Analysis of oxidative stress in zebrafish embryos
| Autor: | Massimo M. Santoro, Annalisa Camporeale, Vera Mugoni |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Male
Programmed cell death Embryo Nonmammalian General Chemical Engineering Context (language use) Oxidative phosphorylation Biology medicine.disease_cause General Biochemistry Genetics and Molecular Biology Animals Genetically Modified In vivo medicine Animals Zebrafish chemistry.chemical_classification Reactive oxygen species General Immunology and Microbiology General Neuroscience Neurodegeneration Anatomy medicine.disease biology.organism_classification Cell biology Oxidative Stress chemistry Female Reactive Oxygen Species Oxidative stress Developmental Biology |
| Popis: | High levels of reactive oxygen species (ROS) may cause a change of cellular redox state towards oxidative stress condition. This situation causes oxidation of molecules (lipid, DNA, protein) and leads to cell death. Oxidative stress also impacts the progression of several pathological conditions such as diabetes, retinopathies, neurodegeneration, and cancer. Thus, it is important to define tools to investigate oxidative stress conditions not only at the level of single cells but also in the context of whole organisms. Here, we consider the zebrafish embryo as a useful in vivo system to perform such studies and present a protocol to measure in vivo oxidative stress. Taking advantage of fluorescent ROS probes and zebrafish transgenic fluorescent lines, we develop two different methods to measure oxidative stress in vivo: i) a "whole embryo ROS-detection method" for qualitative measurement of oxidative stress and ii) a "single-cell ROS detection method" for quantitative measurements of oxidative stress. Herein, we demonstrate the efficacy of these procedures by increasing oxidative stress in tissues by oxidant agents and physiological or genetic methods. This protocol is amenable for forward genetic screens and it will help address cause-effect relationships of ROS in animal models of oxidative stress-related pathologies such as neurological disorders and cancer. |
| Databáze: | OpenAIRE |
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