Quantitative Real-Time Imaging of Glutathione with Sub-Cellular Resolution
Meng C. Wang,
Posted 01 Jul 2018
bioRxiv DOI: 10.1101/360362 (published DOI: 10.1089/ars.2018.7605)
Posted 01 Jul 2018
Aims: Quantitative imaging of glutathione with high spatial and temporal resolution is essential for studying the roles of glutathione in redox biology. To study the long-standing question of compartmentalization of glutathione, especially its distribution between the nucleus and the cytosol, an organelle-targeted quantitative probe is needed. Results: We developed a reversible reaction-based ratiometric fluorescent probe, HaloRT, that can quantitatively measure glutathione dynamics with subcellular resolution in real-time. Using HaloRT, we quantitatively measured the glutathione concentrations in the nucleus and the cytosol of HeLa cells and primary hepatocytes under different treatment conditions and found no appreciable concentration gradients between these two organelles. Innovation and conclusion: We developed the first reversible ratiometric glutathione probe that can be universally targeted to any organelles of interest. Taking advantage of this new tool, we provided a definitive evidence showing that glutathione concentrations are not significantly different between the nucleus and the cytosol, challenging the view of nuclear compartmentalization of glutathione.
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