Versatile Histochemical Approach to Detection of Hydrogen Peroxide in Cells and Tissues Based on Puromycin Staining.
Versatile Histochemical Approach to Detection of Hydrogen Peroxide in Cells and Tissues Based on Puromycin Staining.
复制标题
基于嘌呤霉素染色的细胞和组织中检测过氧化氢的多功能组织化学方法。
DOI：
10.1021/jacs.8b02279
复制
发表时间：
2018-05-16
影响因子：
15
通讯作者：
Chang CJ
中科院分区：
文献类型：
--
作者：
Yik-Sham Chung C;Timblin GA;Saijo K;Chang CJ
过氧化氢(H_2O_2)是一种中枢活性氧物种(ROS)，可导致肥胖、癌症和神经变性等疾病，同时也是一种重要的信号分子。我们现在报告了一种检测过氧化氢的通用组织化学方法，该方法可以在健康和疾病状态下的广泛细胞和组织标本中使用。我们开发了第一代过氧化氢反应类似物Peroxymycin-1，它基于经典的细胞染色分子嘌呤霉素，能够对生物样品进行共价染色，并在固定后保留其信号。H_2O_2介导的硼酸盐裂解使嘌呤霉素氨基核苷发生裂解，从而在处理过的生物标本上留下一个永久的、剂量依赖的标记，通过标准的免疫荧光分析可以高灵敏度和高精度地检测到该标记。Peroxymycin-1具有足够的选择性和敏感性，可以同时显示细胞内和外源性的过氧化氢水平变化，并可以用来分析一组细胞系中静止的过氧化氢水平，根据与ROS状态的相关性，将转移、侵袭性较小的癌细胞与侵袭性较小的癌症和非肿瘤细胞区分开来。此外，我们通过对非酒精性脂肪性肝病小鼠肝组织中过氧化氢水平异常升高的鉴定，证实了过氧霉素-1是体内过氧化氢分析的有效组织化学探针，从而证明了这种方法在研究与过氧化氢相关的疾病状态和进展方面的潜力。这项工作提供了设计原则，应该能够开发更广泛的生物使用的组织化学探针，这些探针通过基于活动的传感进行操作。
Hydrogen peroxide (H2O2) is a central reactive oxygen species (ROS) that contributes to diseases from obesity to cancer to neurodegeneration but is also emerging as an important signaling molecule. We now report a versatile histochemical approach for detection of H2O2 that can be employed across a broad range of cell and tissue specimens in both healthy and disease states. We have developed a first-generation H2O2-responsive analogue named Peroxymycin-1, which is based on the classic cell-staining molecule puromycin and enables covalent staining of biological samples and retains its signal after fixation. H2O2-mediated boronate cleavage uncages the puromycin aminonucleoside, which leaves a permanent and dose-dependent mark on treated biological specimens that can be detected with high sensitivity and precision through a standard immunofluorescence assay. Peroxymycin-1 is selective and sensitive enough to image both exogenous and endogenous changes in cellular H2O2 levels and can be exploited to profile resting H2O2 levels across a panel of cell lines to distinguish metastatic, invasive cancer cells from less invasive cancer and nontumorigenic counterparts, based on correlations with ROS status. Moreover, we establish that Peroxymycin-1 is an effective histochemical probe for in vivo H2O2 analysis, as shown through identification of aberrant elevations in H2O2 levels in liver tissues in a murine model of nonalcoholic fatty liver disease, thus demonstrating the potential of this approach for studying disease states and progression associated with H2O2. This work provides design principles that should enable development of a broader range of histochemical probes for biological use that operate via activity-based sensing.
登录
查看更多内容
影响因子：
15
作者：
Dickinson BC;Huynh C;Chang CJ
通讯作者：
Chang CJ
影响因子：
29
作者：
Cochemé HM;Quin C;McQuaker SJ;Cabreiro F;Logan A;Prime TA;Abakumova I;Patel JV;Fearnley IM;James AM;Porteous CM;Smith RA;Saeed S;Carré JE;Singer M;Gems D;Hartley RC;Partridge L;Murphy MP
通讯作者：
Murphy MP
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：
7.4
作者：
Clark, HA;Kopelman, R;Philbert, MA
通讯作者：
Philbert, MA
影响因子：
21.8
作者：
通讯作者：
--