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>

<ol>
	<li>Capizzi, A., Woo, J., Verduzco-Gutierrez, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Traumatic+brain+injury:+An+overview+of+epidemiology,+pathophysiology,+and+medical+management.">Traumatic brain injury: An overview of epidemiology, pathophysiology, and medical management.</a> The Medical Clinics of North America. 104 (2), 213-238 (2020).</li><li>Bodnar, C. N., Roberts, K. N., Higgins, E. K., Bachstetter, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+review+of+closed+head+injury+models+of+mild+traumatic+brain+injury+in+mice+and+rats.">A systematic review of closed head injury models of mild traumatic brain injury in mice and rats.</a> Journal of Neurotrauma. 36 (11), 1683-1706 (2019).</li><li>Shultz, S. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+potential+for+animal+models+to+provide+insight+into+mild+traumatic+brain+injury:+Translational+challenges+and+strategies.">The potential for animal models to provide insight into mild traumatic brain injury: Translational challenges and strategies.</a> Neuroscience and Biobehavioral Reviews. 76, 396-414 (2017).</li><li>Xiong, Y., Mahmood, A., Chopp, M. <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+traumatic+brain+injury).">Animal models of traumatic brain injury).</a> Nature Reviews Neuroscience. 14 (2), 128-142 (2013).</li><li>Albert-Weissenberger, C., Varrallyay, C., Raslan, F., Kleinschnitz, C., Siren, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+experimental+protocol+for+mimicking+pathomechanisms+of+traumatic+brain+injury+in+mice.">An experimental protocol for mimicking pathomechanisms of traumatic brain injury in mice.</a> Experimental and Translational Stroke Medicine. 4, 1 (2012).</li><li>Chen, Y., Constantini, S., Trembovler, V., Weinstock, M., Shohami, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+experimental+model+of+closed+head+injury+in+mice:+pathophysiology,+histopathology,+and+cognitive+deficits.">An experimental model of closed head injury in mice: pathophysiology, histopathology, and cognitive deficits.</a> Journal of Neurotrauma. 13 (10), 557-568 (1996).</li><li>Dixon, C. E., Clifton, G. L., Lighthall, J. W., Yaghmai, A. A., Hayes, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+controlled+cortical+impact+model+of+traumatic+brain+injury+in+the+rat.">A controlled cortical impact model of traumatic brain injury in the rat.</a> Journal of Neuroscience Methods. 39 (3), 253-262 (1991).</li><li>Schwulst, S. J., Islam, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+model+of+controlled+cortical+impact+for+the+induction+of+traumatic+brain+injury.">Murine model of controlled cortical impact for the induction of traumatic brain injury.</a> Journal of Visualized Experiments. (150), e60027 (2019).</li><li>Cole, J. 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=Craniotomy:+True+sham+for+traumatic+brain+injury,+or+a+sham+of+a+sham.">Craniotomy: True sham for traumatic brain injury, or a sham of a sham.</a> Journal of Neurotrauma. 28 (3), 359-369 (2011).</li><li>Brody, D. 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=Electromagnetic+controlled+cortical+impact+device+for+precise,+graded+experimental+traumatic+brain+injury.">Electromagnetic controlled cortical impact device for precise, graded experimental traumatic brain injury.</a> Journal of Neurotrauma. 24 (4), 657-673 (2007).</li><li>Webster, S. J., Van Eldik, L. J., Watterson, D. M., Bachstetter, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Closed+head+injury+in+an+age-related+Alzheimer+mouse+model+leads+to+an+altered+neuroinflammatory+response+and+persistent+cognitive+impairment.">Closed head injury in an age-related Alzheimer mouse model leads to an altered neuroinflammatory response and persistent cognitive impairment.</a> The Journal of Neuroscience. 35 (16), 6554-6569 (2015).</li><li>Macheda, T., Roberts, K. N., Morganti, J. M., Braun, D. J., Bachstetter, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+and+validation+of+a+modified+radial-arm+water+maze+protocol+using+a+murine+model+of+mild+closed+head+traumatic+brain+injury.">Optimization and validation of a modified radial-arm water maze protocol using a murine model of mild closed head traumatic brain injury.</a> PLoS One. 15 (8), 0232862 (2020).</li><li>Macheda, T., Snider, H. C., Watson, J. B., Roberts, K. N., Bachstetter, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+active+avoidance+behavioral+paradigm+for+use+in+a+mild+closed+head+model+of+traumatic+brain+injury+in+mice.">An active avoidance behavioral paradigm for use in a mild closed head model of traumatic brain injury in mice.</a> Journal of Neuroscience Methods. 343, 108831 (2020).</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=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. 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=The+effects+of+mild+closed+head+injuries+on+tauopathy+and+cognitive+deficits+in+rodents:+Primary+results+in+wild+type+and+rTg4510+mice,+and+a+systematic+review.">The effects of mild closed head injuries on tauopathy and cognitive deficits in rodents: Primary results in wild type and rTg4510 mice, and a systematic review.</a> Experimental Neurology. 326, 113180 (2020).</li><li>Lyons, D. 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=A+mild+traumatic+brain+injury+in+mice+produces+lasting+deficits+in+brain+metabolism.">A mild traumatic brain injury in mice produces lasting deficits in brain metabolism.</a> Journal of Neurotrauma. 35 (20), 2435-2447 (2018).</li><li>Yanckello, L. 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 border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<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&#160;</td>
			<td>Leica BioSystems</td>
			<td>NA&#160;</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&#160;</td>
			<td>Noldus Information Technology</td>
			<td>NA</td>
			<td>RAWM&#160;</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&#160;</td>
			<td>732500000U</td>
			<td>Surgery prep</td>
		</tr>
		<tr>
			<td>HRP conjugated goat anti-rabbit IgG&#160;</td>
			<td>Jackson Immuno Research laboratories</td>
			<td>111-065-144&#160;</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&#160;</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&#160;</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&#160;</td>
			<td>Dako</td>
			<td>Z0334</td>
			<td>IHC</td>
		</tr>
		<tr>
			<td>Rabbit anti-IBA1&#160;</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&#160;</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&#34;&#160;</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

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and mortality. Based on the nature of primary injury following TBI, complex and heterogeneous secondary consequences result, which are followed by regenerative processes 1,2. Primary injury can be induced by a direct contusion to the brain from skull fracture or from shearing and stretching of tissue causing displacement of brain due to movement 3,4. The resulting hematomas and lacerations cause a vascular response 3,5, and the morphological and functional damage of the white matter leads to diffuse axonal injury 6-8. Additional secondary changes commonly seen in the brain are edema and increased intracranial pressure 9. Following TBI there are microscopic alterations in biochemical and physiological pathways involving the release of excitotoxic neurotransmitters, immune mediators and oxygen radicals 10-12, which ultimately result in long-term neurological disabilities 13,14. Thus choosing appropriate animal models of TBI that present similar cellular and molecular events in human and rodent TBI is critical for studying the mechanisms underlying injury and repair.
Various experimental models of TBI have been developed to reproduce aspects of TBI observed in humans, among them three specific models are widely adapted for rodents: fluid percussion, cortical impact and weight drop/impact acceleration 1. The fluid percussion device produces an injury through a craniectomy by applying a brief fluid pressure pulse on to the intact dura. The pulse is created by a pendulum striking the piston of a reservoir of fluid. The percussion produces brief displacement and deformation of neural tissue 1,15. Conversely, cortical impact injury delivers mechanical energy to the intact dura via a rigid impactor under pneumatic pressure 16,17. The weight drop/impact model is characterized by the fall of a rod with a specific mass on the closed skull 18. Among the TBI models, LFP is the most established and commonly used model to evaluate mixed focal and diffuse brain injury 19. It is reproducible and is standardized to allow for the manipulation of injury parameters. LFP recapitulates injuries observed in humans, thus rendering it clinically relevant, and allows for exploration of novel therapeutics for clinical translation 20.
We describe the detailed protocol to perform LFP procedure in mice. The injury inflicted is mild to moderate, with brain regions such as cortex, hippocampus and corpus callosum being most vulnerable. Hippocampal and motor learning tasks are explored following LFP.

Lateral fluid percussion (LFP), an established model of traumatic brain injury in mice, is demonstrated. LFP fulfills three major criteria for animal models: validity, reliability and clinical relevance. The procedure, consisting of surgical craniotomy, fixation of hub followed by induction of injury, resulting in focal and diffuse injuries, is described.

侧向液压冲击:创伤性脑损伤小鼠模型

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and ...

科研

神经科学

Investigations on Alterations of Hippocampal Circuit Function Following Mild Traumatic Brain Injury

Traumatic Brain Injury (TBI) afflicts more than 1.7 million people in the United States each year and even mild TBI can lead to persistent neurological impairments 1. Two pervasive and disabling symptoms experienced by TBI survivors, memory deficits and a reduction in seizure threshold, are thought to be mediated by TBI-induced hippocampal dysfunction 2,3. In order to demonstrate how altered hippocampal circuit function adversely affects behavior after TBI in mice, we employ lateral fluid percussion injury, a commonly used animal model of TBI that recreates many features of human TBI including neuronal cell loss, gliosis, and ionic perturbation 4-6.
	Here we demonstrate a combinatorial method for investigating TBI-induced hippocampal dysfunction. Our approach incorporates multiple ex vivo physiological techniques together with animal behavior and biochemical analysis, in order to analyze post-TBI changes in the hippocampus. We begin with the experimental injury paradigm along with behavioral analysis to assess cognitive disability following TBI. Next, we feature three distinct ex vivo recording techniques: extracellular field potential recording, visualized whole-cell patch-clamping, and voltage sensitive dye recording. Finally, we demonstrate a method for regionally dissecting subregions of the hippocampus that can be useful for detailed analysis of neurochemical and metabolic alterations post-TBI.
	These methods have been used to examine the alterations in hippocampal circuitry following TBI and to probe the opposing changes in network circuit function that occur in the dentate gyrus and CA1 subregions of the hippocampus (see Figure 1). The ability to analyze the post-TBI changes in each subregion is essential to understanding the underlying mechanisms contributing to TBI-induced behavioral and cognitive deficits. 
	The multi-faceted system outlined here allows investigators to push past characterization of phenomenology induced by a disease state (in this case TBI) and determine the mechanisms responsible for the observed pathology associated with TBI.

A multi-faceted approach to investigating functional changes to hippocampal circuitry is explained. Electrophysiological techniques are described along with the injury protocol, behavioral testing and regional dissection method. The combination of these techniques can be applied in similar fashion for other brain regions and scientific questions.

调查轻度创伤性脑损伤海马电路功能的改变

Traumatic Brain Injury (TBI) afflicts more than 1.7 million people in the United States each year and even mild TBI can lead to persistent neurological ...

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Traumatic brain injury (TBI) affects millions of people each year, causing impairment of physical, cognitive, and behavioral functions and death. Studies using Drosophila have contributed important breakthroughs in understanding neurological processes. Thus, with the goal of understanding the cellular and molecular basis of TBI pathologies in humans, we developed the High Impact Trauma (HIT) device to inflict closed head TBI in flies. Flies subjected to the HIT device display phenotypes consistent with human TBI such as temporary incapacitation and progressive neurodegeneration. The HIT device uses a spring-based mechanism to propel flies against the wall of a vial, causing mechanical damage to the fly brain. The device is inexpensive and easy to construct, its operation is simple and rapid, and it produces reproducible results. Consequently, the HIT device can be combined with existing experimental tools and techniques for flies to address fundamental questions about TBI that can lead to the development of diagnostics and treatments for TBI. In particular, the HIT device can be used to perform large-scale genetic screens to understand the genetic basis of TBI pathologies.

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Here we describe a method to inflict closed head traumatic brain injury (TBI) in Drosophila. This method provides a gateway to investigate the cellular and molecular mechanisms that underlie TBI pathologies using the vast array of experimental tools and techniques available for flies.

方法对敌人造成闭合性颅脑创伤性脑损伤<em&gt;果蝇</em

Traumatic brain injury (TBI) affects millions of people each year, causing impairment of physical, cognitive, and behavioral functions and death. Studies ...

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in adult mammalian animals remains elusive. In this study, we describe an experimental procedure for measuring calcium dynamics through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy. This method enables recordings and quantifications of neural activity in bilateral mouse brain regions one at a time in the same experiment for a prolonged period in vivo. Key aspects of this method, which can be completed within an hour, include minimally invasive surgery procedures for creating dual optical windows, and the use of 2P imaging. Although we only demonstrate the technique in the S1 area, the method can be applied to other regions of the living brain facilitating the elucidation of structural and functional complexities of brain neural networks.

Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice

We describe an experimental procedure for measuring neuronal activity through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy in vivo.

胸腺1-gmcmp6 转基因小鼠在原代体血源皮质中的成像神经活性

The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in ...

A Novel In Vitro Model of Blast Traumatic Brain Injury

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the detonation of explosive devices, however, the mechanisms that underlie the brain damage resulting from blast overpressure exposure are not entirely understood and are believed to be unique to this type of brain injury. Preclinical models are crucial tools that contribute to better understand blast-induced brain injury. A novel in vitro blast TBI model was developed using an open-ended shock tube to simulate real-life open-field blast waves modelled by the Friedlander waveform. C57BL/6N mouse organotypic hippocampal slice cultures were exposed to single shock waves and the development of injury was characterized up to 72 h using propidium iodide, a well-established fluorescent marker of cell damage that only penetrates cells with compromised cellular membranes. Propidium iodide fluorescence was significantly higher in the slices exposed to a blast wave when compared with sham slices throughout the duration of the protocol. The brain tissue injury is very reproducible and proportional to the peak overpressure of the shock wave applied.

A Novel In Vitro Model of Blast Traumatic Brain Injury

This paper describes a novel model of primary blast traumatic brain injury. A compressed-air driven shock tube is used to expose in vitro mouse hippocampal slice cultures to a single shock wave. This is a simple and rapid protocol generating a reproducible brain tissue injury with a high throughput.

一种新的脑外伤体损伤模型

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the ...

Effects of Blast-induced Neurotrauma on Pressurized Rodent Middle Cerebral Arteries

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral vascular effects. Impact (i.e., non-blast) traumatic brain injury (TBI) is known to decrease pressure autoregulation in the cerebral vasculature in both humans and experimental animals. The hypothesis that blast-induced traumatic brain injury (bTBI), like impact TBI, results in impaired cerebral vascular reactivity was tested by measuring myogenic dilatory responses to reduced intravascular pressure in rodent middle cerebral arterial (MCA) segments from rats subjected to mild bTBI using an Advanced Blast Simulator (ABS) shock tube. Adult, male Sprague-Dawley rats were anesthetized, intubated, ventilated and prepared for Sham bTBI (identical manipulation and anesthesia except for blast injury) or mild bTBI. Rats were randomly assigned to receive Sham bTBI or mild bTBI followed by sacrifice 30 or 60 min post-injury. Immediately after bTBI, righting reflex (RR) suppression times were assessed, euthanasia at the time points post-injury was completed, the brain was harvested and the individual MCA segments were collected, mounted and pressurized. As the intraluminal pressure perfused through the arterial segments was reduced in 20 mmHg increments from 100 to 20 mmHg, MCA diameters were measured and recorded. With decreasing intraluminal pressure, MCA diameters steadily increased significantly above baseline in the Sham bTBI groups while MCA dilator responses were significantly reduced (p &#60; 0.05) in both bTBI groups as evidenced by the impaired, smaller MCA diameters recorded for the bTBI groups. In addition, RR suppression in the bTBI groups was significantly (p &#60; 0.05) higher than in the Sham bTBI groups. MCA&#39;s collected from the Sham bTBI groups exhibited typical vasodilatory properties to decreases in intraluminal pressure while MCA&#39;s collected following bTBI exhibited significantly impaired myogenic vasodilatory responses to reduced pressure that persisted for at least 60 min after bTBI.

Here, we present a protocol to describe methods for ex vivo vascular reactivity determination following a primary blast traumatic brain injury (bTBI) using isolated, pressurized, rodent middle cerebral arterial (MCA) segments. bTBI induction is accomplished using a shock tube, also known as an Advanced Blast Simulator (ABS) device.

爆炸性神经损伤对压生啮鼠中脑动脉的影响

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral ...

Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also demonstrate that traumatic brain injuries cause changes to the gut microbiota. However, mechanisms underlying the bidirectional regulation of the brain-gut axis remain unknown. Currently, few models exist for studying the changes in gut microbiota after traumatic brain injury. Therefore, the presented study combines protocols for inducing traumatic brain injury using a lateral fluid percussion device and analysis of caecum samples following injury for investigating alterations in the gut microbiome. Alterations of the gut microbiota composition after traumatic brain injury are determined using 16S-rDNA sequencing. This protocol provides an effective method for studying the relationships between enteric microorganisms and traumatic brain injury.

Presented here is a protocol to induce diffuse traumatic brain injury using a lateral fluid percussion device followed by the collection of the caecum content for gut microbiome analysis.

研究小鼠创伤性脑损伤后凯库姆微生物群的改动

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also ...

Advanced Diffusion Imaging in The Hippocampus of Rats with Mild Traumatic Brain Injury

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous variability and heterogeneity (age, gender, type of trauma, other possible pathologies, etc.), animal models play a key role in unraveling factors that are limitations in clinical research. They provide a standardized and controlled setting to investigate the biological mechanisms of injury and repair following TBI. However, not all animal models mimic the diffuse and subtle nature of mTBI effectively. For example, the commonly used controlled cortical impact (CCI) and lateral fluid percussion injury (LFPI) models make use of a craniotomy to expose the brain and induce widespread focal trauma, which are not commonly seen in mTBI. Therefore, these experimental models are not valid to mimic mTBI. Thus, an appropriate model should be used to investigate mTBI. The Marmarou weight drop model for rats induces similar microstructural alterations and cognitive impairments as seen in patients who sustain mild trauma; therefore, this model was selected for this protocol. Conventional computed tomography and magnetic resonance imaging (MRI) scans commonly show no damage following a mild injury, because mTBI induces often only subtle and diffuse injuries. With diffusion weighted MRI, it is possible to investigate microstructural properties of brain tissue, which can provide more insight into the microscopic alterations following mild trauma. Therefore, the goal of this study is to obtain quantitative information of a selected region-of-interest (i.e., hippocampus) to follow up disease progression after obtaining a mild and diffuse brain injury.

The overall goal of this procedure is to obtain quantitative microstructural information of the hippocampus in a rat with mild traumatic brain injury. This is done using an advanced diffusion-weighted magnetic resonance imaging protocol and region-of-interest based analysis of parametric diffusion maps.

轻度创伤性脑损伤大鼠海马学的高级扩散成像

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous ...

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 ...

Low-intensity Blast Wave Model for Preclinical Assessment of Closed-head Mild Traumatic Brain Injury in Rodents

Traumatic brain injury (TBI) is a large-scale public health problem. Mild TBI is the most prevalent form of neurotrauma and accounts for a large number of medical visits in the United States. There are currently no FDA-approved treatments available for TBI. The increased incidence of military-related, blast-induced TBI further accentuates the urgent need for effective TBI treatments. Therefore, new preclinical TBI animal models that recapitulate aspects of human blast-related TBI will greatly advance the research efforts into the neurobiological and pathophysiological processes underlying mild to moderate TBI as well as the development of novel therapeutic strategies for TBI.
Here we present a reliable, reproducible model for the investigation of the molecular, cellular, and behavioral effects of mild to moderate blast-induced TBI. We describe a step-by-step protocol for closed-head, blast-induced mild TBI in rodents using a bench-top setup consisting of a gas-driven shock tube equipped with piezoelectric pressure sensors to ensure consistent test conditions. The benefits of the setup that we have established are its relative low-cost, ease of installation, ease of use and high-throughput capacity. Further advantages of this non-invasive TBI model include the scalability of the blast peak overpressure and the generation of controlled reproducible outcomes. The reproducibility and relevance of this TBI model has been evaluated in a number of downstream applications, including neurobiological, neuropathological, neurophysiological and behavioral analyses, supporting the use of this model for the characterization of processes underlying the etiology of mild to moderate TBI.

We present here a protocol of a blast wave model for rodents to investigate neurobiological and pathophysiological effects of mild to moderate traumatic brain injury. We established a gas-driven, bench-top setup equipped with pressure sensors allowing for reliable and reproducible generation of blast-induced mild to moderate traumatic brain injury.

低强度冲击波模型用于啮齿动物闭头轻度创伤性脑损伤的临床前评估

Traumatic brain injury (TBI) is a large-scale public health problem. Mild TBI is the most prevalent form of neurotrauma and accounts for a large number of ...