这项工作得到了国家神经病学和中风研究所（MPB）的R01 NS067417的支持。

这些研究严格按照"国家卫生研究院实验动物护理和使用指南"中的建议进行。该协议经乔治敦大学机构动物保护和使用委员会批准。将小鼠饲养在温度控制的动物设施中，并保持在12小时光/ 12小时黑暗循环。食物和水随意提供 。 1.制备mTBI仪器注意:mTBI设备包括一个控制冲击参数的数据采集（DAQ）箱，一个执行冲击的高速气动冲击器，以及一个模制的凝胶填充底座，使冲击后头部减速。 <ol><li>打开压缩空气，并将高速气动冲击器初始化为861.85 kPa的压力。 </li><li>将DAQ控制系统校准到以下参数 - 活塞速度为2.35 m / s，表面停留时间e为31.5 ms。 </li><li>将模制的凝胶填充底座（可压缩性64 kPa / mm）放置成中线与冲击器尖端的轨迹垂直。 </li></ol> mTBI冲击<ol><li>记录所有使用的小鼠的体重。 </li><li>在诱导室中麻醉3％异氟烷在氧气中120s的小鼠。 </li><li>将小鼠转移到mTBI装置，通过柔韧的非固定鼻锥继续麻醉。 </li><li>在鼻锥中，将鼠标头放在凝胶垫上，使得颅骨的平坦表面垂直于冲击器尖端呈现。 </li><li>将胶带放在鼠标头上，以创建一个平坦的表面，并将耳朵远离冲击部位。 </li><li>降低聚四氟乙烯冲击器尖端以与头部中心的矢状中线对齐。使用的冲击器尖端直径为10mm，将覆盖从眼睛后面到耳朵中线的头皮区域。调整冲击器，这样就可以了st触摸鼠标头的表面。 </li><li>收回冲击头，并手动拨下所需的偏转深度（7.5 mm）。 </li><li>控制DAQ控制系统，并按下触发按钮来影响鼠标头，或者是单一或重复的影响。重复的影响在每次影响之间不间断地交付。 </li><li>在最终冲击后立即将麻醉和mTBI装置的鼠标移开，并置于仰卧位。 </li><li>使用秒表测量正确反射（仰卧俯仰）返回的延迟，以确定意识时间的消失以及行走时间（无阻碍的行走）。 </li><li>监视鼠标并恢复正常行为，返回到其家中的笼子。 </li><li>假小鼠接受相同的处理和麻醉，但不受影响。 </li></ol>

<ol>
	<li>McMahon, P., 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=Symptomatology+and+functional+outcome+in+mild+traumatic+brain+injury:+results+from+the+prospective+TRACK-TBI+study.">Symptomatology and functional outcome in mild traumatic brain injury: results from the prospective TRACK-TBI study.</a> J Neurotrauma. 31 (1), 26-33 (2014).</li><li>Barkhoudarian, G., Hovda, D. A., Giza, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+molecular+pathophysiology+of+concussive+brain+injury.">The molecular pathophysiology of concussive brain injury.</a> Clin Sports Med. 30 (1), 33-48 (2011).</li><li>Blennow, K., Hardy, J., Zetterberg, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neuropathology+and+neurobiology+of+traumatic+brain+injury.">The neuropathology and neurobiology of traumatic brain injury.</a> Neuron. 76 (5), 886-899 (2012).</li><li>Levin, H. S., Diaz-Arrastia, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnosis,+prognosis,+and+clinical+management+of+mild+traumatic+brain+injury.">Diagnosis, prognosis, and clinical management of mild traumatic brain injury.</a> Lancet Neurol. 14 (5), 506-517 (2015).</li><li>McCrory, P., 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=Consensus+statement+on+concussion+in+sport:+the+4th+International+Conference+on+Concussion+in+Sport,+Zurich,+November+2012.">Consensus statement on concussion in sport: the 4th International Conference on Concussion in Sport, Zurich, November 2012.</a> J Athl Train. 48 (4), 554-575 (2012).</li><li>Talavage, T. 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=Functionally-detected+cognitive+impairment+in+high+school+football+players+without+clinically-diagnosed+concussion.">Functionally-detected cognitive impairment in high school football players without clinically-diagnosed concussion.</a> J Neurotrauma. 31 (4), 327-338 (2014).</li><li>Angoa-Perez, 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=Animal+models+of+sports-related+head+injury:+bridging+the+gap+between+pre-clinical+research+and+clinical+reality.">Animal models of sports-related head injury: bridging the gap between pre-clinical research and clinical reality.</a> J Neurochem. 129 (6), 916-931 (2014).</li><li>Creed, J. A., 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=Concussive+brain+trauma+in+the+mouse+results+in+acute+cognitive+deficits+and+sustained+impairment+of+axonal+function.">Concussive brain trauma in the mouse results in acute cognitive deficits and sustained impairment of axonal function.</a> J Neurotrauma. 28 (4), 547-563 (2011).</li><li>Hamberger, A., 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=Concussion+in+professional+football:+morphology+of+brain+injuries+in+the+NFL+concussion+model--part+16.">Concussion in professional football: morphology of brain injuries in the NFL concussion model--part 16.</a> Neurosurgery. 64 (6), 1174-1182 (2009).</li><li>Kane, M. 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=A+mouse+model+of+human+repetitive+mild+traumatic+brain+injury.">A mouse model of human repetitive mild traumatic brain injury.</a> J Neurosci Methods. 203 (1), 41-49 (2012).</li><li>Prins, M. 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=Repeat+traumatic+brain+injury+in+the+juvenile+rat+is+associated+with+increased+axonal+injury+and+cognitive+impairments.">Repeat traumatic brain injury in the juvenile rat is associated with increased axonal injury and cognitive impairments.</a> Dev Neurosci. 32 (5-6), 510-518 (2010).</li><li>Tang, Y. P., 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+concussive-like+brain+injury+model+in+mice+(II):+selective+neuronal+loss+in+the+cortex+and+hippocampus.">A concussive-like brain injury model in mice (II): selective neuronal loss in the cortex and hippocampus.</a> J Neurotrauma. 14 (11), 863-873 (1997).</li><li>Winston, C. N., 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=Dendritic+Spine+Loss+and+Chronic+White+Matter+Inflammation+in+a+Mouse+Model+of+Highly+Repetitive+Head+Trauma.">Dendritic Spine Loss and Chronic White Matter Inflammation in a Mouse Model of Highly Repetitive Head Trauma.</a> Am J Pathol. 186 (3), 552-567 (2016).</li><li>Johnson, V. E., 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=Inflammation+and+white+matter+degeneration+persist+for+years+after+a+single+traumatic+brain+injury.">Inflammation and white matter degeneration persist for years after a single traumatic brain injury.</a> Brain. 136, 28-42 (2013).</li><li>Barrio, J. 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=In+vivo+characterization+of+chronic+traumatic+encephalopathy+using+[F-18]FDDNP+PET+brain+imaging.">In vivo characterization of chronic traumatic encephalopathy using [F-18]FDDNP PET brain imaging.</a> Proc Natl Acad Sci U S A. 112 (16), 2039-2047 (2015).</li></ol>

作者没有什么可以披露的。

在人类中，mTBI的特征在于没有结构性损伤的功能障碍。这可能与意识丧失有关，或没有意识丧失1 。目前认为接触重复脑震荡是神经变性疾病如CTE 4发展和/或发展的基础。据了解，CTE通常在拳击手和足球运动员中被发现，尽管暴露于重复脑震荡（包括不导致意识丧失的人）被认为是开发CTE的基本要素，但我们还不知道为什么重复脑震荡引起大脑中发生明显变化的机制。 了解这些机制的主要障碍是难以发展模型，准确地概述在维持mTBI影响的个人中观察到的关键症状。这意味着急性和慢性生物拍摄震荡影响的基础设施难以进行调查，不能开发新的治疗手段。特别是，现有的重复脑震荡的动物模型非常严重，仅使用2-3次重复影响，导致大量神经元细胞丢失，颅骨骨折和脑组织损失8,10,11,12。在人类重复脑震荡之后，这种损伤程度不会发生，这表明需要一种单一和重复的轻度头部影响的新模型。 在这里，我们描述了一种新型的mTBI模型，使用了一种专门设计用于通过颅骨和大脑传输能量并远离鼠标的定制弥漫性损伤装置。通过单一影响和重复的影响范式，该模型允许一种手段开始调查脑震荡在生命后期如何造成神经损伤。单身h它的范例概括了人类观察到的轻度创伤性脑损伤的特征，而重复范式允许检查这些看似轻微的损伤如何有助于慢性和持续性变性。该装置允许重复的mTBI头部影响，没有颅骨骨折或结构性脑损伤的风险。 与许多技术一样，该协议的某些方面对于产生准确，可靠的结果非常重要。在腔室诱导和mTBI损伤期间，必须保持每只小鼠一致的麻醉水平。由于权力和行走时间是这种mTBI程序的关键结果测量的事实，研究人员应确保mT​​BI损伤和假动物都暴露于相当水平的麻醉剂。具体来说，在整个手术期间，小鼠只能在麻醉下总共3分钟（2分钟内）用mTBI鼻锥装置1分钟）。这种低水平和麻醉持续时间是超过许多其他mTBI模型的主要优点，并产生一致的反射反应时间（假小鼠20-40秒，mTBI小鼠50-100秒）。另外，当使用重复的mTBI范例时，重要的是在研究期间每天保持小鼠体重记录。这是为了确保在mTBI程序的整个时间过程中对压力和一般动物福利的持续监测。该过程不会导致小鼠被撤回或分离。在手术后的第一个小时内应观察正常的梳理，饮食。 损伤后1个星期的时间过程中单次或多次冲击导致的脑组织没有严重的形态学病理学。在单个mTBI之后，观察到的唯一的细胞反应是兴奋性突触的瞬时减少;日没有炎症，灰色或白质的损失，轴突损伤或细胞死亡。有30例重复mTBI（每天5次，6天以上），有与以前定量相似的水平的视神经的慢性炎症1 。在单次mTBI 14后多年观察到慢性白质炎症，并且可以在具有重复mTBI 15病史的生活运动员中检测到慢性炎症。该模型的局限性在于，它不能用于研究视神经以外的轴突损伤或细胞死亡机制，因为我们的模型中不存在。同样基于我们以前发表的数据，该模型在阿尔茨海默病13小鼠模型中，在mTBI后1个月时不诱导淀粉样蛋白或tau病理变化。我们认为，缺乏淀粉样蛋白和tau病理学与我们模型中轴突病理学的缺失有关。我们的模型确实提供了一个调查平台神经元网络的离散变化，突触完整性和组成以及重复脑震荡后的行为变化。基于这些结果，这种新型模型以受控，严格和有效的方式产生临床相关症状。进一步使用该模型将允许调查基础的机制，支持mTBI的急性和慢性病理生理学和脑震荡。

创伤性脑损伤（TBI）被定义为由外部物理力量持续的头部损伤，导致正常脑功能的破坏。它代表了重大的社会经济和公共卫生负担，"国际疾病预防控制中心2015年报告"向国会估计，每年有250万美国人维持TBI。这不仅影响了患者的生活质量，而且对社会造成极高的经济成本，目前估计每年为765亿美元。传达的实际脑损伤的数量和急性期症状是定义轻度，中度和重度TBI。 轻度创伤性脑损伤（mTBI）也被称为脑震荡，占每年报告的TBI的70％以上1 。参加拳击和足球2高风险接触运动的运动员最常见。不像中度或严重形式的TBI，与mTBI相关的直接损害和症状有时不明显3 。相比之下，mTBI的长期影响可能与中度和重度形式的长期影响一样虚弱。患有重复mTBI的患者已被证明会发展为慢性创伤性脑病（CTE），以及其他认知和退行性疾病。因此，重要的是要进一步了解有助于mTBI之后产生的短期症状和总体长期损害的机制。 在人类中，第四届国际体育冲击大会（苏黎世2012）定义的脑震荡定义5指出，体育震荡的伤害程度轻微，不会造成严重的病理变化，但确实造成短期神经功能缺损这是自发解决的。事实上，研究调查了使用头撞式遥测系统，mTBI对高中足球运动员认知障碍的影响。这项研究显示，在一个赛季，玩家持续头盔影响&gt; 20g的次数从226人（平均每场4.7次）的高点到1855年（平均每场38.6次） 6次 。大多数这些影响并不导致脑震荡的临床诊断;但是使用fMRI 6可以观察到脑功能的功能变化的证据。导致这些功能变化的大脑变化是未知的，因此迫切需要有一个可靠和可重复的模型，以便于研究震荡和次冲击性mTBI的影响。 尽管以前尝试在小鼠和老鼠中建模mTBI 7 ，但许多报告不良反应。特别是，大多数啮齿动物模型的重复性质受到限制5 mTBIs的影响，以及不良的病理事件，包括脑内出血，颅骨骨折，严重的轴突损伤，神经元细胞死亡和增加的死亡率8,9,10,11,12。在这里，我们描述了更接近人类脑震荡真正定义的mTBI的小鼠模型。该模型概括了人类mTBI中观察到的许多症状，例如导致意识暂时丧失的机械力，没有明显的大脑病理学。此外，有利的是，它可以用于长时间段的单次冲击和重复冲击范例，如前所述13 。

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		<col />
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			<td height="21" style="height:21px;width:323px;">Powerlab 8SP data acquisition (DAQ) control box&#160;</td>
			<td style="width:203px;">(AD instruments)</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:323px;">VIP 3000 calibrated vaporizer</td>
			<td style="width:203px;">Matrx</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane</td>
			<td style="width:203px;">Henry Schein Animal Health</td>
			<td style="width:119px;">29405</td>
			<td></td>
		</tr>
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			<td height="21" style="height:21px;width:323px;">Oxygen</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td>Commercially available</td>
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			<td height="21" style="height:21px;width:323px;">Compressed Air</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td>Commercially available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Masking Tape</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td>Commercially available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stop Watch</td>
			<td style="width:203px;">Fisher Scientific</td>
			<td>02-261-840</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">C57 Bl/6 Mice</td>
			<td style="width:203px;">Jackson Laboratories</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">Digital Scale and weigh container</td>
			<td style="width:203px;">Fisher Scientific</td>
			<td>20031</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:323px;">anti-Iba1 antibody</td>
			<td style="width:203px;">Wako</td>
			<td style="width:119px;">019-19741</td>
			<td></td>
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			<td height="21" style="height:21px;width:323px;">HRP labelled secondary&#160;</td>
			<td style="width:203px;">Jackson Immunoresearch</td>
			<td style="width:119px;">111-035-003</td>
			<td></td>
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a mouse model of single and repetitive mild traumatic brain injury

轻度创伤性脑损伤（mTBI）可导致脑功能急性丧失，包括混乱期，意识丧失（LOC），局部神经功能缺损甚至健忘症。参加联系运动的运动员面临着大量mTBI的高风险。在体育运动员的伤害程度方面，mTBI被定义为不会引起严重病理改变的轻度损伤，但是会导致自发解决的短期神经功能缺损。尽管以前曾尝试在小鼠和老鼠中建模mTBI，但许多人报告了总体不良反应，包括颅骨骨折，脑内出血，轴突损伤和神经元细胞死亡。在这里，我们描述了我们高度重复性的mTBI动物模型，其复制临床相关症状。该型号使用定制的气动冲击器装置来传递闭合头部的创伤。这种影响是在精确的速度和变形参数下进行的，创建一个可靠和可重复的模型来检查有助于单个或重复的震荡mTBI的影响的机制。

使用这种新颖的mTBI装置允许单次和重复的轻度头部损伤，没有颅骨骨折或结构性脑损伤的风险。该型号采用定制的气动特氟龙冲击器装置，可提供封闭的头部机械能量冲击。在精确的速度和变形参数下产生影响，创建可靠和可重复的模型来检查有助于单个或重复的震荡mTBI的影响的机制（ 图1 ）。 LOC的存在或不存在是在动物模型中评分脑震荡严重性的有用工具。正确反射时间的返回是对单次和重复mTBI（7.5 mm深度， 图2 ）之后我们用于量化LOC的损伤严重程度的急性神经学评估。在该过程中，小鼠接受总共3分钟的异氟烷<html>在氧气中，因此包括假手术的所有小鼠在麻醉撤除后具有LOC周期。与假手术小鼠相比，单个mTBI导致LOC显着增加（36.4±1.6 sv 64.2±7.7 s，n = 5，** p &lt;0.01， 图2A ）。这也与单次mTBI后增加的行走时间相关（52.0±4.5 sv 140.0±21.1 s，n = 5，** p &lt;0.01， 图2B ）。在重复伤害范式（共30次影响，每天5次影响6天）中，所有测试日均LOC和行走时间显着升高（重复测量双向分析方差损伤效应F 1,14 = 22.92， p &lt;0.0003）。在整个研究期间，6天内的平均LOC为Sham:35.5±1.4 sv mTBI:64.9±1.7 s，n = 8，p &lt;0.01， 图2C和平均行走时间Sham:64.3±3.3 sv mTBI:160.8 ±5.3 s，n = 8，p &lt;0.01，lass ="xfig"&gt;图2D。小胶质细胞/巨噬细胞的Iba1染色显示假手术和单个mTBI小鼠之间无变化，但在重复mTBI小鼠中视神经中广泛的Iba1免疫反应性（ 图3 ）。重复mTBI小鼠在皮层（ 图3 ）或其他脑区没有显示灰质炎症的迹象。 <img alt="图1" class="xfigimg" src="/files/ftp_upload/55713/55713fig1.jpg" /> 图1 :mTBI鼠标模型的说明性表示。 （ A ）设置执行mTBI所需的所有材料。 （ B ）将小鼠放置在异氟烷诱导室中2分钟。 （ C ）在mTBI装置上，将小鼠放置在非固定鼻锥体中以继续麻醉。 （ D ）轻轻的磁带鼠标头，创建一个平坦的表面，并保持耳朵。 IMPACTOr尖端降低以仅接触头部的表面。 （ E ）使用拨盘（插图）降低冲击器尖端并要求偏转深度。通过按下触发按钮执行mTBI。 （ F ）由正反射返回所需的时间（仰卧位）测量的意识丧失。 <a href="http://ecsource.jove.com/files/ftp_upload/55713/55713fig1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图2" class="xfigimg" src="/files/ftp_upload/55713/55713fig2.jpg" /> 图2 :单次和重复的mTBIs增加了瞬间的意识丧失并返回到行动时间。接受单个mTBI的小鼠与假手术对照小鼠相比显着增加（ A ）意识丧失（LOC）和（ B ）移动时间（每组n = 5，** p &lt;0.01）。 （ C - D ）与假手术小鼠相比，重复mTBI（每天5次/ 6天）显着增加每日LOC和行走时间（n = 8 /组* p &lt;0.05，** p &lt;0.01，*** p &lt; 0.001）。数据表示为平均值±SEM，通过双因重复测量方差分析进行Bonferroni事后检验。 <a href="http://ecsource.jove.com/files/ftp_upload/55713/55713fig2large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图3" class="xfigimg" src="/files/ftp_upload/55713/55713fig3.jpg" /> 图3 :重复mTBI诱导光学细胞中的小胶质细胞/巨噬细胞激活。 （ A ）假小鼠Iba1染色显示皮层（a1-a3）中静止的小神经胶质细胞/巨噬细胞和视神经中有限的染色（a4-a6）。 （ B ）接收单个mTBI的小鼠（7.5mm）在皮质（b1-b3）和视神经（b4-b6），冲击后24小时具有与假手术小鼠相似的Iba1染色曲线。 （ C ）重复mTBI小鼠（连续6天每天5次）在视力（c4-c6）中显示强烈的炎症反应，但在最后一次冲击后24小时（c1-c3）不显示（第7天） ）。扩大的图像分别显示了假皮，单次和重复mTBI小鼠的皮层（a3，b3，c3）和视神经纤维（a6，b6，c6）中Iba-1阳性小胶质细胞/巨噬细胞的形态。刻度棒=200μm（A，B，C）; 50μm（a1-a5，b1-b5，c1-c5）; 20μm（a3，a6，b3，b6，c3，c6，放大图）。每组n = 6的图像。皮质（cx），视神经（opt），轻度创伤性脑损伤（mTBI）。 <a href="http://ecsource.jove.com/files/ftp_upload/55713/55713fig3large.jpg" target="_blank">请点击此处查看此图的较大版本。</a>

Watch this Scientific Journal Video about A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury at JoVE.com

A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury

运动员每年吸收数百次轻度创伤性脑损伤（mTBI）/脑震荡;然而，这些对大脑的后果不甚了解。因此，单次和重复性mTBI的动物模型始终如一地复制临床相关症状，为进一步研究mTBI和脑震荡提供了手段。

单一和重复轻度创伤性脑损伤的小鼠模型

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal ...

a-mouse-model-of-single-and-repetitive-mild-traumatic-brain-injury

Nanyang Technological University

Research

JoVE Journal

Medicine

11.0K 次观看.  Georgetown University Medical Center. 运动员每年吸收数百次轻度创伤性脑损伤（mTBI）/脑震荡;然而，这些对大脑的后果不甚了解。因此，单次和重复性mTBI的动物模型始终如一地复制临床相关症状，为进一步研究mTBI和脑震荡提供了手段。 

视频: A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused by whole-body hypoperfusion.5,6 Successful reproduction of whole-body hypoperfusion in rodent models has been fraught with 
difficulty.7-9,9,10 Models which employ focal ischemia have repeatedly demonstrated results which do not translate to the clinical setting, and larger 
animal models which allow for whole body hypoperfusion lack access to the full toolset of genetic manipulation possible in the mouse.11,12 However, in recent 
years a mouse model of cardiac arrest and cardiopulmonary resuscitation has emerged which can be adapted to model AKI.13 This model reliably reproduces 
physiologic, functional, anatomic, and histologic outcomes seen in clinical AKI, is rapidly repeatable, and offers all of the significant advantages of a murine surgical 
model, including access to genetic manipulative techniques, low cost relative to large animals, and ease of use. Our group has developed extensive experience 
with use of this model to assess a number of organ-specific outcomes in AKI.14,15

A powerful model for perioperative and critical care related acute kidney injury is presented. Using whole body hypoperfusion induced by cardiac arrest it is possible to nearly replicate the histologic and functional changes of clinical AKI.

常温心脏骤停和心肺复苏术:缺血再灌注损伤的小鼠模型

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

科研

医学

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

在动物模型揭示神经，血液，创伤后应激障碍和行为相关因素的生物标志物

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

Stretch in Brain Microvascular Endothelial Cells (cEND) as an In Vitro Traumatic Brain Injury Model of the Blood Brain Barrier

Due to the high mortality incident brought about by traumatic brain injury (TBI), methods that would enable one to better understand the underlying mechanisms involved in it are useful for treatment. There are both in vivo and in vitro methods available for this purpose. In vivo models can mimic actual head injury as it occurs during TBI. However, in vivo techniques may not be exploited for studies at the cell physiology level. Hence, in vitro methods are more advantageous for this purpose since they provide easier access to the cells and the extracellular environment for manipulation.
Our protocol presents an in vitro model of TBI using stretch injury in brain microvascular endothelial cells. It utilizes pressure applied to the cells cultured in flexible-bottomed wells. The pressure applied may easily be controlled and can produce injury that ranges from low to severe. The murine brain microvascular endothelial cells (cEND) generated in our laboratory is a well-suited model for the blood brain barrier (BBB) thus providing an advantage to other systems that employ a similar technique. In addition, due to the simplicity of the method, experimental set-ups are easily duplicated. Thus, this model can be used in studying the cellular and molecular mechanisms involved in TBI at the BBB.

Stretch in Brain Microvascular Endothelial Cells cEND as an In Vitro Traumatic Brain Injury Model of the Blood Brain Barrier

In vitro traumatic brain injury models are being developed to reproduce in vivo brain deformation. Stretch-induced injury has been employed for astrocytes, neurons, glial cells, aortic, and brain endothelial cells. However, our system uses a blood brain barrier (BBB) model that possesses properties constituting a legitimate model of the BBB to establish an in vitro&#160;TBI model.

作为脑微血管内皮细胞（CEND）的伸展<em&gt;体外</em创伤性脑损伤模型血脑屏障

Due to the high mortality incident brought about by traumatic brain injury (TBI), methods that would enable one to better understand the underlying ...

An Alkali-burn Injury Model of Corneal Neovascularization in the Mouse

Under normal conditions, the cornea is avascular, and this transparency is essential for maintaining good visual acuity. Neovascularization (NV) of the cornea, which can be caused by trauma, keratoplasty or infectious disease, breaks down the so called &lsquo;angiogenic privilege' of the cornea and forms the basis of multiple visual pathologies that may even lead to blindness. Although there are several treatment options available, the fundamental medical need presented by corneal neovascular pathologies remains unmet. In order to develop safe, effective, and targeted therapies, a reliable model of corneal NV and pharmacological intervention is required. Here, we describe an alkali-burn injury corneal neovascularization model in the mouse. This protocol provides a method for the application of a controlled alkali-burn injury to the cornea, administration of a pharmacological compound of interest, and visualization of the result. This method could prove instrumental for studying the mechanisms and opportunities for intervention in corneal NV and other neovascular disorders.

Neovascularization (NV) of the cornea can complicate multiple visual pathologies. Utilizing a controlled, alkali-burn injury model, a quantifiable level of corneal NV can be produced for mechanistic study of corneal NV and evaluation of potential therapies for neovascular disorders.

角膜新生血管在小鼠碱烧伤模型

Under normal conditions, the cornea is avascular, and this transparency is essential for maintaining good visual acuity. Neovascularization (NV) of the ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

控制皮层影响模型创伤性脑损伤

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable during renal transplantation. Experimental models that accurately and reproducibly recapitulate renal IRI are crucial in dissecting the pathophysiology of AKI and the development of novel therapeutic agents. Presented here is a mouse model of renal IRI that results in reproducible AKI. This is achieved by a midline laparotomy approach for the surgery with one incision allowing both a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia of the left kidney. By careful monitoring of the clamp position and body temperature during the period of ischaemia this model achieves reproducible functional and structural injury. Mice sacrificed 24 hr&#160;following surgery demonstrate loss of renal function with elevation of the serum or plasma creatinine level as well as structural kidney damage with acute tubular necrosis evident. Renal function improves and the acute tissue injury resolves during the course of 7 days following renal IRI such that this model may be used to study renal regeneration. This model of renal IRI has been utilized to study the molecular and cellular pathophysiology of AKI as well as analysis of the subsequent renal regeneration.

The mouse model of renal ischaemia reperfusion injury described here comprises of a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia that results in acute kidney injury. This model uses a midline laparotomy approach with all steps performed via one incision.

肾缺血再灌注损伤:损伤的小鼠模型和再生

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable ...

A Novel Model of Mild Traumatic Brain Injury for Juvenile Rats

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor vehicle accidents, and falls, a reliable animal model of mTBI had previously not been developed for this important aspect of development. The modified weight-drop technique employs a glancing impact to the head of a freely moving rodent transmitting acceleration, deceleration, and rotational forces upon the brain. When applied to juvenile rats, this modified weight-drop technique induced clinically relevant behavioural outcomes that were representative of post-concussion symptomology. The technique is a rapidly applied procedure with an extremely low mortality rate, rendering it ideal for high-throughput studies of therapeutics. In addition, because the procedure involves a mild injury to a closed head, it can easily be used for studies of repetitive brain injury. Owing to the simplistic nature of this technique, and the clinically relevant biomechanics of the injury pathophysiology, the modified weight-drop technique provides researchers with a reliable model of mTBI that can be used in a wide variety of behavioural, molecular, and genetic studies.

The modified weight-drop technique is an easy, cost-effective procedure used for the induction of mild traumatic brain injury in juvenile rats. This novel technique produces clinically relevant symptomology that will advance the study of mild traumatic brain injury (mTBI) and concussion.

轻度脑外伤的幼年大鼠一种新模式

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

新生儿小鼠脊髓压迫损伤模型

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

A Repetitive Concussive Head Injury Model in Mice

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of knowledge on the injury and its effects. To develop a better understanding of concussions, animals are often used because they provide a controlled, rigorous, and efficient model. Studies have adapted traditional animal models to perform mTBI to stimulate mild injury severity by changing the injury parameters. These models have been used because they can produce morphologically similar brain injuries to the clinical condition and provide a spectrum of injury severities. However, they are limited in their ability to present the identical features of injuries in patients. Using a traditional impact system, a repetitive concussive injury (rCHI) model can induce mild to moderate human-like concussion. The injury degree can be determined by measuring the period of loss of consciousness (LOC) with a sign of a transient termination of breathing. The rCHI model is beneficial to use for its accuracy and simplicity in determining mTBI effects and potential treatments.

Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.

重复震荡颅脑损伤模型小鼠

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of ...

A Tissue Displacement-based Contusive Spinal Cord Injury Model in Mice

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between experimental animals. Several contusive SCI models have been developed to produce injuries using different mechanisms. The severity of the SCI is based on the height that a given weight is dropped, the injury force, or the spinal cord displacement. In the current study, we introduce a novel mouse contusive SCI device, the Louisville Injury System Apparatus (LISA) impactor, which can create a displacement-based SCI with high injury velocity and accuracy. This system utilizes laser distance sensors combined with advanced software to produce graded and highly-reproducible injuries. We performed a contusive SCI at the 10th thoracic vertebral (T10) level in mice to demonstrate the step-by-step procedure. The model can also be applied to the cervical and lumbar spinal levels.

We introduce a tissue displacement-based contusive spinal cord injury model that can produce a consistent contusive spinal cord injury in adult mice.

基于组织位移的小鼠脊髓损伤模型

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between ...