Xiqian Jiang1, Chengwei Zhang1, Jianwei Chen1, Sungwoo Choi2, Ying Zhou3,4, Mingkun Zhao1,4, Xianzhou Song1, Xi Chen3, Mirjana Maletić-Savatić5,6,7, Timothy Palzkill1,7, David Moore3,2, Meng C Wang8,9,10, Jin Wang1,3,7. 1. 1 Department of Pharmacology and Chemical Biology,Baylor College of Medicine, Houston, Texas. 2. 2 Program in Developmental Biology, Baylor College of Medicine, Houston, Texas. 3. 4 Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas. 4. 3 Integrative Molecular and Biomedical Sciences Graduate Program, Baylor College of Medicine, Houston, Texas. 5. 5 Department of Pediatrics, and Baylor College of Medicine, Houston, Texas. 6. 6 Jan and Dan Duncan Neurological Research Institute at Texas Children's Hospital, Houston, Texas. 7. 7 Center for Drug Discovery, Baylor College of Medicine, Houston, Texas. 8. 8 Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas. 9. 9 Huffington Center on Aging, Baylor College of Medicine, Houston, Texas. 10. 10 Howard Hughes Medical Institute, Houston, Texas.
Abstract
AIMS: Quantitative imaging of glutathione (GSH) with high spatial and temporal resolution is essential for studying the roles of GSH in redox biology. To study the long-standing question of compartmentalization of GSH, especially its distribution between the nucleus and cytosol, an organelle-targeted quantitative probe is needed. RESULTS: We developed a reversible reaction-based ratiometric fluorescent probe-HaloRT-that can quantitatively measure GSH dynamics with subcellular resolution in real time. Using HaloRT, we quantitatively measured the GSH concentrations in the nucleus and 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 GSH probe that can be universally targeted to any organelle of interest. Taking advantage of this new tool, we provided definitive evidence showing that GSH concentrations are not significantly different between the nucleus and cytosol, challenging the view of nuclear compartmentalization of GSH.
AIMS: Quantitative imaging of glutathione (GSH) with high spatial and temporal resolution is essential for studying the roles of GSH in redox biology. To study the long-standing question of compartmentalization of GSH, especially its distribution between the nucleus and cytosol, an organelle-targeted quantitative probe is needed. RESULTS: We developed a reversible reaction-based ratiometric fluorescent probe-HaloRT-that can quantitatively measure GSH dynamics with subcellular resolution in real time. Using HaloRT, we quantitatively measured the GSH concentrations in the nucleus and 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 GSH probe that can be universally targeted to any organelle of interest. Taking advantage of this new tool, we provided definitive evidence showing that GSH concentrations are not significantly different between the nucleus and cytosol, challenging the view of nuclear compartmentalization of GSH.
Authors: Georgyi V Los; Lance P Encell; Mark G McDougall; Danette D Hartzell; Natasha Karassina; Chad Zimprich; Monika G Wood; Randy Learish; Rachel Friedman Ohana; Marjeta Urh; Dan Simpson; Jacqui Mendez; Kris Zimmerman; Paul Otto; Gediminas Vidugiris; Ji Zhu; Aldis Darzins; Dieter H Klaubert; Robert F Bulleit; Keith V Wood Journal: ACS Chem Biol Date: 2008-06-20 Impact factor: 5.100
Authors: G Bellomo; M Vairetti; L Stivala; F Mirabelli; P Richelmi; S Orrenius Journal: Proc Natl Acad Sci U S A Date: 1992-05-15 Impact factor: 11.205