Name: establishment of acute pontine infarction in rats by electrical stimulation
Uploaded: 2020-08-27T13:00:00
Description: Watch this Scientific Journal Video about Establishment of Acute Pontine Infarction in Rats by Electrical Stimulation at JoVE.com

这项研究得到了中国自然科学基金（81471181和81870933）和江苏省自然科学基金（第81601011号）的财政支持（BK20160345）给朱J和广州市卫生局科学计划（20191A011083）给邱Z.

该协议经广州医科大学第二附属医院动物护理与使用委员会审核通过。该委员会是AAACI认可的机构。这些老鼠是由南方医科大学动物中心提供的。
1. 动物
<ol><li>使用体重为250~10克的成年雄性斯普拉格-道利大鼠。</li>
	<li>运输时，在控制环境条件下，在环境温度为25°C、相对湿度为65%和12小时/12小时光/暗循环的受控环境条件下，将大鼠在手术前至少1周。</li>
	<li>提供食物和?...

<ol>
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本研究为生成急性庞汀梗死大鼠模型提供了一种方案。该模型可用于研究蓬汀中风患者的预后和康复（包括中风后慢性疼痛）。
这种方法有几个优点。首先，它为今后的研究提供了急性庞汀梗死的老鼠模型。如上所述，庞汀梗塞是一种常见的中风亚型，受到的关注较少。中风研究的一个主要缺点是缺乏特定的庞汀梗死模型。其次，与BA15、16的结扎现有庞汀?...

自 20 世纪 80 年代以来，硅胶导的中脑动脉闭塞（ MCAO ）模型在基础中风研究中得到了广泛的应用。还使用了其他方法（即：MCA2的一个分支的苏化和光化学诱发的焦梗死）。这些模型被人们从中可看做为基于MCA的中风模型，对研究中风和潜在治疗的病理生理机制大有贡献。虽然这些实验模型3、4,有局限性，但这些方法已被许多实验室5、6,使用。基于 MCA 的笔画模型表示前循环中的笔画;然而，很少有报道调查了模型模仿中风的后循环7。
前循环中风的病因、机理、临床表现和预后有显著差异。因此，前循环笔触模型得出的结果不能应用于后循环笔画。例如，前循环的再灌注时间窗口已延长至6小时，根据成像结果9，研究部分延长至24小时。然而，根据之前的报告10 和我们自己的临床经验，后循环的时间窗口可能超过24小时。这个拉长的再灌注时间窗口必须在实验模型中进一步研究和确认。
关于后循环中风，庞汀梗塞是最常见的亚型，占所有缺血性中风病例的71例，12例。根据梗塞地形，庞汀梗塞分为隔离和非隔离的庞汀梗塞13。孤立的庞汀梗塞根据基本机制分为三种类型：大动脉疾病（LAD）、罗勒动脉分支病（BABD）和小动脉疾病（SAD）。对14例庞汀梗死的临床调查，对庞汀梗死的机制、表现和预后有一些了解。然而，模仿庞汀梗死的啮齿动物模型却较少受到研究。
在以前的研究中，涉及庞的扩散脑干分段损伤已被探索7。一组试图通过连接罗勒动脉（BA）15，创建一个庞汀梗死模型。另一组使用10-0尼龙单丝缝合有选择地将近极BA的四点选择性地（16）的四点。此模型模仿 LAD， 但大多数庞廷梗死源于 BABD 和 SAD。此外，BA的选择性结扎是一个复杂的手术，具有很高的死亡率。
此处提供了一个详细的协议，用于通过电刺激进行易于执行、易于复制和成功的大鼠模型急性庞汀梗死。

<table border="0" cellpadding="0" cellspacing="0" style="width:664px;" width="663">
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	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">4-0 sucture</td>
			<td style="width:121px;">Shanghai Jinzhong</td>
			<td style="width:161px;"></td>
			<td style="width:167px;">Surgical instruments</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Adhesive tape</td>
			<td style="width:121px;">Shanghai Jinzhong</td>
			<td style="width:161px;"></td>
			<td>Surgical instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Animal anesthesia system</td>
			<td style="width:121px;">RWD</td>
			<td style="width:161px;"></td>
			<td>Wear mask when using the system</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Bone cement</td>
			<td style="width:121px;">Shanghai Jinzhong</td>
			<td style="width:161px;"></td>
			<td>Surgical instrument</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Cured clamp</td>
			<td style="width:121px;">Shanghai Jinzhong</td>
			<td style="width:161px;"></td>
			<td>Surgical instrument</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">General tissue scissors</td>
			<td style="width:121px;">Shanghai Jinzhong</td>
			<td style="width:161px;"></td>
			<td>Surgical instrument</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">IndoPhors</td>
			<td style="width:121px;">Guoyao of China</td>
			<td style="width:161px;"></td>
			<td>Sterilization</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Isoflurane</td>
			<td style="width:121px;">RWD</td>
			<td align="right" style="width:161px;">217181101</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Lesion Making Device</td>
			<td style="width:121px;">Shanghai Yuyan</td>
			<td style="width:161px;"></td>
			<td>Making a lesion</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">MRI system</td>
			<td style="width:121px;">Bruker Biospin</td>
			<td style="width:161px;"></td>
			<td>Confirmation of infarction in vivo</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Needle holder</td>
			<td style="width:121px;">Shanghai Jinzhong</td>
			<td style="width:161px;"></td>
			<td>Surgical instrument</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Penicilin</td>
			<td style="width:121px;">Guoyao of China</td>
			<td style="width:161px;"></td>
			<td>Infection Prevention</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Probe</td>
			<td style="width:121px;">Anke</td>
			<td style="width:161px;"></td>
			<td>Need some modification</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Q-tips</td>
			<td style="width:121px;">Shanghai Jinzhong</td>
			<td style="width:161px;"></td>
			<td>Surgical instrument</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Shearing scissors</td>
			<td style="width:121px;">Shanghai Jinzhong</td>
			<td style="width:161px;"></td>
			<td>Surgical instrument</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">Stereotaxic apparatus</td>
			<td style="width:121px;">RWD</td>
			<td style="width:161px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Suture needle</td>
			<td style="width:121px;">Shanghai Jinzhong</td>
			<td style="width:161px;"></td>
			<td>Surgical instrument</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:215px;">Tissue holding forcepts</td>
			<td style="width:121px;">Shanghai Jinzhong</td>
			<td style="width:161px;"></td>
			<td>Surgical instrument</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:215px;">TTC</td>
			<td style="width:121px;">Sigma-Aldrich</td>
			<td style="width:161px;">BCBW5177</td>
			<td>For infarction confirmation in vitro</td>
		</tr>
	</tbody>
</table>

establishment of acute pontine infarction in rats by electrical stimulation

庞汀梗死是后循环中最常见的中风亚型，而缺乏模仿庞汀梗死的啮齿动物模型。提供此为成功建立急性庞汀梗死大鼠模型的协议。使用重约250克的老鼠，使用立体气器将带绝缘护套的探针注入到庞体内。病变是由单个脉冲的电刺激产生的。Longa 分数、伯德森分数和梁平衡测试用于评估神经缺陷。此外，粘合去除体感测试用于确定感官运动功能，肢体放置测试用于评估自感知。然后使用 MRI 扫描来评估体内的梗塞，TTC 染色用于确认体外梗死。在这里，一个成功的梗死被识别，位于栖息的龙的外端基础。最后，提出了一种建立急性庞汀梗死大鼠模型的新方法。

六只动物接受了上述手术方案。图 4 所示的 对照组 由六只大鼠组成。图 4 所示的大脑 切片来自 每组一只大鼠。
MRI扫描显示，梗死位于庞的基座（图4A）。由于探头被注射到中线左侧 2 mm，因此梗塞被横向定位。这种梗死模仿了患者的异端蓬汀梗塞（图4A）。由于使用了绝缘护套，因此?...

Watch this Scientific Journal Video about Establishment of Acute Pontine Infarction in Rats by Electrical Stimulation at JoVE.com

Establishment of Acute Pontine Infarction in Rats by Electrical Stimulation

此处介绍的是通过单个脉冲的电刺激在大鼠模型中建立急性庞汀梗死的协议。

电刺激在大鼠中建立急性庞廷梗死

Pontine infarction is the most common stroke subtype in the posterior circulation, while there lacks a rodent model mimicking pontine infarction. Provided ...

我们提供一种协议，在一只老鼠中建立梗死。这种庞廷梗死大鼠模型易于执行，易于复制，并且非常成功的模型。电刺激可用于在任何其他脑区，如丘脑区进行梗死。想要执行此模型的人应在实验前学习庞廷自主性。手术前，根据随附文本中描述的行为测试，对大鼠进行称重，并评估其神经表现。麻醉前将加热垫预热至 37 摄氏度，然后将头骨钻头连接到立体轴架上的支架上。确认第一只麻醉大鼠对踏板反射缺乏反应，将大鼠安装在加热垫上容易位置的立体轴上。将耳杆放在耳道上方以固定头部，注意保持头骨水平，以避免注射出现任何倾斜。将软膏涂抹在动物的眼睛上，并使用微型剃须刀从头骨上去除头发。使用标记沿头骨的中线绘制一条三厘米的线，从双边侧管线绘制到后侧方体后面 0.5 厘米，并使用手术刀沿标记线进行中线切口。使用棉签去除任何血液，并在皮瓣的两侧放置一块手术胶带以暴露头骨。使用浸有 0.9% 氯化钠的新拭子轻轻取出头骨上的任何结缔组织，并找到骨骼。使用细尖笔指示啤酒的中心点作为原点，将钻头放在前六毫米，中侧侧两毫米处。使用钻头仔细执行一毫米直径的颅骨切除术。接下来，将钻头替换为 22 表带探头，并采用绝缘护套，尖端位于护套的近端上方两毫米。将纸张插入大脑七毫米，然后沿着护套推进探针，直到探针顶部低于大脑表面九毫米。将电极连接到电气刺激器，然后将阳极连接到探头。然后连接到阴极到老鼠的耳朵。接下来，打开电气刺激器，将单脉冲宽度设置为 4，050 毫秒，将电压设置为 50 伏，将电流设置为 4 毫安。在刺激后将探针留在位置五分钟，然后从大脑中取出设备。用骨水泥盖住颅骨切除术，让水泥在使用4.0聚酰胺缝合丝关闭伤口之前变干。三或四针后，打2-1-1标准手术结。然后内向大鼠注射250微升青霉素，每15分钟监测一次，直到完全清醒，然后将动物返回到笼子。在这项分析中，六只动物接受了手术，六只动物被当作对等控制动物。这些图像中的大脑切片来自每组一只大鼠。MRI扫描显示，梗死位于模拟人类患者异端梗塞的龙的基础上。因为使用了绝缘护套，所以在皮层、小脑或中脑中没有观察到梗塞。扩散加权图像也揭示了急性庞汀梗死。TTC染色用于确认手术后24小时梗死。正如预期的那样，与对照组相比，手术组的梗塞量要高得多。手术前和手术后的行为得分被测量。与对照组相比，有庞汀梗塞的老鼠在左边盘旋。在 Longa、Berderson、肢体放置、梁平衡和胶粘剂去除躯体感官测试分数中观察到显著差异，这些分数与庞汀梗死大鼠对照组大鼠的咆哮。任何特定的大脑区域将被电刺激摧毁。

Research

JoVE Journal

Neuroscience

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

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

Assessment of Neuromuscular Function Using Percutaneous Electrical Nerve Stimulation

神经肌肉功能的应用经皮神经电刺激评估

Percutaneous electrical nerve stimulation is a non-invasive method commonly used to evaluate neuromuscular function from brain to muscle (supra-spinal, ...

Caspase-3 Activity in the Rat Amygdala Measured by Spectrofluorometry After Myocardial Infarction

Caspase-3活性在鼠杏仁核测量用荧光分析法心肌梗死后

Myocardial infarction (MI) has dramatic mid- and long-term consequences at the physiological and behavioral levels, but the mechanisms involved are still ...

Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays

时间依赖性增加对微电极阵列刺激神经元细胞培养物的网络响应

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network ...

Transcranial Electrical Brain Stimulation in Alert Rodents

警惕啮齿动物的经颅电脑刺激

Transcranial electrical brain stimulation can modulate cortical excitability and plasticity in humans and rodents. The most common form of stimulation in ...

Chronic Transcranial Electrical Stimulation and Intracortical Recording in Rats

大鼠慢性经颅电刺激及 Intracortical 记录

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a ...

Sub-acute Cerebral Microhemorrhages Induced by Lipopolysaccharide Injection in Rats

脂多糖注射液致大鼠亚急性脑 Microhemorrhages

Cerebral microhemorrhages (CMHs) are common in aged patients and are correlated to various neuropsychiatric disorders. The etiology of CMHs is complex, ...

Assessing Pupil-linked Changes in Locus Coeruleus-mediated Arousal Elicited by Trigeminal Stimulation

评估由三角刺激引起的由Locus Coeruleus介导的觉醒中与学生相关的变化

Current scientific literature provides evidence that trigeminal sensorimotor activity associated with chewing may affect arousal, attention, and cognitive ...

A Preclinical Model to Assess Brain Recovery After Acute Stroke in Rats

评估大鼠急性中风后脑恢复的临床前模型

The purpose of this study was to establish and validate an animal brain ischemia model in the recovery and sequela stages. A middle cerebral artery ...

Preparation of Peripheral Nerve Stimulation Electrodes for Chronic Implantation in Rats

大鼠慢性植入的外周神经刺激电极的制备

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. ...