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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Knockin mice expressing fluorescent δ-opioid receptors uncover G protein-coupled receptor dynamics in vivo
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Knockin mice expressing fluorescent δ-opioid receptors uncover G protein-coupled receptor dynamics in vivo

机译：表达荧光δ阿片受体的敲敲小鼠在体内揭示G蛋白偶联受体的动力学

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摘要

The combination of fluorescent genetically encoded proteins with mouse engineering provides a fascinating means to study dynamic biological processes in mammals. At present, green fluorescent protein (GFP) mice were mainly developed to study gene expression patterns or cell morphology and migration. Here we used enhanced GFP (EGFP) to achieve functional imaging of a G protein-coupled receptor (GPCR) in vivo. We created mice where the δ-opioid receptor (DOR) is replaced by an active DOR-EGFP fusion. Confocal imaging revealed detailed receptor neuroanatomy throughout the nervous system of knockin mice. Real-time imaging in primary neurons allowed dynamic visualization of drug-induced receptor trafficking. In DOR-EGFP animals, drug treatment triggered receptor endocytosis that correlated with the behavioral response. Mice with internalized receptors were insensitive to subsequent agonist administration, providing evidence that receptor sequestration limits drug efficacy in vivo. Direct receptor visualization in mice is a unique approach to receptor biology and drug design.
机译：荧光基因编码蛋白与小鼠工程学的结合为研究哺乳动物中的动态生物学过程提供了一种有趣的方法。目前，主要开发绿色荧光蛋白（GFP）小鼠来研究基因表达模式或细胞形态和迁移。在这里，我们使用增强的GFP（EGFP）在体内实现G蛋白偶联受体（GPCR）的功能成像。我们创建了小鼠，其中δ阿片受体（DOR）被活性DOR-EGFP融合蛋白替代。共聚焦成像揭示了敲入小鼠整个神经系统的详细受体神经解剖结构。初级神经元的实时成像可以动态观察药物诱导的受体转运。在DOR-EGFP动物中，药物治疗触发了与行为反应相关的受体胞吞作用。具有内在受体的小鼠对随后的激动剂给药不敏感，这提供了受体隔离在体内限制药物功效的证据。小鼠中直接受体可视化是受体生物学和药物设计的独特方法。

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