Imaging of Intracellular Hydrogen Peroxide Production with HyPer upon Stimulation of HeLa Cells with EGF.

Autor: Markvicheva KN; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russian Federation., Bogdanova EA; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russian Federation., Staroverov DB; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russian Federation., Lukyanov S; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russian Federation.; Pirogov Russian National Research Medical University, Moscow, Russian Federation., Belousov VV; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russian Federation. vsevolod.belousov@gmail.com.; Pirogov Russian National Research Medical University, Moscow, Russian Federation. vsevolod.belousov@gmail.com.
Jazyk: angličtina
Zdroj: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2019; Vol. 1990, pp. 85-91.
DOI: 10.1007/978-1-4939-9463-2_7
Abstrakt: Reactive oxygen species (ROS) regulate both normal cell functions by activating a number of enzymatic cascades and pathological processes in many diseases by inducing oxidative stress. For many years since the discovery of ROS in biological systems there were no adequate methods of detection and quantification of these molecules inside the living cells. We developed the first genetically encoded fluorescent indicator for intracellular detection of hydrogen peroxide, HyPer, that can be used for imaging of H 2 O 2 production by cells under various physiological and pathological conditions. Unlike most known ROS indicators, HyPer allows for the generation of real-time image series that give precise information about the time course and intensity of H 2 O 2 changes in any compartment of interest. In this chapter we describe the method of confocal imaging of hydrogen peroxide production in HeLa cells upon stimulation with epidermal growth factor. The technique described may be accepted with minimal variations for the use in other cell lines upon various conditions leading to H 2 O 2 production.
Databáze: MEDLINE
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