我们感谢贝丝Rauch和美国威斯康星大学的小动物磁共振成像成像设施。在美国威斯康星 - 麦迪逊大学的神经外科部为这项工作提供了资金。

缺血大鼠模型术前准备工作应使用无菌操作技术，手术过程中的所有生存。商业消毒剂与消毒手术的工作表面，并准备通过高压灭菌的无菌手术仪器，窗帘，纱布，棉签，缝合线，手术刀片包。应戴口罩，头发的帽子和无菌手套。一个Germinator干珠灭菌也可用于手术器械重新消毒程序之间，如果有多个大鼠手术将在一次会议。 Prewarm一个水套homeothermic毯和吸水垫下，为了防止在手术过程中对大鼠的低温。 <ol><li>放入感应室自发性高血压大鼠（或其他的选择大鼠株）和诱导麻醉与5％的异氟醚（麻醉机应设置为1.0 L / min的O 2和1.0升/分钟的N 2 O） 。降低至1-2％异氟醚维持麻醉。 </li><li>应用人工泪液软膏双眼。 </li><li>剃须超出预期的切口，用快船队（A5＃10刀片奥斯特），喉咙和左颈部区域。 </li><li>优碘纱布垫和皮肤消毒，从手术区域的中心，螺旋式向外。用无菌纱布垫含有70％的乙醇冲洗，移动垫了类似的模式。重复这两个步骤，共3个周期。 </li><li> 0.5％布比卡因沿准切口部位皮下注入0.2毫升。 </li><li>将大鼠在无菌stockinette或盖用无菌手术悬垂。 </li></ol>马华的瞬态闭塞<ol start="7"><li>在手术显微镜下，腹正中切口进行解剖浅筋膜。 </li><li>下面的浅筋膜，腺体组织，形成一个三角形的胸骨，这是在气管中线;二腹肌（轻松地确定其闪亮的白色的肌腱部分）左和三个肌肉;最后，胸锁乳突肌肌肉。 </li><li>仔细尖锐，钝性分离内执行的三角形，以确定颈内动脉（独家钝性剥离可能是由一些首选，以尽量减少意外的组织损伤）。外部，内部，共同的颈总动脉（ECA，ICA CCA）的暴露。颈动脉是大和明显的脉冲。迷走神经是沿着双方共同和颈内动脉外侧窃喜，并大幅关闭的共同和颈内动脉解剖。 </li><li>两个非洲经委会分支，然后急剧解剖，标题内侧首支和第二支横向标题。烧灼和削减两个分支，这使得更易于调动较大的船只。 </li><li>非洲经委会现在更rostrally解剖。舌骨可能会遇到，这将限制喙清扫的程度。颈外动脉结扎尽可能远端用6-0丝线缝合。 </li><li>另外6-0丝线缝合与ICA分叉处经委会周围附近放置松散。确保不堵塞线栓闭塞将经历在后面的步骤船只。 </li><li>显微剪辑放置常见的颈内动脉分叉附近。 </li><li>创建一个初步的部分动脉切开术颈外动脉的缝合关系两者之间的丝绸。 </li><li>介绍了4-0单丝尼龙缝合2.2 -3.0厘米的长度，其浑圆，硅涂层的提示与扭结2.0厘米到非洲经委会流明，对显微剪辑所在的CCA。这些缝线市售（Doccol公司，雷德兰兹，CA，USA）。切非洲经委会的剩余部分（在网站的部分动脉切开术），自由的残端和位置非洲经委会和ICA分叉处以下的残端，这会更容易让线栓进入ICA幻灯片。 </li><li>丝线缝合收紧经委会残端腔内安全的尼龙缝线，防止出血，然后删除从ICA微血管剪辑。慢慢打开剪辑，再取出检查出血。 </li><li>继续推进尼龙缝线从ECA到ICA流明大脑中动脉（MCA）。这个长度一般为18-20毫米，是放置在缝合前扭结插入的原因。后插入一个可变长度的尼龙缝线，可能会感觉到阻力。如果发生这种尼龙缝线还在外面的船只目前，它表明，缝合很可能进入翼腭动脉（见下文附注）。拉回来，并缝合轻微，继续沿ICA，这将运行更内侧的曲线。此外，剥离的PPA的起源可能会做更好的可视化的路径腔内的灯丝。继续插入尼龙缝线，直到阻力和2厘米扭结位置后感到。此时，腔内缝合阻止马华的起源。 MCA闭塞可以确认通过监测局部脑血流减少，使用激光多普勒流量计（见一个源设备材料表）。另外还要注意的ICA翼腭动脉的起源，可以直接挂钩，如果愿意，为了避免这种船只的意外插管腔内缝合。 </li><li>启动定时器和记录闭塞的开始时间。 </li><li>删除从CCA microclip。 </li><li>关闭切口，用3-0丝线缝合（一个简单的连续图案，将有助于重新再灌注）（真皮，panniculus carnosus，皮下组织层）迅速和小心地在回收笼的老鼠。检查鼻子和嘴周围的笼地板的铺垫材料和监测从麻醉中复苏。 </li></ol> MCA的血流恢复（灌注法） <ol start="21"><li>前不久闭塞期应该结束，再次麻醉大鼠，用优碘和70％的乙醇（3个周期，如前）的消毒切口部位，然后重新打开删除关闭缝合切口。 </li><li>广场上共同国家评估microclip，像以前一样。 </li><li>咬合缝合中途撤回从ICA直到缝合结束是通过ICA可见不完全删除从ICA /非洲经委会缝合！ </li><li>广场上的ICA以上腔内缝合结束microclip。 </li><li>完全去除咬合缝合，并紧密配合经委会残端。闭塞再灌注开始时间记录的结束。 </li><li>删除从ICA microclip。 </li><li>删除从CCA microclip。 </li><li>数滴无菌生理盐水湿润的地区，并使用一个简单的中断模式用3-0丝线缝合关闭切口层（真皮，皮下组织，panniculus carnosus）。 </li><li> 0.05毫克/公斤丁丙诺啡（以下您的机构准则或其他适当的手术后镇痛药）管理进入腹膜空间。 </li><li>预热生理盐水腹腔注射5CC。在恢复阶段，这将为水化。 </li><li>监测麻醉大鼠的复苏。 </li><li>大鼠后已经恢复，测试大鼠的尾巴，并观察大鼠是否可以把双方梗死的行为指示。冰壶到一边，不仅是预期。简单的评分尺度可以用来记录战后初期梗死behavorial运作（见讨论）。 </li><li>丁丙诺啡（如上述相同剂量）的额外注资应给予每6至8小时，24小时疼痛缓解。如果动物出现不适的迹象，延长镇痛。 </li></ol>代表性的成果<img src="/files/ftp_upload/1978/1978fig1.jpg" alt="图1" /> 图1。的中间脑动脉血管腔内缝合闭塞 。咬合马华的起源与硅涂层腔内缝合的大鼠颅循环系统的简化框图。 OA和ST左经委会已结扎，并围绕非洲经委会残端缝合领带持有腔内缝合到位。 ACA，大脑前动脉; BA，基底动脉，颈总动脉CCA;非洲经委会，颈外动脉; ICA，颈内动脉;马华，大脑中动脉; OA，枕动脉，大脑后动脉PCA; PComA，后交通动脉;苯丙醇胺，翼腭动脉，ST，甲状腺上动脉。图佐佐木等 4和李3适应。 <img src="/files/ftp_upload/1978/1978fig2.jpg" alt="图2" /> 图2。代表24小时大鼠脑冠状切片的磁共振成像后1小时短暂缺血 。伴随局灶性脑缺血脑水肿在T2加权MRI图像的可视化。梗死区出现高信号（亮）在T2图像。类似梗死模式可以看出，用更先进的磁共振成像技术，如扩散加权成像和表观扩散系数映射5。 <img src="/files/ftp_upload/1978/1978fig3.jpg" alt="图3" /> 图3。染色与TTC 24小时后1小时短暂缺血。TTC染色显示白色（未染色）在如图2所示的相同大鼠大脑皮层和纹状体梗死区域，大鼠脑组织的代表冠状切片。请注意，局灶性缺血性病变的一般模式是在MRI T2加权像看到类似，虽然最后确定组织梗死面积可能略比T2病变。

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作者有没有利益冲突的披露。 该协议被批准机构动物护理和使用委员会在美国威斯康星 - 麦迪逊大学的医学和公共健康，遵守国家机构使用实验动物的健康指引的学校。

脑缺血啮齿动物模型可分为全球性或局灶性和可逆或不可逆的。我们使用一个可逆的局灶性脑缺血模型，以模仿人类的​​中风患者可以出现在下面的血管再通再灌注损伤。虽然大鼠股类似颅流通的人力，广泛颅内侧支循环，由于威利斯和软脑膜吻合结果不一致梗死卷 6圈的优势。我们使用的自发性高血压大鼠（SHR）的菌株，因为梗死面积比 7-10血压正常大鼠局灶性脑缺血的高血压。高血压中风感应也比较一致，可能会产生较少的死亡率比其他常用菌株11。此外，生理参数进行监测，在整个手术，因为波动可以添加到脑梗死体积的变化，这些参数包括血压，动脉氧和二氧化碳含量，低温，和高血糖，可加重缺血性损伤2,12 。低温是特别关注，因为它可以减少神经元缺血性损伤的程度，因此，放置在体温的加热垫的大鼠在手术过程中是必不可少的。 我们聘请商业化准备，企图以增加梗死面积13-15的一致性，在此过程中硅涂层缝合。传统上，用于闭塞马华的腔内缝合已准备火焰围绕3-0尼龙缝线尖或使用热胶枪大衣与缝合的小胶珠 16提示。这些程序产生不同的尖端直径，从而缝线，可能需要多次尝试在闭塞前找到相匹配的一个给定的大鼠血管直径的一期缝合。在这个视频中所描述的缺血方法上的其他变化包括更远端闭塞，为了有一个纯皮质梗死17，MCA及同侧颈总动脉同时闭塞减少侧支血流6,9不遗余力豆纹分行。 可以使用多种行为测试，在赤字的具体任务是体现在一个简单的评分量表评估中风的功能效应。例如，Bederson 等18开发的一个神经功能评分分级大鼠前肢屈曲程度升高身体的位置，不对称性横向推，和开放的领域，盘旋行为。大鼠升高，表现出没有其他的赤字时，两个前肢扩展到地板被评为"正常（0）"，而对侧前肢屈曲大鼠只与"温和的赤字（1）"分类。 "严重级"老鼠显示对侧阻力下降到侧向力和"严重级"老鼠另外，在盘旋的行为。总是观察到，在未来更高档次低年级的行为缺陷（例如，2级和3只动物，也显示前肢屈曲），这些成绩是在预测梗死面积的18差异。因此，这样一个简单的评分系统，可以快速，一致的半定量评估中风的功能赤字。 通常是通过使用TTC，甲酚紫（尼氏染色）或苏木精和曙红的组织损伤的组织学染色测量脑梗死的程度。磁共振成像技术，包括扩散加权MRI可视化在早期的时间点梗死闭塞后，允许在每一个人的大鼠梗死体积的决心没有收获组织的需要。旨在确定神经保护治疗中风后的疗效的研究，这是特别有价值的成像MCAO后大鼠，但之前的治疗管理，使研究者的信心，所有实验组和对照动物都经历过类似的招启动，减少变异的混杂影响中风的感应。

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		<tbody>
		
<tr>
<td> 4-0 Nylon suture</td>
<td>&nbsp;</td>
<td>Doccol Corporation</td>
<td>&nbsp;</td>
<td>2-3 mm silicon coating length, 0.39 mm tip diameter</td>
</tr>
<tr>
<td>Spontaneously hypertensive rats</td>
<td>&nbsp;</td>
<td>Charles River Laboratories</td>
<td>&nbsp;</td>
<td>290-300 g, male</td>
</tr>
<tr>
<td>Gaymar T/Pump thermoregulatory pump and pads</td>
<td>&nbsp;</td>
<td>Gaymar Industries</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Vetroson V-10 bipolar electrosurgical unit</td>
<td>&nbsp;</td>
<td>Summit Hill Laboratories</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Germinator 500 glass bead sterilizer</td>
<td>&nbsp;</td>
<td>CellPoint Scientific </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Vasamedics Bpm2Laserflo blood perfusion monitor</td>
<td>&nbsp;</td>
<td>TSI Inc.</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

focal cerebral ischemia model by endovascular suture occlusion of the middle cerebral artery in the rat

中风是导致残疾和死亡的第三大导致全世界成年人1。在人类中风，存在一个高度可变的临床状态;局灶性脑缺血动物模型的发展，然而，实现实验诱导的梗塞体积的可重复性是必不可少的。老鼠是一个广泛应用于中风动物模型，由于其相对较低的畜牧业成本和颅流通 2,3人类的相似性。对于人类来说，大脑中动脉（MCA）是最常见的中风综合征和马华闭塞（MCAO）的多种方法的影响已在动物模型来模拟这一临床综合征。由于血管再通，通常会出现以下在人力，再灌注的急性脑卒中后一段闭塞已包括在许多这些模型。在这段视频中，我们展示了在自发性高血压大鼠（SHR）的瞬态腔内缝合MCAO模型。灯丝与硅尖端涂层置于intraluminally马华在60分钟的起源，再灌注。需要注意的是最佳的闭塞期可能有所不同，如只Wistar或只Sprague - Dawley大鼠株，。在大鼠中风的几个行为指标显示。局灶性脑缺血是使用T2加权磁共振图像证实，与2,3,5 - 三苯基氯化MCAO后24小时（TTC）染色脑切片。

Watch this Scientific Journal Video about Focal Cerebral Ischemia Model by Endovascular Suture Occlusion of the Middle Cerebral Artery in the Rat at JoVE.com

Focal Cerebral Ischemia Model by Endovascular Suture Occlusion of the Middle Cerebral Artery in the Rat

在大鼠缺血性脑损伤的手术诱导是中风的研究中被广泛使​​用的模型。在这里，我们展示了大脑中动脉闭塞局灶性脑缺血诱导。还显示导致心肌梗死组织学染色和磁共振成像的可视化。

焦距脑缺血模型大鼠大脑中动脉的血管内缝合闭塞

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable ...

focal-cerebral-ischemia-model-endovascular-suture-occlusion-middle

Research

JoVE Journal

Neuroscience

79.5K 次观看.  University of Wisconsin-Madison. 在大鼠缺血性脑损伤的手术诱导是中风的研究中被广泛使​​用的模型。在这里，我们展示了大脑中动脉闭塞局灶性脑缺血诱导。还显示导致心肌梗死组织学染色和磁共振成像的可视化。

视频: Focal Cerebral Ischemia Model by Endovascular Suture Occlusion of the Middle Cerebral Artery in the Rat

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs through activation of their ion channels and neurotransmitter receptors, and process information in part through activity-dependent release of gliotransmitters. Furthermore, astrocytes constitute the main uptake system for glutamate, contribute to potassium spatial buffering, as well as to GABA clearance. These cells therefore constantly monitor synaptic activity, and are thereby sensitive indicators for alterations in synaptically-released glutamate, GABA and extracellular potassium levels. Additionally, alterations in astroglial uptake activity or buffering capacity can have severe effects on neuronal functions, and might be overlooked when characterizing physiopathological situations or knockout mice. Dual recording of neuronal and astroglial activities is therefore an important method to study alterations in synaptic strength associated to concomitant changes in astroglial uptake and buffering capacities. Here we describe how to prepare hippocampal slices, how to identify stratum radiatum astrocytes, and how to record simultaneously neuronal and astroglial electrophysiological responses. Furthermore, we describe how to isolate pharmacologically the synaptically-evoked astroglial currents.

The preparation of acute brain slices from isolated hippocampi, as well as the simultaneous electrophysiological recordings of astrocytes and neurons in stratum radiatum during stimulation of schaffer collaterals is described. The pharmacological isolation of astroglial potassium and glutamate transporter currents is demonstrated.

双突触诱发的星形胶质细胞和神经元的反应，在急性海马脑片电生理记录

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

科研

神经科学

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

Focal cerebral ischemia (i.e., ischemic stroke) may cause major brain injury, leading to a severe loss of neuronal function and consequently to a host of motor and cognitive disabilities. Its high prevalence poses a serious health burden, as stroke is among the principal causes of long-term disability and death worldwide1. Recovery of neuronal function is, in most cases, only partial. So far, treatment options are very limited, in particular due to the narrow time window for thrombolysis2,3. Determining methods to accelerate recovery from stroke remains a prime medical goal; however, this has been hampered by insufficient mechanistic insights into the recovery process. Experimental stroke researchers frequently employ rodent models of focal cerebral ischemia. Beyond the acute phase, stroke research is increasingly focused on the sub-acute and chronic phase following cerebral ischemia. Most stroke researchers apply permanent or transient occlusion of the MCA in mice or rats. In patients, occlusions of the MCA are among the most frequent causes of ischemic stroke4. Besides proximal occlusion of the MCA using the filament model, surgical occlusion of the distal MCA is probably the most frequently used model in experimental stroke research5. Occlusion of a distal (to the branching of the lenticulo-striate arteries) MCA branch typically spares the striatum and primarily affects the neocortex. Vessel occlusion can be permanent or transient. High reproducibility of lesion volume and very low mortality rates with respect to the long-term outcome are the main advantages of this model. Here, we demonstrate how to perform a chronic cranial window (CW) preparation lateral to the sagittal sinus, and afterwards how to surgically induce a distal stroke underneath the window using a craniotomy approach. This approach can be applied for sequential imaging of acute and chronic changes following ischemia via epi-illuminating, confocal laser scanning, and two-photon intravital microscopy.

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

Surgical occlusion of a distal middle cerebral artery branch (MCAo) is a frequently used model in experimental stroke research. This manuscript describes the basic technique of permanent MCAo, combined with the insertion of a lateral cranial window, which offers the opportunity for longitudinal intravital microscopy in mice.

横向慢性颅窗口准备启用<em&gt;在体内</em&gt;观察继末节小鼠大脑中动脉闭塞

Focal cerebral ischemia (i.e., ischemic stroke) may cause major brain injury, leading to a severe loss of neuronal function and consequently to a host of ...

Experimental Models to Study the Neuroprotection of Acidic Postconditioning Against Cerebral Ischemia

Stroke is one of the leading causes of mortality and disability worldwide, with limited therapeutic approaches. As an endogenous strategy for neuroprotection, postconditioning treatments have proven to be promising therapies against cerebral ischemia. However, complicated procedures and potential safety issues limit their clinical application. To overcome these disadvantages, we have developed acidic postconditioning (APC) as a therapy for experimental focal cerebral ischemia. APC refers to the mild acidosis treatment by inhaling CO2 during reperfusion following ischemia. Here we present two models to execute APC in vitro and in vivo, respectively. The oxygen-glucose deprivation (OGD) treatment of mice and the corticostriatal occlusion and middle cerebral artery occlusion (MCAO) of mice were employed to mimic cerebral ischemia. APC can be simply achieved by transferring brain slices to acidic buffer bubbled with 20% CO2, or by mice inhaling 20% CO2. APC showed significant protective effects against cerebral ischemia, as reflected by tissue viability and brain infarct volume.

Acidic postconditioning protects against cerebral ischemia. Here we present two models to execute APC. They are achieved respectively by transferring corticostriatal slices to acidic buffer after oxygen-glucose deprivation in vitro and by inhaling 20% CO2 after middle cerebral artery occlusion in vivo.

实验模型研究酸性后处理对脑缺血的神经保护作用

Stroke is one of the leading causes of mortality and disability worldwide, with limited therapeutic approaches. As an endogenous strategy for ...

A Rat Model of Mild Intrauterine Hypoperfusion with Microcoil Stenosis

Intrauterine hypoperfusion/ischemia is one of the major causes of intrauterine/fetal growth restriction, preterm birth, and low birth weight. Most studies of this phenomenon have been performed in either models with severe intrauterine ischemia or models with gradient degree of intrauterine hypoperfusion. No study has been performed in a model on uniform mild intrauterine hypoperfusion (MIUH). Two models have been used for studies of MIUH: a model based on suture ligation of either side of the arterial arcade formed with the uterine and ovarian arteries, and a transient model based on clipping the bilateral ovarian arteries and aorta having patency. Those two rodent models of MIUH have some limitations, e.g., not all fetuses are subjected to MIUH, depending on their position in the uterine horn. In our MIUH model, all fetuses are subjected to a comparable level of intrauterine hypoperfusion. MIUH was achieved by mild stenosis of all four arteries feeding the uterus, i.e., the bilateral uterine and ovarian arteries.
Arterial stenosis was induced by metal microcoils wrapped around the feeding arteries. Producing arterial stenosis with microcoils allowed us to control, optimize, and reproduce decreased blood flow with very little inter-animal variability and a low mortality rate, thus enabling accurate evaluation. When microcoils with an inner diameter of 0.24 mm were used, the blood flow in both the placenta and fetus was mildly decreased (approximately 30% from the pre-stenosis level in the placenta). The offspring of our MIUH model clearly demonstrates long-lasting alterations in neurological, neuroanatomical and behavioral test results.

Mild intrauterine hypoperfusion was produced by artery stenosis with metal microcoils wrapped around the uterine and ovarian arteries in rats at embryonic day 17. This procedure produced prenatal hypoperfusion and intrauterine growth restriction.

轻度宫内灌注圈狭窄的大鼠模型

Intrauterine hypoperfusion/ischemia is one of the major causes of intrauterine/fetal growth restriction, preterm birth, and low birth weight. Most studies ...

A Volumetric Method for Quantification of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor determining patient outcome and is therefore frequently taken as a study endpoint. However, in small animal studies on SAH, quantification of cerebral vasospasm is a major challenge. Here, an ex vivo method is presented that allows quantification of volumes of entire vessel segments, which can be used as an objective measure to quantify cerebral vasospasm. In a first step, endovascular casting of the cerebral vasculature is performed using a radiopaque casting agent. Then, cross-sectional imaging data are acquired by micro computed tomography. The final step involves 3-dimensional reconstruction of the virtual vascular tree, followed by an algorithm to calculate center lines and volumes of the selected vessel segments. The method resulted in a highly accurate virtual reconstruction of the cerebrovascular tree shown by a diameter-based comparison of anatomical samples with their virtual reconstructions. Compared with vessel diameters alone, the vessel volumes highlight the differences between vasospastic and non-vasospastic vessels shown in a series of SAH and sham-operated mice.

The goal of this article is to present a method that allows a 3-dimensional reconstruction of the cerebrovascular tree in mice after micro computed tomography and determination of volumes of entire vessel segments that can be used to quantify cerebral vasospasm in murine models of subarachnoid hemorrhage.

小鼠蛛网膜下腔出血模型脑血管痉挛量化的容积法

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor ...

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 ...

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to intracranial delivery, intra-arterial administration of NSCs is less invasive and produces a more diffuse distribution of NSCs within the brain parenchyma. Further, intra-arterial delivery allows the first-pass effect in the brain circulation, lessening the potential for trapping of cells in peripheral organs, such as liver and spleen, a complication associated with peripheral injections. Here, we detail the methodology, in both mice and rats, for delivery of NSCs through the common carotid artery (mouse) or external carotid artery (rat) to the ipsilateral hemisphere after an ischemic stroke. Using GFP-labeled NSCs, we illustrate the widespread distribution achieved throughout the rodent ipsilateral hemisphere at 1 d, 1 week and 4 weeks after postischemic delivery, with a higher density in or near the ischemic injury site. In addition to long-term survival, we show evidence of differentiation of GFP-labeled cells at 4 weeks. The intra-arterial delivery approach described here for NSCs can also be used for administration of therapeutic compounds, and thus has broad applicability to varied CNS injury and disease models across multiple species.

A method for delivering neural stem cells, adaptable for injecting solutions or suspensions, through the common carotid artery (mouse) or external carotid artery (rat) after ischemic stroke is reported. Injected cells are distributed broadly throughout the brain parenchyma and can be detected up to 30 d after delivery.

神经干细胞向大鼠和小鼠大脑的动脉内传递:对脑缺血的应用

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to ...

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options are limited to a small proportion of stroke patients within the first hours after stroke. Novel therapeutic strategies are being extensively investigated, especially to prolong the therapeutic time window. These current investigations include the study of important pathophysiological pathways after stroke, such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Over the last decade, there has been increasing concern about the poor reproducibility of experimental results and scientific findings among independent research groups. To overcome the so-called &#8220;replication crisis&#8221;, detailed standardized models for all procedures are urgently needed. As an effort within the &#8220;ImmunoStroke&#8221; research consortium (https://immunostroke.de/), a standardized mouse model of transient middle cerebral artery occlusion (MCAo) is proposed. This model allows the complete restoration of the blood flow upon removal of the filament, simulating the therapeutic or spontaneous clot lysis that occurs in a large proportion of human strokes. The surgical procedure of this &#8220;filament&#8221; stroke model and tools for its functional analysis are demonstrated in the accompanying video.

Different models of middle cerebral artery occlusion (MCAo) are used in experimental stroke research. Here, an experimental stroke model of transient MCAo via the external carotid artery (ECA) is described, which aims to mimic human stroke, in which the cerebrovascular thrombus is removed due to spontaneous clot lysis or therapy.

模拟小鼠中风:通过颈外动脉的短暂性脑中动脉闭塞

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options ...

A Model for Encephalomyosynangiosis Treatment after Middle Cerebral Artery Occlusion-Induced Stroke in Mice

There is no effective treatment available for most patients suffering with ischemic stroke, making development of novel therapeutics imperative. The brain's ability to self-heal after ischemic stroke is limited by inadequate blood supply in the impacted area. Encephalomyosynangiosis (EMS) is a neurosurgical procedure that achieves angiogenesis in patients with moyamoya disease. It involves craniotomy with placement of a vascular temporalis muscle graft on the ischemic brain surface. EMS has never been studied in the setting of acute ischemic stroke in mice. The hypothesis driving this study is that EMS enhances cerebral angiogenesis at the cortical surface surrounding the muscle graft. The protocol shown here describes the procedure and provides initial data supporting the feasibility and efficacy of the EMS approach. In this protocol, after 60 min of transient middle cerebral artery occlusion (MCAo), mice were randomized to either MCAo or MCAo + EMS treatment. The EMS was performed 3-4 h after occlusion. The mice were sacrificed 7 or 21 days after MCAo or MCAo + EMS treatment. Temporalis graft viability was measured using nicotinamide adenine dinucleotide reduced-tetrazolium reductase assay. A mouse angiogenesis array quantified angiogenic and neuromodulating protein expression. Immunohistochemistry was used to visualize graft bonding with brain cortex and change in vessel density. The preliminary data here suggest that grafted muscle remained viable 21 days after EMS. Immunostaining showed successful graft implantation and increase in vessel density near the muscle graft, indicating increased angiogenesis. Data show that EMS increases fibroblast growth factor (FGF) and decreases osteopontin levels after stroke. Additionally, EMS after stroke did not increase mortality suggesting that protocol is safe and reliable. This novel procedure is effective and well-tolerated and has the potential to provide information of novel interventions for enhanced angiogenesis after acute ischemic stroke.

The protocol aims to provide methods for encephalomyosynangiosis-grafting of a vascular temporalis muscle flap on the pial surface of ischemic brain tissue-for the treatment of non-moyamoya acute ischemic stroke. The approach's efficacy in increasing angiogenesis is evaluated using a transient middle cerebral artery occlusion model in mice.

小鼠脑中动脉闭塞诱导脑卒中后脑肌粘连血管病治疗模型

There is no effective treatment available for most patients suffering with ischemic stroke, making development of novel therapeutics imperative. The ...

建立经颅大脑中动脉闭塞小鼠模型

A Modified Transcranial Middle Cerebral Artery Occlusion Model to Study Stroke Outcomes in Aged Mice

In experimental stroke research, middle cerebral artery occlusion (MCAO) with an intraluminal filament is widely used to model ischemic stroke in mice. The filament MCAO model typically exhibits a massive cerebral infarction in C57Bl/6 mice that sometimes includes brain tissue in the territory supplied by the posterior cerebral artery, which is largely due to a high incidence of posterior communicating artery atresia. This phenomenon is considered a major contributor to the high mortality rate observed in C57Bl/6 mice during long-term stroke recovery after filament MCAO. Thus, many chronic stroke studies exploit distal MCAO models. However, these models usually produce infarction only in the cortex area, and consequently, the assessment of post-stroke neurologic deficits could be a challenge. This study has established a modified transcranial MCAO model in which the MCA at the trunk is partially occluded either permanently or transiently via a small cranial window. Since the occlusion location is relatively proximal to the origin of the MCA, this model generates brain damage in both the cortex and striatum. Extensive characterization of this model has demonstrated an excellent long-term survival rate, even in aged mice, as well as readily detectable neurologic deficits. Therefore, the MCAO mouse model described here represents a valuable tool for experimental stroke research.

作者聚焦:中风研究的创新技术和未来方向 

This protocol demonstrates a unique mouse stroke model with a medium-sized infarct and an excellent survival rate. This model allows preclinical stroke researchers to extend the ischemia duration, use aged mice, and assess long-term functional outcomes.

改进的经颅大脑中动脉闭塞模型研究老年小鼠脑卒中结局

In experimental stroke research, middle cerebral artery occlusion (MCAO) with an intraluminal filament is widely used to model ischemic stroke in mice. ...