Measuring Blood-brain-barrier Permeability Using Evans Blue in Mice
伊文思蓝检测小鼠中血脑屏障的渗透性
发布: 2015年08月05日第5卷第15期 DOI: 10.21769/BioProtoc.1548 浏览次数: 24599
评审: Soyun KimTifany DesprezEmmanuelle Berret
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参见作者原研究论文
The authors used this protocol in:
Jan 2015
Abstract
The blood–brain barrier (BBB) is a highly selective permeability barrier that separates the circulating blood from the brain extracellular fluid in the central nervous system. The blood–brain barrier allows the passage of water, some gases, and lipid soluble molecules by passive diffusion, as well as the selective transport of molecules such as glucose and amino acids that are crucial to neural function. This protocol provides a full, detailed method for measuring blood-brain-barrier permeability of mice with Evans blue (EB), that leakage was used to assess blood-brain barrier (BBB) permeability (sample: Anterior Cingulate Cortex, ACC).
Materials and Reagents
- Adult male Kunming mice (the Experimental Animal Center of Xuzhou Medical College, 20-22 g, 5-7 weeks old)
- Evans blue (EB) (Sigma-Aldrich, catalog number: E2129-10G )
- Trichloroacetic acid (Sigma-Aldrich, catalog number: T6399-100G )
- Chloral hydrate (Sigma-Aldrich, catalog number: 15307-500G-R )
- Paraformaldehyde (Sigma-Aldrich, catalog number: 158127-500G )
- NaCl (Sigma-Aldrich, catalog number: S7653-1KG )
- 10% chloral hydrate (see Recipes)
- 4% paraformaldehyde-freshly-prepared (see Recipes)
- 0.9% NaCl (see Recipes)
- 100% trichloroacetic acid solution (see Recipes)
Equipment
- Spectrophotometer (Thermo Fisher Scientific, catalog number: 1510 )
- 96-well plate (Corning Incorporated, catalog number: 3599 )
- 1 and 20 ml syringe (local drugstore)
- Infusion apparatus (local drugstore)
Procedure
文章信息
版权信息
© 2015 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
- Yang, J., Jiang, Y. and Guo, Y. (2015). Measuring Blood-brain-barrier Permeability Using Evans Blue in Mice. Bio-protocol 5(15): e1548. DOI: 10.21769/BioProtoc.1548.
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Yang, J. X., Hua, L., Li, Y. Q., Jiang, Y. Y., Han, D., Liu, H., Tang, Q. Q., Yang, X. N., Yin, C., Hao, L. Y., Yu, L., Wu, P., Shao, C. J., Ding, H. L., Zhang, Y. M. and Cao, J. L. (2015). Caveolin-1 in the anterior cingulate cortex modulates chronic neuropathic pain via regulation of NMDA receptor 2B subunit. J Neurosci 35(1): 36-52.
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分类
神经科学 > 神经解剖学和神经环路 > 动物模型
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