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

  • Joshua Bakhsheshian
  • Bih-Rong Wei
  • Matthew D. Hall
  • R. Mark Simpson
  • Michael M. GottesmanEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1461)


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.

Key words

d-Luciferin BCRP ABCG2 Optical imaging Neuroimaging 



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


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Joshua Bakhsheshian
    • 1
  • Bih-Rong Wei
    • 2
  • Matthew D. Hall
    • 1
  • R. Mark Simpson
    • 2
  • Michael M. Gottesman
    • 1
    Email author
  1. 1.Laboratory of Cell Biology, Center for Cancer Research, National Cancer InstituteNational Institutes of HealthBethesdaUSA
  2. 2.Laboratory of Cancer Biology and Genetics, Center for Cancer Research, National Cancer InstituteNational Institutes of HealthBethesdaUSA

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