Recognizing that current methods do not allow for real-time measurements of glutathione (a natural antioxidant that protects cells from damage) levels inside cells, a team of researchers at the Baylor College of Medicine, Texas Children's Hospital, and Rice University (all in Houston, TX) has developed a fluorescent probe that can track these changes in real time. The new probe, called RealThiol, could help to investigate the roles glutathione plays in aging, health, and diseases such as cancer, Alzheimer's and Parkinson's, cardiovascular conditions, and diabetes, among others.
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Previous methods are based on irreversible chemical reactions that capture all the glutathione that is inside the cells, providing a one-time snapshot of its amount. So Jin Wang, associate professor of pharmacology and chemical biology and of molecular and cellular biology at Baylor, and his colleagues looked for reversible chemical reactions that would capture and release glutathione, allowing for multiple measurements inside the same cell.
The method requires very small amounts of RealThiol, which results in little toxicity and poses minimal perturbance of the antioxidant capacity in the cells, Wang says. The probe can be used in various applications, from microscopy to cell sorting experiments, he adds.
Using RealThiol, the researchers measured enhanced antioxidant capability of activated neurons and dynamic glutathione changes during ferroptosis, a form of cell death. Wang and his group is currently developing glutathione probes with different subcellular specificities. This new tool set can potentially generate knowledge that could help develop new strategies to treat diseases involving glutathione-mediated processes.
Full details of the work appear in the journal Nature Communications.