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Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors
Published on: February 7, 2018
Live-cell imaging approaches for the investigation of xenobiotic-induced oxidant stress
Phillip A Wages1, Wan-Yun Cheng2, Eugene Gibbs-Flournoy3
1Curriculum in Toxicology, University of North Carolina at Chapel Hill, NC, USA.
Background:
Oxidant stress is arguably a universal feature in toxicology. Research studies on the role of oxidant stress induced by xenobiotic exposures have typically relied on the identification of damaged biomolecules using a variety of conventional biochemical and molecular techniques. However, there is increasing evidence that low-level exposure to a variety of toxicants dysregulates cellular physiology by interfering with redox-dependent processes.
Scope Of Review:
The study of events involved in redox toxicology requires methodology capable of detecting transient modifications at relatively low signal strength. This article reviews the advantages of live-cell imaging for redox toxicology studies.
Major Conclusions:
Toxicological studies with xenobiotics of supra-physiological reactivity require careful consideration when using fluorogenic sensors in order to avoid potential artifacts and false negatives. Fortunately, experiments conducted for the purpose of validating the use of these sensors in toxicological applications often yield unexpected insights into the mechanisms through which xenobiotic exposure induces oxidant stress.
General Significance:
Live-cell imaging using a new generation of small molecule and genetically encoded fluorophores with excellent sensitivity and specificity affords unprecedented spatiotemporal resolution that is optimal for redox toxicology studies. This article is part of a Special Issue entitled Air Pollution, edited by Wenjun Ding, Andrew J. Ghio and Weidong Wu.
Insights
Live-cell imaging offers advanced methods for studying oxidant stress in toxicology. This technique detects subtle cellular changes caused by toxicant exposure, improving our understanding of redox-dependent processes.
Area of Science:
- Toxicology
- Cellular Biology
- Biochemistry
Background:
- Oxidant stress is a universal feature in toxicology.
- Conventional methods for studying oxidant stress often rely on detecting damaged biomolecules.
- Emerging evidence suggests low-level toxicant exposure disrupts cellular physiology via redox-dependent pathways.
Purpose of the Study:
- To review the advantages of live-cell imaging for redox toxicology.
- To highlight methodologies for detecting transient modifications in redox toxicology.
- To discuss the application of fluorogenic sensors in toxicological studies.
Main Methods:
- Live-cell imaging utilizing small molecule and genetically encoded fluorophores.
- Detection of transient modifications at low signal strengths.
- Validation of fluorogenic sensors in toxicological applications.
Main Results:
- Live-cell imaging provides high spatiotemporal resolution for redox toxicology.
- New generations of fluorophores offer enhanced sensitivity and specificity.
- Experiments with fluorogenic sensors can reveal mechanisms of toxicant-induced oxidant stress.
Conclusions:
- Live-cell imaging is optimal for redox toxicology studies.
- Careful use of fluorogenic sensors is necessary to avoid artifacts.
- Understanding xenobiotic interactions with redox processes is crucial.
