮的基础, 以管道胶带底部的笼子, 以确保稳定。不要让任何磁带贴出来, 因为老鼠会嚼在暴露的胶带上。</li>
	<li>选择一个没有腐蚀磁性片的运行轮盘。</li>
	<li>将车轮放在底座上, 旋转车轮, 以验证从笼壁和光栅的间隙。</li>
	<li>旋转光盘特定数量的轮换, 以确保车轮运行软件正确计数轮换。</li>
	<li>防止在床上用品中丢失的部分, 或通过管道把食物罐的底部贴在笼子地板上, 远离跑步轮和水瓶而弄湿。 注: 湿周增加食物摄入量, 比干周30。</li>
	<li>把5块的食物放进罐子里。 注意: 在实验前阶段的周是可用的ad 随意, 不需要重置.在驯化期间接触运行车轮可能会加剧 ABA21,31。</li>
	<li>将一个完整的水瓶放在笼子里, 并提供水的ad 随意。</li>
	<li>重复, 直到所有笼子都有一个无线连接运行车轮, 食物和水。</li>
	<li>单独安置老鼠入他们适当的笼子。</li>
	<li>在轮管理软件中, 转到 "文件" 并单击 "开始获取"。</li>
	<li>观看软件更新, 以确保车轮轮换正在计算和系统运行良好。</li>
	<li>使小鼠适应实验性住房条件, 共2天。</li>
	<li>评估小鼠在这个阶段, 以确保他们足够健康, 以处理 ABA 实验。</li>
	<li>每天检查水壶里是否有水淹。如果瓶子进水了, 用新笼子代替旧笼子, 重复必要的步骤, 把轮子和食物粘在笼子里, 不受干扰。</li>
	<li>在2天, 检查笼子的食物水平。如果食物很低, 再加2片。在不干扰老鼠的情况下这样做。 注: 如果在实验前的驯化阶段, 任何鼠标显示 hypophagia 或体重不足, 从研究中删除鼠标。</li>
</ol>7. 实验基线阶段-1 天
<ol><li>选择 "结束", 然后在车轮管理软件中 "开始获取"。</li>
	<li>测量出9克周在一个小塑料烧杯的规模与0.00 克敏感性。记下确切的重量。</li>
	<li>在一个大的塑料烧杯中, 用0.0 克灵敏度轻轻地移除并称量鼠标。记录重量。 注意: 在处理时限制小鼠的压力是至关重要的, 因此要耐心地处理它们。</li>
	<li>处理所有的驯化食物。</li>
	<li>搜索床上用品的任何食物, 可能已经从罐子和处置, 以及。</li>
	<li>把水瓶放在原处, 但要确保它足够充分。</li>
	<li>清洁脏运行车轮与70% 异丙醇和纸巾。</li>
	<li>如果清洗是必要的, 在异丙醇干燥后, 将鼠标返回笼子, 并更换机架上的笼。</li>
	<li>重复这些步骤, 直到每只老鼠的重量和9克的食物被测量出来, 并放置在笼子里。</li>
</ol>8. 实验基线阶段-天 2-7
<ol><li>每天在同一时间返回动物设施。不要因测量超过或低于10分钟而变化。</li>
	<li>测量出9克周在一个小塑料烧杯的规模与0.00 克敏感性。记下确切的重量。</li>
	<li>在一个大的塑料烧杯中, 用0.0 克灵敏度轻轻地移除并称量鼠标。记录重量。</li>
	<li>把食物罐从笼子里拉出, 把所有的食物倒入0.00 克刻度上的小烧杯中。<ol>
<li>如果粪便和床上用品都在食品罐中, 请挑选出所有的床上用品和粪便。用一个小的过滤器把粉状食品揉入称重烧杯中。用镊子 (如有必要) 清除任何粪便。 注: 床上用品不会分解成小块, 以适应通过过滤器。</li>
		<li>在笼子里寻找任何残留的食物, 并将它们添加到烧杯中进行权衡。记录剩下的食物数量。 注: 如果必须搜索床上用品和笼子, 最好这样做, 而鼠标在称量烧杯或转移笼, 以减轻压力。</li>
		<li>处理所有的食物。</li>
	</ol>
</li>
	<li>把罐子擦干净, 放回笼子里。必要时更换磁带。</li>
	<li>把以前测量过的9克新食物放进罐子里。</li>
	<li>清洁的运行车轮与70% 异丙醇, 然后烘干车轮。一次做一个笼子, 不要意外地互换车轮。</li>
	<li>在车轮运行程序中, 每隔一天重新开始购置。</li>
	<li>重复这些步骤, 直到限制阶段开始。</li>
</ol>9. 试验性限制阶段-1 天
<ol><li>结束和开始收购的车轮管理软件。</li>
	<li>衡量老鼠, 旧的, 新的食物每基线协议。</li>
	<li>
现在不要清理车轮.这可能会分散老鼠在喂食期有限的时间。</li>
	<li>允许访问新食品, 9 克的周, 为6 h。记录食物提供的时间。 注意: 在光循环期间, 食物是可利用的时间将影响 ABA 的发展。由于小鼠通常在光照周期内睡觉, 因此在这段时间内获得食物会加剧 ABA 的发育, 因为小鼠通常不会在此时消耗大部分的日常食物。相反, 在黑暗周期中给予食物进入的老鼠会慢慢地发展 ABA32。调整食物访问的持续时间将导致生存时间更长或更短 (图 2)。食品预期活动 (FAA) 是多动症, 在提供食物之前立即发生32,33,34。联邦航空局可能影响 ABA 的发展, 因为它已经表明, 拒绝轮运行访问期间, 联邦航空局变好 ABA 行为 32.<ol>
<li>为每只老鼠取出食物, 6 小时后, 测量剩余的数量。记录食物被移除的时间。</li>
	</ol>
</li>
	<li>清洁车轮, 如果有必要, 并试图这样做, 而不落后于从后面的笼子拉食物。</li>
	<li>权衡食物, 计算食物的差异, 以确定每只老鼠吃的食物量。</li>
	<li>计算每个鼠标的辍学重量。 注意: 辍学重量是当一个小鼠必须从实验中删除时, 它达到其基线体重的最终测量的75%。例如, 如果鼠标在限制阶段开始时重20克, 当它达到15克时, 它必须从实验中移除。</li>
	<li>在动物设施中保留手头的辍学重量的复印件。 注: 评估从 ABA 恢复的实验可能会尝试;食品是提供的ad 随意,或车轮锁定研究的过程中, 老鼠返回健康体重34。</li>
</ol>10. 试验性限制阶段-天2到14
<ol><li>重复限制日1协议中的所有度量步骤。不要重新启动轮收购。</li>
	<li>如果鼠标到达下降重量 (其基线体重的 75%), 从笼子里取出轮子和食物罐, 在笼子底部提供充足的食物, 然后继续其他老鼠。一旦所有的笼子都被照顾, 弄死所有已经达到下降重量的老鼠。</li>
	<li>在实验期间为每只鼠标留出轮子。</li>
	<li>每天检查每只鼠标是否有任何损伤。如果需要, 夹子损坏的脚趾甲, 治疗疮,等如果不确定如何进行损伤, 请向动物护理人员请教。</li>
</ol>11. 结束研究
<ol><li>如果这项研究持续了14天, 那么就在15天对所有的老鼠进行权衡。限制日1重量被认为是基线重量。 注意: 研究的长度可以修改, 以解决被测试的假说。长时间的食物存取期可用于延长研究长度。</li>
	<li>必要时弄死老鼠。</li>
	<li>把脏设备带到合适的房间。</li>
	<li>15天结束轮数购置, 提供10天的完整数据。</li>
	<li>关闭无线轮集线器。</li>
	<li>提取数据。</li>
	<li>分析数据。</li>
</ol>

<ol>
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J., 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=Olanzapine,+but+not+fluoxetine,+treatment+increases+survival+in+activity-based+anorexia+in+mice.">Olanzapine, but not fluoxetine, treatment increases survival in activity-based anorexia in mice.</a> Neuropsychopharmacology. 37 (7), 1620-1631 (2012).</li><li>Ho, E. V., Klenotich, S. J., McMurray, M. S., Dulawa, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activity-Based+Anorexia+Alters+the+Expression+of+BDNF+Transcripts+in+the+Mesocorticolimbic+Reward+Circuit.">Activity-Based Anorexia Alters the Expression of BDNF Transcripts in the Mesocorticolimbic Reward Circuit.</a> PLoS ONE. 11 (11), 0166756 (2016).</li></ol>

作者没有什么可透露的。

ABA 的实验可以由实验者修改, 以测试不同的菌株, 年龄, 药物, 和各种其他变量的. 铭记遗传变异, 对跑步轮或食物进入期的调整将增加或减少严重程度症状和辍学率。这可以用来增加或减少实验的长度, 这取决于实验的兴趣问题。
准确测量食物摄入量是 ABA 范式的一个关键部分, 但往往会造成问题。在食品罐中的床上用品、粪便和尿液, 加上食物被移入笼子空间, 这为精确称量剩余食物提供了困难。老鼠还在罐子里和整个笼子里留下小块食物和粉状食物。滤网有助于缓解通过床上用品筛选出适当重量记录的食物残留物的问题, 但这种方法并非万无一失。处理老鼠获得他们的重量是另一个关键, 但混杂因素在 ABA 范式。随着研究者的干扰, 小鼠更易受到压力, 这会使 ABA 的病情恶化。
ABA 是一种有用的评估方法, 然而这项技术确实有局限性。首先要考虑的是 ABA 模型并不模仿厌食症的所有方面。例如, 获得高脂肪饮食的啮齿类动物在典型实验条件下不会发展 ABA30。然而, 几乎没有动物模型, 概括了精神紊乱的所有方面。因此, ABA 仍可能对相关行为产生重要的洞察力。
药物治疗也可以在 ABA 范式中实施。药物管理局通过饮用水是理想的, 因为它避免每天的注射, 可能会有不必要的影响, 包括局部刺激注射部位和短期镇静的某些药物。然而, 每天调整药物浓度以保持一致的剂量是至关重要的, 因为在 ABA 的过程中, 每天的水摄入量发生了显著的变化, 首先是饮酒的增加, 随后是液体消耗的减少。使用皮下微型泵连续提供药物也可能与 ABA 范式;然而, 必须注意的是, 迷你泵是小到足以让老鼠进出食品罐和运行车轮不畅通。此外, optogenetic 光纤电缆的使用在理论上是可行的, 但需要修改的设置。或者, 使用由设计药物 (DREADD) 专门激活的设计受体来操纵特定电路的活动, 可以与 ABA 范式一起使用, 而不需要留置绳子的并发症。因此, 可以结合多种现代神经科学工具, 研究 ABA 行为的神经机制。

自1953年以来, 据报道, 啮齿类动物在运行车轮时表现出一种自相矛盾的过度活跃状态, 当食物可用性被限制为1时, 它们可以自由进入车轮, 而在接受自愿 hypophagia 时。相反 , 啮齿类动物不会迅速下降体重时, 没有运行车轮或与运行车轮, 并提供食物ad 随意1,2,3, 在一个时间表提供的食物。ABA 模型可靠地导致体重急剧下降, hypophagia, 体温过低, 发情丧失和增加刺激的 HPA 轴4。最终, ABA 会导致死亡, 除非主题被从范例5中删除。ABA 范式为研究人员提供了一种动物模型, 一种复杂的饮食紊乱, 其特点是食欲行为严重失调, 影响大约 1 100 女性, 而男性的比例则更小6。经常出现多动症的患者, 包括极多的运动, 和/或一般躁动不安的7,8。死亡率约为 10%, 是所有精神疾病中死亡率最高的9。目前对 a 的治疗仅限于认知疗法, 因为对于那些患有1011的患者, 没有经过批准的药理治疗。
a 通常被认为是一种影响主要是女性的障碍。作为女性的10倍于男性的诊断, 女性受试者传统上是12中的焦点。但是, 应特别考虑将男性排除在研究之外。虽然诊断的一个遗骸较低的男性, 肌肉 dysmorphia (MD) 是一个条件, 有很多相似之处, 因为身体图像扭曲, 饮食往往紊乱。有支持的概念, MD 和 a 可以归类为类似的方式13,14,15。这可能意味着一些 MD 的案例代表了 a 的 "男性版本"。在动物模型方面, 一些报告指出, 男性比女性更容易受到 ABA 范式的影响。例如, 最近的一项研究显示, 与 C57Bl/6 小鼠16中的雌性相比, 死亡率较高, 食物摄入量减少。对 ABA 易感性的一个预测因子是自发的身体活动 (SPA)。高或低 SPA 的大鼠更容易在 ABA 范式中减肥, 雄性大鼠比雌性老鼠表现出更强的效果17。相反, 在 ABA18的限制阶段, 雌性啮齿动物的运动比男性多。此外, 与 Balb/cJ 小鼠的研究显示了 C57Bl/6 小鼠的相反效果, 在那里雌性小鼠的死亡率较高, 食物摄取量减少, 与男性相比 (图 1)6。由于两性在 ABA 范式中的不同结果和对饮食模式紊乱的男性的认识不断提高, 因此应该对男性和女性的研究对象进行测试。
除了 ABA 范式中的性别差异外, 在选择主语时必须考虑年龄和应变。青春期老鼠可以更准确地模拟 a, 因为通常出现在青春期, 观察老鼠和老鼠19,20,21,22。在基线水平上比其他活动更活跃的菌株具有更高的易感性和更严重的 ABA23。已知有较高的焦虑水平的菌株, 如 DBA/2, 增加了车轮运行活动, 这表明 ABA 范式24中的辍学率更快。根据实验设计的不同, 可根据选择的应变来优化 ABA 的持续时间。
ABA 悖论不是老鼠独有的。其他哺乳动物, 包括老鼠, 仓鼠, 沙土鼠, 猪, 花栗鼠, 和豚鼠已经证明了这种现象6。在哺乳动物物种之间的 aba 现象的保护表明, aba 范式可以提供一个翻译工具, 以调查在人类的厌食行为的机制。老鼠特别适合研究 ABA 的机制。老鼠可以被密集地安置和世代时间是相对地短的。老鼠有一个完整的序列基因组, 和许多近亲, outbred 和特殊菌株, 如 congenics, 是可利用的。已经产生了大量的基因操纵线, 这使它们成为研究评估遗传对疾病的影响的理想. 根据手头的问题, 研究人员可能会操纵复杂的神经回路和/或基因表达评估 ABA 范式中的行为, 有可能回答关于基因影响的问题, 在研究人类时是不可能的。
目前存在数量有限的动物模型。应力模型利用尾捏、新奇诱发 hypophagia、冷游泳和脑刺激等方法诱导啮齿类动物 hypophagia。通过诱发压力, HPA 轴的变化减少食欲, 从而减少体重25。然而, 完全也刺激了 HPA 轴, 这也包含了一些额外的功能, 如多动症。另一种在研究中考虑的模型是慢性食品限制模型。通过将食物限制在ad 随意的40到60% 范围内, 你可以模仿营养不良的26的生理反应。虽然这种方法对研究喂养不足的影响是有效的, 但它并没有重现一个核心问题, 即自愿食物限制。在 ABA 范式, 动物被剥夺了一天的食物进入, 但也自愿减少食物摄入, 如果车轮也存在。遗传模型也被用来检查的病因. 研究人员发现了神经化学和遗传因素, 如基因 BDNF 和递质多巴胺和血清素27。基因模型的使用对于理解背后的神经机制是至关重要的。然而, 基因组广泛的协会研究尚未产生显著的影响, 并没有发现在一个罕见的变种。未来的研究应该结合一个遗传方法与 ABA 模型, 以增加对一个相关的表型的理解。
由于人类疾病的复杂性和异质性, 发展整个精神疾病的动物模型几乎是不可能的。然而, 通过建模特定的, 明确定义的精神障碍的组成部分, 可以获得对潜在神经生物学或病理生理学的独特洞察力。这样的生物洞察力可以用来识别新的治疗方法。因此, 啮齿动物的 ABA 范式提供了一个临床前的工具, 以研究类似行为的机制, 不能道德地研究人类, 如遗传操作的影响, 神经回路的扰动, 和某些环境因素的后果。

<table><tbody><tr><td>Wireless Low Profile Running Wheels, Hubs, Software</td><td>Med Associates</td><td>ENV-047</td><td></td></tr><tr><td>Standard Teklad Rodent Chow</td><td>Envigo</td><td>8604</td><td></td></tr><tr><td>8 Week Old Mice</td><td>Jackson Laboratories</td><td>Balb/cJ</td><td>Strain used in our study - Can use other strains to assess ABA</td></tr><tr><td>Scout Pro Scale 200 g</td><td>Ohaus</td><td>SPE202</td><td>Used to weigh mice</td></tr><tr><td>Scout Pro Scale 400 g</td><td>Ohaus</td><td>SPE402</td><td>Used to weight food</td></tr></tbody></table>

assessing activity-based anorexia in mice

啮齿类动物在受到限制的喂养计划时, 会产生活动性厌食症 (ABA), 并允许自由进入跑步轮。这些情况导致体重减少危及生命。然而, 只接触到这些条件之一的啮齿类动物最终会适应重新建立正常体重。尽管在 aba 条件下, 随着自愿食物摄入量的增加而减少, 这种现象似乎自相矛盾, 但在许多哺乳动物物种中都有 ABA 的行为。
ABA 范式为神经性厌食症 (a) 提供了一种动物模型, 这种饮食紊乱具有严重的失调食欲行为。受试者单独安置在一个运行的车轮的自由访问。每一天, 这个主题在有限的时间内提供食物。在实验过程中, 一个人的体重从高活性和低卡路里摄入量中减少。研究的持续时间根据每天提供的食物的长短、提供的食物种类、老鼠的应变、如果药物被测试以及环境因素而定。
对患者缺乏有效的药理治疗, 其生活质量低下, 治疗费用高, 死亡率高, 表明了进一步研究的紧迫性。我们提供了一个基本的大纲, 以执行 ABA 实验小鼠, 提供一种方法来调查类似的行为, 以发展新的疗法。该协议是优化用于 Balb / cJ 小鼠 , 但可以很容易地纵其他菌株 , 提供了极大的灵活性 , 在处理不同的问题 , 特别是有关 ABA 的遗传因素。

脑源性神经营养因子 (BDNF), 一种有助于调节喂养和体重维持的蛋白质, 在35患者的血清中减少。该实验评估了定时喂养, 运行轮访问, 或两者都对 BDNF 表达的海马 (HPC), 腹侧盖区 (VTA), 伏隔核 (NAc) 和内侧前额皮质 (mPFC) 的影响。还评估了神经细胞黏附分子 1 (NCAM1) 的表达, 探讨了 mesocorticolimbic 通路中对 BDNF 影响的特异性。
小鼠被单独安置在一个12:12 光黑暗的循环中, 在一个动物护理设施的气候控制室。网箱配备了无线运行轮, 每三十年代将数据传递给笔记本电脑。标准周被提供了在一个罐子大约 2 "直径与墙壁大约 1.5" 高在基线和制约期间。
四实验组被分配伪随机: ad 随意(ad Lib), 定时喂养 (STV), 自由车轮运行和食物访问 (运行), 或自由轮运行与食品限制 (ABA) (图 3)。小组分配由最初的体重决定, 以形成四重量相等的小组。在驯化期间, 小鼠被单独安置并且哺养了ad 随意以恒定的进入奔跑的轮子2天, 并且在基线期间的7天。
在预定的12天的食物进入期内, STV 和 ABA 组的食物进入时间仅为每天6小时 (09:00-15:00), 如果他们满足其起始体重25% 的计算出重量, 则进行监测和清除。每天进行调整, 以限制 STV 组的食物进入, 以确保 STV 组的辍学率与 ABA 组相似, 其成员通常会较快地减轻体重。每个运行的鼠标被绑在 ABA 的鼠标, 以去除从研究, 每一个 Ad Lib 鼠标与 STV 鼠标。国内兴起确保所有小鼠都在类似的时间点从研究中移除。这种设计可以比较在同一时间内经历各种实验条件的小鼠组间的基因表达。移除的天数被解释为生存的衡量标准。另一种可能性是包括一个额外的运行组, 它不限于与 ABA 组少吃, 但在6小时内提供过量的食物。此外, 可以使用5个实验组, 包括运行组的两种形式。小组的选择取决于研究的目标。
在从 ABA 范式中去除后, 小鼠被斩首以快速提取大脑。组织从 mPFC, HPC, NAc, 和 VTA 被删除使用脑基质和组织冲床, 并立即冻结使用干冰。在 RNA 提取之前, 样品保存在-80 摄氏度的冷藏库中。
对数据进行统计分析, 采用方差分析法。对于基线数据, 方差分析被应用到每个相关变量 (体重, 食物摄入量和车轮运行)。对方差或交互进行了后特别分析。在应用了对方差的后向分析后进行了 Bonferroni 调整。
分析数据的限制, 一般线性模型 (进程 GLIMMIX;SAS v9.2;根据请求提供相应作者的代码, 用于评估每个变量。自组后分析用于使用 false 发现率方法解析交互。使用 Logrank (壁炉-Cox) 和 Peto-Peto 魏氏后特别测试的卡普兰-梅尔测试进行生存分析。意义在p < 0.05 中设置。
车轮的运行不影响体重在基线, 而食物消耗增加时, 运行车轮可在基线天 2-7。在定时喂养之前, 运行轮上的活动与 ABA 组之间没有什么不同。
由于 Ad Lib 和运行小鼠分别与 STV 和 ABA 小鼠, 在预定喂养期间, 实验组之间的生存没有差异 (图 4a)。如果没有国内兴起, ABA 小鼠的存活率与跑步和 STV 组相比显著减少 35.然而, 两组暴露于定时喂养 (STV 和 ABA 组) (图 4b) 的天数中, 体重减少了 1-5 天。定时喂养也减少了两组中 1-5 天的食物摄取量 (图 4c), 但对轮运行活动没有影响 (图 4d)。在这项研究中, aba 组的多动症是短暂的 (天 1-3) 和显著相比, 运行组, 也许是由于健康迅速下降的小鼠在 ABA 的情况下。
车轮在 VTA 中显著增加了 BDNF 表达式 (图 5a), 而食物限制在 VTA (图 5b) 中增加了 NCAM1 mrna 表达式, 但没有改变 bdnf mRNA 表达式。在 NAc (图 6) 中没有车轮或食物进入的主要影响, 也没有对 BDNF 或 NCAM1 mRNA 表达式的交互作用。在 mPFC 中, 定时喂养降低了 BDNF mrna 的表达, 但不影响 NCAM1 mrna 表达式 (图 7)。车轮的进入并没有改变基因表达, 也没有发现食物限制和车轮接触的相互作用。同样, 在 HPC 中, 由于食物限制增加了 NCAM1 mrna 表达式 (图 8), 因此没有食物限制和车轮进入对 BDNF 或 NCAM1 mrna 表达的相互作用。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57395/57395fig1.jpg"> 图 1: ABA 范式中的性别差异.(a) 随着时间的推移, 雄性和雌性老鼠之间的累积存活率。(b) 雄性和雌性小鼠在限制日 (c) 之间的体重超过限制日的雄性和雌性小鼠的食物摄入量。(d) 在限制天数内在雄性和雌性小鼠之间运行的车轮。此数字已从原始出版物6中修改。<a href="https://cloudflare.jove.com/files/ftp_upload/57395/57395fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57395/57395fig2.jpg"> 图 2: 喂养时间对 ABA 持续时间的影响.这个数字显示了多少天老鼠将保持在限制, 取决于多久食物是可利用的。这是显示的小鼠有和没有运行车轮。此数字已从原始出版物35中修改。<a href="https://cloudflare.jove.com/files/ftp_upload/57395/57395fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/57395/57395fig3.jpg"> 图 3: ABA 范式实验设计.该图像显示了 ABA 实验的建立。此数字已从原始出版物36中修改。<a href="https://cloudflare.jove.com/files/ftp_upload/57395/57395fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/57395/57395fig4.jpg"> 图 4: ABA 范式的影响.(a) 生存期、(b) 每日体重、(c) 食用的食物, 以及 (d) 在食物限制期间运行的车轮。数字斜体描绘了在 ABA 范式中残留的老鼠数量。结果表示为平均值。此数字已从原始出版物36中修改。<a href="https://cloudflare.jove.com/files/ftp_upload/57395/57395fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/57395/57395fig5.jpg"> 图 5: ABA 对 VTA 基因表达的影响.(a) BDNF (b) NCAM1 在 ABA 条件下的小鼠 VTA 中的表达。在对所描述的独立测度进行限制时, 嵌入表明了平均 BDNF 和 NCAM1 表达。结果显示为 log2 (RQ) 的 SEM p < 0.05。此数字已从原始出版物36中修改。<a href="https://cloudflare.jove.com/files/ftp_upload/57395/57395fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/57395/57395fig6.jpg"> 图 6: ABA 对伏 Nucelus 基因表达的影响.(a) BDNF, (b) NCAM1 在暴露于 ABA 条件下的小鼠的 NAc 表达。结果显示为 log2 (RQ) 的 SEM p < 0.05。此数字已从原始出版物36中修改。<a href="https://cloudflare.jove.com/files/ftp_upload/57395/57395fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/57395/57395fig7.jpg"> 图 7: ABA 对 mPFC 基因表达的影响.(a) BDNF (b) NCAM1 在 ABA 条件下的小鼠 mPFC 中的表达。在对所描述的独立测度进行限制时, 该嵌入表示平均 BDNF 表达式。结果显示为平均 log2 (RQ) 的 SEM p < 0.05。此数字已从原始出版物36中修改。<a href="https://cloudflare.jove.com/files/ftp_upload/57395/57395fig7large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/57395/57395fig8.jpg"> 图 8: ABA 对 HPC 基因表达的影响.(a) BDNF (b) NCAM1 在 ABA 条件下的小鼠 HPC 中的表达。在对所描述的独立测度进行限制时, 插入表示平均 BDNF 和 NCAM1 表达。结果显示为平均 log2 (RQ) 的 SEM p < 0.05。此数字已从原始出版物36中修改。<a href="https://cloudflare.jove.com/files/ftp_upload/57395/57395fig8large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Assessing Activity-based Anorexia in Mice at JoVE.com

Assessing Activity-based Anorexia in Mice

老鼠单独安置与一个奔跑的轮子, 当获得食物的有限的获取开发减少食物消耗量和增加活动在奔跑的轮子。这种实验性的现象叫做活动性厌食症。这个范式为研究神经性厌食症的生物学和行为提供了一个实验工具。

小鼠活动性厌食症的评估

Rodents develop activity-based anorexia (ABA) when exposed to a restricted feeding schedule and allowed free access to a running wheel. These conditions ...

assessing-activity-based-anorexia-in-mice

Nanyang Technological University

Research

JoVE Journal

Behavior

12.4K 次观看.  University of California, San Diego. 老鼠单独安置与一个奔跑的轮子, 当获得食物的有限的获取开发减少食物消耗量和增加活动在奔跑的轮子。这种实验性的现象叫做活动性厌食症。这个范式为研究神经性厌食症的生物学和行为提供了一个实验工具。

视频: Assessing Activity-based Anorexia in Mice

Trace Fear Conditioning in Mice

In this experiment we present a technique to measure learning and memory. In the trace fear conditioning protocol presented here there are five pairings between a neutral stimulus and an unconditioned stimulus. There is a 20 sec trace period that separates each conditioning trial. On the following day freezing is measured during presentation of the conditioned stimulus (CS) and trace period. On the third day there is an 8 min test to measure contextual memory. The representative results are from mice that were presented with the aversive unconditioned stimulus (shock) compared to mice that received the tone presentations without the unconditioned stimulus. Trace fear conditioning has been successfully used to detect subtle learning and memory deficits and enhancements in mice that are not found with other fear conditioning methods. This type of fear conditioning is believed to be dependent upon connections between the medial prefrontal cortex and the hippocampus. One current controversy is whether this method is believed to be amygdala-independent. Therefore, other fear conditioning testing is needed to examine amygdala-dependent learning and memory effects, such as through the delay fear conditioning.

In the following experiment we describe a protocol for trace fear conditioning in mice. This type of associative memory includes a trace period that separates the neutral stimulus and the unconditioned stimulus.

跟踪恐惧条件小鼠

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

科研

行为学

Using the Activity-based Anorexia Rodent Model to Study the Neurobiological Basis of Anorexia Nervosa

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A challenging illness to treat, due to the lack of understanding of the underlying neurobiology, AN has the highest mortality rate among psychiatric illnesses. To address this need, neuroscientists are using an animal model to study how neural circuits may contribute toward vulnerability to AN and may be affected by AN. Activity-based anorexia (ABA) is a bio-behavioral phenomenon described in rodents that models the key symptoms of anorexia nervosa. When rodents with free access to voluntary exercise on a running wheel experience food restriction, they become hyperactive &#8211; running more than animals with free access to food. Here, we describe the procedures by which ABA is induced in adolescent female C57BL/6 mice. On postnatal day 36 (P36), the animal is housed with access to voluntary exercise on a running wheel. After 4 days of acclimation to the running wheel, on P40, all food is removed from the cage. For the next 3 days, food is returned to the cage (allowing animals free food access) for 2 hr daily. After the fourth day of food restriction, free access to food is returned and the running wheel is removed from the cage to allow the animals to recover. Continuous multi-day analysis of running wheel activity shows that mice become hyperactive within 24 hr following the onset of food restriction. The mice run even during the limited time during which they have access to food. Additionally, the circadian pattern of wheel running becomes disrupted by the experience of food restriction. We have been able to correlate neurobiological changes with various aspects of the animals&#8217; wheel running behavior to implicate particular brain regions and neurochemical changes with resilience and vulnerability to food-restriction induced hyperactivity.

Here we present a protocol to induce activity-based anorexia (ABA) in female adolescent mice. ABA is a condition of hyperactivity evoked by imposing food restriction on rodents with access to a running wheel. This phenomenon is being used as a model to study the underlying neurobiology of anorexia nervosa.

使用基于活动的厌食症啮齿动物模型来研究神经性厌食症的神经生物学基础

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A ...

Assessing Burrowing, Nest Construction, and Hoarding in Mice

Deterioration in the ability to perform "Activities of daily living" (ADL) is an early sign of Alzheimer's disease (AD). Preclinical behavioural screening of possible treatments for AD currently largely focuses on cognitive testing, which frequently demands expensive equipment and lots of experimenter time. However, human episodic memory (the most severely affected aspect of memory in AD) is different to rodent memory, which seems to be largely non-episodic. Therefore the present ways of screening for new AD treatments for AD in rodents are intrinsically unlikely to succeed. A new approach to preclinical screening would be to characterise the ADL of mice. Fortuitously, several such assays have recently been developed at Oxford, and here the three most sensitive and well-characterised are presented.
Burrowing was first developed in Oxford13. It evolved from a need to develop a mouse hoarding paradigm. Most published rodent hoarding paradigms required a distant food source to be linked to the home cage by a connecting passage. This would involve modifying the home cage as well as making a mouse-proof connecting passage and food source. So it was considered whether it would be possible to put the food source inside the cage. It was found that if a container was placed on the floor it was emptied by the next morning., The food pellets were, however, simply deposited in a heap at the container entrance, rather than placed in a discrete place away from the container, as might be expected if the mice were truly hoarding them. Close inspection showed that the mice were performing digging ("burrowing") movements, not carrying the pellets in their mouths to a selected place as they would if truly hoarding them.6



Food pellets are not an essential substrate for burrowing; mice will empty tubes filled with sand, gravel, even soiled bedding from their own cage. Moreover, they will empty a full tube even if an empty one is placed next to it8.



Several nesting protocols exist in the literature. The present Oxford one simplifies the procedure and has a well-defined scoring system for nest quality5. 



A hoarding paradigm was later developed in which the mice, rather than hoarding back to the real home cage, were adapted to living in the "home base" of a hoarding apparatus. This home base was connected to a tube made of wire mesh, the distal end of which contained the food source. This arrangement proved to yield good hoarding behaviour, as long as the mice were adapted to living in the "home base" during the day and only allowed to enter the hoarding tube at night.

Burrowing, nesting, and hoarding are species-typical activities that mice readily perform in the laboratory. This article describes how they can be easily and cheaply assessed. These protocols are extremely sensitive to mouse strain, brain lesions and diseases. Moreover they constitute &#x201C;environmental enrichment&#x201D; for the mice, and embody the &#x201C;Refinement&#x201D; aspect of the &#x201C;3 Rs&#x201D;18.

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 ...

神经科学

Assessment of Social Transmission of Food Preferences Behaviors

Olfactory recognition deficits are suggested to be able to serve as clinical marker to differentiate Alzheimer&#39;s disease (AD) subjects from healthy aging groups. For example, olfactory dysfunction in AD can present as impairment in olfactory recognition, emerging during early stages of the disease and worsening while the disease progresses. The social transmission of food preferences (STFP) task is based on a rudimentary form of communication between rodents concerning distant foods dependent on the transmission of olfactory cues. Healthy wild-type mice would prefer to eat a novel, flavored food that was previously cued by a conspecific, and this food preference would be hampered in transgenic AD mice, such as the APP/PS1 model. Indeed, a strong preference for the cued food in C57Bl6/J mice of 3 months of age was found, and this was reduced in 3 months old transgenic APP/PS1 mice. In summary, STFP task could be a powerful measure to be integrated in present subclinical detection assays of AD.

This paper presents a protocol for the investigation of social transmission of food preference in mice. The advantages and possible applications for this procedure, for instance, in detecting early changes in AD mouse models, are highlighted. To conclude, interpretation of the results in light of critical details are discussed.

食品偏好行为的社会传播评价

Olfactory recognition deficits are suggested to be able to serve as clinical marker to differentiate Alzheimer&#39;s disease (AD) subjects from healthy ...

Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia

In keeping with the 3R Principle (Replacement, Reduction, Refinement) developed by Russel and Burch, scientific research should use alternatives to animal experimentation whenever possible. When there is no alternative to animal experimentation, the total number of laboratory animals used should be the minimum needed to obtain valuable data. Moreover, appropriate refinement measures should be applied to minimize pain, suffering, and distress accompanying the experimental procedure. The categories used to classify the degree of pain, suffering, and distress are non-recovery, mild, moderate, or severe (EU Directive 2010/63). To determine which categories apply in individual cases, it is crucial to use scientifically sound tools.
The well-being-assessment protocol presented here is designed for procedures during which general anesthesia is used. The protocol focuses on home cage activity, the Mouse Grimace Scale, and luxury behaviors such as burrowing and nest building behavior as indicators of well-being. It also uses the free exploratory paradigm for trait anxiety-related behavior. Fecal corticosterone metabolites as indicators of acute stress are measured over the 24-h post-anesthetic period.
The protocol provides scientifically solid information on the well-being of mice following general anesthesia. Due to its simplicity, the protocol can easily be adapted and integrated in a planned study. Although it does not provide a scale to classify distress in categories according to the EU Directive 2010/63, it can help researchers estimate the degree of severity of a procedure using scientifically sound data. It provides a way to improve the assessment of well-being in a scientific, animal-centered manner.

We developed a protocol to assess well-being in mice during procedures using general anesthesia. A series of behavioral parameters indicating levels of well-being as well as glucocorticoid metabolites were analyzed. The protocol can serve as a general aid to estimate the degree of severity in a scientific, animal-centered manner.

全身麻醉下小鼠幸福度的系统评价

In keeping with the 3R Principle (Replacement, Reduction, Refinement) developed by Russel and Burch, scientific research should use alternatives to animal ...

<meta charset="utf8"></head><body>小鼠模型中自闭症谱系障碍的行为检测

Strategies for Assessing Autistic-Like Behaviors in Mice

Autism spectrum disorder (ASD) is a neurobiologically complex condition with a heterogeneous genetic etiology. Clinically, ASD is diagnosed by social communication impairments and restrictive or repetitive behaviors, such as hand flapping or lining up objects. These behavioral patterns can be reliably observed in mouse models with ASD-linked genetic mutations, making them highly useful tools for studying the underlying cellular and molecular mechanisms in ASD. Understanding how genetic changes affect the neurobiology and behaviors observed in ASD will facilitate the development of novel targeted therapeutic compounds to ameliorate core behavioral impairments. Our lab has employed several protocols encompassing well-described training and testing procedures that reflect a wide range of behavioral deficits related to ASD. Here, we detail two assays to study the core features of ASD in mouse models: self-grooming (a measure of repetitive behavior) and the three-chamber social interaction test (a measure of social interaction approach and preference for social novelty).

<meta charset="utf8"></head><body>评估小鼠自闭症样行为的策略

Rodent models are valuable tools for studying core behaviors related to autism spectrum disorder (ASD). In this article, we expound on two behavioral tests for modeling the core features of ASD in mice: self-grooming, which assesses repetitive behavior, and the three-chamber social interaction test, which documents social impairments.

评估小鼠自闭症样行为的策略

Autism spectrum disorder (ASD) is a neurobiologically complex condition with a heterogeneous genetic etiology. Clinically, ASD is diagnosed by social ...

<meta charset="utf8"></head><body>使用可口的食物获取小鼠模型研究暴饮暴食症

Intermittent Binge-Intake Model in Mice

Obesity affects one in eight individuals globally. Overeating, especially high-calorie foods, plays a significant role in obesity. Binge eating disorder (BED) is a complex condition caused by a combination of biological, psychological, and social factors. It involves excessive consumption of high-calorie foods in a short period and is often linked to anxiety and cravings.
Here, we present a protocol employing continuous and intermittent access to a new highly palatable food (HPF) -- M&amp;M&#39;s -- to develop a mouse model for investigating binge eating disorder (BED). The HPF, chosen for its high fat and sugar content, represents an optimal food source to study due to its strong palatability, which makes it particularly suited for examining compulsive eating behaviors. Using C57BL/6 mice, we provided continuous or intermittent access to M&amp;M&#39;s while allowing unrestricted access to standard chow and water. By the 8th day, the mice in the intermittent access group exhibited pronounced binge-eating behaviors, which persisted through the 26th day. These mice also consumed significantly more calories -- predominantly from the confectionary -- than those in the continuous access and control groups. The contrast between continuous and intermittent access underscores the critical role of feeding schedules in promoting the overconsumption of palatable foods.
Furthermore, behavioral assessments revealed intermittent access to the HPF-induced anxiety-like behaviors, highlighting the psychological impact of access patterns on both eating behavior and emotional states. By incorporating these two distinct feeding paradigms, this study offers valuable insights into how the availability of highly palatable foods can exacerbate binge-eating tendencies. This model provides a more realistic approach to studying binge-eating behaviors and their metabolic consequences. It highlights the importance of standardized models in biomedical research. Our findings offer insights into the physiological and neural mechanisms underlying BED, which could pave the way for developing effective therapeutic interventions for BED and obesity.

<meta charset="utf8"></head><body>作者聚焦:用于研究暴食症的基于 M&M 的可访问小鼠模型 - 对饮食行为、焦虑和神经机制的见解

A binge-eating model was established in mice using intermittent access to M&amp;M's (a highly palatable food made of fat and sugar). Mice with intermittent access to this high-fat, high-sugar food showed stress-like behaviors, increased caloric intake, and a preference for that food over standard chow compared to mice with continuous or no access to the food.

小鼠间歇性暴饮暴食模型

Obesity affects one in eight individuals globally. Overeating, especially high-calorie foods, plays a significant role in obesity. Binge eating disorder ...

小鼠活动性厌食症的评估

摘要

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跟踪恐惧条件小鼠

使用基于活动的厌食症啮齿动物模型来研究神经性厌食症的神经生物学基础

评估挖洞，在小鼠的鸟巢建设和囤积

食品偏好行为的社会传播评价

全身麻醉下小鼠幸福度的系统评价

评估小鼠自闭症样行为的策略

评估小鼠自闭症样行为的策略

评估小鼠自闭症样行为的策略

小鼠间歇性暴饮暴食模型

小鼠间歇性暴饮暴食模型