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General Research Article

β2-肾上腺素受体动力学的单细胞分析 荧光定量显微镜

卷 20, 期 6, 2020

页: [488 - 493] 页: 6

弟呕挨: 10.2174/1566524020666191216125825

价格: $65

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摘要

背景：G蛋白偶联受体（GPCR）代表最大的表面蛋白家族，并参与关键生理过程的调节。 GPCR的特征在于七个跨膜结构域，一个胞外N端和一个胞内C端。这些受体对其配体的细胞反应在很大程度上取决于它们的表面表达和激活后行为，包括表达，脱敏和再敏化。 目的：建立定量荧光显微镜检测法，研究β2-肾上腺素能受体的表达和脱敏。 方法：设计β2-肾上腺素能受体cDNA，将HA标签置于细胞外N端，将GFP标签置于细胞内C端。 GFP荧光可作为细胞总表达的量度；而用CY3结合的抗HA抗体染色而不渗透细胞则代表β2-AR的表面表达。量化图像并使用图像处理软件确定每个细胞的CY3（表面）和GFP（总）荧光量。 结果：该方法灵敏，可同时测量β2-AR的表面和总表达。 结论：描述了一种基于单细胞定量免疫荧光测量β2-AR表面和总表达的高精度方法。该方法可用于确定激动剂诱导的脱敏和再敏化过程，以及受体动力学，例如β2-肾上腺素能受体的胞吞作用和胞吐作用，并且基本上可用于任何其他GPCR。

关键词: G蛋白偶联受体，受体脱敏，定量免疫荧光，荧光显微镜，C末端，显微镜检测。

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