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Reliable Isolation of Central Nervous System Microvessels Across Five Vertebrate Groups
10:35

Reliable Isolation of Central Nervous System Microvessels Across Five Vertebrate Groups

Published on: January 12, 2020

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Straightforward method for singularized and region-specific CNS microvessels isolation

Jacquelyn Rose Dayton1, Marissa Cindy Franke1, Yinyu Yuan1

  • 1University of California, Davis. Anatomy, Physiology & Cell Biology, 1089 Veterinary Medicine Drive, Davis, CA, 95616, United States.

Journal of Neuroscience Methods
|February 25, 2019
PubMed
Abstract

Insights

This study introduces a new method for isolating mouse central nervous system (CNS) microvessels from specific regions, enabling individual analysis without pooling samples. This technique also extends to non-human primate CNS, advancing blood-brain barrier research.

Area of Science:

  • Neuroscience
  • Vascular Biology
  • Biotechnology

Background:

  • Current murine brain microvasculature isolation methods necessitate pooling samples and exclude non-cortical tissues, hindering single-individual analysis.
  • Existing techniques are limited to specific CNS regions, preventing comprehensive neurovascular studies.

Purpose of the Study:

  • To develop a region-specific method for isolating murine central nervous system (CNS) microvessels.
  • To enable comparative analysis of neurovascular units within and between individual animals and treatment groups.
  • To adapt the isolation technique for non-human primate CNS tissue.

Main Methods:

  • A novel approach using a double-pronged pick, elution, and filtration through cell strainers after centrifugation was employed.
  • This method avoids enzymatic dissociation and ultracentrifugation.
  • The technique was successfully adapted for macaque CNS tissue.

Main Results:

  • Region-specific isolation of murine CNS microvessels was achieved, allowing for analysis without pooling.
  • The method facilitates comparisons of neurovascular units across different CNS regions within the same individual.
  • Adaptation to macaque CNS tissue was successful, offering translational potential.

Conclusions:

  • Region-specific microvessel isolation reveals molecular and cellular differences previously obscured by pooling or exclusion of non-cortical tissues.
  • This approach unmasks individual variations, improving study robustness.
  • Application to non-human primate CNS enhances the translational relevance for blood-brain barrier (BBB) research.
Keywords:
Astrocytic end-feetBlood-brain barrierEndothelial cellJunctional proteinsMicrovasculatureNeurovascular unitPericyte

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