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Isolation of Capillaries from Small Amounts of Mouse Brain Tissue

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Neuroprotection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2761))

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Abstract

The integrity of the blood-brain barrier (BBB) is essential for the normal functioning of the central nervous system (CNS). Isolated brain capillaries are essential for analyzing changes in protein and gene expression at the BBB under physiological and pathological conditions. The standard methods for isolating brain capillaries require the use of at least one or more mouse brains in order to obtain sufficient quantity and purity of brain capillaries. Here, we describe an optimized protocol for isolating and purifying capillaries from tiny amounts of mouse cerebral cortex using manual homogenization, density gradient centrifugation, and filtration while preserving the structural integrity and functional activity of microvessel fragments. Western blotting showed that proteins expressed at the BBB were enriched in mouse brain capillaries isolated by the optimized method compared to cerebral cortex protein homogenates. This approach can be used for the analysis of a variety of rare mouse genetic models and can also help the investigators to understand regional differences in susceptibility to pathological phenomena such as ischemia and traumatic brain injury. This will allow the investigators to better understand the physiology and pathology of the BBB.

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Acknowledgments

We thank Elisabeth Wilken for her excellent technical assistance.

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Authors and Affiliations

  1. Department of Anaesthesiology, Intensive Care, Emergency and Pain Medicine, University Hospital Würzburg, Würzburg, Germany

    Junqiao Mi, Aili Sun, Laura Härtel, Christina Dilling, Patrick Meybohm & Malgorzata Burek

  2. Graduate School of Life Sciences, Julius-Maximilians-University Würzburg, Würzburg, Germany

    Junqiao Mi

Authors
  1. Junqiao Mi
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  2. Aili Sun
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  3. Laura Härtel
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  4. Christina Dilling
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  5. Patrick Meybohm
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  6. Malgorzata Burek
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Corresponding author

Correspondence to Malgorzata Burek .

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Editors and Affiliations

  1. Department of Pathology, Microbiology, and Immunology, University of South Carolina School of Medicine, Columbia, SC, USA

    Swapan K. Ray

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Mi, J., Sun, A., Härtel, L., Dilling, C., Meybohm, P., Burek, M. (2024). Isolation of Capillaries from Small Amounts of Mouse Brain Tissue. In: Ray, S.K. (eds) Neuroprotection. Methods in Molecular Biology, vol 2761. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3662-6_2

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  • DOI: https://doi.org/10.1007/978-1-0716-3662-6_2

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3661-9

  • Online ISBN: 978-1-0716-3662-6

  • eBook Packages: Springer Protocols

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