Bakhsheshian Joshua, Wei Bih-Rong, Hall Matthew D, Simpson R Mark, Gottesman Michael M
Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
Laboratory of Cancer Biology and Genetics, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
Methods Mol Biol. 2016;1461:227-39. doi: 10.1007/978-1-4939-3813-1_19.
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.
我们提供了一个详细的方案，用于在转基因小鼠的血脑屏障（BBB）处对ATP结合盒亚家族G成员2（ABCG2）的功能进行成像。D-荧光素由血脑屏障处内皮细胞顶端侧发现的ABCG2特异性转运。荧光素酶-荧光素酶促反应产生生物发光，这使得能够直接测量血脑屏障处ABCG2的功能。因此，生物发光成像（BLI）与血脑屏障处ABCG2的功能相关，这可以通过在脑实质中表达荧光素酶的小鼠模型中给予荧光素来测量。与正电子发射断层扫描等其他策略相比，BLI为体内研究转运蛋白功能提供了一种成本相对较低的替代方法。这种在血脑屏障处对ABCG2功能进行成像的方法可用于研究该转运蛋白的药代动力学抑制作用。
