Reynolds D S, Morton A J
Department of Pharmacology, University of Cambridge, UK.
J Neurosci Methods. 1998 Jan 31;79(1):115-21. doi: 10.1016/s0165-0270(97)00168-4.
A simple method for measuring changes in blood-brain barrier (BBB) permeability following neurotoxic lesions is described. In the brains of animals perfused transcardially with a trypan blue solution at the time of sacrifice, the presence of trypan blue staining correlated with changes in BBB function seen with more traditional markers, such as albumin staining. Thus, trypan blue appears to be useful as a marker for changes in BBB permeability. We have used this method to show increases in BBB permeability in striatal lesions induced by three different neurotoxins: chronic systemic injection of 3-nitropropionic acid (3-NP) and intrastriatal injection of either quinolinic or kainic acid. Trypan blue staining was seen in all three types of lesion, with both the neuropil and some neurones being stained. In the kainic acid lesioned animals, trypan blue also stained hippocampal and cortical neurones which are known to degenerate. Our findings suggest that trypan blue makes a more sensitive marker than albumin for both BBB integrity changes and degenerating neurones. Furthermore, this method has the advantages over others of being quick, economic and compatible with most subsequent histological and immunocytochemical staining.
本文描述了一种用于测量神经毒性损伤后血脑屏障（BBB）通透性变化的简单方法。在处死动物时经心内灌注台盼蓝溶液，台盼蓝染色的存在与用更传统的标记物（如白蛋白染色）所观察到的血脑屏障功能变化相关。因此，台盼蓝似乎可用作血脑屏障通透性变化的标记物。我们已使用该方法显示了由三种不同神经毒素诱导的纹状体损伤中血脑屏障通透性的增加：慢性全身注射3-硝基丙酸（3-NP）以及纹状体内注射喹啉酸或 kainic 酸。在所有三种类型的损伤中均可见台盼蓝染色，神经纤维网和一些神经元均被染色。在 kainic 酸损伤的动物中，台盼蓝还对已知会发生变性的海马和皮质神经元进行了染色。我们的研究结果表明，对于血脑屏障完整性变化和变性神经元，台盼蓝是比白蛋白更敏感的标记物。此外，该方法比其他方法具有快速、经济且与大多数后续组织学和免疫细胞化学染色兼容的优点。
