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C57BL/6小鼠脑脊液抽取——毛细玻璃虹吸管真空吸引技术

张鲁琳¹, 袁玉¹, 梁美钰¹, 刘明欣¹, 王冬霞^1,2, 谢俊霞¹, 宋宁^1,*

¹青岛大学基础医学院生理学与病理生理学系，脑科学与疾病研究院，青岛 266071；²青岛市即墨区中医医院CT中心，青岛 266200

摘要

本研究旨在建立一种适合C57BL/6小鼠脑脊液(cerebrospinal fluid, CSF)抽取的方法。用膜片钳电极拉取仪拉制毛细玻璃虹吸管，用脑立体定位仪将小鼠头部固定，使之与身体之间角度为135°。在立体显微镜下分离小鼠颅脑背部皮肤和肌肉组织，暴露出小脑延髓池处硬脑膜，用毛细玻璃虹吸管(与硬脑膜平面之间角度为20°~30°)在Y形脊椎背侧动脉内侧1 mm处穿刺，进行CSF采样。第一次抽取后将毛细玻璃虹吸管与1 mL无菌注射器相连，构成负压装置，进行第二次抽取。结果显示，CSF抽取成功率为83.33% (30/36)，CSF平均抽取量为(7.16 ± 0.43) μL/只。另外，经鼻给予C57BL/6小鼠枸橼酸铁胺(ferric ammonium citrate, FAC)建立脑内高铁模型，采用本研究建立的CSF抽取技术进行采样。结果显示，生理盐水对照组CSF铁含量没有检出，FAC处理组CSF铁含量为(76.24 ± 38.53) μmol/L，组间差异具有显著性。以上结果提示，本研究建立的小鼠CSF毛细玻璃虹吸管负压吸引技术具有简单易行、成功率高、抽取量多、出血率低的优点，适合在C57BL/6小鼠神经疾病模型上的相关研究。

关键词： 脑脊液; C57BL/6小鼠; 枸橼酸铁胺

A glass micropipette vacuum technique of cerebrospinal fluid sampling in C57BL/6 mice

ZHANG Lu-Lin¹, YUAN Yu¹, LIANG Mei-Yu¹, LIU Ming-Xin¹, WANG Dong-Xia^1,2, XIE Jun-Xia¹, SONG Ning^1,*

¹Department of Physiology and Pathophysiology, School of Basic Medicine, Institute of Brain Science and Disease, Qingdao University, Qingdao 266071, China；²CT Center, Qingdao Jimo District Hospital of Traditional Chinese Medicine, Qingdao 266200, China

Abstract

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.

Key words: cerebrospinal fluid; C57BL/6 mice; ferric ammonium citrate

收稿日期：　　录用日期：

通讯作者：宋宁　　E-mail: ningsong@qdu.edu.cn

DOI: 10.13294/j.aps.2023.0025

引用本文：

张鲁琳, 袁玉, 梁美钰, 刘明欣, 王冬霞, 谢俊霞, 宋宁. C57BL/6小鼠脑脊液抽取——毛细玻璃虹吸管真空吸引技术[J]. 生理学报 2023; 75 (2): 197-204.

ZHANG Lu-Lin, YUAN Yu, LIANG Mei-Yu, LIU Ming-Xin, WANG Dong-Xia, XIE Jun-Xia, SONG Ning. A glass micropipette vacuum technique of cerebrospinal fluid sampling in C57BL/6 mice. Acta Physiol Sin 2023; 75 (2): 197-204 (in Chinese with English abstract).