Label-Free Imaging of Histamine Mediated G Protein-Coupled Receptors Activation in Live Cells.
Label-Free Imaging of Histamine Mediated G Protein-Coupled Receptors Activation in Live Cells.
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活细胞中组胺介导的 G 蛋白偶联受体激活的无标记成像
DOI：
10.1021/acs.analchem.6b02677
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发表时间：
2016-12-06
影响因子：
7.4
通讯作者：
Wang, Shaopeng
中科院分区：
文献类型：
--
作者：
Lu, Jin;Yang, Yunze;Wang, Wei;Li, Jinghong;Tao, Nongjian;Wang, Shaopeng
G蛋白偶联受体(GPCRs)是细胞内最大的信号转导蛋白家族，是重要的药物靶点。传统的无标记分析方法缺乏空间信息来处理GPCRs激活后单个细胞的异质性反应。在这里，我们报告了一项基于等离子体的电化学阻抗显微镜在活细胞中的GPCRs研究。这种无标记光学成像平台能够以亚细胞分辨率解析来自单个细胞的响应。利用这一平台，我们研究了组胺介导的GPCRs激活，揭示了细胞下游反应的时空异质性。观察到单个HeLa细胞对组胺刺激的三相反应。在不到10个S中出现一个快速峰值P_1归因于GPCRs触发的钙释放。蛋白激酶C(PKC)激活后，细胞与基质的黏附发生改变，在1min内出现倒置的P2期。组胺处理后3~6min的主峰(P3)为动态质量再分布，呈剂量依赖性，半数有效浓度(EC_(50))为3·9±1·2μM。不同细胞间P3反应不均，尤其是在高浓度组胺时，表明细胞群体中组胺H1受体表达水平不同。
G protein-coupled receptors (GPCRs) are the largest protein family for cell signal transduction, and most of them are crucial drug targets. Conventional label-free assays lack the spatial information to address the heterogeneous response from single cells after GPCRs activation. Here, we reported a GPCRs study in live cells using plasmonic-based electrochemical impedance microscopy. This label-free optical imaging platform is able to resolve responses from individual cells with subcellular resolution. Using this platform, we studied the histamine mediated GPCRs activation and revealed spatiotemporal heterogeneity of cellular downstream responses. Triphasic responses were observed from individual HeLa cells upon histamine stimulation. A quick peak P1 in less than 10 s was attributed to the GPCRs triggered calcium release. An inverted P2 phase within 1 min was attributed to the alternations of cell–matrix adhesion after the activation of Protein Kinase C (PKC). The main peak (P3) around 3–6 min after the histamine treatment was due to dynamic mass redistribution and showed a dose-dependent response with a half-maximal effective concentration (EC50) of 3.9 ± 1.2 μM. Heterogeneous P3 responses among individual cells were observed, particularly at high histamine concentration, indicating diverse histamine H1 receptor expression level in the cell population.
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影响因子：
7.4
作者：
Liang, Wenbin;Wang, Shaopeng;Festa, Fernanda;Wiktor, Peter;Wang, Wei;Magee, Mitchell;LaBaer, Joshua;Tao, Nongjian
通讯作者：
Tao, Nongjian
影响因子：
29.4
作者：
Shan, Xiaonan;Chen, Shan;Tao, Nongjian
通讯作者：
Tao, Nongjian
单细胞动态和瞬时钙信号传导的无标记成像
影响因子：
16.6
作者：
Lu, Jin;Li, Jinghong
通讯作者：
Li, Jinghong
影响因子：
21.8
作者：
Wang, Wei;Foley, Kyle;Shan, Xiaonan;Wang, Shaopeng;Eaton, Seron;Nagaraj, Vinay J.;Wiktor, Peter;Patel, Urmez;Tao, Nongjian
通讯作者：
Tao, Nongjian
影响因子：
3.3
作者：
Zabner, J;Winter, M;Shasby, M
通讯作者：
Shasby, M