Efficient isolation of brain capillary from a single frozen mouse brain for protein expression analysis

Seiryo Ogata, Shingo Ito, Takeshi Masuda, Sumio Ohtsuki

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)


Isolated brain capillaries are essential for analyzing the changes of protein expressions at the blood–brain barrier (BBB) under pathological conditions. The standard brain capillary isolation methods require the use of at least five mouse brains in order to obtain a sufficient amount and purity of brain capillaries. The purpose of this study was to establish a brain capillary isolation method from a single mouse brain for protein expression analysis. We successfully isolated brain capillaries from a single frozen mouse brain by using a bead homogenizer in the brain homogenization step and combination of cell strainers and glass beads in the purification step. Western blot and proteomic analysis showed that proteins expressed at the BBB in mouse brain capillaries isolated by the developed method were more enriched than those isolated from a pool of five mouse brains by the standard method. By using the developed method, we further verified the changes in expression of BBB proteins in Glut1-deficient mouse. The developed method is useful for the analysis of various mice models with low numbers and enables us to understand, in more detail, the physiology and pathology of BBB.

Original languageEnglish
Pages (from-to)1026-1038
Number of pages13
JournalJournal of Cerebral Blood Flow and Metabolism
Issue number5
Publication statusPublished - 2021 May
Externally publishedYes


  • Blood–brain barrier
  • GLUT1 deficiency syndrome
  • absolute protein expression
  • brain capillary isolation
  • quantitative protein expression profile

ASJC Scopus subject areas

  • Neurology
  • Clinical Neurology
  • Cardiology and Cardiovascular Medicine


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