Department of Pain Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000 Guizhou, China.
Guizhou Key Laboratory of Anesthesia and Organ Protection, Zunyi Medical University, Zunyi, 563002 Guizhou, China.
Biomed Res Int. 2020 Mar 1;2020:4860735. doi: 10.1155/2020/4860735. eCollection 2020.
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 ~200 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.
蓝斑核（LC）是一个多功能核团，也是大脑中去甲肾上腺素的来源。迄今为止，还没有一种简单易用的方法可以在脑切片中定位到小的 LC。在这里，我们报告了一种快速、准确且易于遵循的 protocol，用于定位冷冻切片中的小鼠 LC。固定和脱水后，将成年小鼠的完整大脑放置在水平表面上，并沿着双侧大脑皮质的后缘垂直切割。在冠状切面中，可以用肉眼轻松看到中脑导水管。将小脑部分用最佳切割温度（OCT）复合物包埋后，从切面中切取冠状脑切片，在 1mm 内，中脑导水管消失，第四脑室出现，然后收集包含 LC 的脑切片。从第一次收集开始，在~200μm 处，可以收集到去甲肾上腺素能神经元最丰富的脑切片。酪氨酸羟化酶免疫荧光染色证实了该方法定位 LC 的准确性，并可以用该方法确定去甲肾上腺素能神经元最丰富的切片。
