This works was supported by funding from a Florida Health grant (Brain and spinal cord injury research fund) (KKW).

根据协议＃201207692由佛罗里达大学的机构动物护理和使用委员会，并按照卫生指南实验动物的护理和使用国家认可的机构进行所有的程序。 1.动物护理<ol><li>使用3-4个月大的雄性C57BL / 6J小鼠。提供床上用品，排料，食品， 自由采食和饮水 。保持小鼠控制在20环境温度 - 22℃，恒定的12小时光照/ 12小时黑暗的周期。 </li></ol> 2.预嵌塞准备<ol><li>安装一个特制的硅橡胶涂层的金属尖的电磁影响立体设备。确保尖端的平底平行于探头尖端（ 图1A）的表面上。 </li><li>麻醉，用4％的异氟醚后2.5％异氟烷维持麻醉的小鼠。检查经由流量计的麻醉。 MonitoR中的麻醉水平，直至达到动物展示踏板撤回反射消失手术水平的麻醉。 </li><li>把鼠标在一个加热垫俯卧。使用一个漏斗形鼻锥保持鼠标麻醉下。使用微调完全剃光头。使用鼠标的眼睛凡士林眼药膏，以防止干燥时的麻醉下。 </li></ol> 3.影响参数设置注:冲击系统包括控制箱设置冲击参数，以执行嵌塞的致动器，并用3-运动轴的数字立体定位框架。 <ol><li>冲击装置的速度预先设定为4米/秒和停留时间，以在控制箱240毫秒。 </li></ol> 4.定位影响中心<ol><li>把动物的身下软加热垫，以保持身体的温度接近39℃。安装鼠标立体定位框架中PRON用钝头耳条E的位置。 </li><li>降低的影响，通过尖端移动Z驱动器靠近鼠标的头部。通过向目标坐标矢状缝上述中途移动所述X和Y驱动调整平面冲击尖端（9毫米直径的）。 </li><li>确保冲击尖端的一个边缘被垂直地平行于两个耳朵（ 图 1C）之间画的假想水平线。冲击的中心对应于interfrontal和人字缝之间的中心矢状缝中途（耳间9毫米到耳间0毫米，横向4.5毫米）。 </li></ol> 5.影响深度设定<ol><li>要正确设置的影响深度，使用额外的探头尖端更换绝缘硅橡胶涂层的影响一角。 </li><li>为了确保有切换提示后无冲击中心的转变，切换提示前，数字立体控制面板上的X和Y通道设置为零。 </li><li>移动概率E喷嘴的冲击区域的中心手动移动X和Y驱动器。 </li><li>夹子接触式传感器鼠标的尾巴。 </li><li>移动至冲击（Z驱动），直到探针尖端接触的冲击部位的表面上。 </li><li>立体定位控制面板上的Z通道设置为零。 </li><li>通过手动调节X轴和Y驱动器（未在数字立体控制面板上的零的按钮），直到X和Y驱动移动至冲击尖端回到冲击面积是零（其中冲击尖端先前定位）。 </li><li>由控制箱移动收缩开关缩回致动器。手动4 mm将撞击向下（Z驱动器）。 </li></ol> 6.影响<ol><li>通过点击在控制箱的冲击开关触发的影响，并实现4mm的变形深度。 </li></ol> 7.后嵌塞<ol><li>直到使用定时器鼠标的第一次呼吸测量从影响的时间。 </li><li&gt;从仪器上取下动物，并将其放置在一个温暖的垫来维持自己的体温。不要离开无人看管的动物，直到它已经恢复了足够的意识，以保持胸骨斜卧。 </li><li>返回动物放回一个干净的笼子前，允许进行恢复。不要动物重返公司其他动物，直到完全康复。 </li><li>观察并每天称重小鼠。如果小鼠显示疼痛的迹象，腹膜内以1美洛昔康注射他们 - 2毫克/公斤，每12 - 24小时。 </li></ol> 8.重复嵌塞<ol><li>给老鼠额外伤害7日4，和10最初的损伤（影响之间72小时间隔）之后。 </li></ol> 9.免疫组织化学（IHC） <ol><li> 穿心灌注 <ol><li>麻醉经由腹膜内注射的小鼠的200米/公斤戊巴比妥。 </li><li>评估并通过脚趾捏保证手术面麻醉。二段URE在仰卧位鼠标的前爪和后爪轻轻的录音，以聚苯乙烯泡沫塑料工作表面化学通风柜内。 </li><li>通过沿着刚刚剑突至锁骨下胸腔中线的皮肤做一个切口。使两个附加的皮肤切口在剑突和沿腹胸腔横向的底部进行。 </li><li>打开胸腔，并通过胸部肌肉和胸腔切割暴露心脏。 </li><li>固定跳动的心脏钝钳，并进行1 - 左心室2毫米切口。 </li><li>立即插入一个蝴蝶针入右心房。通过推动注射器缓慢开始20毫升盐水的输注。 </li><li>从切换到生理盐水4％多聚甲醛。继续与多聚甲醛20ml的灌注。 </li><li>斩首鼠标并用剪刀去除皮肤。使用骨刀头骨隔离大脑。 </li></ol></li><li> Cryostat切片<ol><li>在最佳切削温度（OCT）制定和冻结嵌入脑组织在-80℃。放置在低温恒温器的大脑矢状方向。切脑切片5微米厚。 </li></ol></li><li> 染色 <ol><li>干燥在常温下的冷冻切片1小时。 </li><li>用100μl2％山羊血清和0.1％的Triton X-100的磷酸盐缓冲盐水（PBS），在室温1小时孵育的幻灯片。 </li><li>用300微升PBS洗幻灯片的3倍。在4℃下:（200 1）分别过夜:然后孵育用抗GFAP（1 200）的幻灯片或抗铁蛋白抗体。 </li><li>用300微升PBS洗幻灯片的3倍。然后孵育载玻片在室温下与生物素缀合的二级抗体2小时。 </li><li>用300微升PBS洗幻灯片的3倍。然后孵育与抗生物素蛋白 - 生物素复合物（ABC）溶液（1:50）在室温下30分钟的幻灯片。 </li><li>用300微升PBS洗幻灯片的3倍。 8分钟-然后在3,3&#39;-二氨基联苯胺为5（DAB）底物溶液（50毫升的PBS，10微升的 H 2 O 2，10毫克的DAB丸，在使用前过滤器）孵育。观察载玻片，在显微镜下，直到阳性细胞出现。 </li><li>冲洗在缓慢的流动自来水将载玻片5分钟。干净的玻片用实验室擦拭。然后装入带安装介质和盖玻片的章节。 </li></ol></li></ol>

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The authors have no financial interest to disclose.

模仿脑损伤形态相似的临床病症，预期脑震荡后症状。脑震荡的症状后一般包括头痛，头晕，眼花，乏力，记忆力和睡眠障碍，注意力无法集中以及焦虑，情绪低落。由于躯体症状可能还不是在动物模型衡量，运动和认知功能和情感行为的变化作为标准，在动物模型中合理评估脑震荡。在先前的报道的研究中，它表明，在rCHI小鼠模型诱导缺陷在空间学习，记忆和焦虑8。更重要的是，在此...

脑震荡，也被称为轻度颅脑损伤（MTBI），是创伤性脑损伤（TBI）的最频繁发生，影响数百万人在美国。震荡可能会非常棘手的诊断并没有为脑震荡没有具体的治疗。有越来越多的认可和一些证据表明，运动损伤，军事打击，以及其他物理引人入胜的追求导致轻微的机械性损伤可能有累积性，慢性神经后果1,2。然而，仍然有关于脑震荡及其影响的认识不足。现行方法限制病理和治疗的人类研究的，因为只有神经评估和成像评估可用于临床诊断。动物模型提供了学习的脑震荡与进一步的诊断和治疗MTBI的希望高效，严谨，可控的方式的一种手段。 有研究适应传统的TBI车型如控制皮质影响（CCI），液压冲击影响（FPI），体重下降损伤，爆炸伤执行MTBI通过改变参数的伤害低刺激伤害的严重性。这些模型，由于其复制脑外伤形态上类似于临床状况的能力是有益的使用;不过，他们也有自己的局限性。由加速度损伤（重量滴）诱导损伤的严重程度常常是高度可变的。轻度CCI的两种结果 - 蛛网膜下腔出血和挫伤焦点 - 不典型的脑震荡的人可比的。 CCI和FPI需要开颅手术，这不是临床相关，而爆伤是曝光3-6期间关于不同的曝光位置和峰值压力测量一个比较有争议的模型，以及可变继发性损伤。一个更新的震荡动物模型可以转化临床前研究的临床settiNG是必要的研究。 在模拟轻度TBI的关键问题是确定实验损伤严重，其中最密切复制在临床受伤。最近，不同的研究小组开发出闭合性颅脑损伤或震荡头部损伤（CHI）模型7-10。 CHI是CCI的不开颅的修改，但它依然采用的是传统的电子磁场影响系统产生头部撞击。阴气可以通过调节参数的影响引起了脑震荡，从轻度到中度。意识（LOC）的损失可立即通过检测呼吸速率或呼吸的瞬时终止的降低的冲击之后观察到。 LOC的期间被用于确定损伤的严重程度。本文包括小鼠重复CHI（rCHI）模型略有改善和更新版本，以及一个详细的一步一步的协议和代表的搜索结果。该rCHI模型研究战略重新确定MTBI效应和潜在的治疗有益的，特别是因为有能够模仿所有的脑震荡引起的病理变化没有个体动物模型。

<table border="0" cellpadding="0" cellspacing="0" width="831">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">anesthesia machine</td>
			<td style="width:188px;">Eagle Eye Anesthesia, Inc</td>
			<td style="width:137px;">Model 150&#160;</td>
			<td style="width:291px;">anesthesia</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Electromagnetic Impactor</td>
			<td style="width:188px;">LeicaBiosystems</td>
			<td style="width:137px;">Impact One Stereotaxic Impactor</td>
			<td>perform impaction</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Digital Stereotaxic instrument</td>
			<td style="width:188px;">LeicaBiosystems</td>
			<td style="width:137px;">39462501</td>
			<td>mount mouse and positioning tips</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Sicilone rubber-coated metal tip</td>
			<td style="width:188px;">Precision Tool &#38; Engineering, Gainesvill FL</td>
			<td style="width:137px;">custom-made</td>
			<td>impact tip</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Lithium Ion All-in-One Trimmer</td>
			<td style="width:188px;">WAHL Home Products</td>
			<td style="width:137px;">9854-600</td>
			<td>shave mouse hair</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:216px;">paper clips</td>
			<td style="width:188px;">custom-made</td>
			<td style="width:137px;"></td>
			<td>probe tip</td>
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			<td height="21" style="height:21px;width:216px;">Cotton tipped applicators</td>
			<td style="width:188px;">MEDLINE</td>
			<td style="width:137px;">MDS202055</td>
			<td>scrub head with saline</td>
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			<td height="21" style="height:21px;">Tissue Tek O.C.T.</td>
			<td style="width:188px;">ASKURA FINETEK USA INC</td>
			<td style="width:137px;">4583</td>
			<td>tissue embedding</td>
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			<td height="21" style="height:21px;width:216px;">anti-GFAP</td>
			<td style="width:188px;">Dako</td>
			<td style="width:137px;">CA93013</td>
			<td>antibody for IHC</td>
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			<td height="21" style="height:21px;width:216px;">anti Ferritin</td>
			<td style="width:188px;">Sigma</td>
			<td style="width:137px;">F6136</td>
			<td>antibody for IHC</td>
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			<td height="21" style="height:21px;">VECTASTAIN Elite ABC&#160; kit</td>
			<td style="width:188px;">Vector laboratories</td>
			<td style="width:137px;">PK-6100</td>
			<td>IHC detection system</td>
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			<td height="21" style="height:21px;width:216px;">Permount Mounting Medium</td>
			<td style="width:188px;">Fisher Scientific</td>
			<td style="width:137px;">SP15-100</td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">Aperio XT ScanScope scanner</td>
			<td style="width:188px;">Leica Microsystems Inc,</td>
			<td style="width:137px;"></td>
			<td>slides scanning</td>
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			<td height="42" style="height:42px;width:216px;">Leica AutoStainer XL</td>
			<td style="width:188px;">Leica the pathology Company</td>
			<td style="width:137px;">ST2010</td>
			<td>H&#38;E staining</td>
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			<td height="21" style="height:21px;width:216px;">DAB&#160;</td>
			<td style="width:188px;">sigma</td>
			<td style="width:137px;">D3939</td>
			<td>IHC detection system</td>
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</table>

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.

在这个模型中（ 图1 AC），有喘息和呼吸浅的短暂。意识（无意识）的损耗被定义为呼吸率或恢复正常的呼吸呼吸之前短暂终止下降。在头部的中心的冲击造成的短期无意识（7.5±4.7，7.8±5.5，10.2±8.8，在每个冲击9.5±8.0秒分开， 图1D）。小鼠的大脑表现出通过H＆E染色的组织学，这表明来自冲击（ 图2A）产生没有明显的结构损害或组织...

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A Repetitive Concussive Head Injury Model in Mice

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

重复震荡颅脑损伤模型小鼠

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

a-repetitive-concussive-head-injury-model-in-mice

Research

JoVE Journal

Medicine

11.8K 次观看.  University of Florida. 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.

视频: A Repetitive Concussive Head Injury Model in Mice

The Spared Nerve Injury (SNI) Model of Induced Mechanical Allodynia in Mice

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared nerve injury (SNI) model induces symptoms of neuropathic pain such as mechanical allodynia i.e. pain due to tactile stimuli that do not normally provoke a painful response [1]. 
The SNI mouse model involves ligation of two of the three branches of the sciatic nerve (the tibial nerve and the common peroneal nerve), while the sural nerve is left intact [2]. The lesion results in marked hypersensitivity in the lateral area of the paw, which is innervated by the spared sural nerve. The non-operated side of the mouse can be used as a control. The advantages of the SNI model are the robustness of the response and that it doesn&#x2019;t require expert microsurgical skills.
The threshold for mechanical pain response is determined by testing with von Frey filaments of increasing bending force, which are repetitively pressed against the lateral area of the paw [3], [4]. A positive pain reaction is defined as sudden paw withdrawal, flinching and/or paw licking induced by the filament. A positive response in three out of five repetitive stimuli is defined as the pain threshold. 
As demonstrated in the video protocol, C57BL/6 mice experience profound allodynia as early as the day following surgery and maintain this for several weeks.

The Spared Nerve Injury SNI Model of Induced Mechanical Allodynia in Mice

The Spared Nerve Injury animal model is described here as a mouse model of peripheral neuropathic pain following partial denervation of the sciatic nerve by lesioning the tibial and common peroneal nerve branches, leaving the remaining sural nerve intact. Behavioral modification resulting from mechanical allodynia is quantified by von Frey filaments.

幸免神经损伤（SNI）模型引起的机械触感小鼠

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared ...

科研

医学

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

控制宫颈裂伤损伤小鼠

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

挫伤模型脊髓严重损伤的保护作用

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

急性肾损伤和伤后纤维化小鼠缺血再灌注模型

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

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

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

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

肾缺血再灌注损伤:损伤的小鼠模型和再生

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

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

一个简单的临界尺寸股骨缺损模型小鼠

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

Facial Nerve Axotomy in Mice: A Model to Study Motoneuron Response to Injury

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and either cut or crush it to induce peripheral nerve injury. Advantages of this surgery are its simplicity, high reproducibility, and the lack of effect on vital functions or mobility from the subsequent facial paralysis, thus resulting in a relatively mild surgical outcome compared to other nerve injury models. A major advantage of using a cranial nerve injury model is that the motoneurons reside in a relatively homogenous population in the facial motor nucleus in the pons, simplifying the study of the motoneuron cell bodies. Because of the symmetrical nature of facial nerve innervation and the lack of crosstalk between the facial motor nuclei, the operation can be performed unilaterally with the unaxotomized side serving as a paired internal control. A variety of analyses can be performed postoperatively to assess the physiologic response, details of which are beyond the scope of this article. For example, recovery of muscle function can serve as a behavioral marker for reinnervation, or the motoneurons can be quantified to measure cell survival. Additionally, the motoneurons can be accurately captured using laser microdissection for molecular analysis. Because the facial nerve axotomy is minimally invasive and well tolerated, it can be utilized on a wide variety of genetically modified mice. Also, this surgery model can be used to analyze the effectiveness of peripheral nerve injury treatments. Facial nerve injury provides a means for investigating not only motoneurons, but also the responses of the central and peripheral glial microenvironment, immune system, and target musculature. The facial nerve injury model is a widely accepted peripheral nerve injury model that serves as a powerful tool for studying nerve injury and regeneration.

We present a surgical protocol detailing how to perform a cut or crush axotomy on the facial nerve in the mouse. The facial nerve axotomy can be employed to study the physiological response to nerve injury and test therapeutic techniques.

面神经干切断小鼠:模型，研究运动神经元损伤反应

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and ...

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week protocol for repetitive hypoxic preconditioning (RHP) induces long-term protection against central nervous system (CNS) injury in a mouse model of focal ischemic stroke. RHP consists of 9 stochastic exposures to hypoxia that vary in both duration (2 or 4 hr) and intensity (8% and 11% O2). RHP reduces infarct volumes, blood-brain barrier (BBB) disruption, and the post-stroke inflammatory response for weeks following the last exposure to hypoxia, suggesting a long-term induction of an endogenous CNS-protective phenotype. The methodology for the dual quantification of infarct volume and BBB disruption is effective in assessing neurovascular protection in mice with RHP or other putative neuroprotectants. Adult male Swiss Webster mice were preconditioned by RHP or duration-equivalent exposures to 21% O2 (i.e. room air). A 60 min transient middle cerebral artery occlusion (tMCAo) was induced 2 weeks following the last hypoxic exposure. Both the occlusion and reperfusion were confirmed by transcranial laser Doppler flowmetry. Twenty-two hr after reperfusion, Evans Blue (EB) was intravenously administered through a tail vein injection. 2 hr later, animals were sacrificed by isoflurane overdose and brain sections were stained with 2,3,5- triphenyltetrazolium chloride (TTC). Infarcts volumes were then quantified. Next, EB was extracted from the tissue over 48 hr to determine BBB disruption after tMCAo. In summary, RHP is a simple protocol that can be replicated, with minimal cost, to induce long-term endogenous neurovascular protection from stroke injury in mice, with the translational potential for other CNS-based and systemic pro-inflammatory disease states.

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

神经血管保护定量继在小鼠重复缺氧预处理和瞬态大脑中动脉闭塞

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

Photothrombosis引起的局部脑缺血脊髓损伤的小鼠模型

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

小鼠加速动脉硬化的诱导:"电线损伤"模型

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...