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Bioluminescence pp 227–239Cite as

In Vivo Bioluminescent Imaging of ATP-Binding Cassette Transporter-Mediated Efflux at the Blood–Brain Barrier

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1461))

Abstract

We provide a detailed protocol for imaging ATP-binding cassette subfamily G member 2 (ABCG2) function at the blood–brain barrier (BBB) of transgenic mice. d-Luciferin is specifically transported by ABCG2 found on the apical side of endothelial cells at the BBB. The luciferase–luciferin enzymatic reaction produces bioluminescence, which allows a direct measurement of ABCG2 function at the BBB. Therefore bioluminescence imaging (BLI) correlates with ABCG2 function at the BBB and this can be measured by administering luciferin in a mouse model that expresses luciferase in the brain parenchyma. BLI allows for a relatively low-cost alternative for studying transporter function in vivo compared to other strategies such as positron emission tomography. This method for imaging ABCG2 function at the BBB can be used to investigate pharmacokinetic inhibition of the transporter.

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Acknowledgments

We thank Mr. George Leiman for editorial assistance. This research was supported by the Intramural Research Program of the National Institutes of Health (NIH), National Cancer Institute. Joshua Bakhsheshian is a NIH Medical Research Scholar. The Medical Research Scholars Program is a public–private partnership supported jointly by the NIH and contributions to the Foundation for the NIH from Pfizer, the Leona M. and Harry B. Helmsley Charitable Trust, the Howard Hughes Medical Institute, as well as other private donors (listed at www.fnih.org/work/programs-development/medicalresearch-scholars-program).

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Authors and Affiliations

  1. Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA

    Joshua Bakhsheshian, Matthew D. Hall & Michael M. Gottesman

  2. Laboratory of Cancer Biology and Genetics, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA

    Bih-Rong Wei & R. Mark Simpson

Authors
  1. Joshua Bakhsheshian
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  2. Bih-Rong Wei
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  3. Matthew D. Hall
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  4. R. Mark Simpson
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  5. Michael M. Gottesman
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Corresponding author

Correspondence to Michael M. Gottesman .

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Editors and Affiliations

  1. Natl Inst of AIST Res Inst for Envt Mgt Tech, Tsukuba, Japan

    Sung Bae Kim

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Bakhsheshian, J., Wei, BR., Hall, M.D., Simpson, R.M., Gottesman, M.M. (2016). In Vivo Bioluminescent Imaging of ATP-Binding Cassette Transporter-Mediated Efflux at the Blood–Brain Barrier. In: Kim, S. (eds) Bioluminescence. Methods in Molecular Biology, vol 1461. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3813-1_19

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  • DOI: https://doi.org/10.1007/978-1-4939-3813-1_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3811-7

  • Online ISBN: 978-1-4939-3813-1

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