Fast Localization and Sectioning of Mouse Locus Coeruleus
Fast Localization and Sectioning of Mouse Locus Coeruleus
复制标题
小鼠蓝斑的快速定位和切片。
DOI：
10.1155/2020/4860735
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发表时间：
2020-03-02
影响因子：
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通讯作者：
Yu, Tian
中科院分区：
文献类型：
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作者：
Cao, Song;Li, Juan;Yu, Tian
轨迹核（LC）是一个多功能核团，也是脑内去甲肾上腺素的来源。迄今为止，还没有简单易行的方法在脑切片中定位小LC。在这里，我们报告了一个快速，准确，易于遵循的协议，在冷冻切片小鼠LC的本地化。将成年小鼠的完整大脑固定并脱水后，置于水平表面上，沿双侧大脑皮质后缘沿着垂直切割。在冠状切面上，中脑导水管肉眼可见。采用最佳切割温度（OCT）复合物包埋小脑部分后，从切面切取冠状面脑片，在1mm以内，中脑导水管消失，第四脑室出现，收集含LC的脑片。从第一次收集开始，在近似200 μ m的条件下，可以收集去甲肾上腺素能神经元最富集的脑切片。酪氨酸羟化酶免疫荧光染色证实该方法对LC的定位准确，并且可以确定去甲肾上腺素能神经元最丰富的切片。
The locus nucleus (LC) is a multifunctional nucleus which is also the source of norepinephrine in the brain. To date, there is no simple and easy method to locate the small LC in brain sectioning. Here we report a fast, accurate, and easy-to-follow protocol for the localization of mice LC in frozen sectioning. After fixation and dehydration, the intact brains of adult mice were placed on a horizontal surface and vertically cut along the posterior margin of the bilateral cerebral cortex. In the coronal cutting plane, the aqueduct of midbrain can be seen easily with the naked eyes. After embedding the cerebellum part with optimal cutting temperature (OCT) compound, coronal brain slices were cut from the cutting plane, within 1 mm, the aqueduct of midbrain disappeared and the fourth ventricle appeared, then the brain slices contained LC and were collected. From the first collection, at similar to 200 mu m, the noradrenergic neurons' most enriched brain slices can be collected. The tyrosine hydroxylase immunofluorescence staining confirmed that the localization of LC with this method is accurate and the noradrenergic neuron most abundant slices can be determined with this method.