Name: inducing post-traumatic epilepsy in a mouse model of repetitive diffuse traumatic brain injury
Uploaded: 2020-02-10T12:30:00
Description: Watch this Scientific Journal Video about Inducing Post-Traumatic Epilepsy in a Mouse Model of Repetitive Diffuse Traumatic Brain Injury Video at JoVE.com

这项工作得到了R01 NS105807/NS/NINDS NIH HHS/美国和CURE的支持，其依据是美国陆军医学研究和物资司令部（国防部）通过"心理健康和创伤性脑损伤研究计划"获得的第1号奖励。W81XWH-15-2-0069。伊万·祖伊德霍克对手稿进行校对，非常感谢他。

本议定书中描述的所有动物程序均按照弗吉尼亚理工大学机构动物护理和使用委员会（IACUC）执行，并符合美国国家卫生研究院的《实验室动物护理和使用指南》。.
1. 动物处理协议
注：本协议旨在将从供应商订购的动物在抵达后安排到设施，并规定它们由实验者处理。这通过减少压力和焦虑来改善动物福祉，并简化某些需要处理动物的程序，包括诱导TBI?...

<ol>
	<li>Christensen, 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=Long-term+risk+of+epilepsy+after+traumatic+brain+injury+in+children+and+young+adults:+a+population-based+cohort+study.">Long-term risk of epilepsy after traumatic brain injury in children and young adults: a population-based cohort study.</a> Lancet. 373 (9669), 1105-1110 (2009).</li><li>Lowenstein, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epilepsy+after+head+injury:+an+overview.">Epilepsy after head injury: an overview.</a> Epilepsia. 50, 4-9 (2009).</li><li>Ferguson, P. 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=A+population-based+study+of+risk+of+epilepsy+after+hospitalization+for+traumatic+brain+injury.">A population-based study of risk of epilepsy after hospitalization for traumatic brain injury.</a> Epilepsia. 51 (5), 891-898 (2010).</li><li>Abou-Abbass, H., 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=Epidemiology+and+clinical+characteristics+of+traumatic+brain+injury+in+Lebanon:+A+systematic+review.">Epidemiology and clinical characteristics of traumatic brain injury in Lebanon: A systematic review.</a> Medicine (Baltimore). 95 (47), 5342 (2016).</li><li>Management of Concussion/mTBI Working Group. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VA/DoD+Clinical+Practice+Guideline+for+Management+of+Concussion/Mild+Traumatic+Brain+Injury.">VA/DoD Clinical Practice Guideline for Management of Concussion/Mild Traumatic Brain Injury.</a> The Journal of Rehabilitation Research and Development. 46 (6), 1-68 (2009).</li><li>Piccenna, L., Shears, G., O'Brien, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+post-traumatic+epilepsy:+An+evidence+review+over+the+last+5+years+and+future+directions.">Management of post-traumatic epilepsy: An evidence review over the last 5 years and future directions.</a> Epilepsia Open. 2 (2), 123-144 (2017).</li><li>Loscher, W., Brandt, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevention+or+modification+of+epileptogenesis+after+brain+insults:+experimental+approaches+and+translational+research.">Prevention or modification of epileptogenesis after brain insults: experimental approaches and translational research.</a> Pharmacological Reviews. 62 (4), 668-700 (2010).</li><li>Ostergard, T., Sweet, J., Kusyk, D., Herring, E., Miller, J. <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+post-traumatic+epilepsy.">Animal models of post-traumatic epilepsy.</a> Journal of Neuroscience Methods. 272, 50-55 (2016).</li><li>Shandra, O., 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=Repetitive+Diffuse+Mild+Traumatic+Brain+Injury+Causes+an+Atypical+Astrocyte+Response+and+Spontaneous+Recurrent+Seizures.">Repetitive Diffuse Mild Traumatic Brain Injury Causes an Atypical Astrocyte Response and Spontaneous Recurrent Seizures.</a> Journal of Neuroscience. 39 (10), 1944-1963 (2019).</li><li>Foda, M. A., Marmarou, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+model+of+diffuse+brain+injury+in+rats.+Part+II:+Morphological+characterization.">A new model of diffuse brain injury in rats. Part II: Morphological characterization.</a> Journal of Neurosurgery. 80 (2), 301-313 (1994).</li><li>Marmarou, 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=A+new+model+of+diffuse+brain+injury+in+rats.+Part+I:+Pathophysiology+and+biomechanics.">A new model of diffuse brain injury in rats. Part I: Pathophysiology and biomechanics.</a> Journal of Neurosurgery. 80 (2), 291-300 (1994).</li><li>Paxinos, G., Keith, B. J., Franklin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Mouse Brain in Stereotaxic Coordinates. , (2007).</li><li>Shandra, O., Robel, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+and+Manipulating+Astrocyte+Function+In+Vivo+in+the+Context+of+CNS+Injury.">Imaging and Manipulating Astrocyte Function In Vivo in the Context of CNS Injury.</a> Methods in Molecular Biology. 1938, 233-246 (2019).</li><li>Pitkanen, A., Immonen, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epilepsy+related+to+traumatic+brain+injury.">Epilepsy related to traumatic brain injury.</a> Neurotherapeutics. 11 (2), 286-296 (2014).</li><li>Kharatishvili, I., Nissinen, J. P., McIntosh, T. K., Pitkanen, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+model+of+posttraumatic+epilepsy+induced+by+lateral+fluid-percussion+brain+injury+in+rats.">A model of posttraumatic epilepsy induced by lateral fluid-percussion brain injury in rats.</a> Neuroscience. 140 (2), 685-697 (2006).</li><li>Pitkanen, A., Bolkvadze, T., Immonen, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anti-epileptogenesis+in+rodent+post-traumatic+epilepsy+models.">Anti-epileptogenesis in rodent post-traumatic epilepsy models.</a> Neuroscience Letters. 497 (3), 163-171 (2011).</li><li>Gades, N. M., Danneman, P. J., Wixson, S. K., Tolley, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+magnitude+and+duration+of+the+analgesic+effect+of+morphine,+butorphanol,+and+buprenorphine+in+rats+and+mice.">The magnitude and duration of the analgesic effect of morphine, butorphanol, and buprenorphine in rats and mice.</a> Journal of the American Association for Laboratory Animal Science. 39 (2), 8-13 (2000).</li></ol>

与 CCI 和 FPI 模型诱导焦力或焦点和漫反射损伤的组合不同，本协议中描述的重复扩散 TBI 模型允许在没有焦点脑损伤的情况下诱导扩散损伤，并且不需要头皮或颅开口和相关炎症。该模型中没有颅切除术的一个附加好处是，它不仅允许植入用于慢性连续脑电图记录的电极，而且还可以创建一个薄颅骨窗口，用于在动物之前、紧接之后、之后、再重复几天、几周甚至几个月，如Sandra和Robel 2019<sup clas...

每年，TBI 影响全世界大约 6000 万人。受影响的个人患癫痫的风险较高，癫痫发作后数年可能显现出来。虽然严重的TBI与较高的癫痫风险有关，即使是轻微的TBI也会增加个人患癫痫的几率1，2，3，4。所有 TI 都可以分类为焦距、漫反射或两者的组合。扩散性脑损伤，存在于许多（如果不是全部）的TbIs中，是由于加速减速和旋转力导致不同密度的脑组织相互剪切的结果。根据定义，扩散损伤只发生在轻度/脑震荡非穿透性脑损伤中，其中在计算机断层扫描5中看不到脑损伤。
目前，在管理患有或有发生创伤后癫痫（PTE）风险的患者方面，存在两个关键问题。第一，一旦PTE出现，癫痫发作对现有的抗癫痫药物（AEDs）有抗药性6。其次，AED在预防癫痫发作方面同样无效，没有有效的替代治疗方法。为了解决这个缺陷，并找到更好的治疗目标和治疗候选，有必要在PTE6的根部探索新的细胞和分子机制。
创伤后癫痫的一个突出特征是初始创伤事件与自发、无端、复发性发作之间的潜伏期。在这个时间窗口内发生的事件是研究人员的自然焦点，因为这个时间窗口可能允许治疗和预防PTE完全。动物模型最常用于这项研究，因为它们提供了几个截然不同的好处，其中最重要的是，在如此长的时间跨度内，持续监测人类患者既不切实际又昂贵。此外，只有在动物模型中才能探索癫痫发生根源的细胞和分子机制。
与在TBI后通过不太相关的生理相关手段（如通过化学惊厥或急性、慢性或点燃引起电刺激）诱导癫痫发作的模型比较，具有自发创伤后癫痫的动物模型更可取。自发的创伤后癫痫发作模型测试TBI如何改变导致癫痫发作的健康大脑网络。在TBI后使用额外刺激的研究评估接触TBI如何降低癫痫发作阈值并影响癫痫发作的易感性。具有化学或电刺激诱导发作的动物模型的优点是测试耐火性对AED的具体机制以及现有和新型AED的功效。然而，这些数据对人类的相关性和翻译程度可能不明确7，原因如下：1） 癫痫发作机制可能与仅由TBI引起的发作机制不同;1） 癫痫发作机制可能不同于TBI单独诱发的机制。2） 并非所有这些模型导致自发性发作7;3） 由抽搐剂本身产生的病变，其传递所需的管状，或通过刺激电极放置在深度结构（例如，海马或杏仁核）已经可能导致增加的癫痫易感性，甚至海马癫痫表场电位7。此外，一些抽搐剂（即开酸）产生直接的海马病变和硬化症，这在扩散TBI后并不常见。
直到最近，只有两种创伤后癫痫的动物模型存在：受控皮质撞击（CCI，焦）或流体打击损伤（FPI，焦和扩散）8。这两种模型导致大的焦点病变，同时组织损失，出血，和胶质瘤在啮齿动物8。这些模型模仿由大焦点病变引起的创伤后癫痫。最近的一项研究表明，重复（3倍）扩散TBI足以在小鼠中发展自发性发作和癫痫，即使没有焦点病变9，增加了第三个啮齿动物PTE模型与确认的自发性复发性发作。这种新模型模仿由漫射TBI引起的细胞和分子变化，更好地代表轻度脑震荡TBI人群。在此模型中，癫痫发作前三周或以上的潜伏期以及晚期、自发性、复发性发作的出现，有助于调查创伤后癫痫发生的根本原因，测试预防方法的有效性和癫痫发作后的新治疗候选者，并有可能开发创伤后癫痫发生的生物标志物，因为大约一半的动物会发展成创伤后癫痫。
研究创伤后癫痫的动物模型的选择取决于科学问题、所调查的脑损伤类型以及将用什么工具来确定潜在的细胞和分子机制。最终，任何创伤后癫痫模型都必须证明TBI后自发性癫痫发作的出现和TBI动物的初期潜伏期，因为并非所有发生TBI的患者都会发展成癫痫。为此，该协议使用同时视频采集的脑电图（EEG）。了解数据采集硬件和方法背后的技术方面对于准确解释数据至关重要。关键硬件方面包括记录系统的类型、电极类型（螺钉或导线引线）及其材料、同步视频采集（作为 EEG 系统的一部分或第三方）以及计算机系统的属性。根据研究目标、感兴趣的脑电图事件、进一步分析方法和数据存储的可持续性，在任何类型的系统中设置适当的采集参数势在必行。最后，必须考虑电极配置（蒙太奇）的方法，因为每种方法都有优缺点，会影响数据解释。
该协议详细介绍了如何使用经过改进的Marmarou降压模型10，11诱导扩散损伤，导致小鼠自发的、无端的、复发性发作，描述了使用单极、双极或混合蒙太奇获得单通道和多通道连续和同步视频EEG的手术方法。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.10&#34; screw</td>
			<td>Pinnacle Technology Inc., KS, USA</td>
			<td>8209</td>
			<td>0.10 inch long stainless steel</td>
		</tr>
		<tr>
			<td>0.10&#34; screw</td>
			<td>Pinnacle Technology Inc., KS, USA</td>
			<td>8403</td>
			<td>0.10 inch long with pre-soldered wire lead</td>
		</tr>
		<tr>
			<td>0.12&#34; screw</td>
			<td>Pinnacle Technology Inc., KS, USA</td>
			<td>8212</td>
			<td>0.12 inch long stainless steel</td>
		</tr>
		<tr>
			<td>1EEG headmount</td>
			<td>Invitro1 (subsidiary of Plastics One), VA, USA</td>
			<td>MS333/8-A/SPC</td>
			<td>3 individually Teflon-insulated platinum iridium wire electrodes (twisted or untwisted, 0.005 inch diameter) extending below threaded plastic pedestal</td>
		</tr>
		<tr>
			<td>2EEG/1EMG headmount</td>
			<td>Pinnacle Technology Inc., KS, USA</td>
			<td>8201</td>
			<td>2EEG/1EMG channels</td>
		</tr>
		<tr>
			<td>3% hydrogen peroxide</td>
			<td></td>
			<td></td>
			<td>Pharmacy</td>
		</tr>
		<tr>
			<td>3EEG headmount</td>
			<td>Pinnacle Technology Inc., KS, USA</td>
			<td>8235-SM-C</td>
			<td>custom 6-Pin Connector for 3EEG channels</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Par Pharmaceuticals, Cos. Inc., Spring Valley, NY, USA</td>
			<td>060969</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Par Pharmaceuticals, Cos. Inc., Spring Valley, NY, USA</td>
			<td>060969</td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6 mice</td>
			<td>Harlan/Envigo Laboratories Inc</td>
			<td></td>
			<td>male, 12-16 weeks old</td>
		</tr>
		<tr>
			<td>C57BL/6 mice</td>
			<td>The Jackson Laboratory</td>
			<td></td>
			<td>male, 12-16 weeks old</td>
		</tr>
		<tr>
			<td>Carprofen</td>
			<td>Zoetis Services LLC, Parsippany, NJ, USA</td>
			<td>026357</td>
			<td>NOTE: this drug is added during weight drop only if stereotactic electrode implantation will be performed on the same day</td>
		</tr>
		<tr>
			<td>Chlorhexidine antiseptic</td>
			<td></td>
			<td></td>
			<td>Pharmacy</td>
		</tr>
		<tr>
			<td>Dental cement and solvent kit</td>
			<td>Stoelting Co., USA</td>
			<td>51459</td>
			<td></td>
		</tr>
		<tr>
			<td>Drill</td>
			<td>Foredom</td>
			<td>HP4-917</td>
			<td></td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>Meisinger USA, LLC, USA</td>
			<td>HM1-005-HP</td>
			<td>0.5 mm, Round, 1/4, Steel</td>
		</tr>
		<tr>
			<td>Dry sterilizer</td>
			<td>Cellpoint Scientific, USA</td>
			<td></td>
			<td>Germinator 500</td>
		</tr>
		<tr>
			<td>EEG System 1</td>
			<td>Biopac Systems, CA, USA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>EEG System 2</td>
			<td>Pinnacle Technology Inc., KS, USA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol &#8805;70%</td>
			<td>VWR, USA</td>
			<td>71001-652</td>
			<td>KOPTEC USP, Biotechnology Grade (140 Proof)</td>
		</tr>
		<tr>
			<td>Eye ointment</td>
			<td>Pro Labs Ltd, USA</td>
			<td></td>
			<td>Puralube Vet Ointment Sterile Ocular Lubricant available in general online stores and pharmacies</td>
		</tr>
		<tr>
			<td>Fluriso liquid for inhalation anesthesia</td>
			<td>MWI Veterinary Supply Co., USA</td>
			<td>502017</td>
			<td></td>
		</tr>
		<tr>
			<td>Hair removal product</td>
			<td>Church &#38; Dwight Co., Inc., USA</td>
			<td></td>
			<td>Nair cream</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>MWI Veterinary Supply Co., USA</td>
			<td>502017</td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone-iodine surgical solution</td>
			<td>Purdue Products, USA</td>
			<td>004677</td>
			<td>Betadine</td>
		</tr>
		<tr>
			<td>Rimadyl/Carprofen</td>
			<td>Zoetis Services LLC, Parsippany, NJ, USA</td>
			<td>026357</td>
			<td></td>
		</tr>
		<tr>
			<td>Solder</td>
			<td></td>
			<td></td>
			<td>Harware store</td>
		</tr>
		<tr>
			<td>Soldering iron</td>
			<td>Weller, USA</td>
			<td>WP35</td>
			<td>ST7 tip, 0.8mm</td>
		</tr>
		<tr>
			<td>Stainless steel disc</td>
			<td></td>
			<td></td>
			<td>Custom made</td>
		</tr>
		<tr>
			<td>Sterile cotton swabs</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile gauze pads</td>
			<td>Fisher Scientific, USA</td>
			<td>22362178</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile poly-lined absorbent towels pads</td>
			<td>Cardinal Health, USA</td>
			<td>3520</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue adhesive</td>
			<td>3M Animal Care Products, USA</td>
			<td>1469SB</td>
			<td></td>
		</tr>
	</tbody>
</table>

inducing post-traumatic epilepsy in a mouse model of repetitive diffuse traumatic brain injury

创伤性脑损伤（TBI）是后天性癫痫的主要原因。TBI 可导致焦点或扩散性脑损伤。焦点损伤是直接机械力的结果，有时穿透颅骨，在脑组织中产生直接病变。这些在脑成像过程中可见，作为有挫伤、撕裂和出血的区域。焦点病变导致神经元死亡和胶质疤痕的形成，并且存在于所有发生 TBI 的 20% -25% 的人的中。然而，在大多数TBI病例中，损伤是由加速减速力和随后的组织剪切引起的，导致非焦、漫反射损伤。TBI患者的亚群在几个月或几年的潜伏期后继续发展创伤后癫痫（PTE）。目前，无法预测哪些患者会患上PTE，PTE患者的癫痫发作难以控制，需要进一步研究。直到最近，该领域仅限于两个经过验证的自发创伤后发作的动物/啮齿动物模型，两者都出现大的焦点病变，皮层和有时皮下结构组织大量丧失。与这些方法相反，确定使用改良的降重模型诱导的漫射TBI足以启动自发性惊厥和非抽搐发作的发展，即使在没有焦点病变或组织损失的情况下也是如此。与获得性创伤后癫痫的人类患者类似，该模型在发作前呈现损伤后的潜伏期。在该协议中，将为社区提供创伤后癫痫的新模型，详细说明如何诱导扩散性非病变TBI，然后持续进行持续的长期视脑动物监测，持续几个月。该协议将详细说明动物处理、重量下降程序、两个采集系统的电极放置，以及手术、术后监控和数据采集过程中遇到的频繁挑战。

此处概述的协议描述了使用重复扩散 TBI 的小鼠模型（图 1）隔离诱导漫射损伤的方法（例如，在没有焦点病变的情况下）。图 1A描述了用于在此模型中诱导 TBI 的降重装置及其组件（图1A、a1_a5）以及过程中的关键步骤（图 1B，b1_b5）。
该模型的特?...

Watch this Scientific Journal Video about Inducing Post-Traumatic Epilepsy in a Mouse Model of Repetitive Diffuse Traumatic Brain Injury Video at JoVE.com

Inducing Post-Traumatic Epilepsy in a Mouse Model of Repetitive Diffuse Traumatic Brain Injury Video (Video) | JoVE

此系统协议描述了重复性轻度创伤性脑损伤后创伤后癫痫的新动物模型。第一部分详细介绍了使用修改后的降重模型进行创伤性脑损伤诱导的步骤。第二部分对单通道和多通道脑电图数据采集系统的手术方法进行了说明。

在重复性扩散创伤性脑损伤小鼠模型中诱导创伤后癫痫

Traumatic brain injury (TBI) is a leading cause of acquired epilepsy. TBI can result in a focal or diffuse brain injury. Focal injury is a result of ...

我们的协议是第一个详细描述由轻度创伤性脑损伤引起的创伤性癫痫的小鼠模型。使用轻度扩散创伤性脑损伤模型模仿人类创伤性脑损伤病理学的一个重要方面，该病理学几乎总是具有扩散成分。为了开发生物标志物和治疗方法，需要使用适当再现人类病理学方面的模型进行研究。我们的创伤性脑损伤模型也有潜力提供对轻度创伤性脑损伤或脑震荡后导致脑功能损伤的机制的洞察。在确认对疼痛反射缺乏反应后，将鼠标放在泡沫垫上，将鼠标头放在重量下降管下。将一个直径1.3厘米、厚880毫克的不锈钢盘放在动物头部中心的眼睛和耳朵之间。拆下降压管中的销，将 100 克重量杆从 50 厘米的高度释放。要对照小鼠造成虚假伤害，请从管子上取下重量杆，以防止在重量下降期间意外释放销。重量下降诱导后，将动物放在其背上的加热垫上，用无菌的多衬里吸水毛巾覆盖，并监测直至完全卧放。然后将鼠标放在一个干净的笼子里，用回收凝胶和几块湿润的周片加热，45分钟。要准备 EEG 电极植入的手术场，请将温度传感器放在垫上，然后将重新麻醉的脑损伤小鼠放在立体定向装置加热垫顶部的无菌窗帘上。用耳条固定头部后，在动物的眼睛上涂抹润滑膏，并使用脱毛霜将头发从头皮上去除。用三个20秒交替拭子的波维酮碘手术防腐剂溶液和70%乙醇在圆周运动中消毒暴露的皮肤。去除头皮后，在开放的皮肤边框上涂抹小护肤面，以扩大暴露区域。接下来，使用手术刀轻轻取出周长，用过氧化氢擦拭颅骨。当颅骨被清除组织并表现出白色外观时，用无菌纱布或拭子擦干骨质。对于电极植入，使用高速钻头，以每分钟 5，000 到 6，000 次旋转的速度进行 0.5 毫米的圆形钢位，为螺钉电极创建 6 个毛刺孔。钻好所有孔后，将六个螺钉电极放入毛刺孔中，将半勺牙科水泥粉与几滴溶剂混合。使用铲子搅拌，直到牙科水泥混合物是俗气的，但可塑和僵硬，足以适当浓缩时，放置在动物的颅骨。将水泥涂抹在颅骨和每个螺杆电极的整个外露表面。一到两分钟后，打开焊接熨斗，将三个 EEG 头安装到立体定向支架臂中。将头安装在头骨上，使六根导线位置与每个螺钉电极的导线位置相匹配。降低设备，使其腹部放在牙科水泥的顶部。用头安装的相应导线从每个螺钉电极上扭动每个引线，焊接每双绞线，以进行正确的信号传导，并使用剪刀修剪任何多余的导线。将每对焊接的导线绕在头部支座周围弯曲。当所有对都弯曲时，用新鲜的牙科水泥盖住所有电线，只留下头座的黑色部分暴露。释放持有皮瓣的血肉，在植入物周围区域涂抹氯己丁防腐剂，以避免感染。然后将动物从仪器转移到一个平衡，以测量动物的重量，作为未来监测的参考，然后把动物放在一个干净的笼子里，放在一个温暖的加热垫上，有监测，恢复凝胶和一些湿润的周片。在这个数字中，一个自发的癫痫发作在鼠标97天后重复减肥创伤性脑损伤记录使用三个EEG头安装配置，如本视频协议所示显示。在这里，一个自发的非抽搐电性癫痫发作记录在小鼠65天后，反复减肥创伤性脑损伤显示。在这个数字中，在老鼠的自发性癫痫发作23天后，反复减肥创伤性脑损伤记录使用一个EEG头部安装。典型癫痫发作的光谱功率表示最高密度在 10 到 40 赫兹之间，峰值为 15 赫兹。该协议演示了重复体重下降创伤性脑损伤后动物癫痫发作发作的时间表，表明某些动物的癫痫发作聚集的发生率，并强调获得连续而不是间歇性记录的重要性。正确放置钢盘、小心地植入螺钉电极和仔细焊接导线，都是长期试验成功的关键。脑电图监测后，脑损伤导致癫痫发作的动物可以从那些不用于随后分析其细胞、分子和生理差异的动物进行分层。将创伤后癫痫动物从创伤性脑损伤中分层，但没有出现癫痫发作，对于干预措施和生物标志物的发展至关重要。

Research

JoVE Journal

Behavior

Inducing Post-Traumatic Epilepsy in a Mouse Model of Repetitive Diffuse Traumatic Brain Injury

Compensatory Limb Use and Behavioral Assessment of Motor Skill Learning Following Sensorimotor Cortex Injury in a Mouse Model of Ischemic Stroke

Compensatory Limb Use and Behavioral Assessment of Motor Skill Learning Following Sensorimotor Cortex Injury in a Mouse Model of Ischemic Stroke Video (Video) | JoVE

补偿肢使用和运动技能学习行为评估以下感觉运动皮层损伤缺血性脑卒中的小鼠模型

Mouse models have become increasingly popular in the field of behavioral neuroscience, and specifically in studies of experimental stroke. As models ...

科研

行为学

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

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats Video (Video) | JoVE

修改后的孔板 - 测量行为，认知和社会互动小鼠和大鼠

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

Exploring the Use of Isolated Expressions and Film Clips to Evaluate Emotion Recognition by People with Traumatic Brain Injury

Exploring the Use of Isolated Expressions and Film Clips to Evaluate Emotion Recognition by People with Traumatic Brain Injury Video (Video) | JoVE

探索孤立的表达式和电影剪辑的使用与创伤性脑损伤的人评价情感识别

The current study presented 60 people with traumatic brain injury (TBI) and 60 controls with isolated facial emotion expressions, isolated vocal emotion ...

Repetitive Transcranial Magnetic Stimulation to the Unilateral Hemisphere of Rat Brain

Repetitive Transcranial Magnetic Stimulation to the Unilateral Hemisphere of Rat Brain Video (Video) | JoVE

重复经颅磁刺激对大鼠大脑的单边半球

Previous rodent models of repetitive transcranial magnetic stimulation (rTMS) adopted whole-brain stimulation instead of unilateral hemispheric rTMS, ...

Wheel Running and Environmental Complexity as a Therapeutic Intervention in an Animal Model of FASD

Wheel Running and Environmental Complexity as a Therapeutic Intervention in an Animal Model of FASD Video (Video) | JoVE

车轮运行环境的复杂性在胎儿酒精中毒综合症的动物模型干预治疗

Aerobic exercise (e.g., wheel running (WR) extensively used in animal research) positively impacts many measures of neuroplastic potential in the brain, ...

Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze Video (Video) | JoVE

使用径向水痕迷宫评估外伤性脑损伤小鼠模型中的空间记忆障碍

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury Video (Video) | JoVE

颅脑损伤后大鼠行为缺陷的检测

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, ...

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat Video (Video) | JoVE

激光多普勒成像与监测在大鼠脊髓微循环分析中的应用

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory ...

A New Method for Inducing a Depression-Like Behavior in Rats

A New Method for Inducing a Depression-Like Behavior in Rats Video (Video) | JoVE

一种诱导大鼠抑郁症状行为的新方法

Contagious depression is a phenomenon that is yet to be fully recognized and this stems from insufficient material on the subject. At the moment, there is ...

Stress-Enhanced Fear Learning, a Robust Rodent Model of Post-Traumatic Stress Disorder

Stress-Enhanced Fear Learning, a Robust Rodent Model of Post-Traumatic Stress Disorder Video (Video) | JoVE

压力增强恐惧学习--创伤后应激障碍的健壮啮齿动物模型

Fear behaviors are important for survival, but disproportionately high levels of fear can increase the vulnerability for developing psychiatric disorders ...