我们感谢新余李博士在准备稿件的帮助。 SW是由杜兰大学，总统的研究委员会新研究者奖从得克萨斯大学西南医学中心，美国国立卫生研究院资助EY021862，从研究职业发展奖，以防止盲目性基础，一个明亮的焦点奖，年龄相关性黄斑变性研究一个启动基金资助。

注：以下协议和代表性的成果使用HDAC抑制剂SAHA作为一个例子化合物。然而，该协议是决不限于使用SAHA，并且被推荐为测试可溶性化合物对角膜新生血管形成的影响的一般方法。将需要小的修改进行稀释的程度，以及频率和应用的持续时间来进行。另外，是易溶于水的化合物将能够在没有二甲基亚砜的待施用。 道德声明：所有的动物实验只应在遵守国家法律和制度规定进行。这个协议被批准用于由美国杜兰大学实验动物管理和使用委员会使用。 1。材料的制备（按使用顺序排列） <ol><li>切一块纤维素滤纸（11微米）到2毫米圈。 </li><li>通过制备的NaOH中的1M溶液在500毫升蒸馏水H 2 O：将20克NaOH的在室温下储存。注意：NaOH溶液的腐蚀性，能引起碱烧伤。 </li><li>通过在37.5毫升的1×PBS中的20毫克/毫升氯胺酮的终浓度和4毫克/毫升的甲苯噻嗪稀释加入10ml 20毫克/毫升的甲苯噻嗪的100 mg / ml的氯胺酮和2.5ml制备的麻醉剂鸡尾酒。 </li><li>通过在100毫升过滤的PBS中溶解500毫克盐酸丙美卡因的制备盐酸丙美卡因0.5％的溶液（用于局部镇痛）。 </li><li>通过溶解8克氯化钠，0.2克的KCl，1.44克Na 2 HPO 4，和0.23克的NaH 2 PO 4在900ml蒸馏水H 2 O的制备1×PBS中调整至7.4的pH值。使溶液至1,000 ml的终体积用蒸馏水H 2 O和过滤器，以确保无菌。 </li><li>通过在1ml的DMSO中溶解26.4毫克SAHA的制备在〜100毫米1000×储液SAHA在二甲亚砜（DMSO）中。稀释至1倍（〜10微米）溶液在过滤的PBS中为每个使用。存储该原液，在-20℃下长达一个月。注意：DMSO是一种已知的毒素和致突变物。 </li><li>制备多聚甲醛（PFA）固定为4％的溶液。在化学罩，溶解4克煤灰，90毫升的PBS。使混合物达到65℃并滴定至pH为7.4。一旦PFA溶解，使溶液至100毫升的最终体积。储存于4℃最多一个月。注意：PFA是一种已知的过敏，致癌性和毒性。 </li><li>通过稀释5毫升山羊血清和500微升的Triton X-100的进94.5毫升PBS中制备1X封闭缓冲液。 </li><li>通过稀释500微升的Triton X-100的进99.5毫升PBS中制备1X洗涤缓冲液。 </li></ol> 2。碱烧伤与复合治疗<ol><li>浸泡一个圆形滤纸，〜2毫米的直径，在1M的NaOH溶液。 </li><li>麻醉小鼠注射100毫克/千克氯胺酮和5毫克/公斤甲苯噻嗪，WHICH是〜每25克鼠标步1.3 100微升麻醉鸡尾酒。麻醉应约1-2分钟，以设置英寸麻醉深度可轻轻捏动物的脚趾或尾巴来确定，如果麻醉足以应该没有回应。注意事项：应注意确保鼠标不屈服于麻醉诱导低温。 </li><li>使用滴管，降幅达0.5％盐酸丙美卡因局部适用于角膜表面进行局部麻醉。 </li><li>用无菌镊子，拿起一块氢氧化钠浸泡滤纸。如果您发现过量的NaOH抱住或滤纸滴落，简单地点击在干燥的滤纸浸泡过的滤纸吸收多余的。将一块NAOH浸泡滤纸上的角膜中央。搁置30秒，以产生急性碱烧伤〜2×2平方毫米的面积。手术显微镜是有助于正确放置滤纸。注意：只有一只眼睛的小鼠的Should与其他作为控制受伤。 </li><li>取出滤纸。使用10毫升注射器，轻轻冲洗眼睛10毫升的1X PBS洗两次，洗去残留的1 M氢氧化钠。 </li><li>立即申请一滴1X萨哈工作液（或车辆控制由PBS稀释DMSO不含萨哈）局部角膜。重复应用3x/day 14天。注意：在此期间，局部用抗生素软膏不推荐，因为它可能与损伤的发展，并且该化合物的递送干扰。使用液体抗生素溶液，如3％庆大霉素溶液代替。 </li><li>直接进行临床评估（以下协议3）或牺牲鼠标和去核的眼睛角膜平山（协议4以下）或常规石蜡/冰冻病理。 </li></ol> 3。临床评估<ol><li>以盲法进行小鼠每天检查手术显微镜下和点Corneal NV基于角膜混浊，内华达州，和容器的大小。使用至少两个观测并记录最后的得分，它是2的平均值。 <ol><li>分数角膜混浊的规模为0-4。 0 =完全清楚; 1 =轻微混浊，虹膜和瞳孔清晰可见; 2 =轻微不透明，虹膜和瞳孔仍检测; 3 =不透明，学生几乎检测不到; 4 =完全不透明的，没有考虑到学生的。 </li><li>分数NV的规模为0-3。 0 =无新生血管; 1 =新血管在角膜缘; 2 =新生血管跨越角膜缘，接近角膜中心; 3 =新生血管跨过角膜中心。 </li><li>分数容器大小的规模为0-3。 0 =无新生血管; 1 =新生血管的手术显微镜下检测; 2 =新生血管手术显微镜下很容易看到; 3 =新生血管很容易看到没有显微镜。 </li></ol></li><li>使用数码相机，带眼的代表性图像在7天和14天。 </li></ol> 4。角膜染色和平板支架<ol><li>修复去核的眼睛在4％PFA至少1小时，在4°C。 </li><li>转眼球为1x PBS中，使用镊子小心取出多余的组织。 </li><li>与手术显微镜的辅助下，可以使用一针（18号）或微刀穿孔眼的pericorneal区域（注意：这应该从眼睛释放流体）。 <ol><li>从在第4.3.1节提出的穿孔使用一对外科剪刀从后部部分切开眼（角膜）的前部。 </li></ol></li><li>传输角膜放回4％PFA供固定过夜，4℃。 </li><li>丢弃4％PFA（注意：PFA是危险的，必须按照制度规定进行报废处理）和冲洗角膜三次PBS洗涤，去除残留的煤灰。 </li><li>孵育在1x封闭缓冲液中至少2小时，在室温下透化的组织，并防止第一抗体的非特异性结合。 <ol><li>在1x封闭液适用于初级抗体，在100-500倍稀释。 （ 例如大鼠抗小鼠PECAM-1，用于检测血管1:100，兔抗小鼠-LYVE-1为用于检测巨噬细胞中检测淋巴管在1:500和/或大鼠anti-mouse-F4/80在1:100）。孵育在4℃下过夜。 </li><li>在室温下，每次洗6次用1×洗涤缓冲液1小时，以充分除去未结合的一抗。 </li><li>在1x封闭缓冲液中以500-1000倍稀释应用的二次抗体。 （ 例如，488nm的荧光标记的山羊抗大鼠IgG抗体在1:800或594 nm的荧光标记的山羊抗兔IgG抗体在1:800）。孵育在4℃下过夜。 </li><li>用1×PBS，每次1小时，在室温下洗涤​​3次，以完全除去未结合的第二抗体。 </li></ol></li><li>转移角膜到新鲜的1X PBS，并与手术显微镜的帮助下，仔细并使四片刀口从外围向仙呃。这应该划分角膜分成四个象限约等于大小（由此产生的形状看起来应该很像蝴蝶），并允许它平躺在一张幻灯片。 </li><li>安装带有安装介质适用于荧光成像。为获得最佳结果，样品应立即成像和拍摄的数字照片，但也可以存储于避光，4℃几周</li></ol>

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作者什么都没有透露。

这里介绍的协议导致hemangiogenesis，淋巴管生成和炎症的重现性水平，使其成为理想的系统来研究这三个（相互）的进程。虽然这种方法产生了集中角膜NV，已开发造成更多的定向NV，即缝合角膜17和植入表达丸粒18的生长因子的几种方法，也可能有兴趣。我们的协议是专为在成年小鼠的使用，提供了一个易于使用的动物模型，同时还允许一个实验室采取了一系列尚未公布的大型哺乳?...

角膜盲是导致失明的第四个最常见的原因，负责所有案件1约4％。角膜新生血管（NV）在很多这些疾病，包括疱疹性角膜炎（失明的西方主导的感染原因）和沙眼（感染性失明的主要原因之一）2的显著作用。目前的治疗包括类固醇，非类固醇消炎药（NSAID），抗-VEGF治疗药物，以及环孢菌素A以及常规或激光外科手术技术3。然而，角膜基于NV的病症的严重衰弱的性质，能够治疗角膜NV的手术设施，以及缺乏一个表现强劲的药理选项的缺乏导致最近专家圆桌会议的结论是，尽管现存的治疗，最根本的医疗需求通过这些疾病仍然呈现未满足4。 人的角膜共有5层，3层细胞（上皮，基质和内皮细胞）和2接口（鲍曼膜和弹力膜）。它作为机械屏障和折射面的眼睛。其透明的本质是其组成部分的一种微妙的平衡的结果，是不可或缺的正常功能5。通常无血管，角膜接收血液从沿其外缘从睫状体和眼动脉供给运行微血管。当刺激促进这些船只的血管新生，让他们成长对角膜的中心，因此限制了视力6角膜NV的发生。角膜血管生成包括hemangiogenesis和淋巴管生成，从而导致血管和淋巴管从对角膜的中心角膜缘血管街机向内生长。这导致角膜“血管生成特权”，增加角膜混浊及纤维化，角膜拉破坏明细Y一代和水肿7。角膜NV的精确触发很多，从传染病的响应，如沙眼的化学诱导的状态造成的传统药物，工业化学品，甚至是化学战剂。 这一过程的分子机制都没有，迄今为止，充分的特点;然而，一些关键球员已经确定。在正常条件下的角膜具有独特的“血管生成特权&#39;通过抗血管生成因子（例如可溶性VEGF-R1）8冗余阵列保持。然而，响应于外部刺激（例如，损伤），会有的促血管生成因子（ 例如 VEGF-A）的局部表达上调。这提示促和抗血管生成的因素的基础角膜的血管生成的特权，并导致hemangiogenesis，lymphangeogenesis和炎症，因此造成角膜病变，甚至失明9的平衡。 &lt;P类=“j​​ove_content”&gt;鉴于这种高度衰弱的病理未满足的医疗需求，这是感兴趣的领域有角膜NV的可靠动物模型。在这里，我们提出这样一个模式：控制碱烧伤。基于使用滤纸圈各种眼损伤模型自20世纪70年代10已被使用。 1989年，一群哈佛医学院眼科特点的中央角膜碱烧伤的标准模型兔基于浸泡一块圆形滤纸用氢氧化钠（NaOH），并把它应用到角膜在一个特定的范围内浓度11。自那时以来，这种技术已被改编为在小鼠12-14使用。近日，王实验室研究组蛋白去乙酰化酶（HDAC）抑制剂SAHA在使用鼠标角膜碱烧伤模型角膜15内华达州的发病的治疗效果。这里介绍的小鼠角膜碱烧伤模型的方法是建主要是对其他两篇论文14,16的前期工作。

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			<td height="21" style="height:21px;width:216px;">1 mL Syringe</td>
			<td style="width:96px;">BD</td>
			<td align="right" style="width:119px;">309659</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">18 Guage Needle</td>
			<td style="width:96px;">BD</td>
			<td align="right" style="width:119px;">305918</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">10 mL Syringe</td>
			<td style="width:96px;">BD</td>
			<td align="right" style="width:119px;">306575</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">25 Guage Needle</td>
			<td style="width:96px;">BD</td>
			<td align="right" style="width:119px;">305916</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Anti-F4/80 (rat anti-mouse)</td>
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			<td>MCA497RT</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Anti-LYVE-1 (rabbit anti-mouse)</td>
			<td style="width:96px;">Abcam</td>
			<td style="width:119px;">ab14917</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Anti-PECAM-1 (rat anti-mouse)</td>
			<td style="width:96px;">BD</td>
			<td align="right" style="width:119px;">553370</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Anti-IgG Alexa488 (goat anti-rat)</td>
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			<td style="width:119px;">12-548-B</td>
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			<td style="width:96px;">Sigma</td>
			<td style="width:119px;">D4540-1L</td>
			<td>Caution: Mutagenic, Toxic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Forceps (Blunt), Iris</td>
			<td style="width:96px;">WPI</td>
			<td align="right" style="width:119px;">15915</td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">Forceps (Sharp), Dumont #4</td>
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			<td>Controlled substance, proper license required for use.</td>
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			<td height="21" style="height:21px;width:216px;">Microscope Slides</td>
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			<td height="21" style="height:21px;width:216px;">Xylazine Solution</td>
			<td style="width:96px;">MedVet</td>
			<td style="width:119px;">RXANASED-20</td>
			<td></td>
		</tr>
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an alkali-burn injury model of corneal neovascularization in the mouse

在正常条件下，角膜没有血管，这透明度是保持良好的视觉敏锐度是必不可少的。新生血管的角膜，其可通过创伤，角膜移植术或感染性疾病引起的，的（NV）的分解角膜的所谓的“血管生成特权&#39;和形成了多个视觉病症甚至可能导致失明的基础。虽然有几个可用的治疗方案，基本医疗需要角膜新生血管的病症仍然呈现未得到满足的。为了开发安全，有效，有针对性的治疗，需要角膜NV和药物干预的可靠模型。在这里，我们描述了在小鼠碱烧伤角膜新生血管模型。这个协议规定了一个控制的碱烧伤角膜损伤，施用感兴趣的药理化合物，其结果的可视化应用程序的方法。这种方法可以证明拉琴TAL为研究角膜NV和其他新生血管性疾病的机制和机会进行干预。

经过碱烧伤，角膜NV发生在一个可预见的，时间依赖性。 图1展示了无论是在未经处理的动物（ 图1A），并与HDAC抑制剂SAHA（ 图1B对待动物之间的新生血管和角膜混浊形成了鲜明的差异）在7天的时间点。 图2A和2B展示未处理的对照眼原发性PECAM-1和LYVE-1染色和二抗Alexa Fluor 488和594染色（分别）的角膜平面安装。 <stro...

Watch this Scientific Journal Video about An Alkali-burn Injury Model of Corneal Neovascularization in the Mouse at JoVE.com

An Alkali-burn Injury Model of Corneal Neovascularization in the Mouse

新生血管的角膜（内华达州）可复杂多视觉病症。利用控制，碱烧伤模型，角膜NV的量化级别可以生产为新生血管性疾病的潜在治疗角膜NV和评价机理研究。

角膜新生血管在小鼠碱烧伤模型

Under normal conditions, the cornea is avascular, and this transparency is essential for maintaining good visual acuity. Neovascularization (NV) of the ...

an-alkali-burn-injury-model-of-corneal-neovascularization-in-the-mouse

Research

JoVE Journal

Medicine

16.4K 次观看.  Tulane University. 新生血管的角膜（内华达州）可复杂多视觉病症。利用控制，碱烧伤模型，角膜NV的量化级别可以生产为新生血管性疾病的潜在治疗角膜NV和评价机理研究。 

视频: An Alkali-burn Injury Model of Corneal Neovascularization in the Mouse

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 ...

科研

医学

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

新生儿小鼠脊髓压迫损伤模型

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

A Mouse Model for Laser-induced Choroidal Neovascularization

The mouse laser-induced choroidal neovascularization (CNV) model has been a crucial mainstay model for neovascular age-related macular degeneration (AMD) research. By administering targeted laser injury to the RPE and Bruch&#8217;s membrane, the procedure induces angiogenesis, modeling the hallmark pathology observed in neovascular AMD. 
First developed in non-human primates, the laser-induced CNV model has come to be implemented into many other species, the most recent of which being the mouse. Mouse experiments are advantageously more cost-effective, experiments can be executed on a much faster timeline, and they allow the use of various transgenic models. The miniature size of the mouse eye, however, poses a particular challenge when performing the procedure. Manipulation of the eye to visualize the retina requires practice of fine dexterity skills as well as simultaneous hand-eye-foot coordination to operate the laser. However, once mastered, the model can be applied to study many aspects of neovascular AMD such as molecular mechanisms, the effect of genetic manipulations, and drug treatment effects. 
The laser-induced CNV model, though useful, is not a perfect model of the disease. The wild-type mouse eye is otherwise healthy, and the chorio-retinal environment does not mimic the pathologic changes in human AMD. Furthermore, injury-induced angiogenesis does not reflect the same pathways as angiogenesis occurring in an age-related and chronic disease state as in AMD.
Despite its shortcomings, the laser-induced CNV model is one of the best methods currently available to study the debilitating pathology of neovascular AMD. Its implementation has led to a deeper understanding of the pathogenesis of AMD, as well as contributing to the development of many of the AMD therapies currently available.

Here, we present the mouse laser-induced choroidal neovascularization (CNV) protocol, an experimental model that re-creates the vascular hallmarks of neovascular age-related macular degeneration (AMD). Once mastered, it can reliably and effectively induce CNV as a model system to test various experimental measures.

小鼠模型的激光诱导脉络膜新生血管

The mouse laser-induced choroidal neovascularization (CNV) model has been a crucial mainstay model for neovascular age-related macular degeneration (AMD) ...

A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury

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

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

单一和重复轻度创伤性脑损伤的小鼠模型

Mild Traumatic Brain Injury (mTBI) can result in the acute loss of brain function, including a period of confusion, a loss of consciousness (LOC), focal ...

Puncture-Induced Iris Neovascularization as a Mouse Model of Rubeosis Iridis

We describe a model of puncture-induced iris neovascularization as a general model for noninvasive evaluation of angiogenesis. The model is also relevant for targeting neovascular glaucoma, a sight-threatening complication of diabetic retinopathy. This method is based on the induction of iris vascular response by a series of self-sealing uveal punctures on BALB/c mice and takes advantage of the postpartum maturation of mouse ocular vasculature. Mouse pups undergo uveal punctures from postnatal day 12.5, when the pups naturally open their eyes, until postnatal day 24.5. Due to the transparency of the cornea, iris vasculature can be analyzed easily through time by noninvasive in vivo methods. Furthermore, the semitransparent iris of BALB/c mice can be flatmounted for detailed immunohistologic analysis with minimal non-specific background staining. In this model, angiogenesis is mainly driven by the inflammatory and plasminogen activating systems. The puncture-induced model is the first to induce iris neovascularization in small rodents, and has the advantage of allowing direct noninvasive in vivo analysis of the angiogenic process. Moreover, the model can be combined with angiogenic modulating substances, which highlights its potential in the study of angiogenesis with an in vivo perspective.

Iris neovascularization, a common complication of ischemic retinal disease, may lead to sight-threatening neovascular glaucoma. Here, we describe a murine protocol for inducing experimental iris neovascularization that may be used for noninvasive evaluation of angiogenesis-modulating substances.

穿刺诱导的虹膜新生血管虹膜虹膜小鼠模型的建立

We describe a model of puncture-induced iris neovascularization as a general model for noninvasive evaluation of angiogenesis. The model is also relevant ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

眼毛刺角膜上皮磨损与角膜创面愈合的模型研究

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

A Novel Approach to Monitoring Graft Neovascularization in the Human Gingiva

Laser speckle contrast imaging (LSCI) is a novel method for measuring superficial blood perfusion over large areas. Since it is non-invasive and avoids direct contact with the measured area, it is suitable for monitoring blood flow changes during wound healing in human patients. Vestibuloplasty is periodontal surgery to the oral vestibule, aiming to restore vestibular depth with simultaneous enlargement of the keratinized gingiva. In this special clinical case, a split thickness flap was elevated at the first upper premolar and a xenogenic collagen matrix was adapted to the resulting recipient bed. LSCI was used to monitor the re- and neovascularization of the graft and the surrounding mucosa for one year. A protocol is introduced for the correct adjustment of microcirculation measurement in the oral mucosa, highlighting difficulties and possible failures.
The clinical case study presented demonstrated that &#8212; following the appropriate protocol &#8212; LSCI is a suitable and reliable method for following up microcirculation in a healing wound in the human oral mucosa and gives useful information on graft integration.

This study introduces a protocol for measuring microcirculation in human oral mucosa by laser speckle contrast imaging. The monitoring of wound healing after vestibuloplasty combined with a xenogenic collagen graft is presented on a clinical case.

一种监测人牙龈移植新生血管的新方法

Laser speckle contrast imaging (LSCI) is a novel method for measuring superficial blood perfusion over large areas. Since it is non-invasive and avoids ...

Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the established technique for a modified blood circuit reconstruction in a heterotopic abdominal heart transplantation model is presented. This method uses the intrathoracic inferior vena cava (IIVC) instead of the pulmonary artery of the donor heart for the anastomosis to the inferior vena cava of the recipient. It is facilitating and improving success rates for abdominal heart transplantation in mice.

A novel technique for blood circuit reconstruction in a heterotopic abdominal mouse heart transplantation model is demonstrated.

腹部小鼠心脏移植模型中的血液回路重建

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the ...

An Effective Mouse Model of Unilateral Renal Ischemia-Reperfusion Injury

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models are the only available resources that mimic the complexities of the IRI-associated damage encountered in vivo. This paper describes an effective mouse model of unilateral renal IRI that delivers highly reproducible data. Ischemia is induced by occluding the right renal pedicle for 30 min followed by reperfusion. In addition to the surgical procedure, a sequential overview of the expected physiological and histopathological changes following renal IRI will be provided by comparing data from seven different reperfusion times (4 h, 8 h, 16 h, 1 day, 2 days, 4 days, and 7 days). Critical data for planning experiments ahead, such as mean surgical time, average anesthetic consumption, and body weight changes over time, will be shared. This work will help researchers implement a reliable renal IRI model and select the appropriate reperfusion time that aligns with their intended investigative goals.

Renal ischemia-reperfusion injury is associated with high morbidity and mortality in hospitalized patients. Here, we present a simple and effective mouse model of unilateral renal ischemia-reperfusion injury and provide a sequential overview of representative pathological changes observed in the kidney.

单侧肾缺血再灌注损伤的有效小鼠模型

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models ...