Zhang Lu-Lin, Yuan Yu, Liang Mei-Yu, Liu Ming-Xin, Wang Dong-Xia, Xie Jun-Xia, Song Ning
Department of Physiology and Pathophysiology, School of Basic Medicine, Institute of Brain Science and Disease, Qingdao University, Qingdao 266071, China.
CT Center, Qingdao Jimo District Hospital of Traditional Chinese Medicine, Qingdao 266200, China.
Sheng Li Xue Bao. 2023 Apr 25;75(2):197-204.
The purpose of this study was to establish a suitable method for extracting cerebrospinal fluid (CSF) from C57BL/6 mice. A patch clamp electrode puller was used to draw a glass micropipette, and a brain stereotaxic device was used to fix the mouse's head at an angle of 135° from the body. Under a stereoscopic microscope, the skin and muscle tissue on the back of the mouse's head were separated, and the dura mater at the cerebellomedullary cistern was exposed. The glass micropipette (with an angle of 20° to 30° from the dura mater) was used to puncture at a point 1 mm inboard of Y-shaped dorsal vertebral artery for CSF sampling. After the first extraction, the glass micropipette was connected with a 1 mL sterile syringe to form a negative pressure device for the second extraction. The results showed that the successful rate of CSF extraction was 83.33% (30/36). Average CSF extraction amount was (7.16 ± 0.43) μL per mouse. In addition, C57BL/6 mice were given intranasally ferric ammonium citrate (FAC) to establish a model of brain iron accumulation, and the CSF extraction technique established in the present study was used for sampling. The results showed that iron content in the CSF from the normal saline control group was not detected, while the iron content in the CSF from FAC-treated group was (76.24 ± 38.53) μmol/L, and the difference was significant. These results suggest that glass micropipette vacuum technique of CSF sampling established in the present study has the advantages of simplicity, high success rate, large extraction volume, and low bleeding rate, and is suitable for the research on C57BL/6 mouse neurological disease models.
本研究的目的是建立一种从C57BL/6小鼠中提取脑脊液（CSF）的合适方法。使用膜片钳电极拉制器拉制玻璃微吸管，并使用脑立体定位装置将小鼠头部固定在与身体呈135°角的位置。在立体显微镜下，分离小鼠头部背部的皮肤和肌肉组织，暴露小脑延髓池处的硬脑膜。使用玻璃微吸管（与硬脑膜呈20°至30°角）在Y形背侧椎动脉内侧1 mm处穿刺以采集脑脊液样本。首次提取后，将玻璃微吸管与1 mL无菌注射器连接形成负压装置进行第二次提取。结果显示，脑脊液提取成功率为83.33%（30/36）。每只小鼠的平均脑脊液提取量为（7.16±0.43）μL。此外，给C57BL/6小鼠鼻内注射柠檬酸铁铵（FAC）以建立脑铁蓄积模型，并使用本研究建立的脑脊液提取技术进行采样。结果显示，生理盐水对照组脑脊液中未检测到铁含量，而FAC处理组脑脊液中铁含量为（76.24±38.53）μmol/L，差异有统计学意义。这些结果表明，本研究建立的玻璃微吸管真空脑脊液采样技术具有操作简单、成功率高、提取量较大、出血率低等优点，适用于C57BL/6小鼠神经疾病模型的研究。
