Microscopic visualization of metabotropic glutamate receptors on the surface of living cells using bifunctional magnetic resonance imaging probes.
Microscopic visualization of metabotropic glutamate receptors on the surface of living cells using bifunctional magnetic resonance imaging probes.
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使用双功能磁共振成像探针对活细胞表面的代谢型谷氨酸受体进行显微可视化。
DOI：
10.1021/cn400175m
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发表时间：
2014
影响因子：
5
通讯作者：
Mishra A
中科院分区：
文献类型：
--
作者：
Mishra A
基于已建立的竞争性代谢型Glu受体亚型5（mGluR5）拮抗剂，开发并评价了一系列双峰代谢型谷氨酸受体靶向MRI造影剂。为了直接观察这些试剂在活星形胶质细胞表面上的mGluR5结合，进行了核心结构的变化。相应地制备了一组含有花青染料或荧光素部分的钆缀合物，以允许在细胞中通过光学显微镜观察。在每种情况下，表面受体结合受损，并观察到细胞内化。另一种方法，通过敏化发射检查铽类似物的位置，也表现出神经元细胞系模型中的非特异性细胞摄取。最后，制备了两种先导化合物的生物素衍生物，并利用全内反射荧光（TIRF）和共聚焦显微镜，借助其荧光标记的抗生物素蛋白缀合物证明了与mGluR5细胞表面受体结合的特异性。
A series of bimodal metabotropic glutamate-receptor targeted MRI contrast agents has been developed and evaluated, based on established competitive metabotropic Glu receptor subtype 5 (mGluR5) antagonists. In order to directly visualize mGluR5binding of these agents on the surface of live astrocytes, variations in the core structure were made. A set of gadolinium conjugates containing either a cyanine dye or a fluorescein moiety was accordingly prepared, to allow visualization by optical microscopyin cellulo. In each case, surface receptor binding was compromised and cell internalization observed. Another approach, examining the location of a terbium analogue via sensitized emission, also exhibited nonspecific cell uptake in neuronal cell line models. Finally, biotin derivatives of two lead compounds were prepared, and the specificity of binding to the mGluR5cell surface receptors was demonstrated with the aid of their fluorescently labeled avidin conjugates, using both total internal reflection fluorescence (TIRF) and confocal microscopy.