Imaging the Redox States of Live Cells with the Time-Resolved Fluorescence of Genetically Encoded Biosensors

被引：10

作者：

Li, Lei ^{[1
,4
]}

Zhang, Changcheng ^{[2
,3
]}

Wang, Peng ^{[1
]}

Wang, Aoxue ^{[2
,3
]}

Zhou, Jiasheng ^{[1
]}

Chen, Guoqing ^{[4
]}

Xu, Jianhua ^{[1
]}

Yang, Yi ^{[2
,5
]}

Zhao, Yuzheng ^{[2
,3
]}

Zhang, Sanjun ^{[1
,6
,7
]}

Tian, Yang ^{[8
]}

机构：

[1] East China Normal Univ, State Key Lab Precis Spect, 3663 North Zhongshan Rd, Shanghai 200062, Peoples R China

[2] East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg Technol, State Key Lab Bioreactor Engn, Synthet Biol & Biotechnol Lab, 130 Mei Long Rd, Shanghai 200237, Peoples R China

[3] East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab New Drug Design, 130 Mei Long Rd, Shanghai 200237, Peoples R China

[4] Jiangnan Univ, Sch Sci, Wuxi 214122, Peoples R China

[5] Chinese Acad Sci, Shanghai Inst Biol Sci, CAS Ctr Excellence Brain Sci, Optogenet & Synthet Biol Interdisciplinary Res Ct, Shanghai 200031, Peoples R China

[6] Shanxi Univ, Collaborat Innovat Ctr Extreme Opt, Taiyuan 030006, Shanxi, Peoples R China

[7] NYU Shanghai, NYU ECNU Inst Phys, 3663 North Zhongshan Rd, Shanghai 200062, Peoples R China

[8] East China Normal Univ, Shanghai Key Lab Green Chem & Chem Proc, Sch Chem & Mol Engn, Dongchuan Rd 500, Shanghai 200241, Peoples R China

来源：

ANALYTICAL CHEMISTRY | 2019年 / 91卷 / 06期

基金：

美国国家科学基金会;

关键词：

OXIDATIVE STRESS; HYDROGEN-PEROXIDE; PROTEIN; SENSOR; MITOCHONDRIAL; SONAR; H2O2;

D O I：

10.1021/acs.analchem.8b04292

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Redox environments in cells influence many important physiological and pathological processes. In this study, the time-resolved fluorescence of a recently reported thiol redox-sensitive sensor based on vertebrate fluorescent protein UnaG, roUnaG, was studied, along with the application of the time-resolved fluorescence of roUnaG to image the redox states of the mitochondria, cytoplasm, and nucleus in live cells. Time-resolved fluorescence images of roUnaG clearly demonstrated that potent anticancer compound KP372-1 induced extreme oxidative stress. A more stressful redox state observed in activated macrophages further demonstrated the validity of roUnaG with time-resolved fluorescence. For comparison, time-resolved fluorescence images of four other frequently used redox biosensors (roGFP1, HyPer, HyPerRed, and rxRFP) were also captured. The time-resolved fluorescence allows an intrinsically ratiometric measurement for biosensors with one excitation wavelength and provides new opportunities for bioimaging.

引用

页码：3869 / 3876

页数：8

共 29 条

[1] Genetically encoded fluorescent indicator for intracellular hydrogen peroxide [J].

Belousov, VV ;

Fradkov, AF ;

Lukyanov, KA ;

Staroverov, DB ;

Shakhbazov, KS ;

Terskikh, AV ;

Lukyanov, S .

NATURE METHODS, 2006, 3 (04) :281-286

[2] HyPer-3: A Genetically Encoded H2O2 Probe with Improved Performance for Ratiometric and Fluorescence Lifetime Imaging [J].

Bilan, Dmitry S. ;

Pase, Luke ;

Joosen, Linda ;

Gorokhovatsky, Andrey Yu ;

Ermakova, Yulia G. ;

Gadella, Theodorus W. J. ;

Grabher, Clemens ;

Schultz, Carsten ;

Lukyanov, Sergey ;

Belousov, Vsevolod V. .

ACS CHEMICAL BIOLOGY, 2013, 8 (03) :535-542

[3] Using Fractional Intensities of Time-resolved Fluorescence to Sensitively Quantify NADH/NAD+ with Genetically Encoded Fluorescent Biosensors [J].

Chang, Mengfang ;

Li, Lei ;

Hu, Hanyang ;

Hu, Qingxun ;

Wang, Aoxue ;

Cao, Xiaodan ;

Yu, Xiantong ;

Zhang, Sanjun ;

Zhao, Yuzheng ;

Chen, Jinquan ;

Yang, Yi ;

Xu, Jianhua .

SCIENTIFIC REPORTS, 2017, 7

[4] Imaging dynamic redox changes in mammalian cells with green fluorescent protein indicators [J].

Dooley, CT ;

Dore, TM ;

Hanson, GT ;

Jackson, WC ;

Remington, SJ ;

Tsien, RY .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (21) :22284-22293

[5] Free radicals in the physiological control of cell function [J].

Dröge, W .

PHYSIOLOGICAL REVIEWS, 2002, 82 (01) :47-95

[6] Red fluorescent genetically encoded indicator for intracellular hydrogen peroxide [J].

Ermakova, Yulia G. ;

Bilan, Dmitry S. ;

Matlashov, Mikhail E. ;

Mishina, Natalia M. ;

Markvicheva, Ksenia N. ;

Subach, Oksana M. ;

Subach, Fedor V. ;

Bogeski, Ivan ;

Hoth, Markus ;

Enikolopov, Grigori ;

Belousov, Vsevolod V. .

NATURE COMMUNICATIONS, 2014, 5

[7] Monitoring Redox Dynamics in Living Cells with a Redox-Sensitive Red Fluorescent Protein [J].

Fan, Yichong ;

Chen, Zhijie ;

Ai, Hui-wang .

ANALYTICAL CHEMISTRY, 2015, 87 (05) :2802-2810

[8]

Forman Henry Jay, 2001, Molecular Aspects of Medicine, V22, P189

[9] Reactive oxygen species and cell signaling - Respiratory burst in macrophage signaling [J].

Forman, HJ ;

Torres, M .

AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, 2002, 166 (12) :S4-S8

[10] Investigating mitochondrial redox potential with redox-sensitive green fluorescent protein indicators [J].

Hanson, GT ;

Aggeler, R ;

Oglesbee, D ;

Cannon, M ;

Capaldi, RA ;

Tsien, RY ;

Remington, SJ .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (13) :13044-13053

← 1 2 3 →