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Neuroscience
电刺激在大鼠中建立急性庞廷梗死
电刺激在大鼠中建立急性庞廷梗死
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Establishment of Acute Pontine Infarction in Rats by Electrical Stimulation

电刺激在大鼠中建立急性庞廷梗死

Full Text
6,191 Views
05:02 min
August 27, 2020

DOI: 10.3791/60783-v

Ming Luo1, Xiangyue Tang1, Juehua Zhu2, Zhihua Qiu1, Yongjun Jiang1

1The Second Affiliated Hospital of Guangzhou Medical University, 2Department of Neurology,The First Affiliated Hospital of Soochow University

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Summary

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。

Transcript

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。

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