In Vivo Bioluminescent Imaging of ATP-Binding Cassette Transporter-Mediated Efflux at the Blood-Brain Barrier
In Vivo Bioluminescent Imaging of ATP-Binding Cassette Transporter-Mediated Efflux at the Blood-Brain Barrier
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DOI：
10.1007/978-1-4939-3813-1_19
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发表时间：
2016-01-01
期刊：
影响因子：
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通讯作者：
Gottesman, Michael M.
中科院分区：
文献类型：
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作者：
Bakhsheshian, Joshua;Wei, Bih-Rong;Gottesman, Michael M.
我们提供了一个详细的协议成像ATP结合盒亚家族G成员2（ABCG 2）的功能在血脑屏障（BBB）的转基因小鼠。D-荧光素通过在BBB处内皮细胞顶侧发现的ABCG 2特异性转运。在酶促反应中的ABCG 2酶产生生物发光，这允许在BBB直接测量ABCG 2功能。因此，生物发光成像（BLI）与BBB处的ABCG 2功能相关，并且这可以通过在脑实质中表达荧光素酶的小鼠模型中施用蓝蛋白来测量。与其他策略（如正电子发射断层扫描）相比，BLI允许在体内研究转运蛋白功能的相对低成本的替代方案。这种用于在BBB成像ABCG 2功能的方法可用于研究转运蛋白的药代动力学抑制。
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.