Name: electromagnetic controlled closed-head model of mild traumatic brain injury in mice
Uploaded: 2022-09-28T15:00:00.000+00:00
Duration: 9 min 7 s
Description: Watch this Scientific Journal Video about Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice at JoVE.com

这项工作得到了美国国立卫生研究院的部分支持，奖励号为R01NS120882，RF1NS119165和R01NS103785以及国防部奖励号AZ190017。内容完全由作者负责，不代表美国国立卫生研究院或国防部的官方观点。

进行的实验得到了肯塔基大学机构动物护理和使用委员会（IACUC）的批准，并且在研究期间遵循了ARRIVE和实验动物护理和使用指南指南。
1. 手术设置
注意:将小鼠饲养在4-5/笼中，房屋室内湿度保持在43%-47%，温度保持在22-23°C。让小鼠 随意 获得食物和水，并暴露于12小时/ 12小时光照/黑暗循环（上午7点/ 下午7点）。
<ol><li>使用指定的手术区域，例如头罩或专用外科手术室，进行动物手术。</li>
	<li>确保手术区域包括加热垫、配备电磁冲击器的立体定位框架和设计用于施用异氟醚气体的麻醉面罩（见 图1A）。</li>
	<li>确保外科医生或参与手术的人员穿着干净的实验室外套、口罩、手套和手术帽。</li>
	<li>使用无菌手术工具、无菌棉头涂抹器和纱布垫。在手术当天使用热珠灭菌器对小鼠之间的器械进行消毒。</li>
	<li>使用麻醉诱导室为小鼠在术前区域进行手术做准备。</li>
	<li>使用加热垫来维持动物的温度，清洁术后小鼠的笼子，并使用计时器记录手术后小鼠的扶正反射。</li>
</ol>2. 术前程序
<ol><li>准备头部支撑装置（见 图1B）。<ol><li>从 1 mL 乳胶移液器灯泡（充气端）上取下卷起端脊（见 图 1C）。</li>
		<li>使用封口膜将灯泡连接到管道上（见 图1C）。</li>
		<li>使用旋塞阀将管子连接到 10 mL 注射器。用水填充注射器（见 图1C）。 注意:1 mL乳胶移液器灯泡将放置在鼠标头部下方，以将冲击力从耳朵上移开。使用前尽量从灯泡中去除空气，使灯泡大部分充满水而不是空气。</li>
	</ol></li>
	<li>冲击器设置。<ol><li>选择 5 mm 探头尖端，将其拧到致动器底部中心（较大圆柱体内部）的活塞上，然后轻轻拧紧探头，不要用力过大。在撞击之间重新拧紧尖端（见 图 1B）。</li>
		<li>在打开冲击器之前，请确保伸出/缩回开关位于中心关闭位置。然后，将执行器上的电缆连接到冲击器控制箱前面板上的插孔，并将传感器电缆连接到前面板上的插孔。然后，打开后面板上的电源开关（请参阅 图 1D）。 注意: 不使用时，伸展/缩回拨动开关需要保持在中心关闭位置。</li>
		<li>通过旋转控制盒左侧的大旋钮来设置冲击速度，直到显示屏上出现 5.0 ± 0.2 m/s 的冲击速度（见 图 1D）。</li>
		<li>旋转拨盘，直到停留读数为 0.01，将停留计数器设置为 100 ms（参见 图 1D）。 注意:停留是自动缩回发生之前的接触时间。</li>
		<li>将冲击器致动器放在冰袋上以防止塑料气缸膨胀，从而将气缸锁定到位，防止气缸移动和未来冲击的传递（见 图 1E）。</li>
	</ol></li>
	<li>准备手术小鼠。<ol><li>手术前目视检查小鼠，如果观察到以下情况之一，请将小鼠从研究中剔除:4个月大的小鼠的皮毛状况差，嗜睡或体重差（<20克）。</li>
		<li>使用放置在加热垫上的诱导室在100%氧气中用4%-5%异氟醚麻醉小鼠1-2分钟。</li>
		<li>使用电动理发器从手术部位剃掉皮毛。</li>
		<li>在手术开始前至少15分钟用无菌酒精准备垫清洁头部，并在剃光的头皮上涂抹局部麻醉剂。</li>
		<li>手术前将鼠标放回干净的保持笼中。局部麻醉应用至少15分钟后开始手术（诱导时间）。 注意:麻醉时间可能因手术中使用的麻醉剂而异。</li>
	</ol></li>
	<li>再次检查立体定位框架、冲击器和数字立体定位显示器（见 图1F）是否已准备好使用。</li>
	<li>将小鼠放回异氟醚诱导室，用4%-5%异氟醚在100%氧气中约3分钟。</li>
	<li>将鼠标固定在头部阶段。</li>
</ol>3. 外科手术
<ol><li>使用轻质乙缩醛树脂锥形尖头耳杆、咬杆和小鼠麻醉面罩将鼠标固定到立体定位框架中（见 图 1G，H）。异氟醚气体在室内空气中以 2%-3% 的速度以 100-200 mL/min 的速度输送。仔细监测小鼠的呼吸以确保麻醉深度，并根据需要调整气体水平。</li>
	<li>在眼睛上涂抹无菌眼部润滑剂，以防止角膜干燥。</li>
	<li>用聚维酮碘拭子和无菌酒精垫对头皮进行三次消毒。</li>
	<li>通过验证没有捏脚趾反应来确保鼠标已深度麻醉。</li>
	<li>使用手术刀在眼睛和颈部之间切开约1厘米的中线头皮切口，露出头骨（见 图1I）。</li>
	<li>让头骨干燥1-2分钟。</li>
	<li>识别前胛（冠状缝合线和矢状缝线的交点）和λ（矢状面和λ缝合线的交点）（见 图1J）。 注意:小鼠大脑图谱可供参考。</li>
	<li>将头部支撑装置放在头部下方，用水给灯泡充气，直到它压在鼠标头部的底部，但不要将头部从咬杆上抬起。 注意:此步骤对于减少CHI可能引起的耳朵问题至关重要。 任何耳杆对耳朵造成损害，导致滚动或出血的动物，都应从研究中消除并实施安乐死。</li>
	<li>将撞击器移到动物头顶的位置。</li>
	<li>通过将伸展 /缩回 拨动开关（在撞击器控制盒上）放在伸展上来 伸出冲击器。 注意:请务必通过向下拉尖端来检查尖端是否已完全伸展。</li>
	<li>将撞击器对齐，直到它位于前膛中心（见 图1K）。</li>
	<li>将立体定位阅读器中的数字立体定位 x 和 y 坐标重置为 0（在触摸屏控件上）</li>
	<li>通过将探头从前膛移动到目标坐标，将探头对准撞击位置:内侧 = 0.0 mm，前后 = −1.6 mm。</li>
	<li>将接触式传感器夹在动物的耳朵上。<ol><li>用伸出的探头慢慢降低探头尖端，直到第一次与表面接触。在哔哔声处停止。</li>
		<li>将立体定位读取器中的数字立体定位 z 坐标重置为 0。</li>
	</ol></li>
	<li>仔细检查尖端是否与颅骨齐平（内侧-外侧和前后平面）。 注意:定位探头尖端是此过程中最关键的步骤，以防止颅骨骨折和耳朵损伤。</li>
	<li>通过将控制盒上的拨动开关置于缩回位置来缩回撞击器。尖端退出，在撞击之前不再与动物的头部接触。</li>
	<li>通过将背腹深度调整为 −1.2 mm 来设置冲击深度。 注意:撞击的深度会影响伤害的严重程度。应根据不同年龄、体重和品系的小鼠滴度滴度，以达到所需的损伤严重程度。深度可能需要随着时间的推移进行调整/重新滴定，以保持一致的损伤严重程度。严重程度可以从神经病理学上评估:小胶质细胞和星形胶质细胞 （IHC），以及行为学:桡臂水迷宫和主动回避试验。</li>
	<li>仔细监测小鼠的呼吸以确保麻醉深度并根据需要调整气体水平。 注意:通常，异氟醚气体的百分比应在撞击前降低或关闭10-20秒。密切观察呼吸是否略微加速。如果在撞击时呼吸太慢，动物可能会在呼吸暂停撞击后的前 60 秒内死亡。这可以通过在撞击前几秒钟内调整麻醉深度来预防。</li>
	<li>通过按右切换开关进行撞击来引起 撞击。探头尖端以显示的速度下降，然后在设定的停留时间内保持向下并缩回。 注意:假小鼠接受与CHI小鼠相同的处理，但不会产生影响。</li>
	<li>在传递CHI冲击后立即启动计时器以记录校正时间（从侧面位置返回到俯卧位置的时间），或者在将鼠标从假小鼠的立体定位框架中移开时启动计时器。平均矫正反射时间为5-15分钟。 注意:矫正反射时间可能因小鼠应变和年龄而异。</li>
	<li>评估小鼠是否有可见的颅骨骨折、出血和呼吸暂停。从研究中排除颅骨骨折或可见出血的小鼠。 注意:颅骨骨折有分级水平。对颅骨骨折的动物实施安乐死（首先使用CO2 ，斩首作为次要方法）。如果撞击器尖端设置正确，这些类型的颅骨骨折极为罕见。如果确实发生颅骨骨折，更常见的表现是颅骨上有一小滴血和颅骨轻微的触觉粗糙，通常沿着连接鼻骨后尖的缝合线。这些小鼠在记录中被记录为可能的颅骨骨折，但通常不会被排除在研究之外。</li>
	<li>将动物从立体定位框架中取出。</li>
	<li>通过将皮肤缝合在一起来关闭头皮。 注意:可吸收或不可吸收的缝合线可用于闭合头皮，作为订书钉的替代品。</li>
	<li>用无菌棉头涂抹器在闭合的切口上涂抹三重抗生素软膏。</li>
	<li>将鼠标放回干净的保持架进行恢复。恢复笼的一半位于加热垫上（低设置），提供在清醒时远离热量的能力，并在无意识时保持动物的温度（见 图1L）。 注:鼠标侧放在恢复架中。为防止窒息，请将动物放在没有垫料的恢复笼中，如果床上用品在笼子里，则放在纸巾上。</li>
	<li>将伸展/缩回拨动开关返回到中心/关闭位置。 注意: 如果开关处于伸出或缩回位置，电流将继续运行，从而导致活塞膨胀。然后，在活塞冷却之前，撞击器将无法正常工作。</li>
	<li>从支架上取下撞击器，轻轻地将其放在冰袋上。 注意:将撞击器放在冰袋上有助于减少撞击器的潜在膨胀。</li>
	<li>监测动物直到发生矫正反射并记录矫正前的时间（见 图1M）。 注意:右倾反射定义为鼠标返回俯卧位的那一刻。笼子需要不受干扰;如果笼子被触摸、移动或暴露于一些噪音中，鼠标可能会正确。</li>
	<li>当小鼠清醒和警觉时，将它们放回它们的家笼。通常，在受伤后1小时内，动物完全清醒并走动。另外，在笼子底部添加一些潮湿的食物。</li>
</ol>4. 术后护理
<ol><li>手术后监测动物5天。</li>
	<li>记录他们的体重和任何身体/行为变化，如呼吸频率（定性呼吸功能）、步态、身体和毛发状况、进食、饮水、排便和排尿。</li>
	<li>观察小鼠是否有任何不适迹象和手术伤口是否有肿胀，渗出物或红边，裂开。如果动物出现疼痛和不适的迹象（发声，不动，体温过低，不喝或不吃），请联系兽医。</li>
	<li>手术后7-10天在麻醉下和加热垫上取下订书钉。 注意:如果使用不可吸收的缝合线，则必须在手术后7-10天在麻醉下将其移除。</li>
</ol>5. 清洁
<ol><li>清洁和消毒手术区域和工具。</li>
	<li>每次使用后清洁探头尖端，并在一天结束时用酒精制备垫清洁探头。 注意:撞击器在工厂经过校准，据报道随着时间的推移和使用是稳定的。无需常规校准。但是，应常规检查撞击器和立体定位框架。此外，应监测模型终点周长，例如矫正反射时间、死亡率和神经病理学，以评估可能的实验漂移。</li>
</ol>6. 排除标准
<ol><li>手术前排除健康状况不佳的动物，例如体重差<4个月大的小鼠为20克，嗜睡和皮毛状况不佳。</li>
	<li>排除手术期间出现并发症的动物，如凹陷性颅骨骨折、与手术相关的可见出血或耳出血。</li>
	<li>将具有以下术后症状的动物排除在研究之外:无法进食和/或正常移动、异常发声、体重减轻或手术后伤口无法正常愈合。 注意:此模型可用作轻度 TBI 的重复模型。如果小鼠在第一次手术间隔24小时接受第二次手术，则可以移除订书钉或缝合线，并且可以使用相同的切口来暴露头骨。如果手术间隔时间较长，则需要做一个新的切口。</li>
</ol>

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D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attenuation+of+traumatic+brain+injury-induced+cognitive+impairment+in+mice+by+targeting+increased+cytokine+levels+with+a+small+molecule+experimental+therapeutic.">Attenuation of traumatic brain injury-induced cognitive impairment in mice by targeting increased cytokine levels with a small molecule experimental therapeutic.</a> Journal of Neuroinflammation. 12, 69 (2015).</li><li>Bachstetter, A. 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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=Inulin+supplementation+mitigates+gut+dysbiosis+and+brain+impairment+induced+by+mild+traumatic+brain+injury+during+chronic+phase.">Inulin supplementation mitigates gut dysbiosis and brain impairment induced by mild traumatic brain injury during chronic phase.</a> Journal of Cellular Immunology. 4 (2), 50-64 (2022).</li><li>Bachstetter, A. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+stage+drug+treatment+that+normalizes+proinflammatory+cytokine+production+attenuates+synaptic+dysfunction+in+a+mouse+model+that+exhibits+age-dependent+progression+of+Alzheimer's+disease-related+pathology.">Early stage drug treatment that normalizes proinflammatory cytokine production attenuates synaptic dysfunction in a mouse model that exhibits age-dependent progression of Alzheimer's disease-related pathology.</a> The Journal of Neuroscience. 32 (30), 10201-10210 (2012).</li><li>Zvejniece, 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=Skull+fractures+induce+neuroinflammation+and+worsen+outcomes+after+closed+head+injury+in+mice.">Skull fractures induce neuroinflammation and worsen outcomes after closed head injury in mice.</a> Journal of Neurotrauma. 37 (2), 295-304 (2020).</li><li>Flierl, M. 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=Mouse+closed+head+injury+model+induced+by+a+weight-drop+device.">Mouse closed head injury model induced by a weight-drop device.</a> Nature Protocols. 4 (9), 1328-1337 (2009).</li><li>Yang, Z., 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=Temporal+MRI+characterization,+neurobiochemical+and+neurobehavioral+changes+in+a+mouse+repetitive+concussive+head+injury+model.">Temporal MRI characterization, neurobiochemical and neurobehavioral changes in a mouse repetitive concussive head injury model.</a> Scientific Reports. 5, 11178 (2015).</li><li>Petraglia, A. 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=The+spectrum+of+neurobehavioral+sequelae+after+repetitive+mild+traumatic+brain+injury:+a+novel+mouse+model+of+chronic+traumatic+encephalopathy.">The spectrum of neurobehavioral sequelae after repetitive mild traumatic brain injury: a novel mouse model of chronic traumatic encephalopathy.</a> Journal of Neurotrauma. 31 (13), 1211-1224 (2014).</li><li>Laskowitz, D. T., 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=COG1410,+a+novel+apolipoprotein+E-based+peptide,+improves+functional+recovery+in+a+murine+model+of+traumatic+brain+injury.">COG1410, a novel apolipoprotein E-based peptide, improves functional recovery in a murine model of traumatic brain injury.</a> Journal of Neurotrauma. 24 (7), 1093-1107 (2007).</li><li>Lloyd, E., Somera-Molina, K., Van Eldik, L. J., Watterson, D. M., Wainwright, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+acute+proinflammatory+cytokine+and+chemokine+upregulation+by+post-injury+administration+of+a+novel+small+molecule+improves+long-term+neurologic+outcome+in+a+mouse+model+of+traumatic+brain+injury.">Suppression of acute proinflammatory cytokine and chemokine upregulation by post-injury administration of a novel small molecule improves long-term neurologic outcome in a mouse model of traumatic brain injury.</a> Journal of Neuroinflammation. 5, 28 (2008).</li><li>Lillie, E. M., Urban, J. E., Lynch, S. K., Weaver, A. A., Stitzel, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+skull+cortical+thickness+changes+with+age+and+sex+from+computed+tomography+scans.">Evaluation of skull cortical thickness changes with age and sex from computed tomography scans.</a> Journal of Bone and Mineral Research. 31 (2), 299-307 (2016).</li><li>Kawakami, M., Yamamura, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cranial+bone+morphometric+study+among+mouse+strains.">Cranial bone morphometric study among mouse strains.</a> BMC Evolutionary Biology. 8, 73 (2008).</li></ol>

作者没有什么可透露的。

使用所描述的模型重新创建一致的伤害模型涉及几个步骤。首先，将动物正确固定在立体定位框架中至关重要。动物的头部不应能够横向移动，头骨应完全平坦，前膛和λ读取相同的坐标。正确放置耳杆是这种手术中最困难的方面，这只能通过实践来学习。如果颅骨不水平，应在诱发CHI之前调整头部。 未能调整头部位置会导致颅骨骨折。要评估头骨是否扁平，应该从尖端周围的各个角度查看头骨?...

创伤性脑损伤 （TBI） 是由大脑上的外力引起的，通常与跌倒、运动损伤、身体暴力或交通事故有关。2014 年，美国疾病控制和预防中心确定，有 253 万美国人前往急诊室寻求 TBI 相关事故的医疗帮助1.由于轻度 TBI （mTBI） 占 TBI 病例的大多数，在过去的几十年中，已经采用了多种 mTBI 模型，包括体重下降、活塞驱动的闭合性头部损伤和控制皮质冲击、旋转损伤、轻度液体叩诊损伤和爆炸损伤模型2，3。mTBI模型的异质性有助于解决与人类中观察到的mTBI相关的不同特征，并帮助评估与脑损伤相关的细胞和分子机制。
在常用的闭合性头部损伤模型中，最早也是最广泛使用的模型之一是重量下降法，其中物体从特定高度掉落到动物的头部（麻醉或清醒）2，4。在减重法中，损伤的严重程度取决于几个参数，包括是否进行开颅术，头部固定或自由，以及坠落物体的距离和重量2，4。该模型的一个缺点是损伤严重程度的高度可变性和与呼吸抑制相关的高死亡率5，6。一种常见的替代方案是使用气动或电磁装置进行冲击，这可以直接在暴露的硬脑膜（受控皮质冲击:CCI）或闭合颅骨（闭合性头部损伤:CHI）上进行。活塞驱动损伤的优势之一是其高可重复性和低死亡率。然而，CCI 需要开颅术7，8，而开颅术本身会诱发炎症9。相反，在CHI模型中，不需要开颅手术。如前所述，每个模型都有局限性。本文中描述的CHI模型的局限性之一是使用立体定位框架进行手术，并且动物的头部是固定的。虽然完全头部固定可确保可重复性，但它不考虑撞击后可能导致与mTBI相关的损伤的运动。
该协议描述了在鼠标10中用市售电磁撞击器装置10执行CHI撞击的基本方法。该协议详细说明了实现高度可重复损伤所涉及的确切参数。特别是，研究者可以精确控制参数（损伤深度、停留时间和撞击速度），以精确定义损伤严重程度。如前所述，该CHI模型产生的损伤导致双侧病理，包括弥漫性和微观（即，神经胶质细胞的慢性激活，轴突和血管损伤）和行为表型11，12，13，14，15。此外，所描述的模型被认为是温和的，因为它即使在受伤后 1 年也不会诱导基于 MRI 的结构性脑损伤或病理学上的大体病变16，17。

<table><tbody><tr><td>9 mm Autoclip Applier</td><td>Braintree scientific</td><td>ACS- APL</td><td>Surgery</td></tr><tr><td>9 mm Autoclip Remover</td><td>Braintree scientific</td><td>ACS- RMV</td><td>Surgery</td></tr><tr><td>9 mm Autoclip, Case of 1,000 clips</td><td>Braintree scientific</td><td>ACS- CS</td><td>Surgery (Staples)</td></tr><tr><td>Aperio ImageScope software&amp;nbsp;</td><td>Leica BioSystems</td><td>NA&amp;nbsp;</td><td>IHC</td></tr><tr><td>BladeFLASK Blade Remover</td><td>Fisher Scientific</td><td>22-444-275</td><td>Surgery</td></tr><tr><td>Cotton tip applicator</td><td>VWR</td><td>89031-270</td><td>Surgery</td></tr><tr><td>Digitial mouse stereotaxic frame</td><td>Stoelting</td><td>51730D</td><td>Surgery</td></tr><tr><td>Dumont #7 Forceps</td><td>Roboz</td><td>RS-5047</td><td>Surgery</td></tr><tr><td>Ear bars</td><td>Stoelting</td><td>51649</td><td>Surgery</td></tr><tr><td>EthoVision XT 11.0&amp;nbsp;</td><td>Noldus Information Technology</td><td>NA</td><td>RAWM&amp;nbsp;</td></tr><tr><td>Fiber-Lite</td><td>Dolan-Jeffer Industries</td><td>UN16103-DG</td><td>Surgery</td></tr><tr><td>Fisherbrand Bulb for Small Pipets</td><td>Fisher Scientific</td><td>03-448-21</td><td>Head support apparatus</td></tr><tr><td>Gemini Avoidance System</td><td>San Diego Instruments</td><td>NA</td><td>Active avoidance</td></tr><tr><td>Heating Pad</td><td>Sunbeam&amp;nbsp;</td><td>732500000U</td><td>Surgery prep</td></tr><tr><td>HRP conjugated goat anti-rabbit IgG&amp;nbsp;</td><td>Jackson Immuno Research laboratories</td><td>111-065-144&amp;nbsp;</td><td>IHC</td></tr><tr><td>Induction chamber</td><td>Kent Scientific</td><td>VetFlo-0530XS</td><td>Surgery prep</td></tr><tr><td>Isoflurane, USP</td><td>Covetrus</td><td>NDC: 11695-6777-2</td><td>Surgery</td></tr><tr><td>Mouse gas anesthesia head holder</td><td>Stoelting</td><td>51609M</td><td>Surgery</td></tr><tr><td>Neuropactor Stereotaxic Impactor</td><td>Neuroscience Tools</td><td>n/a</td><td>Surgery: Formally distributed by Lecia as impact one</td></tr><tr><td>NexGen Mouse 500</td><td>Allentown&amp;nbsp;</td><td>n/a</td><td>Post-surgery, holding cage</td></tr><tr><td>Parafilm</td><td>Bemis</td><td>PM992</td><td>Head support apparatus</td></tr><tr><td>Peanut - Professional Hair Clipper</td><td>Whal</td><td>8655-200&amp;nbsp;</td><td>Surgery prep</td></tr><tr><td>Povidone-Iodine Solution USP, 10% (w/v), 1% (w/v) available Iodine, for laboratory</td><td>Ricca</td><td>3955-16</td><td>Surgery</td></tr><tr><td>Puralube Vet Oinment,petrolatum ophthalmic ointment, Sterile ocular lubricant</td><td>Dechra</td><td>17033-211-38</td><td>Surgery</td></tr><tr><td>Rabbit anti-GFAP&amp;nbsp;</td><td>Dako</td><td>Z0334</td><td>IHC</td></tr><tr><td>Rabbit anti-IBA1&amp;nbsp;</td><td>Wako</td><td>019-19741</td><td>IHC</td></tr><tr><td>8-arm Radial Arm Water Maze</td><td>MazeEngineers</td><td>n/a</td><td>RAWM&amp;nbsp;</td></tr><tr><td>Scale</td><td>OHAUS CS series</td><td>BAL-101</td><td>Surgery prep</td></tr><tr><td>Scalpel Handle #7 Solid 6.25&quot;&amp;nbsp;</td><td>Roboz</td><td>RS-9847</td><td>Surgery</td></tr><tr><td>Sterile Alcohol Prep Pads (isopropyl alcohol 70% v/v)</td><td>Fisher Brand</td><td>22-363-750</td><td>Surgery prep</td></tr><tr><td>SumnoSuite low-flow anesthesia system</td><td>Kent Scientific</td><td>SS-01</td><td>Surgery</td></tr><tr><td>10 mL syringe Luer-Lok Tip</td><td>BD Bard-Parker</td><td>302995</td><td>Head support apparatus</td></tr><tr><td>Timers</td><td>Fisher Scientific</td><td>6KED8</td><td>Surgery</td></tr><tr><td>Topical anesthetic cream</td><td>L.M.X 4</td><td>NDC 0496-0882-15</td><td>Surgery prep</td></tr><tr><td>Triple antibiotic ointment</td><td>Major</td><td>NDC 0904-0734-31</td><td>Post-surgery</td></tr><tr><td>Tubing</td><td>MasterFlex</td><td>96410-16</td><td>Head support apparatus</td></tr><tr><td>Vaporizer Single Channel Anesthesia System</td><td>Kent Scientific</td><td>VetFlo-1210S</td><td>Surgery prep</td></tr></tbody></table>

electromagnetic controlled closed-head model of mild traumatic brain injury in mice

需要高度可重复的创伤性脑损伤（TBI）动物模型，具有明确的病理，用于测试治疗干预和了解TBI如何改变大脑功能的机制。TBI的多种动物模型的可用性对于模拟人类中TBI的不同方面和严重程度是必要的。本手稿描述了使用中线闭合性头部损伤 （CHI） 开发轻度 TBI 小鼠模型。该模型被认为是轻微的，因为它不会产生基于神经影像学或严重神经元丢失的结构性脑损伤。然而，一次撞击会产生足够的病理，以至于认知障碍在受伤后至少 1 个月是可测量的。论文中定义了使用立体定向引导的电磁撞击器在小鼠中诱导CHI的分步方案。轻度中线CHI模型的优点包括损伤引起的变化的可重复性和低死亡率。该模型在损伤后长达 1 年内对神经影像学、神经化学、神经病理学和行为变化进行了时间表征。该模型与使用相同的撞击装置的受控皮质撞击的开放颅骨模型相辅相成。因此，实验室可以使用相同的撞击器模拟轻度弥漫性 TBI 和局灶性中度至重度 TBI。

这种立体定位电磁冲击器装置用途广泛。它用于开放性颅骨控制皮质冲击 （CCI） 或闭合性头部损伤 （CHI） 手术。此外，可以通过改变损伤参数（如冲击速度、停留时间、撞击深度、撞击器尖端和损伤目标）来调节损伤严重程度。本文描述了使用5.0毫米钢头冲击器的CHI手术。这种损伤被认为是轻微的，因为没有结构性脑损伤。成年小鼠的死亡率低于0.9%11，14，<sup class="xr...

Watch this Scientific Journal Video about Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice at JoVE.com

Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

该协议描述了小鼠模型中的轻度创伤性脑损伤。特别是，充分解释了诱导轻度中线闭合性头部损伤的分步方案和动物模型的特征。

小鼠轻度创伤性脑损伤的电磁控制闭头模型

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and ...

electromagnetic-controlled-closed-head-model-mild-traumatic-brain

Research

JoVE Journal

Neuroscience

2.0K 次观看.  University of Kentucky. 该协议描述了小鼠模型中的轻度创伤性脑损伤。特别是，充分解释了诱导轻度中线闭合性头部损伤的分步方案和动物模型的特征。

视频: Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

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 ...

科研

医学

A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury

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 neurological deficits and even amnesia. Athletes participating in contact sports are at high risk of exposure to large number of mTBIs. In terms of the level of injury in a sporting athlete, a mTBI is defined as a mild injury that does not cause gross pathological changes, but does cause short-term neurological deficits that are spontaneously resolved. Despite previous attempts to model mTBI in mice and rats, many have reported gross adverse effects including skull fractures, intracerebral bleeding, axonal injury and neuronal cell death. Herein, we describe our highly reproducible animal model of mTBI that reproduces clinically relevant symptoms. This model uses a custom made pneumatic impactor device to deliver a closed-head trauma. This impact is made under precise velocity and deformation parameters, creating a reliable and reproducible model to examine the mechanisms that contribute to effects of single or repetitive concussive mTBI.

Athletes absorb several hundred mild traumatic brain injuries (mTBI)/concussions every year; however, the consequence of these on the brain is poorly understood. Therefore, an animal model of single and repetitive mTBI that consistently replicates clinically relevant symptoms provides the means to advance the study of mTBI and concussion.

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 ...

Murine Model of Controlled Cortical Impact for the Induction of Traumatic Brain Injury

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the United States. In fact, TBI is a contributing factor to over a third of all injury-related mortality. Nonetheless, the cellular and molecular mechanisms underlying the pathophysiology of TBI are poorly understood. Thus, preclinical models of TBI capable of replicating the injury mechanisms pertinent to TBI in human patients are a critical research need. The controlled cortical impact (CCI) model of TBI utilizes a mechanical device to directly impact the exposed cortex. While no model can full recapitulate the disparate injury patterns and heterogeneous nature of TBI in human patients, CCI is capable of inducing a wide range of clinically applicable TBI. Furthermore, CCI is easily standardized allowing investigators to compare results across experiments as well as across investigative groups. The following protocol is a detailed description of applying a severe CCI with a commercially available impacting device in a murine model of TBI.

Here we describe a protocol for the induction of murine traumatic brain injury via an open-head controlled cortical impact.

创伤性脑损伤诱导受控皮质冲击的母模型

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the ...

神经科学

Assessing Changes in Synaptic Plasticity Using an Awake Closed-Head Injury Model of Mild Traumatic Brain Injury

Mild traumatic brain injuries (mTBIs) are a prevalent health issue in North America. There is increasing pressure to utilize ecologically valid models of closed-head mTBI in the preclinical setting to increase translatability to the clinical population. The awake closed-headed injury (ACHI) model uses a modified controlled cortical impactor to deliver closed-headed injury, inducing clinically relevant behavioral deficits without the need for a craniotomy or the use of an anesthetic.
This technique does not normally induce fatalities, skull fractures, or brain bleeds, and is more consistent with being a mild injury. Indeed, the mild nature of the ACHI procedure makes it ideal for studies investigating repetitive mTBI (r-mTBI). Growing evidence indicates that r-mTBI can result in a cumulative injury that produces behavioral symptoms, neuropathological changes, and neurodegeneration. r-mTBI is common in youths playing sports, and these injuries occur during a period of robust synaptic reorganization and myelination, making the younger population particularly vulnerable to the long-term influences of r-mTBI.
Further, r-mTBI occurs in cases of intimate partner violence, a condition for which there are few objective screening measures. In these experiments, synaptic function was assessed in the hippocampus in juvenile rats that had experienced r-mTBI using the ACHI model. Following the injuries, a tissue slicer was utilized to make hippocampal slices to evaluate bidirectional synaptic plasticity in the hippocampus at either 1 or 7 days following the r-mTBI. Overall, the ACHI model provides researchers with an ecologically valid model to study changes in synaptic plasticity following mTBI and r-mTBI.

Here, it is demonstrated how an awake closed-head injury model can be used for examining the effects of repeated mild traumatic brain injury (r-mTBI) on synaptic plasticity in the hippocampus. The model replicates important features of r-mTBI in patients and is used in conjunction with in vitro electrophysiology.

使用轻度创伤性脑损伤的清醒闭合性头部损伤模型评估突触可塑性的变化

Mild traumatic brain injuries (mTBIs) are a prevalent health issue in North America. There is increasing pressure to utilize ecologically valid models of ...

在 mTBI 大鼠模型中验证损伤反应和行为评估

Rat Model of Closed-Head Mild Traumatic Injury and its Validation

Animal models are crucial for advancing our understanding of mild traumatic brain injury (mTBI) and guiding clinical research. To achieve meaningful insights, developing a stable and reproducible animal model is essential. In this study, we report a detailed description of a closed-head mTBI model and a representative validation method using Sprague-Dawley rats to verify the modeling effect. The model involves dropping a 550 g mass weight from a height of 100 cm directly onto the head of a rat on a destructible surface, followed by a 180-degree turn. To assess the injury, rats underwent a series of neurobehavioral assessments 10 min post-injury, including time of loss of consciousness, first seeking-behavior time, escape ability, and beam balance ability test. During the acute and subacute stages following the injury, behavioral tests were conducted to assess motor coordination ability (Beam task), anxiety (Open Field test), and learning and memory abilities (Morris Water Maze test). The closed-head mTBI model produced a consistent injury response with minimal mortality and replicated real-life situations. The validation method effectively verified the model development and ensured the stability and consistency of the model.

作者聚焦:推进创伤性脑损伤研究 - 用于准确复制和快速评估的闭头模型 

Here, we present a closed-head mild traumatic brain injury (mTBI) rat model and its validation exhibiting remarkable similarity to human mTBI concerning behavioral manifestations during the acute and subacute stages.

闭合头轻度创伤大鼠模型及其验证

Animal models are crucial for advancing our understanding of mild traumatic brain injury (mTBI) and guiding clinical research. To achieve meaningful ...

通过闭合头部损伤法在小鼠中诱导 rmTBI

Modified Mouse Model of Repetitive Mild Traumatic Brain Injury Incorporating Thinned-Skull Window and Fluid Percussion

Mild traumatic brain injury is a clinically highly heterogeneous neurological disorder. Highly reproducible traumatic brain injury (TBI) animal models with well-defined pathologies are urgently needed for studying the mechanisms of neuropathology after mild TBI and testing therapeutics. Replicating the entire sequelae of TBI in animal models has proven to be a challenge. Therefore, the availability of multiple animal models of TBI is necessary to account for the diverse aspects and severities seen in TBI patients. CHI is one of the most common methods for fabricating rodent models of rmTBI. However, this method is susceptible to many factors, including the impact method used, the thickness and shape of the skull bone, animal apnea, and the type of head support and immobilization utilized. The aim of this protocol is to demonstrate a combination of the thinned-skull window and fluid percussion injury (FPI) methods to produce a precise mouse model of CHI-associated rmTBI. The primary objective of this protocol is to minimize factors that could impact the accuracy and consistency of CHI and FPI modeling, including skull bone thickness, shape, and head support. By utilizing a thinned-skull window method, potential inflammation due to craniotomy and FPI is minimized, resulting in an improved mouse model that replicates the clinical features observed in patients with mild TBI. Results from behavior and histological analysis using hematoxylin and eosin (HE) staining suggest that rmTBI can lead to a cumulative injury that produces changes in both behavior and gross morphology of the brain. Overall, the modified CHI-associated rmTBI presents a useful tool for researchers to explore the underlying mechanisms that contribute to focal and diffuse pathophysiological changes in rmTBI.

作者聚焦：优化啮齿动物模型以研究反复轻度创伤性脑损伤的机制和康复

This protocol presents a modified mouse model of repetitive mild traumatic brain injury (rmTBI) induced via a closed-head injury (CHI) method. The approach features a thinned-skull window and fluid percussion to reduce the inflammation commonly caused by meninges exposure, along with improved reproducibility and accuracy in modeling rmTBI in rodents.

结合颅骨窗变薄和液体叩击的重复性轻度创伤性脑损伤改良小鼠模型

Mild traumatic brain injury is a clinically highly heterogeneous neurological disorder. Highly reproducible traumatic brain injury (TBI) animal models ...

小鼠轻度创伤性脑损伤的电磁控制闭头模型

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闭合头轻度创伤大鼠模型及其验证

闭合头轻度创伤大鼠模型及其验证

闭合头轻度创伤大鼠模型及其验证

结合颅骨窗变薄和液体叩击的重复性轻度创伤性脑损伤改良小鼠模型

结合颅骨窗变薄和液体叩击的重复性轻度创伤性脑损伤改良小鼠模型

结合颅骨窗变薄和液体叩击的重复性轻度创伤性脑损伤改良小鼠模型