A palette of fluorescent probes with varying emission colors for imaging hydrogen peroxide signaling in living cells.
A palette of fluorescent probes with varying emission colors for imaging hydrogen peroxide signaling in living cells.
复制标题
DOI：
10.1021/ja1014103
复制
发表时间：
2010-04-28
影响因子：
15
通讯作者：
Chang CJ
中科院分区：
文献类型：
--
作者：
Dickinson BC;Huynh C;Chang CJ
我们提出了一个新的家庭的荧光探针与不同的发射颜色选择性成像过氧化氢（H2O2）产生的生理细胞信号水平。这种结构上同源的基于荧光素和玫瑰红的报告基因系列依赖于化学特异性硼酸酯至苯酚的开关，以通过一组生物学相关的活性氧物质（ROS）对H2O2作出响应，该活性氧物质具有可调的激发和发射最大值以及对内源性产生的H2O2信号的敏感性，如在RAW 264.7巨噬细胞在吞噬呼吸爆发期间和A431细胞响应EGF刺激的研究所示。我们进一步证明了这些试剂的实用性，通过使用一种新的H2O2特异性探针，过氧橙子1（PO 1），与绿色荧光高活性氧（hROS）探针，APF在荧光成像实验。这种双探针方法允许选择性区分活RAW 264.7巨噬细胞中H2O2和次氯酸（HOCl）水平的变化。此外，当用PO1和APF标记的巨噬细胞被刺激以诱导免疫应答时，我们发现了三种不同类型的吞噬体：主要产生hROS的吞噬体，主要产生H2O2的吞噬体，以及同时具有两种ROS的吞噬体。使用不同颜色的荧光探针的调色板同时监测多个ROS通量的能力开辟了新的机会，通过分子成像来解开氧化生物学对生命系统的复杂贡献。
We present a new family of fluorescent probes with varying emission colors for selectively imaging hydrogen peroxide (H2O2) generated at physiological cell signaling levels. This structurally homologous series of fluorescein- and rhodol-based reporters relies on a chemospecific boronate-to-phenol switch to respond to H2O2 over a panel of biologically relevant reactive oxygen species (ROS) with tunable excitation and emission maxima and sensitivity to endogenously produced H2O2 signals, as shown by studies in RAW 264.7 macrophages during the phagocytic respiratory burst and A431 cells in response to EGF stimulation. We further demonstrate the utility of these reagents in multicolor imaging experiments by using one of the new H2O2-specific probes, Peroxy Orange 1 (PO1), in conjunction with the green-fluorescent highly reactive oxygen species (hROS) probe, APF. This dual-probe approach allows for selective discrimination between changes in H2O2 and hypochlorous acid (HOCl) levels in live RAW 264.7 macrophages. Moreover, when macrophages labeled with both PO1 and APF were stimulated to induce an immune response, we discovered three distinct types of phagosomes: those that generated mainly hROS, those that produced mainly H2O2, and those that possessed both types of ROS. The ability to monitor multiple ROS fluxes simultaneously using a palette of different colored fluorescent probes opens new opporunities to disentangle the complex contributions of oxidation biology to living systems by molecular imaging.
登录
查看更多内容
DOI：
10.1073/pnas.1834314100
发表时间：
2003-09-30
影响因子：
11.1
作者：
Avshalumov, MV;Rice, ME
通讯作者：
Rice, ME
影响因子：
48
作者：
Gutscher, Marcus;Pauleau, Anne-Laure;Dick, Tobias P.
通讯作者：
Dick, Tobias P.
微芯片电泳简单快速测定RAW264.7巨噬细胞中过氧化氢的新途径
影响因子：
2.9
作者：
Gong, Xiaocong;Li, Qingling;Zhong, Hongbo
通讯作者：
Zhong, Hongbo
影响因子：
2.9
作者：
Cline, DJ;Thorpe, C;Schneider, JP
通讯作者：
Schneider, JP
影响因子：
4.6
作者：
Burdette, SC;Lippard, SJ
通讯作者：
Lippard, SJ