Novel modified method for injection into the cerebrospinal fluid via the cerebellomedullary cistern in mice
Novel modified method for injection into the cerebrospinal fluid via the cerebellomedullary cistern in mice
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DOI：
10.55782/ane-2013-1938
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发表时间：
2013-01-01
影响因子：
1.4
通讯作者：
Saito, Nobuhito
中科院分区：
文献类型：
--
作者：
Chen, Yili;Imai, Hideaki;Saito, Nobuhito
本文介绍了一种改良的小鼠小脑延髓池注射方法，即在吸入麻醉下用特制面罩固定小鼠，暴露颅骨矢状缝和项中线，使注射点直观可见。通过脑池内注射6 μ l亚甲基蓝水溶液，利用染料在颅内的时间和空间分布评价改良方法的准确性。注射后1小时，在CM池、腹侧池和主要脑神经的颅内近端部分发现高浓度的染料。CM池和腹侧池中的染料颜色较浅，并且在注射后6小时染料已到达主要颅神经的颅内远端部分。在注射后24小时，通过脑脊液（CSF）循环完全消除染料。在20只小鼠中没有发现严重的脑损伤。根据染料分布的确认，在注射后1小时或6小时处死的所有14只小鼠中，脑池内注射均成功。本实验观察了内皮素-1（ET-1）中枢给药对小鼠脑血供、脑动脉收缩和呼吸变化的影响。研究了三种剂量的ET 1：2 μ g（0.8 nmol）、4 μ g（1.6 nmol）和6 μ g（2.4 nmol）。注射后使用激光多普勒探头监测脑血流量（CBF）60分钟。脑池内注射ET 1可引起剂量依赖性的CBF减少、脑动脉收缩和呼吸抑制。这种在直接可视化下注射到CM池中的改进方法提供了准确和可重复的注射到CSF中，并且可用于研究各种化学物质对小鼠中枢神经系统的影响。
A modified method of injection into the cerebellomedullary (CM) cistern of mice was developed based on fixation of the mouse with a special mask under inhalation anesthesia, and exposure of the sagittal suture of the cranium and midline of the nape to allow us to visualize injection point directly. The accuracy of the modified method was evaluated using the temporal and spatial intracranial distribution of dye by intracisternal injection of 6-mu l methylene blue aqueous solution. A high concentration of dye was found in the CM cistern, the ventral cisterns, and intracranial proximal portion of the main cranial nerves at 1 hour after injection. The color of the dye in the CM cistern and the ventral cisterns was lighter, and the dye had reached the intracranial distal portion of the main cranial nerves at 6 hours after injection. The dye was completely eliminated by cerebrospinal fluid (CSF) circulation at 24 hours after injection. No severe brain injury was found in any of the 20 mice. Intracisternal injection was successful in all 14 mice sacrificed 1 hour or 6 hours after injection according to the confirmation of dye distribution. The effects of central administration of endothelin-1 (ET1) were evaluated on cerebral blood supply, constriction of cerebral arteries, and change of respiration in mice. Three doses of ET1 were studied: 2 mu g (0.8 nmol), 4 mu g (1.6 nmol), and 6 mu g (2.4 nmol). Cerebral blood flow (CBF) was monitored for 60 minutes following injection using a laser Doppler probe. Intracisternal ET1 injection induced dose-dependent reduction of CBF, constriction of cerebral arteries, and respiratory depression in mice. This modified method of injection into the CM cistern under direct visualization provides accurate and reproducible injection into the CSF, and can be used to investigate the effects of various chemical substances on the central nervous system in mice.